a racknenih aches anemia a a Tete #2 DOR ny CORNELL UNIVERSITY THE Roswell P. Flower Library THE GIFT OF ROSWELL P. FLOWER FOR THE USE OF THE N. Y. STATE VETERINARY COLLEGE ; eee vel, over app ission ii ‘ommi 83 i R tae i a ral < G £ 2 a 2 2 a SS o —— o oS=== 2 —— e SSS 2 o c = 3 oO 3 1 FE i ‘ti R PART TL. “APPENDIX. 2 VOLUME. TEL, 2 REPORTS | ON” INVESTIGATIONS es = DEALING wires ae Weprisuen “by BY H © RS ge A Ne {be Sei rin LANE, £0. and OLIVER AED BOYD, “Tynspbaun Court, ; =H LeD, 116, N STRE pi 0, [Cd, 5615] Sad. ROYAL COMMISSION ON TUBERCULOSIS (HUMAN AND BOVINE). FINAL REPORT OF THE ROYAL COMMISSION APPOINTED TO INQUIRE INTO THE RELATIONS OF HUMAN AND ANIMAL TUBERCULOSIS. PART IIT. APPENDIX. VOLUME III. REPORTS ON’ INVESTIGATIONS DEALING WITH 1.—Certain Human Viruses of Irregular Type. 2.—The Excretion of Tubercle Bacilli in the Milk of Animals, 3.—Swine Tuberculosis, 4.—Immunity. - A, STANLEY GRIFFITH, M.D, and F. GRIFFITH, MB, TOGETHER WITH Two Reports submitted to the Commission in 1906 by Dr. L. COBBETT, Presented to both Houses of Parliament by Command of His Majesty. LONDON: PUBLISHED BY HIS MAJESTY’S STATIONERY* OFFICE. To be purchased, either directly or through any Bookseller, from: ~ WYMAN anp SONS, LTD., FETTER LANE, H.C., and 32, ABINGDON STREET, 8.W.; or OLIVER anpd BOYD, TWEEDDALE COURT, EDINBURGH; or BE. PONSONBY, Lrp., 116, GRAFTON STREET, DUBLIN. PRINTED BY DARLING AND SON, LTD., BACON STREET, E. 1911. [Cd. 5893.] Price 3s. 3d. 2 Dd a ( ) C+ ae VAY NO in ( Vv ; “i O¢ ¥Y\ ( 24 ee i N S&S 7 : A Ee pe) ' > m g é / elf. / ant CORNELL UNIVERSITY. THE Roswell P. Flower Library THE GIFT OF ROSWELL P. FLOWER FOR THE USE OF THE N. Y. STATE VETERINARY COLLEGE. 1897 os PREFACE. This volume contains the Reports of four separate investigations carried out at Stansted under the direction of the Commission, the first two being the work of Dr. A. Stanley Griffith alone and the others of himself and Dr. F. Griffith jointly. The first Report, dealing with Human Viruses of Irregular Type, is divided into three parts, two of which refer to further investigations of certain viruses that had already been investigated by Dr. Cobbett and placed provisionally by the Commission in a group by themselves. The details of the previous investigation of these viruses will be found in the Commission’s Second Interim Report and Appendix. The third part deals with the cultures isolated since the issue of the Second Interim Report from two human viruses that exhibited similar features to those other irregular ones placed in Group III. The second Report in this volume gives the details of an investigation in connection with the excretion of tubercle bacilli in the milk of animals (goats and cows) after experimental inoculation. The results of Dr. F. Griffith’s investigations into the excretion of tubercle bacilli in the milk of similar animals naturally infected with tuberculosis, but showing no evidence of disease of the udder, have already been published in the Commission’s Third Interim Report and Appendix issued in 1909. The investigation of Swine Tuberculosis, which forms the subject of the third Report contained in this volume, was carried out at Walpole Farm. The experi- ments were commenced by Dr. A. Stanley Griffith and, after he was transferred to Blythwood Farm, continued by Dr. F. Griffith. ‘The whole of the experimental work on which the last Report, namely that on Immunity, is based was carried out by Dr. A. Stanley Griffith and Dr. F. Griffith jointly at Walpole Farm. Two Reports submitted to the Commission in 1906 by Dr. Cobbett on certain investigations carried out by him at Blythwood Farm are also included in this volume. These Reports deal with the Stability of Virulence of Tubercle Bacilli in the Living Body and Experiments with Mixed Viruses. They are referred to in the Second Interim Report of the Commission (pages 41 and 42), but as the experiments were of a preliminary nature the Reports on them were not included in the Appendix to that Report. The two Reports have not been revised by Dr. Cobbett to include subsequent experiments on the same subjects carried out at Blythwood, but are now printed as originally written by him. Epwarp J. STEEGMANN. 2000. Wts. T.L. 277 & 13670. 12/08. 32. 27676D. A 2 LIST OF INVESTIGATIONS CONTAINED IN THIS VOLUME. PAGE. 1.—Investigation of certain Human Viruses of Irregular Type. (A. STANLEY GRIFFITH, M.D.).... ae sia See see ie a (a) Results of the Further Investigation of Cultures from the Viruses placed in Group III. (see 2nd Interim Report, page 30). (b) Results of the Special Investigation of two of the Group Ii. Cultures (H 13. “A.D.” (Calf 301), and H 49. “'T.C.”) which exhibited exceptional features. (c) Results of the Special Investigation of the two Cultures aH 60. “ W.B.” (Bronchial Gland) and H 90. “1-P.” (Retroperitoneal Gland), isolated since the 2nd Interim Report, which exhibited the features of the two Group III. Cultures. 2.—The Excretion of Tubercle Bacilli— (a) In the Milk of Cows and Goats. (b) Into the Undeveloped Udder (A. STANLEY GRIFFITH, M.D.) ... 79 3.—Investigation of Tubercle Bacilli from Cases of Swine Tuberculosis. (F. GrirFiTH, M.B., and A. STANLEY GRIFFITH, M.D.)... xox! AS 4.—Report on Immunity Experiments. (A. STANLEY GRIFFITH, M.D., and ¥. GRIFFITH, M.B.) ... ies ie sais sue vee «6-265 and also 5.—Report (submitted to the Commission in 1906) on the Stability of Tubercle Bacilli in the Living Animal. (L. Copperr, m.p.) ... 297 6.—Report (submitted to the Commission in 1906) on Experiments with Mixed Viruses— when grown (@) In Artificial Culture. (6) In the Body of a Living Animal. (L. COBBETT, M.D.) 341 ‘Results of the Further Investigation of Cultures from the Viruses placed in Group III. (See also 2nd Interim Report, page 30.) Results of the Special Investigation of two of the Group III. Cultures (H 18. “A.D.” (Calf 301) and H 49. “'T.C.” (0.M.)) which exhibited exceptional features. Results of the Special Investigation of the two Cultures H 60. “W.B.” (bronchial gland) and H 90. “LP.” (retroperitoneal gland), isolated since the 2nd Interim Report, which exhibited the features of the two Group III. Cultures. BY A. STANLEY GRIFFITH, M.D. 6 INVESTIGATION OF CERTAIN HUMAN VIRUSES OF IRREGULAR TYPE. TABLE OF CONTENTS. PAGE. Results of the further investigation of cultures from the viruses placed in Group III. Introduction Os aes ORS. Wes Ney. aay OE et, Details of the further investigation of :— Virus H 2. “Sp.A.” wee ee ea a rn te sa bts oy Ade wane TE Virus H 13. “ A.D.” gs ae aa ele se ee sh ei sits so we 8 Virus H 16. “J.H.” we sisi sity ida _ sie see wee ee ie gies 9) Virus H 17. “Sp.B.” sii oe ne8 a ows sie sh ies aes ves .. 10 Virus H 49. “ T.C.” cs be sibs ae sis a za sg ais is -» 10 Summary sas se aia see ts ons sie ave are seis ais wes oe ey deh Results of the special investigation of two of the Group III. cultures (H 13. “A.D.” (Calf 301) and H-49. “1.C.” (0.M.) which exhibited exceptional features. Separation Experiments. Introduction... ei uate woe an wes ald ee ae sas is Bou eee deg 2? Experiments with Virus H 13. “ A.D.” :— Separation by Animal Inoculation ane lad oes ee eu ‘i ns cee ww. «14 Separation by Glycerin Media... aes aise ae see wit aif sis wee ww «14 Separation by Plate Cultures... aes a sins aes See soe is se ww. «14 Experiments with Virus H 49. “TC.” :— Separation by Animal Inoculation ae ses a aha ae oe aes see we 16 Separation by Glycerin Media... we aie is ie eas ane aes ss w= 16 Separation by Plate Cultures... ies tee aise a5 ae ise ae aad soe AE Summary and Conclusions... ee aes a6 a as a et eee et see vox TT Results of the special investigation of the two cultures H 60. “W.B.” (bronchial gland) and H 90. “IP.” (retroperitoneal gland), isolated since the second interim Report, which exhibited the features of the two Group III. cultures. Separation Experiments. Summary of Results ... Ba aie 7 18 Experiments with Virus H 60. “ W.B.” :— Separation by Animal Inoculation 20 Separation by Glycerin Media 20 Separation by Plate Cultures 20 Summary is aes aie 21 Experiments with Virus H 90. “I.P.” :— Separation by Animal Inoculation 1 Separation by Glycerin Media sj 7 a 99 Separation by Plate Cultures 29 Summary 22 Tables, Charts, and Post-Mortem Notes. Tabular Summaries of the Preliminary Inoculation Tests with Viruses H 2. “ Sp.A.”, H 13. “ A.D.” 16.“S.R", HI Spe” and BAG OO" sce ease ea en) ee H 90. “TP.” ‘ mt Full Post-Mortem Notes of the Calves Inoculated Charts of the Separation Experiments with Viruses H 13.“ A.D.”, H 49.“ 'T.C.”, H 60. “W.B.” a ’ . . 7 4 ose aie sive wer 49 Plates. Photographs of Cultures ... jee ss re 77-78 RESULTS OF THE FURTHER INVESTIGATION OF CULTURES FROM THE VIRUSES PLACED IN GROUP IIL. INTRODUCTION. In the 2nd interim report the results are given of experiments with certain viruses, which, on account of their exceptional features, were put together in a separate group (Group III.). The cases differed widely, and included— ae A ae of a character intermediate between Group [. and Group IL. (H 49.“ T.C.”). (2) Viruses in which a eugonic bacillus of low virulence had been replaced by a dysgonic bacillus of high virulence (‘ Passage” experiments H 16.“ J.H.”, H 13.“ A.D.”, H 17.“ Sp.B.’). Two possible explanations of the cases were put forward and discussed by the Commission. The first was that the results obtained were due to a mixture of viruses, of a virus from a human source and a virus from a bovine source, each possessing stable characters. The second was that tubercle bacilli under conditions as yet not definable may manifest instability, and may, when subjected to vital influences, such as are supplied by human or bovine tissues, become modified in character. A definite decision as to which of the two views is the true one was held over pending further work with the cultures obtained at various stages of the experiments. All the work in connection with these original experiments, which are detailed in the Second Interim Report and summarised in this, was done by Dr. Cobbett. The further investigation of the cultures was entrusted to me, Dr. Cobbett being unable to complete the work on account of his appointment to the Chair of Pathology in the University of Sheffield, and I was directed by the Commission first te test the cultural characters and the virulence of the cultures that existed of the various viruses which had been included in this group (Group III), and then to investigate specially those cultures which exhibited any want of accordance between cultural characters and virulence. In order to save the necessity of referring to previous reports for the origin of the cultures, a brief summary, illustrated by marginal diagrams, is given of all the viruses reinvestigated. These viruses are H 13.‘ A.D.”, H 16.“ J.H.”, H 17.“ 5p.B.”, and H 49.“ T.C.”. Cultures from the virus H 2.“ Sp.A.”, the original material of which was mixed sputum, were also investigated. The further investigations with the lupus virus H 53. “ D.H.,” which had been provisionally included in Group III, are recorded in Volume II of the Appendix. VIRUS H 2. “Sp. A.” (Mixed Sputum.) RECAPITULATION. Two heifers, Nos, 11 and 13, were fed every day with sputum from persons suffering from pulmonary tuberculosis, one for about 200, the other for about 300 days. Heifer 11 was killed 209 days after the 8 experiment began ; several mesenteric glands and a few hepatic lymphatic glands were found to be tuber- culous, caseous and calcareous, and there was a pedun- culated growth on the lung. An emulsion from the’ Human Sputum. mesenteric glands, injected subcutaneously into a calf, i or 02.) gave rise to general though not fatal tuberculosis. _ Emulsions from various lymphatic glands of this ial ‘ | second animal injected into six calves, two subcuta-- Heifer 11. Heifer 13. neously, two intraperitoneally, and two intravenously, Fed. Fed. gave rise to general progressive tuberculosis fatal in Killed 209 days. Killed 303 days. Local Tuberculosis. ; the two inoculated intravenously but not in the others Local Tuberculosis. within 90 days. ] In the other heifer, No. 13, calcareous mesenteric glands were found after death. An emulsion from these calcareous glands, injected subcutaneously into Calf 111 gave rise toa small nodule ina prepectoral gland only, while the same emulsion passed through a guinea-pig produced in one calf fatal generalised tuberculosis, and in another slight retrogressive- tuberculosis. A culture isolated from the prescapular gland of Calf 111 through Guinea-pig 690 was found by Dr.. Cobbeit to be eugonic and to have low virulence for the calf and rabbit; it produced in broth a well-marked acid reaction (A.8.G.). A culture from Calf 93, one of the six calves inoculated with the virus from Heifer 11, . grew less well than the culture from Calf 111 and was fully virulent. | Calf 111. Subcut. Killed 138 days. Focus in prepectoral gland. | Calf 79. Subcut. Killed (ill) 71 days. G.T. not severe. Six calves of which Calf 93 (Intrap.) was _ one. G.T. in all. | Guinea-pig 690. RESULTS OF REINVESTIGATION. Two strains of the virus existed, the one derived from Calf 93, the other from Calf 111. : Virulence Tests.—The culture from Calf 93 was retested after three years eight months’ artificial cultivation.. A series of five rabbits was inoculated, two intravenously, two intraperitoneally, and one subcutaneously. The subcutaneous rabbit died of heart disease in five days ; the others died of general tuberculosis in from 18 to 33 days. The culture derived from Calf 111 was tested after three years three months’ artificial cultivation. A series of seven rabbits was inoculated, two intravenously, two intraperitoneally, and three subcutaneously. All were killed after 111 days and showed slight retrogressive tuberculosis only. Cultural Characters—The culture from Calf 93 grew moderately well on glycerin media but not better than the more easy growing cultures of bovine origin. The culture from Calf 111 grew luxuriantly like the human tubercle bacillus. Summary.—The Calf 93 strain exhibited the characters of a bovine tubercle bacillus; the Calf 111 strain. those of a human tubercle bacillus. VIRUS H13. “A.D.” RECAPITULATION. Tuberculous material obtained from the bronchia? glands and spleen of a child, aged 4 years, who died of acute generalised tuberculosis, was mixed and inoculated subcutaneously into two calves and gave rise in each to a very limited retrogressive tuberculosis confined to the seat of inoculation and nearest glands. The virus from the prescapular gland of one of Bronchial Glands the calves was passed through three consecutive series. and Spleen. of guinea-pigs and then inoculated into two calves, | one subcutaneously, the other intraperitoneally. Calf 129. _ The animal inoculated intraperitoneally was killed Subcut. in 87 days and showed tuberculous peritonitis and tuberculosis of the thoracic and a few other glands and no disease of the organs. The other calf No. 301, inoculated subcutaneously, was killed when very ill after 33 days, and showed severe general tuberculosis. Killed 66 days. Local Tuberculosis. 3 series of guinea- igs. i ai An emulsion made from the thoracic glands of Calf 301. Calf 301 gave rise in two calves (of which one was Sutnat, Calf 321), after subcutaneous inoculation, to severe Killed (very ill) 33 days. General Tuberculosis. | Rat 15. K. 167 days. G.T. Two calves of which Calf 321 (Subcut.) was one: G.T. general progressive tuberculosis, and to fatal tubercu- losis in two rabbits. Cultures were isolated from the mediastinal gland of Calf 301, the preseapular gland of Calf 321, and from a rat, No. 15, which had been inoculated with the emulsion from Calf 301. _ The culture from Calf 301 was eugonic and had irregular virulence ; inoculated, after fifteen months’ artificial cultivation, into two calves, in a dose of 50 milligrammes, it produced in one slight general tuberculosis, the lesions in the lungs resembling those produced by a Group I bacillus, and i tuberculosis limited to the site of inoculation and the nearest gland; in wabbits slight disease we ae in one animal and chronic general tuberculosis in two others. ~ P The cultures from Calf 321 and Rat 15 were dysgonic and fully virulent. RESULTS OF REINVESTIGATION, The cultures from Calf 301 and Rat 15 were the only two which bad been preserved The last subculture (24th generation) of the strain from Calf 301 ee about three months old and subcultures from it on to serum failed ; in order to recover it a guinea-pig was inoculated and several tubes of the egg medium were sown at the same time ; the guinea-pig died of tuberculosis, but since growth had taken place on the egg tubes it was unnecessary to isolate a culture from this animal. The strain now grew readily on serum, and after three subcultures on this medium, its virulence was tested. _ _ Virulence Tests.—The strain from Calf 301 was inoculated into a series of six rabbits, two intravenously, two intraperitoneally, and two subcutaneously ; the culture used was the 28th generation, the strain having been three years in artificial cultivation. One rabbit died after two days of causes other than tuberculosis, The remaining five died of general tuberculosis in from 12 to 52 days, the duration of life being a little longer than in rabbits inoculated with equivalent doses of bovine tubercle bacilli. Two cats inoculated at the same time as the rabbits died of acute tuberculosis. The culture derived from Rat 15 was inoculated into a series of four rabbits (two intravenously and two intraperitoneally). 7% months in artificial cultivation. The culture used was the 25th generation and the strain had been two years The rabbits inoculated intravenously (doses 1:0 and 0-1 mg.) died of general tuberculosis in 13 and 25 days, the intraperitoneal rabbits in i101 and 106 days ; the prolonged duration of life in the latter two was due to the injection being in one partly intracoecal and in the other partly subcutaneous and partly intracoecal. Cultural Characters—The culture from Calf 301 grew well on the differential media, the growths obtained closely resembling those produced by the human tubercle bacillus (Group II.). The strain from Rat 15 was dysgonic. Summary.—The Calf 301 strain exhibited the cultural characters of a human tubercle bacillus, but resembled the bovine tubercle bacillus in its virulence. bacillus. The Rat 15 strain was identical with a bovine tubercle VIRUS H 16. “J.H.” RECAPITULATION. Tuberculous synovial membrane from knee joint. Carp! 57. Subcut. K. 67 days. Local Tuberculosis. cals 137, Subcut. K. 81 days. Local Tuberculosis. | Calf 273. K. 89 days. G.T. not severe. | | Calf 337. Calf 355. Subcut. Subcut. K. 80 days. K. (dying) 43 days. G.T. moderately severe. G.T. | Rabbit 66. | Bull 4234. K. (very ill) 42 days. G.T. | Culture. | Calf 559. K. (dying) 39 days. G.T. The original material in this case was the tuber- culous synovial membrane removed from the knee joint of an adult man; the membrane was emulsified and inoculated subcutaneously into two calves. The calves were killed 54 and 67 days after respectively, and showed in each case a small local lesion and slight disease of the nearest gland, and in one case a calcareous focus in a bronchial gland. An emulsion of the prescapular gland of one of the calves, No. 157, inoculated subcutaneously into another calf, No. 187, produced a small calcareous local lesion only. An emulsion of this local lesion and the adjacent gland was made and inoculated subcutaneously into Calf 273 ; this calf was killed in 89 days and showed a local lesion, enlargement and caseation of the pre- scapular gland, many tuberculous nodules in the lungs, some tuberculosis of the liver and spleen, thoracic and hepatic glands, and one parotid lymphatic gland. The prescapular gland of Oalf 273 was emulsified and inoculated into two calves subcutaneously (Nos. 337 and 355) and one rabbit intraperitoneally. The rabbit died of general tuberculosis. One of the calves died of general tuberculosis in 43 days, the other was killed in 80 days and showed general progressive tuberculosis. With an emulsion made from the pre- scapular gland of Calf 355, four calves and two rabbits were inoculated. The two rabbits died of general tuberculosis and all the calves developed general progressive tuberculosis. The virus from one of these rabbits (Rabbit 66) was inoculated into a bull (Bull 4234), which was killed when very ill 42 days later and showed general tuberculosis. The culture isolated from this bull was fully virulent. Cultures isolated from Calf 157, from a guinea-pig inoculated with the original material, and from the prescapular gland of Calf 273 were eugonic and had low virulence for the calf and rabbit; cultures from Calf 337 and Bull 4234 and Calf 559 were dysgonic and virulent. RESULTS OF REINVESTIGATION. Three strains of the virus were alive, those from Calf 273, Bull 423a, and Calf 559. Virulence Tests.—The culture derived from the prescapular gland of Calf 273 was retested after 2 years 114 months’ artificial cultivation ; the 36th generation of culture was used. Six rabbits were inoculated, two intravenously, two intraperitoneally and two subcutaneously. All were killed when well 94 days later and showed slight tuberculosis of a chronic type. The culture derived from the prepectoral gland of Bull 4234 was tested on a series of five rabbits, two intravenously, two intraperitoneally, and one subcutaneously, after 2 years 6 months’ artificial cultivation ; the 29th generation of culture was used. The two intravenous and the two intraperitoneal animals died of general tuberculosis in from 17 to 26 days ; the subcutaneous animal died of severe general tuberculosis in 126 days. The culture derived from the hepatic gland of Calf 559 was tested after 2 years 14 months’ artificial cultivation ; the culture used was the 25th generation. Five rabbits were inoculated, two intravenously, two intraperitoneally, and one subcutaneously. All the rabbits died of general tuberculosis. Cultural Characters—The culture from Calf 273 was eugonic ; the cultures from Bull 4234 and Calf 559 were dysgonic. Summary.—The Calf 273 strain was identical with the human tubercle bacillus, the other two strains with the bovine tubercle bacillus. 97676 B 10 VIRUS H 1%. “Sp. B.” (Mixed Sputum.) RECAPITULATION. Four calves were fed, one for 91, the others for 120 days with the mixed sputum from many patients suffering from pulmonary tuberculosis ; in each animal aretrogressive calcareous tuberculosis was found limited Human Sputum. | | Calf 161. Calf 169. K. 125 days. K. 169 days. practically to the glands of the alimentary tract. i eanlial The tuberculous mesenteric glands of one of the sr ae nn oul ame calves, No. a were emulsified and ee g 3 Be ace two calves subcutaneously ; one of the calves kille een : 2 SPH OE ganeh Ne. after 114 days showed no sign of tuberculosis, the K. 107 days. Calf 339. other (Calf 265) had slight retrogressive tuberculosis. Slight Tuberculosis. Subcut. A culture isolated from the latter calf, injected subeu- K. 69 days. taneously into two calves in 50 mg. doses, produced only slight tuberculosis. This passage did not proceed any further. The virus from two of the other original calves ] was also passed each through a series of animals, one Calf 475 through calves, the other through guinea-pigs, a calf, . and goats, witbout any change in virulence. The tuberculous mesenteric glands from the fourth calf (Calf 169) were emulsified and inoculated into a series of guinea-pigs, and from this series the virus was passed through another series of guinea-pigs and then Local Tuberculosis. 1 series of guinea-pigs. Intrav. K. 88 days. Tuberculosis chiefly of lungs and thoracic aie inoculated into a calf (Calf 339); this calf showed a Calf 539. local lesion and tuberculosis of the nearest glands only. Tatra: The virus after passage through another series of K. (dying) 18 days. guinea-pigs was inoculated intravenously into Calf 475. Early G.T. This calf was killed in 88 days and showed progressing | tuberculosis confined chiefly to the lungs and thoracic | pea rae a poate the lung was | : inoculated intravenously into Calf 539 which was ee ee ee killed when dying of general tuberculosis in 18 days. oa The virus from this calf was inoculated intraperi- D. ee ey ee ae toneally into Rabbit 181 (which died in 22 case of aincucied at eqate ae general tuberculosis), and was passed through a series Gdlaws des cradiccda of of four calves in succession. Bie Gcietane “ees Cultures isolated from Calves 265 and 339 were from the second of these ¢Ugonic and slightly virulent. A culture from Rabbit produced .fiial “eanersl 181 was eugonic and at first virulent ; when it had been Pecans i (exc oe cultivation 16 months it was found to be only slightly ‘aouulated éabeutanéousk virulent. The cultures isolated from calves which Y- died of general tuberculosis were dysgonic and virulent. RESULTS OF REINVESTIGATION. Only two strains of this virus were alive, the one derived from Calf 265, the other from Rabbit 181 ; Virulence Tests.—T he strain from Calf 265 was retested after it had been three years in artificial cultivation Six rabbits were inoculated, two intravenously, two intraperitoneally, and two subcutaneously. All were killed after 97 days and showed slight chronic tuberculosis of the usual type. : The strain derived from Calf 539 through Rabbit 181 was tested when two years and two months old (27th Eee a two intravenously, two intraperitoneally, and two subcu- taneously. were killed after ays and showed chronic tuberculosis of the type inoculation of a Group IT bacillus. ss a aa An earlier generation of this strain, i.e. the 11th, was also tested on rabbits; thi lt preserved in a cupboard and was 253 days old, too old ‘to obtain a culture direct from it ai wes ee inoculated into a guinea-pig and a culture was raised from the latter’s tuberculous organs ; the strain wh i 7k ae _ be Sarena into a ee of rabbits none of which developed progressive tuberculosis = ultura aracters.—The strain from Calf 265 and the two strains from Rabbi : he ee abbit 181 grew luxuriantly on Summary.—The strains from Calf 265 and Rabbit 181 exhibited the cult i Gor hee eee ultural characters and virulence of VIRUS H 49. “T.C.” RECAPITULATION. Original Material (Mesenteric Glands). _ This virus was obtained from a youth aged 18 who ‘ died of tuberculosis of the lungs ; several of the ulture. mesenteric glands were calcareous and as hard as a | stone. A culture was obtained di 1 0 8 a 2 irecti y fr m th | | mesenteric glands : the attempt t t j . ; ; Oo ob alin a culture from Subcut. Snbeut. The culture when two months ol i Died 81 days. K. 83 days. subcutaneously into two eee aa i oe G.T. Slight G:T. each and intraperitoneally into rabbits ; lt gave jah : : . one of the cal gressive tuberculosis and in the other (Calf 797) to a limited siege Meo rabbits to fatal general tuberculosis. , A culture obtained from the prescapular gland of Calf 797 (in which igi a limited retrogressive tuberculosis) when two months old sratuee ena Meee ea produced only 50 ey also in a dose of 10 mg. ; it was fully virulent also for rabbits Mo wallyes mh te-dose oF en nine months old, the original culture (7th subculture) failed tc ; rabbits, and when 14 months old (14th subculture) produced in : calf oo tee Pn tuberculosis in retrogressive tuberculosis and in rabbits slight non-progressive tuberculosis mg. only a very limited The original culture, therefore, when tested soon after its isolation exhibited a virul that of Group I. ; this virulence was increased by passage through the body of the IF t es tubercle bacillus and diminished by subculture on serum to the level of ‘ce a The original culture was, according to Dr. Cobbett, eugonic, rade II. ul generalised pro- it gave rise in the e somewhat below e level of i a human tubercle bacillus. » Bovine and was placed by Dr. Eastwood in his 11 RESULTS OF REINVESTIGATION. Four strains of the virus were in existence, two from the original material which bad been separately ‘subcultivated, and those from Calves 787 and 797. All four strains were tested on rabbits and two on calves. Virulence Tests.—The culture derived from the original material which had been cultivated by Cobbett in the ‘Blythwood laboratory was tested after 20} months’ cultivation (21st generation). Six rabbits were inoculated, two intravenously, two intraperitoneally, and two subcutaneously. One died of pseudo-tuberculosis in 11 days ; the others were all killed after 130 days and showed slight tuberculosis of a chronic type. Later on a culture ‘of the 25th generation was inoculated subcutaneously into two calves (Calf 1191 and Calf 1193) and two rabbits (doses 50 and 100 mg.) ; the calves after 94 and 99 days had only slight retrogressive tuberculosis, and each of the rabbits after 142 days had a local lesion only. The culture from the original material had been given shortly after its isolation to Dr. Eastwood, and had been subcultivated regularly on serum ever since at the Royalcot Jaboratory ; in order to ascertain whether this strain had also lost its virulence, I obtained a culture from him and inoculated it into a series of three rabbits ; the culture had then been 223 months in cultivation and was the 25th generation. Two of the rabbits -were inoculated intravenously and one intraperitoneally ; the former died of general tuberculosis in 16 and 31 days respectively ; the latter was killed after 163 days and showed chronic general tuberculosis, the injection having been partly intracoecal ; the type of disease produced in the three cases was identical with that produced ‘by a bovine tubercle bacillus. The culture derived from Calf 787 was tested after 16 months’ artificial cultivation. Six rabbits were inoculated, two intravenously, two intraperitoneally, and two subcutaneously. All died of general tuberculosis in from 14 to 90 days. The culture derived from Calf 797 was also tested after 16 months’ artificial cultivation. Six rabbits were inoculated, two intravenously, two intraperitoneally, and two subcutaneously. One intraperitoneal animal died in 12 days of acute tuberculosis, complicated with severe psorospermosis of the liver ; the others all died of general tuberculosis in from 13 to 80 days. Cultural Characters.—The Blythwood strain of the original culture and a strain from each of the two -calves inoculated with it (Nos. 1191 and 1193) grew well on glycerin media, as did also the Royalcot strain. The two strains from Calf 787 and 797 did not grow so well as the strains from the original material, and resembled the more easy growing strains of bovine origin. Summary.—The Blythwood strain of the original culture exhibited the cultural characters and virulence of a human tubercle bacillus: the Royalcot strain had the cultural characters of a human tubercle bacillus but resembled a bovine tubercle bacillus in virulence ; the strains derived from Calves 787 and 797 were identical with bovine tubercle bacilli. SUMMARY OF PRELIMINARY EXPERIMENTS. Ten strains from the four viruses (H 13.“ A.D.”, H 16. JS.H.”, H 17.“Sp.B.”, and H 49. T.C.”), which during passage through the animal body underwent a change of character have been retested as to their cultural characters and virulence for rabbits. Two strains from the virus designated H 2.“Sp.A.”, the original material of which was mixed sputum have also been retested [this virus was passed through a series of animals, and from some a slightly virulent, from others a virulent culture was isolated ]. With two exceptions the cultural characters and the virulence of the twelve strains were found strictly in accordance ; those strains which grew well on culture media had low virulence for rabbits, those which grew with difficulty were highly virulent for rabbits. The preliminary tests therefore gave no indicaticn of the presence in any of these ten strains of two different kinds of organisms, and no further investi- gations with them were carried out. Whatever they had been they were now stable. The first exception was the culture isolated from the mediastinal gland of ‘Calf 301 (Virus H 13.“ A.D.”), an animal which died of general tuberculosis after the subcutaneous inoculation (in passage) of an emulsion from the tuberculous organs of guinea-pigs ; the culture was eugonic, and when tested by Dr. Cobbett after 15 months artificial cultivation produced in two rabbits slight chronic general tuber- -culosis, and in two calves inoculated with 50 milligrammes subcutaneously limited tuberculosis in one, and slight general tuberculosis in the other. When retested by me after a period of three years’ artificial cultivation it grew luxuriantly on glycerin media, and produced in two calves generalised tuberculosis, not severe and not apparently progressive, and fatal general progressive tuberculosis in rabbits. It was niore virulent therefore than when tested by Dr. Cobbett, but less virulent than a bovine tubercle bacillus. The second exception was the culture derived directly from the mesenteric glands ‘of a youth aged 18 (Virus H 49.“ T.C.”). This culture had been cultivated in two different laboratories, in the Blythwood laboratory in charge of Dr. Cobbett and. ‘since an early generation, in the Royalcot laboratory in charge of Dr. Eastwood. The former, called the Blythwood strain, had been tested by Dr. Cobbett two and seven months after isolation, and produced in calves and rabbits fatal general _progressive tuberculosis ; it was again tested 13% months after its isolation and was ‘found to have lost its virulence. The latter, or Royalcot strain, was tested by me when it had been in cultivation about two years, and was found to be viruient for the calf and rabbit, though not so virulent as a bovine tubercle bacillus. Both of ‘the strains grew luxuriantly on artificial media, and were in this respect identical. With these two exceptional strains further experiments were made (vide infra). 12 RESULTS OF THE SPECIAL INVESTIGATION OF TWO OF THE GROUP IIL CULTURES [H 13. “A.D.” (CALF 301), AND H 49. “7.0” (0.M.)| WHICH EXHIBITED EXCEPTIONAL FEATURES. SEPARATION EXPERIMENTS. ~ INTRODUCTION. The preliminary experiments with various strains of the viruses which had been placed in Group ILI. showed that two, i¢., the Calf 301 strain of Virus H 13.“ A.D.” and the strain from the original material of Virus H 49. “ TAs though they grew like the slightly viruient human tubercle bacillus, were never- theless able to set up in rabbits generalise] progressive tuberculosis. They behaved in fact like artificial mixtures of human and bovine tubercle bacilli. For purposes of further investigation therefore they were assumed to be mixtures of this nature and attempts were made in each case to separate the component elements. In an artificial mixture of human and bovine tubercle bacilli a separation of the two kinds of bacilli may be effected in three ways. : 1. By passing the culture through the bodies of animals (e.g. calves and rabbits) which permit the multiplication of one kind of bacillus and resist that of the other. 2. By subculture on media containing a substance (glycerin) which is more favourable to the growth of one kind of bacillus than it is to that of the other. 3. By a method of plate culture. All the three methods have been employed in the case of each strain. The first method depends for its success on the fact that the bovine tubercle bacillus is able when inoculated subcutaneously to set up progressive tuberculosis in the calf and rabbit whereas the human tubercle bacillus (Group II.) is not so able. In these animals after subcutaneous inoculation there is with both kinds of bacilli an initial dispersal all over the body but the human tubercle bacillus tends gradually to disappear while the bovine tubercle bacillus multiplies and gives rise to definite tuberculous lesions. The human tubercle bacillus remains alive however in the tissues of the calf and rabbit for a long time and may conceivably be present within lesions produced by the bovine tubercle bacillus. One passage through the animal body may not therefore be sufficient to. eliminate entirely the possibility of cultivating the human tubercle bacillus along with the bovine unless the duration of life of the animal be long and the dose of slightly virulent bacilli be small. The rabbit is more susceptible than the calf to the human tubercle bacillus and after subcutaneous inoculation of even moderate doses there may be a considerable number of tuberculous lesions in the lungs and kidneys, and since the lungs and kidneys are often the only organs affected in rabbits inoculated subcutaneously with bovine tubercle bacilli, elimination of the human tubercle bacillus in a mixture would be expected to be less certain in the rabbit than in the calf; and this experiment has shown to be the case. in the calf there is a greater range of lesions to choose from and the cultures can be raised from regions which only rarely show naked eye evidence of the presence of the human tubercle bacillus : it has been shown in several instances in this investigation that if glands (6.9.5 the mediastinal and portal) are chosen which frequently show lesions after the subcu- taneous ineculation of human bacilli (Group II.) there exists the possibility of obtaining in a culture both kinds of bacilli together. Since one passage, either through the body of the calf or the rabbit could not be relied upon absolutely to. eliminate the human tubercle bacillus the culture was in the majority of cases passed through more than one animal ; in the case of the calves cultures were always isolated from distal lesions as well as the nearest gland. While the method just clescribed eliminates the eugonic element in a mixture, the second method—cultivation on glycerin media—is applied to eliminate the virulent element. On glycerin media bovine tubercle bacilli grow, as a general rule, less well than the human tubercle bacillus, and in a culture containing both human and bovine tubercle baciili would after several generations be expected to be finally overgrown Many bovine tubercle bacilli, however, grow well on glycerii media, and all, even the most dysgonic, grow more luxuriantly when repeatedly subcultured on glycerin media. The method would therefore fail to eliminate the bovine tubercle bacillus if the 18 latter were well adapted from-the outset to grow on glycerin media. It might be at once successful, 7.¢., after one subculture, if the bacillus grew very badly ; there is a certain class of bovine tubercle bacilli which begin to grow on glycerin serum apparently only during the second or third weeks, when they produce discrete colonies ; the portion ‘of culture selected for the second glycerin subculture might within: this period very easily not contain any of these colonies, and the strain would then be found to have lost its virulence ; but if the culture transplanted on to the second glycerin serum tube included any of the colonies, the bacilli which gave rise to them would tend not gradually to disappear but to be per petuated, since the most dysgonic tubercle bacilli are improved in their growth capacity after one subculture on a glycerinated medium. Subculture on glycerin media may be successful therefore, but cannot be relied upon in every case to eliminate the bovine bacillus in a mixture of human and bovine tubercle bacilli. Dr. Cobbett used the method extensively, and was successful in eliminating the virulent element from an artificial mixture and from a natural mixture; in both of his cases the virulent bacillus was a dysgonic bacillus. Though the invariable replacement cf a eugonic virulent strain by a dysgonic virulent strain by passage through the bodies of calves and rabbits, and though the complete loss or diminution of the virulence of the culture by growth on glycerin media would point strongly to the presence originally of two different kinds of bacilli, the only conclusive evidence of mixture would be furnished by the actual isolation from the culture, by means of plate culture, of strains with different and distinctive properties. On account of the tendency of tubercle bacilli to adhere together in masses, and the impossibility of obtaining a homogeneous suspension, the application of the ordi- nary plate methods to a culture is only very rarely likely to be attended with success. Ina carefully prepared suspension of a culture there may be many single bacilli which may give rise to colonies, and these the investigator may be fortunate enough to select for investigation. But to determine whether or no a culture of tubercle bacilli is composed of one kind of bacillus involves a considerable amount of work —the culture must be tested, not only as to its characters on artificial media, but also as to its virulence for rabbits ; naked eye characters alone of colonies are not sufficient to differentiate between the bovine and human bacilli ; ifa glycerin medium were used for the separation experiments, a small non-pigmenjed colony might probably be shown to be composed of virulent bacilli only, but it would be impossible, as the necessity for the present investigation shows, to say from cultural characters alone whether a culture which grows readily had notan admixture of dysgonic bacilli. It was very important, therefore, in order to avoid unnecessary labour, that the bacilli in the material used for sowing the plates should be as isolated as possible. In emulsions of the tuberculous lesions of the guinea-pig, the isolation of individual bacilli, though not absolute, is very much greater than could possibly be secured by any method of emulsification of a culture. In each case, therefore, the culture was inoculated into a guinea-pig, and the plates were sown with an emulsion made from its tuberculous tissues. The guinea-pig is a most suitable animal for the purpose since it is almost equally susceptible both to the human and to the bovine tubercle bacillus, and the relative proportion of the two kinds of bacilli is not therefore likely to be seriously altered by the passage. The medium used for the plates was bovine serum with the addition of 5 per cent. glycerin ; the glycerin was added in order to assist in the differentiation of the colonies, for on glycerinated serum the human tubercle bacillus grows generally with greater ease than the bovine bacillus, and often forms pigmented colonies. The Petri dishes were filled to a depth of about 2 mm., and the serum was coagulated in the usual way ; after sowing, the plates were sealed with paraffin. The method of separation was by surface cultivation, and plates were used ,merely for convenience and to secure as large a surface as possible ; ; large test-tubes containing glycerin serum were always sown at the same time as the plates, and in some instances colonies from these tubes were removed for investigation. The emulsion was diluted to an extent sufficient to secure isolated colonies, and spread thinly over the surface of the plates by means of a sterile camel’s hair brush. It was found to be possible, however, to pour plates in the usual way. This was done in one case, but only one of the colonies produced was investigated : the medium used in this instance was agar with the addition of 10 per cent. serum; the agar was melted and then cooled to 45° C. ; the serum and a loopful of the emulsion was then added, and the mixture poured into the plates. 27676 G 14 VIRUS H 13. “A.D.” (Calf 301 strain.) SEPARATION BY ANIMAL INUCULATION. (See Tables A and B.) Calves,—T wo calves were inoculated subcutaneously on March 23, 1907 ; one, No. 1177, with 50 re eT the other, No. 1175, with 100 milligrammes of the culture derived from the mediastinal gland of Ca Rapiae the culture had then been three years two months in artificial cultivation and had been maintained during n on egg. this period on ith epti tion which was grow g . ; ; Base ante Cree aT Bee Both calves reridaed well and were killed in good health 81 and 110 days later respectively; they Culture from Calf 301. showed at the post-mortem examination generalised tuberculosis, not severe and not apparently pro- | | gressive. Calf 1175. Calf 1177, An emulsion of the caseous prescapular gland of Dose : 100 mg. Dose : 50 mg. Calf 1175 was prepared and 20 cubic centimetres Subcut. Subcut. inoculated subcutaneously into Calf 1237. The calf K. 81 days, K. 110 days. was killed when ill after 52 days and showed severe P.M.—G.T. P.M.—G.7?. general tuberculosis. : | Rabbits were inoculated subcutaneously with Oalf 1237, emulsions of tuberculous tissues from each of the Dose: 20 ce, ° first two calves, the prescapular, a mediastinal, and a Subcut. popliteal gland in one case, the prescapular gland and K, (ill) 52 days. the spleen in the other; all died of general tuber- P.M.—G.T. culosis in periods varying from 47 to 167 days. ! Cultures were isolated from each of the three Culture calves (1175, 1177, and 1237), from the prescapular from bronchial gland. gland as well as from metastatic lesions and from | several of the rabbits; altogether eight strains from Calf 1327. calves wereinvestigated in culture and twofrom rabbits. Dose : 50 mg. With one exception all the strains grew badly on : Subcut. artificial media ; on glycerin serum growth was rather Died 27 days. better than on serum alone, on glycerin agar a patchy P.M.—G.T. grey layer was formed in which discrete colonies subsequently developed, on potato growth was thin grey and uniform, and on broth thin incomplete and broken up into islands. The exceptional strain was obtained from one of the mediastinal glands of Calf 1175; this behaved like the original strain, growing well on artificial media and producivg fatal general tuberculosis in rabbits ; for the further investigation of this strain see under “ Separation by Plate Cultures.” The strain derived by culture from the bronchial gland of Calf 1237, the second calf of the series, was inoculated subcutaneously in a dose of 50 milligrammes into Calf 1327 and in doses of 10 mg. into each of two rabbits ; the calf died of miliary tuberculosis in 27 days and the rabbits of general tuberculosis in 37 and 68 days. This strain was identical with a bovine tubercle bacillus therefore both with respect to its cultural characters and its virulence. Rabbiis—As has already been mentioned, a preliminary series of rabbits was inoculated with the Calf 301 strain to ascertain its virulence. All these animals died of general tuberculosis, and from two of them cultures were isolated ; one of the rabbits, No. 1137, had been inoculated intravenously and the culture was obtained from théspleen ; the other, No. 1140, had been inoculated subcutaneously and the culture was obtained from the lung. In each case the culture grew irregularly ; on most of the tubes sown scanty growths were obtained, but, occasionally, more especially on potato, the growths were very good ; the cultures behaved as if they were composed of poorly growing bacilli with a slight admixture of easy growing bacilli. Each culture when it had been in cultivation in one case 212 days, in the other 188 days, was inoculated subcutaneously into a rabbit. Both died of general tuberculosis, one in 62, the other in 29 days. The cultures isolated from these animals grew badly on all media and resembled precisely the poorly growing strains obtained from the calves. Two rabbits were incculated subcutaneously with the original culture at the same time as Calves 1175 and 1177, one with 50 mg., the other with 10 mg.; the former died in 28 days of general tuberculosis ; the latter was killed when ill after 107 days and showed chronic general tuberculosis. A culture isolated ‘from the latter animal was dysgonic and resembled the poorly growing strains isolated from the calves. The result therefore was the same, whether the culture was passed through the body of the calf or of the rabbit, the eugonic element was eliminated and there was obtained a dysgonic fully virulent bacillus, SEPARATION BY GLYCERIN MEDIA. (See Table C.) The original culture was subcultivated on glycerin serum for two generations, and then inoculated ‘ subcutaneously into a calf in a dose of 100 mg. and two rabbits in doses of 50 and 10mg. Both rabbits died of general tuberculosis in 103 and 100 days respectively. The calf was killed after 92 days, and showed general tuberculosis very similar in extent and distribution to that produced in the two calves inoculated with serum culture. Four rabbits were inoculated with tuberculous tissues from this calf, two from the prescapular gland and two from a mediastinal gland; one of each pair died prematurely; the other two died of ceaaral tuberculosis in 92 and 53 days respectively. Cultures were isolated from the prescapular gland and a mediastinal gland of the calf and from a rabbit inoculated with the prescapular gland. All the strains grew poorly on glycerin media in spite of the fact that they had resided for two generations on glycerin serum. Subculture on glycerin serum therefore failed to eliminate the virulent element from th : virulence was nevertheless reduced, as shown by the long duration of life of the two paphita ted subcutaneously. e SEPARATION BY PLATE CULTURES. (See Tables D, E, and F,) To furnish material for sowing the plates, a guinea-pig, No. 2491, was inoculated int i j emulsion of a large colony from an egg tube sown with the Calf "301 strain ; the ee The guinea-pig was killed 25 days later and found to have early general tuberculosis: an emulsion ad ad : from a caseous sternal gland, and spread over the surface of glycerin serum plates with a camel’s hae brush. After several weeks incubation the plates were removed from the incubator and ined ; found to be covered with growth which in the main consisted of two quite distinct kinds of colonies eee Tied 15 was small a i i ing i ha al ae not pigmented, the other was large yellow and wrinkled, spreading in some cases up Eight of the large colonies and two of the small ones were separately subculti i These strains after subculture were tested on rabbits and on the differential ao eagles The strains raised from the two small colonies (Colonies 4 & 5) were highly virulent for rabbits, and grew poorly on the differential media ; one (Colony 5) was inoculated subcutaneously in a dose of 50 mg. into a calf (1309) which died of acute tuberculosis in 30 days. Both these strains therefore were identical with a bovine tubercle bacillus, The strains from the large colonies grew more luxuriantly than those from the small colonies on all the media used ; on serum they produced a yellow pigment, on potato and agar some of the strains grew very abundantly, others only moderately well. Hach of the strains was tested on rabbits subcutaneously ; the doses were not estimated, but were in every case very large, consisting of the whole or half of the gro wth on a large glycerin serum tube. Three of the rabbits, Nos. 1516, 1519, and 1521, inoculated respectively with culture from Coloni and 8, had when killed 160 days later besides a local lesion only slight fabsreillagis of the lungs and one ote tubercles in the kidneys. The amount and extent of the disease in these animals were not greater than might have been produced by a Group II. bacillus, but on the other hand they were not less than might have been produced by a small dose of a Group I. bacillus. In order, therefore, to ascertain whether the particular strains used for these rabbits were composed of one kind of organism only, it was necessary to isolate and test cultures from metastatic lesions. A culture was accordingly raised from the lung of each of the three rabbits (1516, 1519, and 1521). These cultures grew well on the differential media, and were inoculated intravenously into two rabbits in doses of 0-1 and 0:01 mg. The results showed that in the cultures inoculated there were no bacilli capable of giving rise to progressive tuberculosis in the rabbit. The three colonies were composed solely therefore of eugonic slightly virulent bacilli. The other five strains (from Colonies 1, 2, 7,9, and 10) each produced in the rabbit progressive tuberculosis and from the lesions in two of the rabbits dysgonic bacilli were isolated. Two of the colonies (Colonies 2 and 9) were retested, the former on a calf (Calf 1301) and two rabbits, the latter on a series of three rabbits. The animals in each series died of general tuberculosis, and from the portal gland of the calf (Colony 2 strain) and from one of the rabbits inoculated with Colony 9 strain, dysgonic cultures were isolated. : These five strains resembled therefore the original culture. In the event that the experiments with the strains from Guinea-pig 2491 were not successful in separating the virulent from the eugonic element, an emulsion made from one of the large colonies (Colony 11) on a plate sown from Guinea-pig 2491 was inoculated intraperitoneally into Guinea-pig 2605 and plate cultures were sown with emulsions made from the omentum and spleen; a rabbit inoculated subcutaneously at the same time as the guinea-pig died of general tuberculosis. A culture was raised on egg in the ordinary way from the omentum and spleen of the guinea-pig ; these strains grew luxuriantly on artificial media and produced general tuberculosis in a series of two and four rabbits; the duration of life was however longer than in rabbits inoculated with equivalent doses of a bovine tubercle bacillus. , Numerous colonies appeared on all the plates but on one, sown from the omentum, they were sufficiently discrete to enable one to pick off separate colonies. The colonies were of two kinds; some were small, flat, and of a greyish colour, the others were a little larger, more raised, and of a canary yellow colour. Eleven colonies, four grey and seven yellow, were selected and separately subcultivated. On glycerin potato three of the strains from grey colonies grew poorly, the fourth grew well ; all the strains from yellow colonies produced thick wrinkled pigmented growths. One of the strains from a grey colony (Colony 4) and five from yellow colonies (Colonies 6, 7, 8, 9, and 16) were inoculated each into two rabbits intravenously. The two rabbits inoculated with the Colony 4 strain died of miliary tuberculosis in 18 and 28 days; this strain was therefore dysgonic and fully virulent. Two of the strains, raised from the yellow Colonies 8 and 10, produced progressive tuberculosis in rabbits ; they resembled therefore the original culture. The three remaining strains from the yellow Colonies 6,7, and 9 were inoculated each into two rabbits ; one died in 69 days and showed slight tuberculosis of the lungs and kidneys ; the other five remained well and when killed in from 104 to 119 days showed the type of tuberculosis commonly met with after the intravenous inoculation of slightly virulent human tubercle bacilli. Two of these strains, from Colonies 6 and 7, were each inoculated subcutaneously into a calf, one in a dose of 59 mg., the other in a dose of 78 mg.; they were killed 76 and 84 days later respectively and each showed a local lesion and tuberculosis of the nearest glands only (in one of the calves there was a doubtful focus in a bronchial gland and a doubtful tubercle in the lung). The emulsion from the omentum of Guinea-pig 2605 was also plated in the way ordinarily used for separating the commoner organisms. The medium used was agar, to which had been added 10 per cent. serum. Sparsely scattered colonies were obtained on all the plates, some growing on the surface, some in the depth ; one of the surface colonies was removed and separately cultivated ; the culture grew luxuriantly on glycerin media ; three rabbits (Nos. 1777-1779) were inoculated with it, two intravenously and one subcutaneously. The results showed that the culture was composed of the slightly virulent type of bacillus only. The investigation of the strains from Guinea-pig 2605 has shown therefore that of the seven tested in culture and by inoculation one was dysgonic and virulent and resembled a bovine tubercle bacillus, four were eugonic and slightly virulent and resembled a human Group II baciilus, and two were eugonic and virulent and were like the original culture. The culture isolated from the mediastinal gland of Calf 1175 closely resembled the original culture—it grew well on glycerin media and was virulent for rabbits—and an attempt was made by means of plate cultures to separate a eugonic slightly virulent bacillus and a dysgonic virulent bacillus. A guinea-pig (No. 2621) was inoculated with a 70 days’ old glycerin serum culture and large tubes of glycerin serum were sown from itz tuberculous spleen and sternal gland. On these tubes two kinds of colonies could be distinguished, grey and yellow. Four of the yellow colonies and two of the grey were taken and separately subcultivated. After several subcultures on serum the four strains from yellow colonies (Colonies 1, 2, 3, and 4) were inoculated into rabbits, the two former intravenously each into two rabbits and the two latter subcutanecusly each into one rabbit. The first two colonies produced progressive tuberculosis in rabbits; the two rabbits inoculated with the other two colonies died prematurely in 56 and 41 days and showed slight tuberculosis, The two strains from grey colonies (Colonies 5 and 6) inoculated each into a rabbit subcutaneously, produced fatal general tuberculosis identical with that produced by a bovine tubercle bacillus. The culture raised in the ordinary way from the spleen of Guinea-pig 2621 was eugonic and virulent. From each of three different strains of the Calf 301 culture therefore, cultures raised from separate colonies have been obtained whick are identical some with a bovine tubercle bacillus, others with a human tubercle bacillus. © 27676 C2 16 VIRUS H 49. “TG.” (Strain from the original material.) SEPARATION BY ANIMAL INOCULATION. (See Tables G, H, and L.) Culture from O.M. Calves.—Calf 1213 was inoculated subcutaneously with 50 mg. of (Royalcot strain.) the original culture ; the strain had then been 746 days in artificial cultivation, and had been maintained during this time on serum Calf 1213. alone ; a series of four rabbits was inoculated at the same time as the Subcut. calf. The calf was killed in good health 80 days after inoculation, 50 mg. and showed general progressive tuberculosis of a chronic type. All K. 80 days. the rabbits developed fatal general tuberculosis, but the duration of Chronic G.T. life was prolonged as compared with that of rabbits inoculated with | bovine tubercle bacilli. Calf 1263. An emulsion was prepared from the enlarged caseous prescapular Subcat. gland of Calf 1213, and inoculated subcutaneously into Calf 1263 ; E. of Prescap. Gland. two rabbits were also inoculated subcutaneously with the prescapular K. 91 days. gland emulsion, and two with an emulsion made from a mediastinal Chronic G.T. : gland. : . ; 3 The calf was killed in good health 91 days after inoculation, and Culture from thoracic gland. showed chronic general tuberculosis, the lungs being covered with i “perlsucht ” growths ; all the rabbits developed generalised progres- Calf 1347. sive tuberculosis. Subcut. Two rabbits were inoculated subcutaneously with tuberculous : 50 mg. tissues from Calf 1263, one with an emulsion of the prescapular K. (ill) 52 days. gland, the other with an emulsion from a portal gland; the latter Severe G.T. died prematurely, the former was killed after 232 days and showed chronic progressive tuberculosis. Cultures were isolated from each of the two calves, 1213 and 1263, from the prescapular gland as well as from distal lesions. The strain from the mediastinal gland of Calf 1213 grew luxuriantly ou broth, and moderately well on the other media. ‘he other strains from this calf, as well as all those from Calf 1263 grew much less well than the original strain, and were indistinguishable from a bovine tubercle bacillus. ; ; The culture derived from the mediastinal gland of Calf 1263 was tested subcutaneously on a calf in a dose of 50 mg., and on two rabbits in doses of 10 and 7:3 mg. The calf (1247) was killed when ill 52 days after inoculation and showed generalised progressive tuber- culosis ; tbe rabbits died in 74 and 78 days respectively of general tuberculosis. The culture obtained from the prescapular gland of Calf 1213 was also tested on rabbits ; twelve colonies were taken from an egg tube and separately cultivated for three generations ; the growth from one serum tube in the case of each strain was then inoculated subcutaneously into a rabbit ; nine of the rabbits died in from 23 to 91 days of general tuberculosis, the other three were killed after 108 days, and all showed generalised progressive tuberculosis. Rabbits.—Cultures were isolated from two of the three rabbits inoculated to test the virulence of the origina! strain. One of the rabbits, No. 1181, died 31 days after an intravenous inoculation of 0°1 mg., and cultures were obtained from the lung and a kidney; the other rabbit (No. 1182) had been inoculated intraperitoneally with 1-0 mg. of culture, and died of general tuberculosis in 106 days, the long duration of life being due to the fact that the injection was partly subcutaneous ; the culture in this case was taken from a pleural nodule. The culture from the latter rabbit was dysgonic aud resembled the strains from the calves. The cultures from the other rabbit (Rabbit 1181) were irregular in their growth ; on potato for example very good growths were occasionally obtained, while on other media the growths were usually poor. An emulsion of the lung of Rabbit 1181 was inoculated subcutaneously into two rabbits ; both animals died of general tuberculosis, one in 74, the other in 82 days. A culture isolated from the scapular gland of one of the rabbits, No. 1219, was dysgonic and resembled the strains from the calves. By passage through the body of the calf, as well as through that of the rabbit therefore the eugonic element in the culture has been eliminated, and a dysgonic virulent culture obtained. : SEPARATION BY GLYCERIN MEDIA. (See Table J.) A separation of the virulent element from the easy growing element by means of glycerin unwittingly. A good growth had been obtained on a bbe of Slyeevin agar aud with the Sbject oF eee preponderance of easy growing bacilli in the material for sowing plates this culture was used to inoculate a guinea-pig, No. 2490. The guinea-pig developed tuberculosis and plates of glycerin serum were sown with an emulsion made from the caseous sternal gland. Colonies were obtained sufficiently discrete and subcultures were made from twelve ; the colonies varied in size but uot in type and a selection was made of large and small. , : After two subcultures on serum the strains were tested on glycerin potato ; the growths produced on some of the potatoes were very good, on others poor. This preliminary test seemed to indicate that a separation of a dysgonic and a eugonic bacillus had been effected. Six of the strains, representing both easy growing and poorly growing strains were therefore tested on rabbits, one rabbit being inoculated subcutancousl with on serum culture of each strain. All the rabbits remained well and when killed in periods varying fon 94 to 99 days showed slight and retrogressive tuberculosis only ; in each rabbit there were some miliar tubercles i the lungs and in two very slight tuberculosis of the kidneys. Cultures were isolated from cath ong of th ie avimals, four being nee nae ee from local lesions and one from a scapular gland. All the strains were eugonic. e scapular gland culture was tested on two rabbits i ‘ daly aliehtly viwenk P 8 its intravenously and found to be Subsequently Eek ati the twelve strains were tested on rabbits, subcutaneously, and three of the six that had been inoculated subcutaneously were r i on two rabbits. The results in all these rabbits showed that the strains had Only ett ols nensnelyy race Strain 9 was also inoculated into a guinea-pig (No. 2816) and cultures were obtained from the s lee d sternal gland. Both strains grew well and the spleen strain had only slight virulence for the rabbit reais When it was realised that all the colonies isolated from Guinea-pig 2490 were of one type and onl slight] virulent for the rabbit the culture obtained from the guinea-pig in the ordinary way i.e. ue leasing the daetiee two intravenously and two Ld of an egg tube, and arising therefore from a number of colonies, was inoculated into a series of four rabbits, three intravenously and one subcutaneously. None of the rabbits developed fatal tuberculosis, the disease produced in all being consistent with the inoculation of a Group II. bacillus. _ It is clear from these experiments that there were no virulent bacilli in the culture inoculated into the guinea-pig and that subculture on a medium containing glycerin had resulted in the elimination of the virulent element from the virulent eugonic culture. ! The experiments show further that size of colony alone cannot be relied upon as a means of differentiating two kinds of organisms SEPARATION BY PLATE CULTURES. (See Tables J and K.) As has already been described the first attempt to separate different types of bacilli from the culture bv means of plate cultures was unsuccessful ; it had failed because cultivation on a glycerin medium had caused the elimination of the virulent element. In a second experiment I started with a culture which had not been subjected to the influence of glycerin. The culture in its 35th generation (2 years, 11 months old) was inoculated intraperitoneally into Guinea- pig 2918, and a generation later was tested on a series of four rabbits (see Table G) in order to see whether the original culture maintained on serum had retained its virulence ; all the rabbits died of general tuberculosis, the oe of life being a little prolonged as compared with that after the inoculation of bovine tubercle bacilli. The guinea-pig died in 14 days of tuberculosis, and emulsions were made from the sternal gland and spleen. Four glycerin serum plates were sown with the emulsions as well as a number of egg tubey and large glycerin serum tubes, Numerous colonies grew on the tubes while on the plates the colonies were very sparsely scattered ; they varied in size but did not differ in any other respect. A subculture made in the ordinary way from an egg tube produced luxuriant growths on the glycerin media. : . Eighteen colonies, twelve from the plates and six from the glycerin serum tubes, were taken and separately subcultivated on serum ; after four subcultures all the strains were tested on glycerin potato in order to gain a preliminary impression as to the properties of the strains. . ' Five of the strains grew very well, producing yellow wrinkled or warty layers; of the rest, some grew very poorly, wnile the others produced intermediate amounts of growth. Four of the eugonic and one of the dysgonic strains were selected for inoculation into animals. Each strain was inoculated intravenously into a series of three rabbits. Two of the eugonic strains (from Colonies 5 and 7) produced slight retrogressive tuberculosis in rabbits. Fifty milligrammes of culture from Colony 7 produced in a calf (Calf 1415) inoculated subcutaneously local tuberculosis only. These two strains therefore contained no virulent bacilli. The other two eugonic strains (from Colonies 3 and 13) produced fatal general tuberculosis in all the rabbits inoculated, and one of them (from Colony 3) gave rise in a calf (Calf 1387) inoculated subcu- taneously with 50 mg. to general tuberculosis fatal in 58 days. These two strains exhibited therefore the properties of the original culture. The dysgonic strain (from Colony 17) produced rapidly fatal general tuberculosis in all the rabbits inoculated. It was also inoculated subcutaneously into two calves in doses of 50 mg. (Calf 1413) and 48 mg. (Calf 1501). The former was killed after 107 days and showed slight retrogressive general tuberculosis, the latter died in 45 days of general tuberculosis. Thus by the use of the plate method of separation strains of tubercle bacilli with the properties of the human and of the bovine tubercle bacillus respectively have been isolated from the H 49. “ T.C.” culture. SUMMARY AND CONCLUSION. The second part of this report is concerned with two cultures—H 13. “ A.D.” Calf 301 strain, and H 49. “T.C.,” strain from the original material (mesenteric gland)—which had been shown to possess in common the capacity to grow luxuriantly on artificial media and to produce general progressive tuberculosis in calves and rabbits. Each of the cultures has been specially investigated with the object of ascertain- ing whether the want of that accordance which usually obtains between cultural characters and viruience was due to a mixture of bacilli, with the properties of the human and the bovine tubercle bacillus respectively. To each culture, three methods of separation have been applied, viz. :— (1.) Animal inoculation: (2.) Cultivation on glycerin media: (3.) Plate cultivation.— With the following results :— By passage through the bodies of calves and rabbits the eugonic element in each culture has been eliminated, the culture finally obtained being dysgonic and virulent. Cultivation on glycerin media has been successful in eliminating the virulent element in one case only, H 49. “T.C.” ; in this case a eugonic slightly virulent culture was obtained after one subculture on glycerin agar. By means of plate culture, dysgonic virulent strains and eugonic slightly virulent strains have been obtained at one and the same time, from each of the ‘two cultures. The experiments show conclusively therefore, that each of these cultures is a mixture of two kinds of organisms, one with the properties of the bovine tubercle bacillus, the other with the properties of the human tubercle bacillus. 18 RESULTS OF THE SPECIAL INVESTIGATION OF THE TWO CULTURES H 60. “ W.B.” (bronchial gland) and H 90. “IP.” (retroperitoneal gland), ISOLATED SINCE THE SECOND INTERIM REPORT, WHICH EX- HIBITED THE FEATURES OF THE TWO GROUP III. CULTURES. SEPARATION EXPERIMENTS. SUMMARY OF RESULTS. Among the strains of culture isolated from human tuberculous lesions and investigated since the issue of the second interim report there were two which dis- played the peculiarity of the two Group III. cultures [H 13. “A.D.” (Calf 301) and H 49. “T.C.” (mesenteric gland—Royalcot strain)] of which the results of the reinvestigation are given in the second part of this report. ; This peculiarity was the association of high cultural luxuriance with the power to set up general progressive tuberculosis in calves and rabbits when inoculated subcutaneously. The cultures (i.e. the bronchial gland strain of Virus H 60. “ W.B.” and the retroperitoneal gland strain of Virus H 90. ‘1.P.”) were investigated therefore in exactly the same manner as the .two above-mentioned Group III. cultures, with the object of ascertaining whether or no their exceptional properties were due to the association of virulent tubercle bacilli with eugonic slightly virulent tubercle bacilli. The culture from Virus H 60. ‘“ W.B.” was isolated from a guinea-pig which had been inoculated with an emulsion of the caseous bronchial glands of a child aged four years and seven months who died of general miliary tuberculosis. The culture was equal in virulence to a bovine tubercle bacillus, producing fatal general tuberculosis in calves after subcutaneous inoculation, but grew more readily on artificial media than bovine tubercle bacilli. Cultures from the meninges, lung, and mesenteric gland of the child grew luxuriantly on glycerin media and were only slightly virulent for calves and rabbits.* The strain of Virus H 90. “I.P.” investigated was obtained directly from the retroperitoneal gland of a man aged 70 who died of pneumonia. The culture grew luxuriantly on artificial media like the human tubercle bacillus ; it produced in calves slowly progressing tuberculosis not severe and not fatal within the period of observation, and in rabbits progressive tuberculosis not so quickly fatal as after the inoculation of equivalent doses of bovine tubercle bacilli. The culture was therefore not so virulent as the H 60. ‘“‘ W.B.” strain. A culture isolated from the mesenteric gland of the same patient through the guinea-pig also grew luxuriantly, but was only slightly virulent for the rabbit. Cultures obtained from distal lesions produced in the rabbits as well as the calves. inoculated with the original strains (H 60. ‘‘ W.B.” bronchial gland, and H 90 “I.P.” retroperitoneal gland) grew with difficulty on artificial media and were fully virulent ; the capacity to grow luxuriantly therefore was lost by both cultures in their passage through the bodies of the calf and rabbit, and bacilli were obtained which exactly resembled bovine tubercle bacilli in cultural characters and virulence. * Dr. Eastwood who tested the cultures shortly after their isolation found that the bronchial gland strain did not grow so well as the other three. Dr. Cobbett produced luxuriant growths. on agar and glycerin serum with the 2nd generation of the bronchial gland strain. When I tested them after 10 months cultivation all the strains grew equally well on the glycerin media With a culture which inoculation tests showed to be largely composed of virulent bacilli it is not surprising that irregular results were at first obtained on artificial media; in this case- long cultivation had apparently favoured the multiplication of the easy growing bacilli in the culture and their increased proportion then rendered it impossible to distinguish the culture. from the other strains of the same virus. . 19 Results similar to these had been obtained by passage through the animal body with the two Group ILI. cultures (H 13 and H 49) and with artificial mixtures of human and bovine tubercle bacilli (see Reports 5 and 6 in this Volume). Attempts were made therefore to separate by means of the plate method of cultivation used for the Group III. cultures, the two kinds of tubercle bacilli of which the cultures under consideration were apparently composed. These separation experiments were successful, and strains of culture raised from single colonies were obtained from each of the two cultures which corresponded exactly on the one hand to the human tubercle bacillus and on the other to the bovine tubercle bacillus. The results showed conclusively therefore that the bronchial gland strain of H 60. “W.B.” and the retroperitoneal gland strain of H 90. “LP.” were mixtures containing bovine and human tubercle bacilli. Attempts were also made to eliminate the virulent bacilli from the cultures by growing them for a period on a medium containing glycerin. After subculture on glycerin-serum both cultures were still virulent for rabbits ; in both cases, however, the virulence was reduced. The failure to eliminate the virulent bacilli by this means was due to the fact that the virulent bacilli grew on glycerin serum fairly readily. The details and results of the experiments with each culture are set out under the three headings “ Separation by Animal Inoculation,” “ Separation by Glycerin Media,” and ‘ Separation by Plate Cultures.” A. STANLEY GRIFFITH. 20 VIRUS H 60. “W.B.” (Bronchial gland strain.) SEPARATION BY ANIMAL INOCULATION. (See Tables L and M.) Calves.—Calf 1099 was inoculated subcutaneously with 50 mg. and Calves 1107 and 1109 each with 10 mg. of culture derived from the bronchial gland of H 60. W.B. through guinea-pig 1825. : lised The first calf died of severe generalised tuberculosis in 28 days; the other two died of generalise tuberculosis in 58 and 32 days respectively. Vanda’ of : Cultures were isolated from the spleen of Calf 1099 and from the prescapular and portal glands o alf 1107. : at The strain from Calf 1099 grew well on some media, particularly on potato, on which it produced a layer indistinguishable from that produced by a Group II. bacillus. The strains from Calf 1107 were dysgonic. Rabbits —Cultures have been isolated from a number of rabbits inoculated at various times to test the virulence of the bronchial gland strain. ; ; hacia : : eh The first, No. 908, inoculated intraperitoneally with 1 mg., died in 33 days of general tuberculosis 3 1 culture isolated from the lung grew poorly on most media but well on potato ; it probably contained a sma: proportion of eugonic and a large proportion of dysgonic bacilli. . The second, No. 951, inoculated intraperitoneally with 1 mg., was killed after 103 days and found to have general tuberculosis ; the culturé isolated from a kidney was dysgonic. Rae The third, No. 935, inoculated intraperitoneally with 0-1 mg., died of general tuberculosis in 82 days; a culture isolated from it was dysgonic. A few small colonies appeared on a glycerin serum tube, and it was thought that these might be composed of eugonic slightly virulent bacilli only ; one of the colonies therefore when it was 31 days old was removed and inoculated intraperitoneally into Rabbit 1017 ; this rabbit died of general tuberculosis in 32 days ; cultures were again obtained on a glycerin serum tube, and « single colony was emulsified and inoculated subcutaneously into Rabbit 1121; this rabbit died in 112 days of general tuberculosis, and a culture isolated from it was dysgonic. Passage through the bodies of calves as well as rabbits had eliminated the eugonic bacilli in the culture SEPARATION BY GLYCERIN MEDIA.. (See Tables M and N.) Cobbett attempted by means of glycerin serum to separate the eugonic bacillus from the virulent bacillus when these bad been properly mixed. He took asingle colony from a glycerin serum tube, subcullivated for three generations, and then inoculated two rabbits (Nos. 949 and 950) intraperitoneally, each with 1 mg. ; one died in 31 days of general tuberculosis, the other was killed after 103 days and showed general progressive tuberculosis, the long duration of life being due to the intracoecal inoculation of some of the dose. From each of these rabbits I isolated a culture; the cultures made in the usual way, by scraping the surface of an egg tube, were dysgonic. Numerous colonies were obtained on a glycerin serum tube sown from Rabbit 949 ; since one does not commonly get a good primary growth of virulent bacilli on glycerin serum it was thought that these colonies might be the product of slightly virulent bacilli; one colony was selected, emulsified, and inoculated intraperitoneally into a rabbit (No. 1018); this animal died in 30 days of miliary tuberculosis. Numerous colonies were again obtained on glycerin serum, and when 36 days old, one was removed, emulsified and inoculated subcutaneously into Rabbit 1120; this rabbit died in 246 days of general tuberculosis and a culture obtained from it was dysgonic. Another attempt was equally unsuccessful. The culture, when it was 10 months old (10th generation) and had been grown for two generations on glycerin media, was inoculated intraperitoneally into two rabbits,. each rabbit receiving half a glycerin serum culture; the rabbits died of general tuberculosis in 109 and 128 days respectively. The same strain was subcultivated on serum for two more months and then inoculated into a calf (Calf 1203) in a dose of 48 mg., and into a series of four rabbits. The calf was killed when well after 97 days and showed general tuberculosis of a retrogressive character ; all the rabbits died of general tuberculosis, but the duration of life was longer than in rabbits inoculated with equivalent doses of bovine tubercle bacilli. The residence on glycerin media had diminished but had not eliminated the virulence of the culture. From one of the egg tubes sown with an emulsion from the prescapular gland of the calf (1203), 12 single colonies were taken and separately subcultivated on serum; after two generations on serum each strain was inoculated subcutaneously into a rabbit. One rabbit died prematurely; all the others developed generalised progressive tuberculosis. Two rabbits were inoculated subcutaneously with emulsions of tuberculous tissues from Calf 1203, one from the prescapular gland and one from a bronchial gland ; they died of general tuberculosis in 138 and 111 days respectively. The cultures isolated from the prescapular gland. a bronchial gland, and the spleen of the calf were all dysgonic. Subculture on glycerin serum, therefore, failed in this case to eliminate the virulent bacillus. SEPARATION BY PLATE CULTURES. (See Table 0.) Glycerin serum plates were sown with an emulsion of tbe caseous sternal gland of a guinea-pig No. 2489, which had been inoculated intraperitoneally with the bronchial gland strain of H 60. W.B.: the guinea pig was killed 16 days after inoculation and showed early general tuberculosis. , The plates were incubated for several weeks and only six colonies altogether developed ; three of these were large, one was small, and two were of intermediate size. Separate subcultures were made on glycerin serum from each a the six colonies, a after 19 days’ cultivation, the whole of the growth from one tube- was, in the case of each strain, inoculated subcutaneously into a rabbit: the o r tubes than on others and therefore the doses varied. ? , eS. were: etter. am, sone One of the rabbits (strain from Colony 2) died in 11 days from causes other i , (strairs from Colonies 5 and 4) died in 53 and 96 days of oe tuberculosis ; the shes @hoaee oe 3) were killed 99 days after inoculation ; one had chronic general tuberculosis, another had a local lesion and slight retrogressive tuberculosis of the lungs, and the remaining one had a local lesion only. Later on, three of the strains, those from Colonies 2, 5 and 1, were retested on rabbits ; in each case two rabbits were inoculated intravenously, one with 0-1 mg., the other with 0:01 mg.: the two rabbits inoculated with the strain from Colony 5 which had when first tested produced general tuberculosis = the rabbit iroculated subcutaneously, died in 42 and 67 days of general tuberculosis ; the remaining four (inoculated: with strains 2 and 1) were killed 128 days later and showed a very limited retr noe : ogressive tuberculosis. In the event that it were found that none of the eugonic strains eas composed eolly of slightly. 21 virulent bacilli, one of the strains, that from Colony 1, was inoculated into a guinea-pig (No. 2817) and plate: cultures were made from its tuberculous organs. On all the plates numerous colonies appeared ; they were of one kind and grew to a large size. Separate colonies were not investigated since the result of the first experiment, which by this time had been ascertained, left little room for doubt that the strain was a pure culture of slightly virulent bacilli. A culture obtained from the guinea-pig in the ordinary way but cultivated ae on glycerin serum had slight virulence only for the rabbit and grew luxuriantly on the differential media. Two of the strains, those from Colonies 1 and 5, were subsequently inoculated subcutaneously each into a ealf (dose 50 mg.) and into rabbits. The strain from Colony 1 which grew luxuriantly on artificial media produced slight retrogressive tuberculosis in the calf (Calf 1481) and the rabbits ; the strain from Colony 5, which grew poorly on artificial media, gave rise to fatal generalised progressive tuberculosis in the calf (Calf 1425) and the rabbits. The inoculation experiments with the six strains showed therefore that three of the strains were virulent and three slightly virulent for the rabbit. The three slightly virulent strains (from Colonies 1, 2, and 3) grew well on artificial media and resembled therefore a Group II. bacillus; two of the virulent strains (from Colonies 4 and 5) grew poorly and resembled exactly a bovine tubercle bacillus; the other virulent strain (from Colony 6) grew well and resembled the original culture. An unsuccessful attempt was made to separate a eugonic and a dysgonic bacillus from the original culture directly ; an emulsion of the culture was made, diluted, and sown over the surface of glycerin serum plates. Ten single colonies were chosen, separately subcultivated on serum, and then tested on rabbits ; in the case of each strain a rabbit was inoculated subcutaneously with the growth from one serum tube. Three of the rabbits died prematurely, four died of general tuberculosis. SUMMARY. From the culture derived from the bronchial gland of Virus H 60. ‘ W.B.” the eugonic element was eliminated by passage through the bodies of calves and rabbits, and there remained a dysgonic fully virulent. bacillus ; and by means of the plate method of cultivation strains of culture have been obtained with the properties of the human and the bovine tubercle bacillus respectively. VIRUS H 90. “IP.” (Retroperitoneal Gland Strain.) SEPARATION BY ANIMAL INOCULATION. (See Tables P, Q and R.) Calves.—T wo calves were inoculated subcutaneously when the culture had been 340 days in artificial cultivation, one calf (No. 1383) receiving 50 milligrammes, the other (No. 1471) 80 milligrammes. They were killed in good health 92 and 90 days respectively after inoculation and showed generalised tuberculosis not severe but obviously progressive. Cultures were isolated from the lung, spleen and a pudic gland of Calf 1383 and the prescapular gland and a bronchial gland of Calf 1471. All the strains were identical in their cultural characters and resembled the more difficultly growing bovine viruses ; they produced on serum glycerin-serum and potato thin grey layers, and on agar very scanty growths. The strain from the pudic gland of Calf 1383 produced in a calf inoculated subcutaneously with 50 milligrammes generalised progressive tuberculosis fatal in 42 days. The strain from the bronchial gland of Calf 1471 inoculated subcutaneously into a calf in a dose of 45 milligrammes produced generalised tuberculosis fatal in 21 days. After passage through the calf therefore, the easy growing bacilli in the culture inoculated were eliminated and there remained dysgonic virulent bacilli identical in every respect with bovine tubercle bacilli. Labbits.—Cultures have been recovered from four of the rabbits inoculated with the original strain. Two of the rabbits were killed 111 days after subcutaneous inoculation with 10 milligrammes of culture, and showed general progressive tuberculosis. Cultures were isolated from the scapular gland and a kidney of one (No. 1553) and from one of the kidneys of the other (1554) ; the two strains from the kidneys grew poorly on glycerin, that from the scapular gland produced pigmented layers on serum anda yellow wrinkled layer on potato ; the strain from the scapular gland therefore still contained eugonic bacilli while those from the kidneys were purely dysgonic. The other two cultures were obtained from two rabbits which had been inoculated intravenously one with 0:1, the other with 0°01 milligramme. One of the rabbits (No. 1794, dose 0°1 mg.) died of general tuberculosis in 64 days and the culture was obtained from the lung, the other (No. 1795, dose 0°01 mg.) died of general tuberculosis in 116 days and the culture was obtained from the kidney. The strain from the kidney of rabbit 1795 was dysgonic, that from the lung of rabbit 1794 was lost before its cultural characters had been determined. Both strains were tested on rabbits and found to be fully virulent. Passage through the rabbit therefore had the same effect on the culture as passage through the calf :— the eugonic bacilli were eliminated and dysgonic virulent bacilli alone remained. Goat.—The retroperitoneal gland strain was at first thought to be a pure culture of slightly virulent bacilli like the strain from the mesenteric gland and in this belief it was used in a good many experiments made to compare the effects of slightly virulent and virulent bacilli. One of these experiments was with a goat which was inoculated subcutaneously with the object of ascertaining whether human tubercle bacilli are excreted in the milk of an animal after subcutaneous inoculation. The goat received 50 milligrammes of a glycerin serum culture under the skin of the neck and was killed 108 days later. The milk of the goat withdrawr at intervals of 24 hours during the first eight days and then on the 14th, 21st and 75th days after her inoculation produced tuberculosis in guinea pigs, and from one of these guinea-pigs (one inoculated with the 14th day milk) a culture was isolated ; this culture grew well on artificial media and had low virulence for the rabbit. . The post-mortem examination of the goat showed a small local lesion, caseous nodules in the adjacent prescapular gland, a number of small grey tubercles in the lungs and one in each kidney, soft yellow tubercles in the mesenteric glands, and several miliary fibro-caseous tubercles in the mamma. 27676 D 22 An emulsion of the prescapular gland was inoculated subcutaneously into two rabbits both of which died of general tuberculosis one in 145 the other in 157 days, anda culture was isolated from the lung of each. Beth cultures were dysgonic and one was shown to be fully virulent for the rabbit. et a The culture raised directly from the prescapular gland was eugonic and after 301 days artificial cultivation was found to be slightly virulent for rabbits. The result therefore of the inoculation of the culture into the goat is the exact converse of that following the inoculation into the calf. In the calf dysgonic virulent bacilli only were isolated from the distal lesions while the culture from the nearest gland was composed of virulent bacilli with a slight admixture of easy- growing bacilli. In the goat on the other hand apparently only eugonic slightly virulent, bacilli were disseminated, the culture from a remote part (the mamma) being composed wholly of bacilli of this type, while in the gland nearest the local lesion the bacilli were mainly of the eugonic slightly virulent type, virulent tubercle bacilli being present in very small numbers, as shown by the results in rabbits of the inoculation of an emulsion of the gland. The difference in the results obtained with the goat from those obtained with the calf was probably due to the fact that a glycerin serum and not a serum culture was used Growth on glycerin serum had reduced the proportion of the virulent bacilli by favouring the multiplication of the more easy-growing slightly virulent bacilli. SEPARATION EXPERIMENT ON GLYCERIN MEDIA. (See Table Q.) When the culture had been over 16 months in artificial cultivation on serum, it was sown on glycerin serum and subcultivated continuously on this media for four months. The culture was then tested on a series of rabbits (Nos. 2300-2302) all of which died of general tuberculosis. Cultivation on glycerin serum had not. therefore eliminated the virulent bacilli in the culture. SEPARATION BY PLATE CULTURES. (See Table 8.) re from A guinea-pig, No. 3300, was inoculated intraperitoneally with the retro-peritoneal gland strain and killed 21 a-pig days afterwards. Serum plates and egg tubes were sown with emulsions from a sternal gland and the pyloric gland ; the serum plates became so closely covered with colonies that it was impossible to pick off single colonies. On the egg tubes however sown from the pyloric gland the colonies were in places sufficiently discrete, and three large colonies were taken and separately subcultivated. The three strains produced pigmented growths on serum and they were each tested on a series of rabbits intravenously and found to be slightly virulent. ; re from Separate colonies were also investigated from a rabbit which had been inoculated intravenously with the +2019.| original culture at the same time as the guinea-pig. It died of acute tuberculosis in 20 days. Five colonies were taken from two egg tubes (which had been sown with an emulsion of the spleen) and separately cultivated; three were tested on rabbits, each strain being inoculated intravenously into two rabbits in doses of 0°1 and 0-01 milligramme ; two of the strains produced rapidly fatal tuberculosis in all the animals inoculated and were équal in virulence to a bovine tubercle bacillus; one of the two rabbits inoculated with the third strain died in 17 days of coccidiosis of the liver and showed slight tuberculosis of the lungs, the other was killed after 104 days and showed disseminated tuberculosis of a type not unusual with viruses of low bovine virulence. The two virulent strains grew poorly on glycerin media; the third strain was lost before it was tested on glycerin media, but it produced pigmented growths on serum. From the original culture through the guinea-pig therefore three strains of culture have been isolated which are identical with the human tubercle bacillus, and from a rabbit inoculated with the original culture there have been isolated one strain with the properties of the human tubercle bacillus and two with the properties of the bovine tubercle bacillus. SUMMARY. From the culture derived from the retroperitoneal gland of Virus H 90.“1.P.” bacilli with i of the human and the bovine tubercle bacillus respectively have been obtained by passage ica dee body and by means of the plate method of cultivation. A. STantey GRIFFITH. 23 TABULAR SUMMARIES OF THE PRELIMINARY INOCU- LATION TESTS WITH VIRUSES H 2, H 18, H 16, H 1%, AND H 49. Virus H 2. Virus H 13. Virus H 16. Virus H 17. Virus H 49. “Sp. A.” “ A.D.” “JH.” “Sp. B.” “TC.” eI 28 6 f | 3 | \. ( | 1 | t ( | { | L Strain derived from Calf 93 ? ” Calf.111 Strain derived from Calf 301 ’ ” ” Rat 15 Strain derived from Calf 275 ” " ” ” Bull 4234 Calf 559 Strain derived from Calf 265 ” ” Strain derived from original material (Blythwood Strain ” ” ” | eee eee eee oe eee Calf 539 (through Rabbit 181) Calf 539 (through Rabbit 181 and Guinea pig 2017). ” Strain derived from Calf 797 ” ” Calf 787 (Royalcot Strain) eee 1 | | J PAGE 24 24 Ww 26 27 a4 VIRUS H 2. “Sp. A.” Culture derived from Calf 93 through G.P. 534. Total duration of artificial cultivation—3 years 8 months. when 21 days old. The 30th generation of culture was used ight. Number Mode of Dose in Ween of tou a Milli- Duration of Life. Result. Rabbit. grammes. | Thitial.| Final. 1165 Intrav. ae 1,700 | 1,200 Died 18 days General tuberculosis. Subcut. 10 sf} > , oe 1166 Intrav. O1 1,900 | 1,430 Died 27 ,, General tuberculosis. 1167 Intrap. 10:0 1,750 1,250 Died 20 _,, General tuberculosis. 1168 Intrap. 1:0 2,000 1,420 Died 33 ,, General tuberculosis. 1169 Subcut, 10°0 1,700 1,590 Died 5 ,, No tuberculosis. Death from heart disease. Culture derived from Calf 111 Total duration of artificial cultivation—3 years 3 months. through G.P. 640. The subculture was used when 21 days old. Number | yode of Dose in Weights of : Milli- Duration of Life. Result, Rabbit. Inoculation. rammes é 8 "| Initial. | Final. 1061 Intrav. 10 1,100 | 1,250 Killed 111 days | Very slight tuberculosis of lungs and kidneys. 1062 Intrav. O1 950 | 2,100 Pe 43 Slight tuberculosis of lungs and kidneys. | 1063 Intrap. 10°0 1,100 | 2,400 % a Slight general tuberculosis. | 1064 Intrap. 1-0 850 | 1,750 ‘ 5 Very slight general tuberculosis. 1065 Subcut. 50°0 900 ! 1,900 5 59 Local lesion : one tubercle in lung. 1066 Subcut. 10°0 850 | 2,100 45 ‘ Local tuberculosis only, 1067 Subcut. 1:0 950 2,250 3 a Local tuberculosis only, VIRUS H 13. “A.D.” Culture derived from Calf 301. (Thoracic Gland.) Total duration of artificial cultivation—3 years. The 28th generation of culture was used when 21 days old Number | wroge of Dose in Wee of | tnoculation,| _ Milli- Duration of Life. Result Rabbit. *| grammes, si 3 : Initial. | Final. 1186 Intrav. 1:0 2,950 | 2,300 Died 18 days General tuberculosis, 1137 Intrav. 0'1 2,370 | 1,700 Died 27 ,, General tuberculosis, 1138 Intrap. 10°0 2,070 1,470 Died 12, Acute tuberculosis, 1139 Intrap. 1:0 1,490 1,100 Died 27 ,, General tuberculosis, 1140 Subcut. 10°0 1,620 | 1,220 Died 52 _,, General tuberculosis, 1141 Subcut. 10:0 1,000 850 Died 2 ,, No tuberculosis. The culture was again tested a year later to ascertain whether there had been an The 39th generation of culture was used when 7 days old. 1712 1710 1711 1713 Intrav. Intrav. Intrav. Subcut. 10 - O1 O1 20°0 1,450 2,100 1,700 } 1,200 1,600 1,300 Died 11 days Died 25 n Died 29 y alteration in virulence, General tuberculosis, General miliary tuberculosis, Acute miliary tuberculosis, Total duration of artificial cultivation—2 years 7} months. 25 Culture derived from Calf 301 through Rat 15. when 21 days old. The 25th generation of culture was used Number Mode of Dose in Weight of ae Milli- Duration of Life. Result. Rabbit Inoculation. eames - 8 "| Initial. | Final. 1176 Intrav. 10 1,570 1,300 Died 15 days General tuberculosis, 1177 Intrav. O1 800 750 Died 25 ,, General tuberculosis. 1178 Intrap. 1:0 800 1,100 Died 101 ,, General tuberculosis (injection partly intracoecal), 1179 Intrap. O1 600 900 Died 106 ,, General tuberculosis (injection partly subcutaneous and probably partly in- tracoecal), VIRUS H 16. “J.H.” Culture derived from Calf 273. (Prescapular Gland.) Total duration of artificial cultivation—3 years. The 36th generation of culture was used when 21 days old. 3 Weight. Number Mode of Dose in of Toeulation Milli- Duration of Life. Result. Rabbit. grammes. | Tnitial.| Final. 1106 Intrav. 1:0 1,350 2,300 Killed 94 days Slight tuberculosis of lungs and kidneys. 1107 Intrav. 0-1 1,000 1,850 33 55 Very slight general tuberculosis, 1108 Intrap. 10:0 1,050 1,700 5 a Very slight general tuberculosis, 1109 Intrap. 10 1,020 2,050 i 7 Tuberculosis of peritoneum and kidneys. 1110 Subcut. 30°0 1,100 2,350 o 5 Slight general tuberculosis, 1111 Subcut. 10°0 950 1,850 iy of Local lesion and slight tuberculosis of lungs. Total duration of artificial cultivation—2 years 6 months. Culture derived from Bull 423A. when 21 days old, (Prepectoral Gland.) The 29th generation of culture was used ‘i Weight. Number Mode of Dose in of | tnooulation.| _ Milli- Duration of Life. Result. Rabbit. grammes. | Thitial,| Final. 1131 Intrav, 1:0 1,700 1,380 Died 17 days General tuberculosis. 1182 Iutrav. O1 1,200 1,050 Died 20 ,, General tuberculosis. 1138 Intrap. 10°0 1,150 830 Died 17 ,, General tuberculosis. 1134 Intrap. 1:0 1,250 900 Died 26 ,, General tuberculosis. 1185 Subcut, 10°0 1,550 1,360 Died 126 ,, General tuberculosis. 26 Culture derived from Calf 559. (Hepatic Gland.) Total. duration of artificial cultivation—2 years 1} months. The 25th generation of culture was used when 21 days old. o Weight Number Dose in of gue | Mille Duration of Life. Result. Behbis "| grammes. | qyitial, | Final. 1126 Intrav. 1-0 1,550 | 1,100 Died 20 days General tuberculosis 1127 Intrav. O1 1,950 | 1,190 Died 109 ,, Tuberculosis of lungs and kidneys (severe) and large intestines, : 1128 Intrap. 10-0 8,050 | 1,720 Died 80 ,, General tuberculosis (injection mainly ; intracoecal). 1129 Intrap. 1-0 1,850 |} 1,510 Died 24 ,, General tuberculosis. 11380 Subcut. 10-0 1,700 1,570 Died 119 ,, General tuberculosis. VIRUS H 17. “Sp. B.” Culture derived from Calf 265. (Prescapular Gland.) Total duration of artificial cultivation—3 years. The 30th generation of culture was used when 21 days old. Weight. Number Mode of Dose in eght of | Taoculation Milli- Duration of Life. Result. Rabbit. grammes. | Tyitial.| Final. 1100 Intrav. 10 900 1,750 Killed 97 days Slight general tuberculosis. 1101 Intrav. O1 850 1,950 9 r Slight general tuberculosis. 1102 Intrap. 10-0 700 1,150 33 ss Slight general tuberculosis. 1103 Intrap. 1:0 670 1,550 rr ff Slight general tuberculosis. 1104 Subcut. 50°0 540 1,450 3 3 Local tuberculosis and miliary tuberculosis of the lungs. 1105 Subcut. 10-0 670 1,900 % 43 Local tuberculosis and miliary tuberculosis of the lungs, Total duration of artificial cultivation—2 years 2 months. Culture derived from Calf 539, through Rabbit 181. when 21 days old. The 27th generation of culture was used Number Mode of Dose in Weight of f Milli- Duration of Life. Resul. «, | Inoculation. esult. Rabbit. grammes. | Tnitial.| Final. 1113 Intrav. 1:0 900 1,900 Killed 97 days Tuberculosis of lungs and kidneys. 1112 Intrav. ol 900 2,020 4 3 Slight tuberculosis of lungs and kidneys. 1114 Intrap. 100 410 | 1,300 si ‘i Chronio general tuberculosis, 1115 Intrap. 10 450 | 1,500 ” » ies tuberculosis and slight tuberculosis of lungs. 1116 Succut. 47°5 430 1,750 o a sea: tuberculosis and slight tuberculosi of lungs. 1117 Subcut. 10:0 410 | 1,600 ” ” Local tuberculosis and slight tuberculosis of lungs, , ; 27 Culture derived from Calf 539 through Rabbit 181 and Guinea-pig 2017. Total duration of artificial cultivation—53 days. The 4th generation of culture was used when 20 days old, Number Mode of Dose in noe of | Tioculation: Milli. |-————_-—__—_| Duration of Life. Result, Rabbit. grammes. | y,itial,| Final. 1142 Intrav. 1°0 1,850 1,000 Killed 85 days General tuberculosis complicated with pseudo-tuberculosis. 1143 Intrav. 01 1,450 2,300 Killed 85 ,, Tuberculosis of lungs and kidneys. 1144 Intrap. 10°0 1,750 | “1,350 Died 11 ,, Early tuberculosis of omentum ; no sign of disease elsewhere. 1145 Intrap. 10 1,070 2,300 Killed 85 ,, Slight general tuberculosis, VIRUS H 49. “T.C.” Culture derived from the Mesenteric Glands, direct. Blythwood strain. Total duration of artificial cultivation—l year 8} months. The 2lst generation of culture was used when 22 days old. Weight. Number Mode of Dose in ia of | Thooulation Milli- |————~——|| Duration of Life. Result. Rabbit. grammes. | ritial. | Final. | 1048 Intrav. — 10 1,100 2,600 Killed 130 days | Slight tuberculosis of lungs and kidneys. 1049 Intrav. 01 1,150 1,050 Died 11 ,, Pseudo-tuberculosis. 1051 Intrap. 10:0 1,250 2,500 Killed 130 __,, | Slight tuberculosis of omentum and (?) of lungs. 1050 Intrap. 1-0 1,450 3,050 Killed 130 ,, | Slight tuberculosis of omentum, kidneys { and lungs, 1052 Subcut. 10-0 1,150 2,700 Killed 130 ., ~~ Local lesion and slight tuberculosis of ' lungs. 1053 Subcut. 1:0 1,250 2,800 Killed 130 ,, | Local lesion only. Culture derived from the Mesenteric Glands, direct. Royalcot strain. Total duration of artiticial cultivation—1 year 10} months. The 25th generation of culture was used . when 21 days old. 5 Weight. Number Mode of Dose in of | tnooulation.| _ Blilli- Duration of Life. Result. Bans: grammes. | Titial. | Final. 1180 Intrav. 10 1,000 1,000 Died 16 days General tuberculosis. 1181 Intrav. Or1 1,100 920 Died 31 ,, General miliary tuberculosis. 1182 Intrap. 1:0 750 2,050 | Killed 163 _,, Chronic general tuberculosis (injection intracoecal). Culture derived from Calf 797. (Prescapular Gland.) Total duration of artificial cultivation—1l year 4 months. The 13th generation of culture was used when 22 days old. : Weight. Number Mode of Dose in of Prnenicinad Milli- |-—-~—------_| Duration of Life. Result. Rabbit. grammes. | rnitial. | Final. 1071 Intrav. 1-0 1,300 | 1,240 Died 13 days General tuberculosis. 1072 Intrav. O1 1,450 1,470 Died 16, General tuberculosis. 1073 Intrap. 10:0 1,300 1,110 Died 12, Acute tuberculosis complicated by severe psorospermosis. 1074 Intrap. 1:0 950 770 Died 13, General tuberculosis. 1075 Subeut. 10°0 900 1,060 Died 41 _,, General tuberculosis. 1076 Subcut, 10 900 950 Died 80 ,, General tuberculosis. 28 Culture derived from Calf 787. Total duration of artificial cultivation—1 year 4 months, The 13th generation of culture was used when 21 days old. ight. Number Mode of Dose in Wetght of : Milli- Duration of Life. Result, Rabbit, Inoculation. eames : g "| Initial. | Final. 1079 Intrav. 1:0 1,850 1,460 Died 17 days General tuberculosis. 1080 Intrav. O1 1,800 | 1,790 Died 19 ,, General tuberculosis. 1081 Intrap. 10°0 1,870 1,270 Died 14 ,, General tuberculosis. 1082 Intrap. 10 1,720 1,470 Died 19 ,, General tuberculosis. 1083 Subcut. 10°) 1,720 | 1,050 Died 71 ,, General tuberculosis. 1084 Subcut. 10 1,600 1,800 Died 90 ,, General tuberculosis. VIRUS H 49. “TC.” CULTURE DERIVED FROM THE MESENTERIC GLANDS (BLYTHWooD STRAIN). CULTURE INOCULATIONS. The strain was derived from the original material, and had been in cultivation a total period of 1 year 11 months. The 25th generation of culture was used on Feb. 26, 1907, when 21 days old. CALF 1191. CALF 1193. RABBITS. Subcutaneous. Subcutaneous. - Dose: 50:0 mg. Dose : 100°0 mg. Duration of Killed : May 31,1907. Killed: June 5, 1907, |Number-| Method. | Dose. Life, Result. 94 days. 99 days. P.M.—Local tubercu- P.M.—Local tubereu- losis only. losis only 1198 | Subcut. | 1000 mg. | K. 142 days | Local T. only. 1192 | Subcut. 45:0 mg. | K. 142 ,, Local T. only. | Raszit 1318. Subcut. (Emulsion of prescap- ular gland.) Killed : 129 days. Local lesion only. | -| | | I Sah 29 CHARTS OF SEPARATION EXPERIMENTS WITH VIRUSES H 18.“ A.D.,” H 49.“ T.C..” H 60.“ W.B.,” AND H 90.“ LP.” Virus H 13. “A.D.” (Calf 301 Strain.) Table A.—Separation by animal inoculation (Calf). » B- ‘i a 8 i (Rabbit). » C.—Separation by glycerin media. » D.—Investigation of separate colonies. » ES » Fe ” ” ” ” Virus H 49. “T.C.” (Original Strain). Table G.—Rabbit inoculations with the original culture. » H.—Separation by animal inoculation (Calf). » I—Investigation of separate colonies. » J » K— ” ” ” ” Virus H 60. “W.B.” (Bronchial Gland Strain.) Table L.—Separation by animal inoculation (Calf). Separation by animal inoculation (Rabbit). 1 nen of a single colony on glycerin serum. » N.—Test of original culture after growth on glycerin media. » O.—Investigation of separate colonies. Virus H 90. “I.P.” (Retroperitoneal Gland Strain.) Table P.—Separation by animal inoculation (Calf). ” Q.— ” ” ” ” (Rabbit). ” R.— ” ” ” ” (Goat). }, 8.—Investigation of separate colonies. 27676 Bb H 9L9L3 LO| “ sL'a | Bug, | ymoqng | 619T | os t . : ; ‘99A08 L's) | sfep p2 "a | “Sa 0.01 | “gnoqng ST9T ‘sisofnoreqny [e1ouey — ‘'d ‘shep GG : ‘oxVT : . : | “S06T ‘0S “G90 = TI Woys polly q[usey jo uoremg asoq poqiey | ‘teqarnyy | face (ae reou — ‘snoeueynoqng ‘SLICAVU “APET TIVO j | . ‘ ( rae ‘ d ae 1g wom ‘2061 ‘0g 99d i pesa oO esneo ‘Lo : 1 “‘qnoqn c ® O° is ea | ens AEST ned va a SA ss jo Smeun | 3 . ; ij ; eu, ‘shep eg jo por eatssorsoid ormoiyg | skep zeg‘q| “ynoqug 9EGT deoseid jo 'q @ TOeATI[ND TeOgILE UL teed peg pue ‘pues [eurjserpemt oq} ory WOLF POATLep SVA UIeI4sS oy, “4[Nsoy jo uoryeinq | POMP | “T9dNN = “BUNLTNO “SLIGAVU | rd) “ 98 ‘d = “quoqng LET { “pur [ea ‘Lo | * ser'a}’ — | moqng | gagt |) SetPomt oa - : *aTOAOS 10U ‘SISO[NO ‘9 8h S| 29 0.F “ynoqng 6L4EL1 “purys rr] -Jeqny [e1eues olaolyO— Wd “I'D | s4ep cg qd | 990-7 | ‘ynoqng SET “ndeoseid Jo “qf ‘sftp 16 "LOBT ‘8 “290 + PETES *(snoremint 5 “OsTT i aaiTt é tas Ayaresy -gq'y,) purla aepndeoseid ee JO WOTVeING e8od PEE AUE ||P Jo WOIs[nMe FO ‘09 Y.OT : es0q “snosueynoqng “SLICE VA “S96 ATVO ‘LO | shep ford | “S8ar0-0r | ymoqng | AIST “Ld |steppg() a} “Sur0.-2h =| “ynoqng | 99@T “Ld “0a ‘BUI 1.0 “ACIYUT G9GT ‘ad £4 ora0aryo & Jo stsopno1Eqn4 aalssoisoi1d yersuex) — "' ‘LO | sfep 61'a "SUL 0.1 ‘aerquy | POST ‘skep 0g “LOGT ‘6 Ane 5 oy] ‘ 7 ‘ GIVey S7VIOpOUr UT MoM Pore yy | qjasey jo woreang as0q. pope | azequny Sot Dog : BOG “snoouejnogng ‘SLICG VU “GIGI TIVO “plo skep Gg meq ‘2Z06E ‘0% [4dy uo posn sem ainqino Jo uolye19NEd 446g SUL, ‘syoom Z% savok Z Jo porsod [e404 & MONeATI[NO [eLOgIAe | UI UAE8q PLY PUs ‘[vI9}eOI [eULSII0 OG} WOT] poaliop Se ILI ONL, ‘“NOILVTQOOONI 'IVWINV Ad “HUALTIND NOILVUVdds (NIVULG LOOIVAOY) SANVIQ) OLYAINGSA AHL WOwd GUAINad TUNING «OL, “6% H SNUIA ‘H GQIGVi LE “L Are tay | “ eg -q | SL ‘109 ‘qmoqng | G6PT “G2 a] TL 100 | “moqng | 16%T DL ‘aleaes A10A qou “J, eatsseasord “ue | “ gore | OL 19D | ‘anoqng | O6FT ‘ereaos AroA qou “J, eatsseisoid ‘wey | “ QOL 6 "190 ‘qnoqng 68FL LS] “ 3 ‘a 8 190 ‘qnoqug | 88PhT Lo) “ 16 4 T00 ‘ynoqng | ASPT LTo| “ 3 9 “100 ‘ynoqng | O8PT Lo} “ 8g ¢ “190 ‘qnoqug | GgPI a a a wo) ” $6 “a % 190 ‘gnoqng | F8PI a a a ‘LD| “ 8¢ € 199 =| “ymoqng | €8FT ‘yL Arey 9 | “9g @ 190 “gnoqug 68rI “‘ssuny UL a1eaes “J, oatssoisoad ‘uss | shep BOT T “199 “ynoqng T8h1 * “fk | “qnsey jo ao ae ; aoe "poyeR | “requinyy | “SLIGAVa “LOGL ‘G% WNSNY UO ‘{IqQquy e OFUL po}e[NOOUI SEA SUTLIYS ZT og} JO Yove Jo uoteieues pig oy} Jo aqny Windies ouo WOIZ q}MOIS eq ‘“peyearyqnoqns Ayaqeaedes pue ‘eTZ~I J[eO jo pueps aejndeoseid 04} WOIF MMOS Oqny 58a UL WOdJ PoxeTOST 919A SOIMOTOO O[STIS 9ATOA\ T, “HUALTAO | ‘(skep 08) 2061 ‘6 410 ‘pete “SpUt[S O110}MeSeMl Og} WOIf poallop onzyno Jo “SUL 0.0G qyim A[snooueynoqns peye[noouy ‘e181 dIVO ‘'SGINOION ALVUVdES AO NOLLVOLLSHANT *‘(NIVELG LOOIVAOY) SANVIX) OIUBDLNASAT], AHL WOUd CGAIMIG AANLTAN «OL, ‘6%? H SNUIA IT Gavi 8& 9 “ 96a ‘Bar 10.0 | “derqay 9&6 ‘Lp | sep g-a ‘Ba T.Q | “deaquy Ge6 ‘Lo ouorgg | skep gIT (I) “| “8a T.0 | ‘deauy 86 “O19A08 ‘STSO[ND ws sep 96 “C ‘Ba g.t | ‘dexyuy €86 -Ieqny [eqseu99— Wd "ehep 3g ; ‘ov i : ‘ "9061 ‘p “3dog : por ie go woryean(y eran ee ae “BUI 0.0T : e80q “‘snooueynoqug “SLICGV a “60TT ATVO *a10A0S ‘SISO[ND -daqny [e18ueH—' WT “skep gg “9061 ‘og ‘dog : porq “BU (QT : o80q “snooueynoqug “LOTT ATVO ‘plo sfep ¢% ‘aoTye10N0S YIP 94} SBM POsN oiny[no 94,7, ‘skep [Z] 2 UOeay[No [eloyTAe Jo UOTeVANp [e40,], “9061 ‘g Isanay (9) *4U04X0 FYSI[S B OF ‘ynoqns A,qeqoad uomooluy ‘LZ eaIsseIs0Id o1U0LYO Ls ‘Lo “ ¢g'q_=s| ‘Bur 1.0 “ ee ‘a skep gz ‘O° | 801 0.01 “BUI 0-1 “q[nsey ‘OSUT youonemqg | a , SGNVTS IVIHONOUE AHL WOWd GGATYad GYNLITAO “SLIGAVaA "deaquy 606 ‘deayuy 806 "a19A08 ‘SISO[NT ; -leqny yereuey — "PT dear | 406 ‘skep 92 9 208! ‘et tag . : Burkp wey parry poqjey |toquiny Bea pio 2600 “snosueynoqng “6601 TIVO ‘po skep [zg ‘uoleazoued pag oy} SBM posN ein}[Nd OG, ‘shep ZL JO potsod [e409 @ MOTYRATI[NO UI U9eq pey pur ‘qzgy sid-eaumy ysno1y} spars [eiqouorq 64} MOAy poaltop Se UIvI4s OUT, “906I ‘GT ana (7) ‘worye[nooUT [RUTTY Aq woneredag OM » 09 H SNUIA "Tl WTavii IV TABLE M. VIRUS H 60. W.B. CULTURE DERIVED FROM THE Human BRONCHIAL GLAND THROUGH GuINnEA-PIG 1825. SEPARATION BY ANIMAL INOCULATION. INVESTIGATION OF A SINGLE COLONY ON GLYCERIN-SERUM. RABBIT 935. The first 3 generations of culiure were on serum; the fourth was ConTROL EXPERIMENT. Intraperitoneal. Bparsely: SSW On. glycerin-serum. One of the four of five colonies The 6th generation of culture on serum was used for inoculation on which appeared was removed, cultivated on glycerin-agar, and then on Sept, 26, 1906, when 21 days old August 3, 1906. serum. The serum culture was used for inoculation on Sept. 26, 1906, peers : y 3 . Dose : 0-1 mg. of culture derived from when 21 days old. RABBITS. Guinea-pig 1825. RABBITS. Died : 82 days. G.T. Duration of i Number. | Method. Dose. 3 Result. Number. | Method. Dose. ea of Result. arcu ie oe Life. — 951 Intrap. 1:0 mg. | K.103days| G.T. Injection CULTURE. 949 Intrap. | 1:0 mg. | D. 31 days| G.T. eras ° 7 partly d intra ere caecal. A single colony from a glycerin-serum 950 Intrap. 10mg. |K.103 ,, | G.T. Injection 952 Intrap. 10mg. |D. 35 , | GT. tube sown from Rabbit 935 was probably partly emulsified and inoculated on Nov. 23, intracaecal. 1906, when 31 days old, into : 7 RABBIT 1017. Intraperitoneal. ied : 32 days. G.T. DERE CULTURE. A single colony froma glycerin-serum tube sown from Rabbit 949 was emulsified and inoculated on Nov. 93, 1906, when 27 days old, into : CULTURE: RABBIT 1018. : Intraperitoneal. A single colony from a glycerin-serum : Died: 30 days. Acute miliary Ty, tube sown from Rabbit 1017 was emulsified and inoculated on Jan. 29, 1907, when 35 days old, into : RABBIT 1121. CULTURE. Subcutaneous. Died : 112 days. G,T. A single colony from a glycerin-serum tube sown from Rabbit 1018 was emulsified and inoculated on Jan. 29, 1907, when 36 days old, into ° RABBIT 1120. Subcutaneous. Died ; 246 days. 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POLL -[no $ wor ‘Ep | ‘shep gata yynorg |) -deyquy | €9TT “qnsey yo couninet 280g, ‘pomp | “tequinyy ‘SLIGAVA (‘Uindes TO e10M SIOy}O OY} ‘1vSv UTI04]9 UO Sv TOT}eIOUED TYG OY) plo sep OZ Woy ‘Z06T ‘8 Areniqeg m0 pesn seM ‘MINJos-UIse04[S UO ‘UOI}edeUOS YIOT OY] : UOTeaTy[No yeIOgMae ut skep OTE Ueeq pey uTeA4s onL (2) ‘SNOLLVIQOONT @4OLTOO ‘VIGHW NIQHOATS NO HLMOYUSD BALAV GaLsaL “GZ8T DId-VENIND HONOUHL GNVIQ) TVIHONOUG NVWOW AHL WOUd ATATUTG TAnWIAQ «€M, “09 H SQUIA ‘N Gavi Sv es 46 TABLE Q. VIRUS H 90. “IP.” IRE DERIVED FROM THE RETROPERITONEAL GLAND. ouUL Separation by Animal Inoculation (Rabbits). MBER 5, 1907. Aprin 14, 1909. from the human retroperi- SEPARATION EXPERIMENT ON GLYCERIN MEDIA. een in artificial cultivation a od of 94 days. The strain was derived from the human retroperi- toneal gland, and had been cultivated on glycerin- Derived from the , gth generation, 14 days old: gerum for four months; it was then sown on the July Sine serum. serum and the resulting growth inoculated (33rd generation, 8 days old). BBITS. RABBITS. CALF 1383. e. | Durationof | Result ) ool . . i f | Subcutaneous. ee Number. Method.| Dose. ' a : | Result. { ‘ | 1 i Dope 0) meg. ag.| D. 26 days |G. T. | | Killed : October 8, 1 2301 | Intrav. , 0-1 mg. | D. 19 days General 92 days. ag.| D. 79 ,, | T.oflungs miliary T. P.M.—General tu and kid- 2300 | Intrav. |0-01mg.; D. 35, hae losi : neys. oe ng.|K. 111 ,, |G. T. 2302 | Intrav. |0-‘Olmg.| D.28 ,, |General miliary T. og.|K.111 ,, |G. T. | CULTURE. The strain was der from the pudic gl and had been 48 day artificial cultivation. 4th generation of cul was used for inocula on November, 25, 1 when 12 days old. II.—Marcx 30, 1908. n was derived from the same source and had been in cultivation a total period of 240 days. sulture used was the 16th generation, 13 days old, on bovine serum. CALF 1489. Subcutaneous. Dose : 50°0 mg. Died : January 6, 19 42 days. P.M.—General tu cuiosis. RABBITS. Number. Method. Dose. ae of Result. 1793 | Intrav. | 10 mg.| D. 32 days| G. T. 1794 | Intrav.| O-lmg.| D. 64 , | GT. 1795 | Intrav. |0°Olmg.| D.116 ,, | G. T. sTURE. CULTURE. Rabbit 1794. Inoculated on Derived from the kidney of Rabbit 1795. Inoculated 34 days cultivation. The 5th on October 13, 1908, after 81 days cultivation. The sed when 18 days old. 6th generation was used when 22 days old. BBITS. RABBITS. . pe of Result. Number.| Method.} Dose. pees of Result. ig. | D. 27 days | G. T 2040 | Intrav. | 10mg.| D.lddays |Acute ag.| D.28 ,, | G.T. 2041 | Intrav. | OL mg.| D.21,, Gone ag.| D. 32, : G.T 2042 | Intrav. |0-0lmg.| D. 36 ,, jen “ ye “(qqsys) shoupry FO pus ‘(eevr1opoul) ssuny Joy ‘worseypeoory] “ ZOT “M | "Sur 9.0¢|"noqng | OLEI -soupry pue sSuny “LOL M |'Sar 0.0] ‘AeayUT | 696T JOD MHS APA) © pop -y |-sarz.o | -aemquz | 9961 ‘soupry puessuny ory, |shep 297 “M |*S10.1 | “Aez}UT | 1967 “qpnseqy jo ec enial ‘asoq) =| ‘(poqyeyl | oqumnyy "SLIGAVA —Suyye[noout rog “gQgT ‘F] ysnSny uo posn sea ‘pro sX+p [] ‘erng[no Jo uoereMes 44), OUT, = ; . CL “'y ‘shep 8g Ul FSIp) Joppn oy} Jo Jey yyStr ay} WLoIZ ATE Jo "00 Y.OT THM "L061 ‘8Z JoquissonT WO poxefnoont ‘[Eyg sid-eauIny wMoaz poatsog : “HUOLTAO “L Aseria perouey | “2s | Bar70.0} “AeTyUT | 9108 ‘LD | “ @6'a | 8a T.0 | aeaqal | F106 ‘T, Azerpran aqgnoy | shep gt qd | Sur g.7 | ‘aerjay | E10’ “q[nsey jo one nq ‘es0q =| ‘poqyey | ‘toquin jy “SLIGAVU “806L ‘FZ Tequieydeg uo UoTe[NoOUT Joy pesn sem “plo skep ET ‘WorRieues OIF OUT, “TPLT HaqeY Jo Suny oy} worz poatsog ae ‘BD! “ 24t-a]snsqng| ZPLt ‘L°y | shep cht a |ymoqng | 1PLT ‘SLIGAVU —OjUl pues Iejndeoserd jo uospaug “spurs jeumsar pue shou “pry ‘sBuny jo "Lass | “ get M| Baty. | aera | 6602 ‘A[UO ssunt UE sepaoqng Mey VW) “ Ost'M | SU T-0 | aerjyUy | 860% “sofOSNUL pus ‘skoupry ‘ssuny Joy, |sfep cot “| ‘Sar g.y | ‘avaqyuy | 2608 "4ynsaqy jo ayeati ‘esoq §=| "pom ey | roquinyy - “SLICAV A ‘plo sep ¢] ‘WonedemeS WET OU} SVM posn oAny4pno ey, “WOTyeA -1yno perogtyre skep TOE 101Fe ‘QOGT ‘ZZ Toquiecsq, UO peye[NoOUI sem pue ‘gg 4voxy Fo puels aepndeosead 04} Woy paattop SV UTeI4S SILT, “MUALTAO "(qeoy) woTyeTNoouy TewIUY Aq:uoTyeredag : : “aNV19 (TVANOLIGGAOULTA AHL WOUd GHAINAC AMOLINO «dT, “06 H SNUIA ‘a ATaVio ‘eulIou spurys qdud, Aremmeuresrdng ‘syutod SNO@sed YIM yoseq Yoyed WAY [[VUIs BPSMOUS IUSII oy | 1007 snoesvo Y}IA Se[MpOU pAVY OM} PUB Sor}UED SNOESed TIM sopoJeqny, snorqyg JO JoquInU & pesUiejuCO Jappn oy} Fo Frey yor ey, ‘soyoqed pue soporeqn, £44113 moTjad Jos oTOM o10T} SpUe[s olto}UesoM Oy4 JO ySOU UT = “epoTeqny, suo pouTe}u0D Aoupty Yoo { seporeqny snorqg Aoid [jews snorsunu A[mez pomoys sun ey, ‘sernpou 444123 sno -98e0 pouTeyUoo pues tejndeoserd yor] “WOISsey [BOOT [TeMgG— Wd ‘skep €01 "B06T ‘Gg Arenageg : perty ‘SUL (0.0G : 880q “‘snosueynoqng “Gg ‘ia “LOGI ‘FI tequeAON UO TONL[NOOUT JoF posn sea ‘uInass-uIe04]3 uo ‘pfo skep FT ‘uoMereues yIOT ou ‘skep GOT FO ported [e107 B UOT}eATY[NO UL UEEq Pry pue ‘pues [eouoqiiod-orjer weMINY oy} Wodz Peatsep SV UTeIT4S OTL, “HaNLTND | 48 TABLE §&. VIRUS H 90. “TP. CULTURE DERIVED FROM THE RETROPERITONEAL GLAND. \ Investigation of Separate Colonies. RABBIT 2019. Inoculated intravenously with a fairly large dose of a serum culture (24th generation, 16 days old) derived from the retroperitoueal gland, on September 28, 1908. CULTURE. Single colonies were isolated from two egg tubes sown with an emulsion of spleen. Died of acute tuberculosis on October 18, 1908 (20 days). RABBITS inoculated with cultures derived from these colonies. Date of Number of Age and Number of Daration of Inoculation. | Colony. Generation. Rabbit. Mashed.) Dose. Life. Besule. t Jan! 28, 1909 Col. 1 4th gen.; 21 2136 Intrav. | 0-Lmg.] D. 21 days | General miliary T. [ days old, on serum. 2135 Intrav. |0-‘0lmg.| D. 32 ,, |G. T. 45 Col. 2 4th gen. ; 21 2138 Intrav. | O-lmg.| D. 21 ,, | General miliary T. days old, on serum. 2137 Intrav. |0-Olmg.| D. 33 ,, |G. T. (| 2184 Intrav. | O-lmg. | K.104 ,, | Slight T. of kid- neys, moderately Jan. 27, 1909 Col. 5 4th gen. ; 20 severe disease of days old, on lungs, and severe serum. | T. of eyes and lachrymal glands. L 2133 Intrav. |0‘O0lmg.} D. 17 ,, | Slight T.-of lungs. Death from cocci- | diosis. GUINEA-PIG 3300. Inoculated intraperitoneally with the same culture as Rabbit 2019, on September 28, 1908. Killed 21 days later on October 19, 1908, and showed general tuberculosis. CULTURE. Single colonies were isolated from an egg tube sown from the pyloric gland. RABBITS inoculated with cultures derived from these colonies. Date of Number of Age and Number of D i : Inoculation. | Colony. Generation. Rabbit, | Method.| Dose. eee a Result. ! oe pay Col. 1 ei one [ 2109 Intrav. | 1:0 mg.| D. 121 days | T. _of lungs and a , ae old, 0 4 2110 I eanere: aeres ,on ntrav. | 0-1 mg.| K.170_,, Very slight T. of i : lungsand kidneys. , 2111 Intrav. |0-01 mg.| K.170 ,, vey slight T. Of Jan. 27,1909} Col. 2 a Ren. 5 21 (| 2130 | Intrav. | 0-1 mg. | K. 104, Se) T ays old, on ight T. of lungs serum. l 2129 Intrav. |0-01 mg.) K.104__,, and kidneys. \ : 35 ! Col. 8 "ae a “ 2132 Intrav. | 0-1 mg. | K. 104 5 Bight T. of luogs a : and kidneys. | serum, l 2131 Intrav. | 0-01 mg.| K. 104 ,, Slight T. of lungs only. : 49 FULL POST-MORTEM NOTES OF THE CALVES INOCULATED. PAGE ( Calf 1175 _ a $5 a Sy om a aa BO | Calf 1177 a ee hee a sah 7 ae oe A | Cal 1237 ob = ue a “ue a 3 52 Calf 1327 is ae si ue ‘Y a sig ca BB Virus H13 “A.D.” 2 Cale 1207 sh 2 ef ss mF x5 2 8 | cat 1301 a ee “is re a ae Be iy. Bi | Cate 1309 i om oe bia es i sat oe BB | Calf 1337 x - Os i me a = BY “L Calf 1343 es as 2 = a ie sat .. 60 ae oe a wie oe 3 dee .. 61 oy Calf 1193 S 4 Saf a os im Sas be. GH | Calf 1213 ne si oe on ss ts a .. 62 i} Calf 1263 we me ace Be ss a i (64 Virus H 49. “9.0.” 4 Galg 1347. ae a ne ie si seks oa .. 66 i | ale 1387 a rr af Hi _ Z 7 ue 67 Calf 1415 ee we 22 big fe ba ee . 68 : | Calf 1413 ae tg i 2 is a a 69 { cate 1501 Ly oe a fee Pe WEY 1g os dos, 189 ( Calf 1203 oe bee ee “4 sae a on i 74 Virus H 60. “W.B.” Calf 1481 sa aoe ea Seis aes kins se dene 42. Calf 1425 5 7 si iz mt fh ae wa 93 ; Calf 1489 = bes ow S te ane a 4 Virus H 90. “P| 7 Calf 1493 ee Rae ate 12 me ss ss. ey ae Note.—The full post-mortem notes of three calves, Calves 1099, 1107 and 1109, which were inoculated to. test the virulence of the bronchial gland strain from Virus H 60. “ W.B.”, will be found on pages 279-282 of the Report on Cases of Human Tuberculosis (other than lupus). The full post-mortem notes of Calves 1383 and 1471, which were inoculated to test the virulence of the retroperitoneal gland strain of Virus H 90. “I.P.”, together with the post-mortem notes of Goat 55, will be found on pages 460 and 461 of the same report. 27676 50 CALF 1175. VirusH 13. “A.D.” Subcutaneous inoculation of culture derived from Calf 301 (thoracic gland), direct. Dose—100-0 milligrammes. Date of Inoculation—March 23, 1907. [Age, 13 weeks. ] Killed when in good health—June 12, 1907. [81 days after inoculation. ] Clinical Notes. A large pendulous tumour developed at the seat of inoculation on the left side of the neck ; on the 27th day this measured 18 by 9 cm. The tissues of the dewlap were slightly oedematous, and the left pre- scapular gland was very large, measuring 14 cm. in length. The general health of the animal was unimpaired. Shortly afterwards the tumour opened and dis- charged caseo-purulent matter, subsequently diminish- ing in size. On the 52nd day it was firm and slightly raised, measuring 13 by 6 cm., and showing over the lower part an ulcer 4 cm. in greatest diameter, partly covered with a caseous scab. The prescapular gland was still considerably enlarged, measuring 11 cm. in length. The calf was apparently in good health. On the 81st day the calf was killed. Temperature—During part of the second and third weeks the temperature was a little raised (maximum 39-3° C.) ; subsequently it was normal. Tuberculin Test.—The calf was not tested subsequent to inoculation. Weights. ewt. qrs. ibs. March 23,1907 ... pas we Ll 0 14 June 12,1907 ... et we 1 88 Total gain of weight.—2 qrs. 22 lbs. Average rate of gain per week —6°9 lbs. POST-MORTEM EXAMINATION. The carcass was in good condition. Local Lesion.—The local lesion was firm and measured 10 by 7 by 4 cm.; it showed towards the lower extremity an oval punched-out ulcer 3:5 cm. in length with inverted edges ; this ulcer communicated with a flat space in the centre of the tumour which contained a small amount of yellow tenacious caseo- pus ; the space was surrounded by a zone of fibrous tissue, 1 cm. in greatest thickness, infiltrated with caseous streaks and foci; the muscles beneath this collapsed cavity or space were fibroid and closely beset with caseo-calcareous nodules ranging in size from a millet seed to a pea; the skin was much thickened and was adherent to the wall of the cavity. Left Prescapular Gland—The left prescapular gland measured 8°5 by 5 by 4:5 cm., and weighed 4 ozs.; on section it was tuberculous throughout, the greater portion being composed of dense pinkish yellow caseo-necrotic substance gritty around the periphery, the rest of caseo-calcareous tissue; the capsule was greatly thickened in places. Right Prescapular Gland.—The right prescapular gland measured 5 by 2:3 by 1:4 cm. and showed in the cortex fairly numerous soft caseous nodules similar to those m the precrural glands (which see). Left Prepectoral Glands.—The rounded prepectoral gland, the size of a swallow’s egg, showed about half the cortex dense and caseous, while the rest was studded with caseo-calcareous tubercles. The reniform gland was enlarged and contained a few caseous tubercles. Cervical Glands—One in the middle of the neck on the left side, the size of a large pea, was caseous throughout. Other cervical glands showed discrete yellow caseous nodules up to a hemp-seed in size. A bemolymnh gland on the left side of the neck contained a soft caseous millet-seed sized tubercle. Thorax. Pleura.—On the costal pleura there were two hemp- seed sized tubercles, each with a minute yellow caseous centre. On the visceral pleura were two small pinkish fibrous nodules not caseous nor calcareous. Lungs.—The lungs were crepitant throughout and showed no collapsed areas; they contained fairly numerous evenly distributed tubercles ranging in size from a mere point up to 2 mm. in diameter; the larger tubercles had yellow calcareous centres, the smaller ones were grey and homogeneous througbout. In the thin margin of one of the cephalic lobes there was a larger nodule with a caseo-calcareous centre. Thoracic Glands.—The dorsal mediastinal and the bronchial glands were slightly enlarged and were closely beset with yellow caseo-calcareous tubercles, mainly discrete, but here and there aggregated together ; the long mediastinal measured 3°3 by 2 by 2 cm. The ventral mediastinal glands contained three or four caseous tubercles. Heart.—On the pericardium covering the great vessels and the auricles, and along the interven- tricular sulci, were patches of tuberculous granu- lations, and here and there clusters of grey tubercles ; on the surface of the ventricles there were a few isolated grey tubercles. There were a few patches of tuberculous granulations on the parietal pericardium as well as some discrete tubercles. Heart muscle and valves normal. Larynx and Trachea.—Normal. Abdomen. Omentum.—On the inferior surface there were two loosely pedunculated millet-seed sized grey tubercles, each with a calcareous grain in the centre. Peritoneun.—On the peritoneal surface of the dia- phragm near the falciform ligament there was a slightly raised greyish pink nodule 1 cm. in diameter with a caseo-calcareous centre. On the falciform ligament there were three or four small flattened caseous tubercles with grey margins, Spleen.—The spleen was normal in size, and showed in the pulp fairly numerous evenly distributed soft yellow caseous nodules, varying in size from 1°5 to about 5 mm. in diameter ; the nodules had grey ee margins and the larger ones were lobular in outline. Liver.—The capsule of the liver showed three flattened grey slightly raised tubercles about 1-5 mm. ia diameter, not involving the liver substance. On the anterior surface there were besides a few filmy grey fibrous tufts loosely attached to the capsule. In the substance of the liver two minute opaque points were seen, but no definite tubercles. Portal Glands.—The portal glands, not obviously enlarged, were closely beset with discrete yellow miliary caseo-calcareous tubercles, Coeliac Glands.—The coeliac glands were slightly enlarged, and closely beset with yellow miliary caseo- calcareous tubercles, as in the portal glands. The renal gland was similarly affected. se Kidneys.—In the cortex of the left near the 2 surface were three, and in that of the right four grey a the largest the size of a millet seed: two were calcareous in the centre, the oth t geneous throughout. sacs Suprarenal Bodies. —Normal., Zesticles.—Normal. Alimentary Tract. Tongue, Pharynz, Tonsils.—Normal. Intestines.— All the Peyer’s patches showed scattered about in the lymphoid tissue soft yellow foci, mostly about the size of millet seeds ; they were most con- spicuous from the serous surface, and in several places were aggregated together to form large nodules ; the mucous membrane over them was raised, and in some cases showed a small depression or pit but no actual ulcers. The large intestine was normal. Mesenteric Gilunds.—All the mesenteric ‘glands showed in the cortex fairly numerous yellow caseo- calcareous tubercles, mostly about the size of a millet seed or less, but several attaining the size of a hemp- seed ; they were for the most part discrete, but here and there formed irregular calcareous patches. Lleo-colic Glands.—The ileo-colic glands were most severely affected, and their substance extensively re- placed by irregular caseo-calcareous patches. Various Lymphatic Glands. The Precrural Glands showed in the cortex numerous soft caseous nodules, varying in size up to about 4 mm. in diameter ; the larger nodules were lobular in out- line, and appeared to consist of several smaller ones united together ; no calcareous foci were detected in the caseo-pus. The Popliteal, Gluteai, [schiatic, Iliac, and Lumbar Glands contained similar nodules in varying number. There were similar nodules in the Submazillury, Retropharyngeal, and Parotideal Glands, but these were generally larger, and had deeply congested margins. Microscopical Examination. Emulsion of Prescapular Gland.—Tubercle bacilli numerous. Emulsion of Popliteal Gland. — Tubercle bacilli scanty. Emulsion of Mediastinal Gland.—Tubercle bacilli scanty. Tubercle from Intestine.—One tubercle bacillus seen. Focus from Liver.—No tubercle bacilli seen. Animals Inoculated. Calf 1237 was inoculated subcutaneously with 20 cc. of emulsion made from the prescapular gland. Rabbit 1336 was inoculated subcutaneously with the same emulsion. (Died 78 days, G.T.) Rabbits 1337 and 1338 were inoculated subcu- taneously with an emulsion made from the mediastinal gland, (Died 92 and 47 days, G.T.) CALF 1177. Virus H 13. 66 A.D.” Subcutaneous inoculation of culture derived from Calf 301 (thoracic gland), direct. Dose—50°0 milligrammes. Date of Inoculation—March 23, 1907. Killed when in good health—July 11, 1907. Clinical Notes. A prominent tumour developed at the seat of inoculation on the left side of the neck. On April 19 it measured 14 by 8 cm. The left prescapular gland was then very large—12cm. in length ; the muscle between the tumour and the gland was slightly thickened. The calf was in good health. On May 14 the tumour was pear shaped, measur- ing 14 by 7cm. The gland was 10cm. in length. The calf at this time was a little thinner, and the rate of respiration seemed a little increased ; but it quickly recovered, and during the remaining eight weeks of the experiment looked well and fat. On July 11 the tumour was found to have opened and to be discharging caseo-pus ; the calf was there- fore killed on this day. Temperature.—The temperature rose on the 10th day, reaching a maximum of 40°6°C. on the 14th day. Tt then slowly returned to the normal and remained normal subsequently. Tuberculin Test.—July 5, 1907. [104 days after inoculation.] Reacted. Rise of temperature, 1-8°C. Weights. cwt. qrs. lbs. March 23, 1907 ... ; i 0 3 17 July 11,1907... eee » 1 38 9 Zotal gain of weight.—3 qrs. 20 Ibs. Average rate of gain per week.—6°5 lbs. POST-MORTEM EXAMINATION. The carcass was in very good condition. 27676 [Age about 12 weeks.] {110 days after inoculation.] Local Lesion.—At the seat of inoculation on the left side of the neck there was a firm tumour, measuring 10 by 6°5 by 4°5cm., the skin towards the upper extremity of which showed a small opening at the bottom of a funnel shaped depression through which thick caseo-pus could be expressed. On section the tumour was lenticular in outline (plano-convex), and was composed of a dense pinkish yellow caseous sequestrum, and a fibrous capsule from which the caseous mass was almost completely separated by thick caseo-pus ; the capsule was lined internally with a layer of granulation tissue 1:5 mm. in thickness, containing here and there calcareous grains ; the skin over the tumour was much thickened. Left Prescapular Gland—The left prescapular gland measured 8:2 by 5:1 by 4cm.; on section it was composed almost throughout of dense pinkish yellow caseous tissue, gritty around the peripheral parts; at one extremity there was some normal- looking gland tissue which was very oedematous and contained discrete calcareous tubercles. The capsule was much thickened, especially on the convex surface. Right Prescapular G'land.—The right prescapular gland measured 55 by 2°5 by 1 cm, It was oede- matous (recent tuberculin inoculation) and contained scattered calcareous tubercles. Prepectoral Glands.—On the left side one was very oedematous but free from tubercles ; two others each contained two calcareous tubercles. Two on the right side each contained two or three calcareous tubercles. Cervical Glands.—On the left side a midcervical gland showed a calcareous patch. Other cervical glands on this side and all those on the right contained scattered calcareous tubercles. K 2 Thoraz. Pleura.—Normal.! Lungs.—The lungs were pinkish in colour, crepitant throughout, and collapsed normally ; they contained evenly distributcd though not very numerous tubercles, ranging in size from 1 to 2 mm. in diameter ; the larger ones were yellow and calcareous with grey margins ; the smaller ones were grey and had glassy gritty centres. Thoracic Glands.—The bronchial and mediastinal glands were slightly enlarged and showed around the cortices irregular calcareous patches, composed of aggregated tubercles, and discrete calcareous tubercles. Ventral Mediastinal Glands—There were a few small calcareous tubercles in each. Heart, Larynx, and Trachea.—Normal. Abdomen. Omenium.—On the ventral surface there were about a dozen pedunculated lenticular. tubercles with cal- careous centres, the largest about twice the size of a millet seed. Peritoneum.—Normal. Spleen—The spleen, normal in size, showed on section fairly numerous svenly distributed yellow calcareous tubercles with thin fibrous capsules, ranging from 1 to 2 mm. in diameter. Liver—Very sparsely scattered throughout the substance of the liver were minute pearly or glassy calcareous tubercles which readily shelled out. Portal Glands.—The portal glands, not enlarged, contained fairly numerous irregular calcareous tubercles, here and there aggregated together into small groups. Kidneys.—Normal. Suprarenal Bodies.—In the cortex of the left there was a grey tubercle, calcareous in the centre, 2 mm. in diameter. : The right was normal. Alimentary Tract. Tongue, Pharynz, Tonsils——Normal. Intestines.—Several of the Peyer’s patches in the CALF 1237. Virus H 13. 52 small intestine contained soft yellow submucous tubercles slightly gritty from calcification. Large intestine normal. Mesenteric and Ileo-colic Glands.—These glands were little if at all enlarged, but were rather extensively tuberculous, containing numerous calcareous patches and nodules as well as discrete tubercles ; they were not quite so severely affected as the bronchial and mediastinal glands. Eyes.—Normal. Mammary Gland.—Normal. Various Lymphatic Glands. The Precrural Glands showed in the cortices scat- tered yellow calcareous tubercles, irregular in outline, the largest about the size of a millet seed; they readily shelled out from the gland substance. The Popliteal, Axillary, Submazillary, Hyoid, Retro- pharyngeal, Gluteal, Ischiatic and Pudic, Lumbar and Iliac Glands contained similar tubercles. The Renal Gland contained similar but rather more numerous tubercles. Coeliac Glands.—One contained a calcareous patch and a number of discrete calcareous tubercles ; the rest contained discrete calcareous tubercles as in the peripheral lymphatic glands Microscopical Examination. Emulsion of Prescapular Gland.—Tubercle bacilli moderately numerous. Emulsion of Spleen.—Six tubercle bacilli seen. animals Inoculated. One rabbit, No. 1380. was inoculated subcutaneously with an emulsion made from the prescapular gland. (Died 61 days, G.T.) i Two rabbits, Nos. 1381 and 1382, were inoculated subcutaneously with an emulsion made from the spleen. (Died 167 and 110 days, G.T.) “AD” Subcutaneous inoculation of emulsion of the left prescapular gland from Calf 1175. Dose—20:0 cc. of emulsion containing numerous tubercle bacilli. Date of Inoculation—June 12, 1907. Killed when very ill—August 3, 1907. Clinical Notes. The course of the disease was similar to that usually seen in calves suffering from acute tuberculosis. On [Age about 15 weeks. ] [52 days after inoculation. ] Saturday, August 3, the calf was very ill, emaciated, and breathing rapidly, and appeared likel to die day or two. It was therefore killed. a ee pr o 3° Temperature. WHE JOLY AUG Tuberculin Test.—The calf was not tested sub- sequent to inoculation. Weights. ewt. qrs. lbs, June 12, 1907 ... 1 017 August 3, 1907 ... 1 0 1 Total loss of weight.—16 lbs. Average rate of loss per week.—2°1 lbs. POST-MORTEM EXAMINATION. The carcass was very thin. Local Lesion—At the seat of inoculation on the left side of the neck there was a firm elongated tumour measuring 17 by 9°5 cm. by 5 cm. in thickness (the latter measurement including skin and muscle), and weighing 1 lb. 5 ozs. On section it was composed of three layers, the outer one of skin, 1 cm. in thickness, closely beset with congested caseous tubercles, the inner one of dense pinkish yellow caseo-necrotic substance, the internal one of fibroid muscle tissue closely infiltrated with irregular caseous nodules of various sizes. Left Prescapular Gland.— The left prescapular gland measured 9 by 5 by 5 cm. and weighed 4 ozs., and was composed practically throughout of dense pinkish yellow caseo-necrotic substance, showing many petechial points of hemorrhage ; the capsule was much thickened. Right Prescapular. Gland.—The right prescapular gland measured 4 by 1°5 by 0°8 cm., and appeared normal on section. Prepectoral Glands.—On the left side there were four ; one rounded and about 1°3 cm. in diameter was caseous throughout, another about the same size was indurated and beset with caseous tubercles, a third was very large, 4 cm. in length, oedematous, and showed scattered whitish foci; the fourth, a very small gland, contained a few early tubercles, On the right side one was speckled with minute greyish-white foci; another contained three caseous tubercles. Cervical Glands.—On the left side in the lower part of the neck there was a firm gland 3 cm. in length which was caseous throughout ; in the upper part of the neck a gland was caseating throughout, but was not so advanced as that in the lower ; a gland near the latter showed numerous caseous tubercles in the cortex ; a mid-cervical gland contained one caseous tubercle. On the right side a lower cervical gland had some caseating patches in the cortex ; the rest were normal. Agillary Gilands.— There were three caseous tubercles in the right axillary gland and one in the left. Thoraz. Pieura.—The lymphatic fringes along the margins of the ribs were here and there very slightly hypertrophied and contained a few miliary tubercles. On the diaphragmatic pleura there was a cluster of caseating tubercles and a few patches of tuberculous granulations. There were two or three grey millet- seed sized tubercles on the pericardial pleura. Lungs —The lungs weighed 6 Ibs. 10 ozs, The cephalic, the right middle, and the antero - ventral portions of the caudal lobes were extensively hepatized, very little crepitant lung tissue remaining and this chiefly along the dorsal borders, and their surfaces were closely studded with raised grey nodules, the largest about the size of a hemp seed or more ; the surfaces of the posterior parts of the caudal lobes were smooth and showed under the pleura numerous nodules, many of which were surrounded by hepatized patches, some occupying the whole of a lobule; on section the parenchyma of the lung was closely beset with grey nodules with yellow caseous centres similar to those seen from the surface; in the caudal lobes several lobules were uniformly dense and caseating. Thoracic Glands.—The dorsal mediastinal and bronchial glands were enlarged, weighing together 6 ozs.; the long mediastinal gland measured 11 cm. in length. On section, the cortices were composed almost entirely of firm grey translucent tissue mottled with irregular caseous foci and small patches in places forming a network ; many of the smaller glands were very oedematous. Heart.—Around the margin of the left auricle were three grey miliary tubercles. In the wall of the right auricle there was a millet-seed sized caseous tubercle. The heart was otherwise normal. Laryns and Trachea.—Normal. Abdomen. There was no fluid in the peritoneal cavity. Omentum and Peritoneum.—On the ventral surface of the omentum there were two millet-seed sized grey tubercles, The peritoneum was normal. Spleen.—The pulp was closely beset with firm caseous nodules with grey margins, varying from 1:5 to 3° mm. in diameter ; there were no tubercles on the surface. Liver.—The liver showed on the surface numerous nodules ; some were slightly raised and flattened out up to 3 mm. in diameter, others were situated just beneath the capsule, while others again were dimly seen in the depth; on section the substance was moderately closely beset with tubercles which varied from 1 to 2°5 mm. in diameter ; the smaller ones were greyish white, some being distinctly more opaque in the centre than around the margins, the larger ones had yellowish centres. Portal Glands.—The portal glands were enlarged and closely beset with caseating nodules in places confluent. Kidneys.—Many of the lobules showed atrophied and fibroid patches in the cortex to which the capsule was adherent ; in one of the lobules the greater part of the cortex was replaced by cheesy purulent substances ; the cortex of each kidney contained fairly numerous caseating nodules varying from a pin’s head to a hemp seed. Suprarenal Bodies.—These appeared normal. Coeliac Glands.—Two were slightly enlarged and showed, on section, patches composed of aggregated caseating nocules as well as discrete caseating nodules ; another contained one caseating nodule, irregular in outline, the size of a hemp seed. fenal Gland.—The renal gland was enlarged and closely beset with irregular caseous nodules varying up to 2°5 mm. in diameter. Lumbar Glands.—The lumbar glands resembled the renal. Alimentary Tract. Tongue, Pharynx, Palate, Tonsils—Normal. Submacillary, Retropharyngeal and Parotideal Glands——Each contained a few miliary caseous tubercles. Small Intestines—All the Peyer’s patches contained miliary caseous tubercles in small number. Large Intestine—Normal. Mesenteric Glands.—The mesenteric glands, not enlarged, contained scattered discrete caseating nodules, the largest 5 mm. in diameter. 54 Tleo-colic Glands.—The ileo-colic glands contained: numerous small tubercles. Testes. —Normal. Eyes.—Normal. Various Lymphatic Glands. Precrural Glands—The right was normal; the: left contained three caseous tubercles.* Popliteal, Gluteal and Pudic Glands.—Normal. Microscopical Examination. Emulsion of Left Prescapular Gland.—Tuberele- bacilli moderately numerous. Emulsion of Portal Gland.—Tubercle bacilli scanty. Emulsion of Bronchial Gland. — Tubercle bacilli scanty. Tubercle from Heart.—Tubercle bacilli, two seen. Smear from Right Suprarenal.—No tubercle bacilli: seen. Smear from Faeces—No tubercle bacilli seen. CALF 1827. Virus H 18. “ aD.” Subcutaneous inoculation of culture derived from the bronchial gland of Calf 1237. Dose—50'0 milligrammes. Date of Inoculation—December 30, 1907. Died—January 26, 1908. Clinical Notes. The course of the disease was similar to that usually seen in calves suffering from acute tuber- culosis, Temperature—The temperature rose to 39°6°C. on the third day and reached a maximum of 40°2°C. on the 8th day. It subsequently remained above normal (between 39:0 and 39:9°C.) tor 15 days. During the following three days the temperature fell one degree (to 38-0° C.), and on the morning of the next day the animal was found dead. Weights. grs. Ibs. December 30, 1907 3 20 January 26, 1908 3 13 Total loss of weight.—7 lbs. POST-MORTEM EXAMINATION. The body was thin. Local Lesion—At the seat of inoculation on the ,eft side of the neck there was a caseo-necrotic mass, semilunar on longitudinal section, measuring 13 by 9 by 3cm.; both the skin and the muscles were infiltrated, and the prescapular gland was adherent to it. Left Prescapular Gland.—The left prescapular gland measured 9 by 4°5 by 4:5cm., and was com- posed throughout of dense yellow caseated substance. Prepectoral Glands.—The prepectoral glands on both sides contained discrete miliary caseous tubercles. Right Prescapular Glond.—The right prescapular gland measured 5 by 2 by 1cm., and showed in the cortex numerous discrete miliary caseous tubercles, [Age about 11 weeks.] [27 days after inoculation. ] Thoraz. Pleura.—The costal and diaphragmatic pleura on: the right side was thickly covered with caseous miliary tubercles ; there were numerous similar tubercles on the pericardial pleura; on the left parietal pleura miliary tubercles were very sparsely scattered, but in the fringes here and there were granules. groups of grey Lungs.—The lungs were heavy and weighed 4 Ibs. 13 ozs. The anterior and right middle lobes and the greater portions of the posterior lobes were dark red and solid ; the rest of the caudal lobes was congested and showed irregular diffuse areas of consolidation - the tissue of the lungs was closely and evenly beset with caseating miliary tubercles. At the root of the right lung in front of the bronchus there was a collection of thick greenish pus with a fibrous capsule. It was not connected with a bronchus and appeared to be a suppurating gland. LPhoracie Glands.—The bronchial and mediastinal glands were enlarged; their cortices were closely beset with grey caseating tubercles, the majority of which had coalesced forming irregular patches, Heart.— Muscle and valves normai. There were several clusters of tubercles on the pericardium at the root of the great vessels and a crop of miliary caseous tubercles on the parietal pericardium. Abdomen. Omentum and Peritoneum.—The ventral ; ha face of the omentum was rather thickl fed Gath ak caseating tubercles, ekly covered with miliary The parietal peritoneum was normal. ® Spleen.—The spleen was much enlarged and weighed 14 ozs. ; the pulp was firm and was packed almost as -closely as possible with miliary caseating tubercles with reddish margins, with difficulty differentiated from the small amount of red pulp existing between the tubercles ; the cut surface had a coarsely granular appearance. Liver.—The substance of the liver was closely and evenly beset with greyish-white miliary tubercles. Gall-bladder.—There were a few tubercles in the ‘wall of the gall-bladder. Portal Glands.—The portal glands were enlarged and showed around the cortices irregular grey patches containing caseous foci and here and there a caseous network. Kidneys.—Each kidney showed in the cortex fairly numerous grey miliary tubercles with caseous centres. Suprarenal Bodies.—HEach contained about half-a- -dozen submiliary greyish-white tubercles. The Coeliac, Lumbar, and Renal Glands resembled 5 the portal, showing in the cortex aggregations of caseating miliary tubercles. Testes.—Normal. Alimentary Tract. The pharyngeal, laryngeal and tracheal mucous membranes shewed numerous miliary tubercles. There were tubercles under the mucous membrane of the tongue at the base and margins. Intestines. —There were extremely numerous miliary caseous submucous tubercles throughout the whole length of the small intestine; many of the Peyer's patches were exceptionally closely beset. The large intestine also shuwed numerous sub- mucous caseous tubercles. Various Lymphatic Glands.—All the peripheral lymphatic glands and the mesenteric glands were closely beset with caseous miliary tubercles. CALF 120%. Virus H 13. Subcutaneous inoculation of culture derived from Calf 301, direct. -for two generations on glycerin-serum. | Dose—100°0 milligrammes. Date of Inoculation—March 26, 1907. Killed when in good health—June 26, 1907. Clinical Notes. A tumour of moderate size developed at the seat of “inoculation on the left side of the ueck. On the 24th day it was pear-shaped, measuring 9 by 6 cm.; on the 51st day it measured 11 by 6 cm. and was ‘fluctuating. “ A.D.” [The culture had been growing [Age 13 weeks.] [92 days after inoculation. ] The left prescapular gland became moderately enlarged, and on the 51st day measured 9 em. in length. No changes of importance took place subsequently in the local conditions. The calf showed no signs of illness during tke experiment. Temperature. MARCH MAY Trea June | ts June 56 he [Enptralint ernnind? hik- (eK 35a e Tuberculin Test.—The calf was not tested subse- POST-MORTEM EXAMINATION. - quent to inoculation. The carcass was in fair condition. Weights. Local Lesion.—On the left side of the neck there ewt. qrs. Ibs. was a somewhat pear-shaped fluctuating tumour March 26, 1907 aus ae 1 0 16 measuring 7 by 5 by 3 cm.; on section it was found June 26, 1907... a ees 1 Total gain of weight.—2 qrs. 5 lbs. slverage rate of gain per week.—4'7 lbs. to be a cyst, surrounded by a fibrous wall lined with a thin layer of caseous substance rough and shagg internally, and filled with turbid watery fluid and caseo-necrotic masses and shreds. 2-2 Left Prescapular Gland.—The left prescapular gland measured 7:5 by 45 by 3 cm., and on section showed about three-quarters of the cortex caseous, parts being homogeneous and of a salmon-pink colour, the rest yellow and highly calcareous ; in the remainder of the gland were discrete calcareous tubercles. Right Prescapular Gland.—The right prescapular gland measured 3-5 by 1:8 by 1 cm. and contained a few calcareous tubercles. Prepectoral Glands.—On the left side the rounded gland was slightly enlarged and closely beset with ‘minute calcareous tubercles. Another gland contained one small calcareous tubercle. Those on the right side were normal. Cervical Glands.—One in the middle of the neck on the left side and two near the articulation of the jaw showed each one or two calcareous tubercles. The rest were normal. Thorax. Pleura.—Normal. Lungs.—The lungs were pinkish in colour and were crepitant throughout with the exception of two or three hepatised lobules in the caudal lobes ; the lung parenchyma contained fairly numerous pale grey tubercles somewhat irregular and occasionally inde- finite in outline, varying in size up to 1 mm.; they were most numerous in the caudal lobes, sparsely scattered in the left cephalic and almost absent in the right cephalic. The larger tubercles all bad calcareous centres, but most of the smaller ones were homo- geneous throughout. Thoracic Glands —The mediastinal and bronchial glands were almost twice the normal in size and were closely beset with calcareous tubercles, in places loosely aggregated together, especially in the larger glands, to form irregular calcareous patches. Heart.—Normal. Abdomen. Omentum and Peritoneum.—Normal. Spleen.—The spleen was normal in size and showed attached to the capsule agreyish pink flattened nodule, I cm. in diameter, containing calcareous grains in the centre ; on section the pulp contained fairly numerous small -calcareous tubercles with grey margins, the largest about 1 mm. in diameter ; 20 were counted in an area 5 cm. square. : , Liver.—The liver appeared normal on the surface and on section. Gall-bladder.—Normal. Portal Glands—The portal glands were slightly enlarged, and their cortices were closely beset with irregular yellow calcareous miliary tubercles, forming in places small groups. Kidneys.—In the cortex of each kidney there were about half-a-dozen translucent grey tubercles slightly opaque in the centre, the largest 1 mm. in diameter ; some were situated just under the capsule while others were found in the depth of the cortex. Suprarenal Bodies.—In the cortex of the left sup- rarenal there was a millet seed sized tubercle with a calcareous centre anda fibrous margin. The right was normal. Coeliac Glands.—The cortex of one was rather closely beset with small irregular calcareous tubercles ; in the others tubercles were more sparsely scattered. 36 Renal and Lumbar Glands.—These were slightly enlarged, and resembled on section the first coeliac gland. Iliac Glands.—In the left there were two minute calcareous tubercles. The right contained rather more numerous tubercles, two of which were larger than millet seeds. Alimentary Tract. Tongue, Pharynz, Tonsils—Normal. Submaxillary and Retro-pharyngeal Glands.-—These contained fairly numerous small calcareous tubercles. Parotideal Glands.—In the parotid glands there were similar but not so numerous tubercles. Small Intestines—A Peyer’s patch towards the anterior end contained a yellow slightly gritty tubercle the size of a pin’s head. -In the long terminal patch there were several similar foci and a few small ulcers in the mucous membrane with slightly raised margins but with no caseation in the base. Large Intestine—Normal. Mesenteric Glands. — All showed in the cortex calcareous patches composed of aggregated tubercles, as well as discrete calcareous tubercles. Lleo-colic Glands. — The ileo-colic glands were closely beset with calcareous tubercles and resembled the mediastinal and portal glands. Gastric and Colic Glands.—These contained a few discrete calcareous tubercles, Testicles.—Normal. Various Lymphatic Glands. . Asillary—The right contained a pinhead-sized calcareous tubercle. In the left were two calcareous grains. Popliteal—The right was normal. tained two calcareous foci. Precrural.—In the cortex of each there were scattered tubercles varying up to 1 mm. in diameter ; The left con- a few of tue tubercles were caseous and softened; -but the majority were quite calcareous. : Pudic.—One contained a millet-seed sized caseous tubercle. ae : Ischiatic.—Normal. Giuteal—In one there was an irregular yellow calcareous tubercie ; the other was normal. Microscopical Examination. Emulsion of Prescapular Giland.—Tubercle bacilli numerous. Emulsion of Mediastinal Gland.—Tubercle bacilli fairly numerous. Animals Inoculated. Two rabbits (1354 and 1355) were inoculated sub- cutaneously with the prescapular gland emulsion, and two (1356 and 1357) with the emulsion made from the mediastinal gland. 1354 and 1357 died of G.T. in 92 and 53 days re- spectively ; the others died prematurely. ~ o CALF 1301. Virus H 13. “ A.D.” Subcutaneous inoculation of culture derived from a large (eugonic) colony [No. 2] on a glycerin-serum plate sown from Guinea-pig 2491. This guinea-pig had been injected with the “ Calf 301” strain of culture. Date of Inoculation—November 12, 1907. [Age about five months. ] Dose—100-0 milligrammes. Died—January 2, 1908. [51 days after inoculation. ] Clinical Notes. Twenty-three days after inoculation there was a prominent fluctuating pear-shaped tumour measuring 16 by 9 cm. at the seat of inoculation; the adjacent prescapular gland was enlarged and very hard, measuring 11 cm. in length. The coat was somewhat staring, and the animal appeared to have lost flesh, otherwise the condition was fairly good. The disease subsequently progressed slowly ; the animal gradually became weak and very emaciated, and the respiration laboured, and it died on the 51st day after inoculation. Temperature. NOVEMBER Tuberculin Test.—The calf was not tested subsequent to inoculation. Weights. ewt. qrs. lbs. November 12,1907... wo 1 2 18 January 2, 1908... 0 ow. 1 0 22 Total loss of weight.—1 qr. 24 lbs. Average rate of loss per week.—7 lbs. POST-MORTEM EXAMINATION. The carcass was emaciated. Local Lesion.—The tumour at the seat of inoculation was pear-shaped, prominent, and fluctuating, it weighed 1 Ib. 2 ozs. and measured 16 by 9 by 7 cm. On section a large cavity was seen in the upper part filled with yellow serous fluid containing flakes of caseous material and crossed by fibrous trabeculae. The remainder of the tumour was composed of pinkish-yellow caseo-necrotic substance, breaking down into ragged masses around the cavity, firm elsewhere. The overlying skin and the subjacent muscles were infiltrated to a slight extent. Left Prescapulur Gland.—The left prescapular gland measured 12°55 by 7 by 4 cm., and was composed throughout of dense pinkish yellow caseated tissue showing no sign of calcification. Prepectoral Glands.—The rounded gland on the left side measured 1°5 cm. in diameter, and was packed with confluent yellowish-white caseating tubercles. The kidney-shaped gland was swollen and slightly congested, and showed one small caseating nodule in the cortex. A third gland on this side was normal. The prepectoral glands on the right side were normal, Right Prescapular Gland.—The right prescapular gland measured 5 by 2°4 by lem. It was oedematous 27676 “DECEMBER and contained a number of irregular caseating tubercles, the largest about 2 mm. in diameter. Cervical Glands.—A small mid-cervical gland on the left side contained caseating patches ; one on the right side showed two caseating tubercles. 4 The upper cervical glands on the left side were normal ; one on the right side contained a caseating tubercle. Axillary Glands.—The right axillary gland con- tained a hempseed-sized caseating nodule in the cortex ; the left was normal. 4 Thorax. Lungs.— The lungs were very heavy, weighing 8 Ibs. 14 ozs. ; they almost completely filled the chest. The surface of the whole of the left lung and of the upper lobe of the right was mottled with dark red confluent patches of consolidation ; the surface of the remaining lobes of the right lung was almost entirely dark red in colour, small patches and islands of pink tissue alone remaining. The substance of both lungs under the pleura was thickly and evenly beset with greyish-yellow caseating . nodules sometimes confluent, ranging from 1 to 8 mm. in diameter. The upper lobes on both sides were slightly less severely affected than the rest of the lung. On section of the lung the tubercles appeared even more numerous than they did from the surface, and large caseating patches of various sizes were seen, the largest measuring 1°5 by 0°7 cm. ; the condition of the organ on section resembled that on the surface, the right lung being to a large extent dark red and consolidated, the left showing only small patches of consolidation. Thoracic Glands.—The thoracic glands were very much enlarged and weighed altogether 14 ozs. The long mediastinal gland measured 15 by 6 by 2°7 cm. On section the cortices were firm and composed of grey translucent fibroid tissue closely beset with yellow caseous areas. pp Heart and Pericardium.—Normal. Larynx and Trachea.—Normal. Pleura.—There was one loosely attached caseating flattened nodule measuring 8 by 6 mm. on the pleura just behind the second rib on the right side; no tubercles were seen elsewhere on the parietal pleura. On the pleural surface of the diaphragm there was a group of small loosely attached lenticular tubercles. =.There were two caseating nodules situated close together in the wall of the diaphragm, the larger about a hemp seed in size. The peritoneal surface of the diaphragm was normal. ' Abdomen. Omentum and Peritoneum.—On the omentum there was one loosely attached flattened caseating tubercle 0°5 cm. in greatest diameter. The peritoneum was normal, Spleen—The spleen was not enlarged, and on section showed scattered yellow caseating tubercles, the largest about 2 mm. in diameter. About sixty were counted in a longitudinal section. Liver—The liver was fatty looking and weighed 5 Ibs. 6 ozs. The surface was mottled and showed thinly scattered tubercles with yellow centres and translucent margins varying in size from a mere point up to 4mm. in diameter; one or two of the Jarger projected on the surface, many were dimly seen through the capsule. Sparsely scattered tubercles were seen on section. Portal Glands.—The portal glands were enlarged and oedematous and were packed with coalescing caseating nodules. Kidneys.—About thirty yellowish-white tubercles were counted on the surface of the left kidney varying from 1 to 4 mm. in diameter. Similar tubercles were sparsely scattered throughout the cortex. The right kidney was similarly affected. Suprarenal Bodies.—In the cortex of the left three yellow caseating tubercles were seen, the largest being 3 mm. in diameter. The right was normal. Coeliac Glands. — One coeliac gland Several small caseating nodules. contained 58 Lumbar and Renal Glands.—These glands were enlarged, oedematous, and similar on section to the portal. Iliac Glands.—The left was normal, the right contained two small caseating nodules. Testicles.—Normal. Alimentary Tract. Tongue, Pharynx, Tonsils.—Normal. Retro-pharyngeal Glands.—The left contained a nodule 6 mm. in diameter, the right a single small caseating tubercle. Parotideal Glands.—Each contained several casea- ting nodules. Submacillary Glands.—Normal. Intestines—In one Peyer’s patch two millet-seed sized caseous tubercles were seen, in another there was a single tubercle. Mesenteric Glands.—Nearly all the mesenteric glands. contained one or two irregular caseating nodules in the cortex. Lleo-Colic Glands——Each contained one or two small caseating patches in the cortex. Various Lymphatic Glands. Precrural._—Each contained two caseating nodules, the larger the size of a hemp-seed. Popliteal—_The left contained two similar nodules, the right was normal. Pudic, Ischiatic, Gluteal.—Normal. Microscopical Examination. Emulsion of Left Prescapular Gland.—Tubercle bacilli numerous. Emulsion of Portal Gland. — Tubercle | bacilli numerous. Tubercle from u Peyer's Patch.—Tubercle bacilli very numerous. CALF 13809. Virus H 13. “ A.D.” Subcutaneous inoculation of culture derived from a small (dysgonic) colony [No. 5] on a glycerin- serum plate sown from Guinea-pig 2491. of culture. This guinea-pig had been injected with the ‘Calf 301” strain Date of Inoculation—November 12, 1907. [Age about five months. ] Dose—44°0 milligrammes. Died—December 12, 1907. Clinical Notes. The disease ran a rapid course and was similar to that frequently seen in calves inoculated with 50 mg. of « virulent virus. On December 5 there was a firm prominent tumour at the seat of inoculation infiltrating the muscles and adherent to the skin and measuring 14 by 9cm. The adjacent prescapular gland was much enlarged, measuring 11 cm. in length, The respiration was laboured and much increased in frequency ; the animal stood with head stretched out, and coughed frequently. The appetite was poor, but there was no very obvious loss of flesh. Daring the following week the animai rapidly grew worse ; the respirations became more frequent, the calf lost its appetite and became emaciated. It died on December 12. Tenperalure.—On the 7th day the temperature rose to 40°3° C., and remained high [between 39°6° C. and [30 days after inoculation. ] 40°6 C.] until December 10, a period of 22 days. On December 11 the temperature fell 2-0° C. [to 384° C.] and the following day the calf was dead. ’ Tuberculin Test—The calf was not tested subse- quent to inoculation. Weights. ewt. qrs. lbs. November 12,1907... ears UE = i December 12,1907... ow 1 21 4 Total loss of weight—1 qr. 0 lbs, Average rate of loss per week.—6°5 lbs. POST-MORTEM EXAMINATION. The carcass was emaciated. Local Lesion—In the subcutaneous tissues on the left side of the neck there was a lenticular mass of juicy yellow caseo-necrotie substance measuring 14 by 7em., by 2°5cm. in thickness; the mass was adherent to the skin and muscles, both of which were infiltrated to a slight extent ; the muscles were indu- rated for a depth of about 1cm. and beset with discrete caseous tubercles. Left Prescapular Gland.—The left prescapular gland measured 8 by 5°5 by 3cm., and showed on section about half the cortex dense yellow and caseo- necrotic ; the rest of the cortex was indurated, congested, and beset with discrete caseous tubercles. __ Right Prescapular Gland.—The right prescapular gland measured 4°6 by 15 by 0°8cm., and showed in the cortex a number of greyish-white miliary tubercles with deeply congested margins. Prepectoral Glands.——The left prepectoral glands were slightly enlarged, congested, and oedematous, and showed in the cortex scattered greyish-white tubercles up to a millet-seed in size. The right prepectoral glands contained tubercles similar to those in the right prescapular. Cervical Glands.—All the cervical glands contained tubercles similar to those in the peripheral lymphatic glands (vide infra). Thorax. Pleura.—The fringes along the margins of the ribs were slightly hypertrophied and deeply congested ; no tubercles were visible. Lungs—The lungs were heavy weighing 7 lbs. 11 ozs.; almost the whole of the parenchyma was dark red firm and quite airless ; the only crepitant lung tissue that remained was along the dorsal borders, and this was congested, emphysematous, and showed here and there consolidated lobules. The tissue of the lungs was closely and evenly beset throughout with miliary grey translucent tubercles. The condition of the lungs closely resembled that following an intravenous inoculation. Thoracic G'lands—The dorsal mediastinal and bronchial glands were enlarged and weighed together 8 ozs. ; on section the cortices were composed almost throughout of firm greyish tissue containing caseous foci and here and there a caseous network; the medullary portions of the glands were deeply con- gested. Heart.—The heart was a little enlarged and flabby, and showed numerous petechial haemorrhages under the pericardium ; on the right ventricle a grey miliary tubercle was seen ; the endocardium of the right side of the heart showed a number of small greyish-white tubercles ; none was seen on the left side. Larynx and Trachea——Normal. Abdomen. Omentum and Peritoneum.—On the ventral surface of the omentum there was a number of small tubercles, one or two caseating. The parietal peritoneum was normal. ~ o 9 Spieen.—The spleen was flabby and not enlarged ; on section the pulp was soft, and showed on close inspection fairly numerous grey tubercles, the largest the size of a pin’s head. Liver.—The liver had a bright red colour, and was mottled or speckled with yellow foci (parenchymatous degeneration of the lopuli). Under the capsule a few small grey tubercles were seen ; similar tubercles were sparsely scattered throughout the substance. Portal Glands—The portal glands were enlarged and very oedematous; they showed on section firm greyish patches continuous around the margins of the cortex containing scattered greyish-white foci. Gall-bladder. — There was one minute tubercle under the mucous membrane of the gall-bladder. Kidneys.—In the cortex of each kidney, on the surface as well as in the depth, there were scattered greyish-white tubercles, the largest a millimetre in diameter. Suprarenal Bodies.—In the cortex of each a few minute grey tubercles were seen. Alimentary Tract. Pharyne.—In the corrugated membrane of the vault of the pharynx there were two or three small tubercles. Tongue und Tonsils—Normal. Intestines —The mucous membrane of the small intestine was congested ; the Peyer’s patches contained scattered soft yellow foci and an occasional grey tubercle. The mucous membrane of the large intestine was congested ; no definite tubercles were seen. Testicles—Normal. Various Lymphatic Glands. The Peripheral Lymphatic Glands, the Coeliac, Gastric, Mesenteric and Colic Glands, the Lumbar, Iliae, and Renal Glands, and the Retro-pharyngeal, Submacillary, and Parotideal Glands all contained numerous discrete grey or greyish-white tubercles, which varied in size from a mere point to that of a millet seed ; in many of the glands the tubercles were deeply congested around the margins. The Ischiatic Glands alone were apparently normal. Microscopical Examination. Lung (smear from).—Short tubercle bacilli, moder- ately numerous, in cells. Suprarenal (tubercle from).—One tubercle bacillus seen. Spleen (smear from pulp).—One tubercle bacillus seen. Intestine (tubercle from a Peyer’s patch).—Tubercle bacilli numerous, Intestine (soft focus from a Peyer’s patch).—No tubercle bacilli. CALF 1337. Virus H 13. “A.D.” Subcutaneous inoculation of culture derived from a single colony [No.7] isolated on a glycerin-serum plate sown from the omentum of Guinea-pig 2605 (from Guinea-pig 2491, ‘‘ Calf 301” strain). Dose—59°0 milligrammes. Date of Inoculation—January 17, 1908. [Age about 14 weeks. ] Killed when in good health—April 2, 1908. [76 days after inoculation. ] Clinical Notes. The calf remained perfectly well during the experi- ment. Temperature-—On the seventh day after inoculation the temperature rose to 40°0°C. and reached a 27676 maximum (40°8°C.) on the 11th day; it remained high for ten days; on the 18th day the temperature was again normal, and it remained normal subseqzently. Tuberculin Test.—March 4, 1908. [47 days after inoculation.] Dose, 4°0 ce. of avian tuberculin. No reaction ? Rise of temperature, 0°5° C. L2 Weights. cwt. qrs. lbs. 0.63 «8 January 17, 1908 awe 1 23 April2}1908 .. 00. we Total gain of weight.—2 qrs. 0 Ibs. Average rate of gain per week.—6 lbs. POST-MORTEM EXAMINATION. The carcass was in good condition. Local Leston.—At the site of inoculation on the left side of the neck there was a rounded fluctuating swelling measuring 6 by 5 by 3'5 cm.; it was com- posed of yellow caseo-purulent substance surrounded by a fibrous capsule. Left Prescapular Gland.—The left prescapular gland measured 5°5 by 2°5 by 15 cm.; on section at one extremity there was a group of three dense caseo- calcareous nodules, the largest the size of a pea; at the other there was an irregular caseo-calcareous nodule, 1 cm. in diameter. Right Prescapular Gland.—The right pvescapular gland measured 4°5 by 2 by 1 cm. and was normal on section. Cervical Glands.—On the left side a gland in the middle of the neck was slightly enlarged and the cortex was beset with yellow caseo-calcareous miliary tubercles, in places confluent. Other cervical glands were normal. Thoras. Lungs, Heart—Normal. 60 Thoracic Glands.—The left bronchial gland showed one minute greyish-white point. Other thoracic glands were normal. Pleura and Peritoneum.—On the peritoneum of the floor of the abdomen, and on the surface of the lung, there were several fibrous tags and a few small fibrous nodules. Abdomen. Liver, Spleen, Kidneys, Suprarenal Bodies.—Normal. Portal, Coeliac, Renal, Lumbar, and Iliac Glands.— Normal. Testicles.—Normal. Alimentary Tract. Tongue, Pharyna, Tonsils, Submaxillary and Retro- pharyngeal Glands.—Normal. Intestines and Mesenteric Glands.—Normal. Various Lymphatic Glands. Precrural, Popliteal, Gluteal, Ischiatic, Awillary, Pudic.—Normal. Microscopical Examination. Emulsion of Prescapular Gland.—Tubercle bacilli moderately numerous. CALF 1348. Virus H 13, tau Subcutaneous inoculation of culture derived from a single colony [No. 6] isolated on a glycerin-serum plate sown from the omentum of Guinea-pig 2605 (from Guinea-pig 2491, “Calf 301” strain). Dose—78°0 milligrammes. Date of Inoculation—January 16, 1908. Killed when in good health—April 9, 1908. Clinical Notes. The calf remained well during the experiment. Temperature, On the 8th day after inoculation the temperature rose to 40-0° C., and reached 40°3° C. on the 12th day : it then slowly fell, was normal on the 23rd day, and remained normal subsequently. Tuberculin Zest. March 4, 1908. [48 days after inoculation. ] Dose—8:0 cc. of Leishmann’s Tuberculin. No reaction (?). Rise of temperature—0°6° C. Weights. cwt. qrs. Ibs. January 16, 1908 oes we 1 0 19 April 9,1908 ... ee we CR 62 OF Total gain of weight.—1 qr. 26 lbs. Average rate of gain per week.—4'5 lbs. POST-MORTEM EXAMINATION. The carcass was in good condition. Local Lesion.—At the seat of inoculation there was a prominent circumscribed fluctuating swelling measur- ing 9 by 7 by 5cm. On section, it was a cyst contain- ing light brownish turbid fluid and caseo-purulent masses; the wall was of dense white fibrous tissue lined internally with granulation tissue, and the cavity was crossed by stout fibrous trabeculae. Left Prescapular Gland.—The left prescapular gland measured 5 by 2°5 by 1°7.cm, and showed in the cortex five dense caseo-calcareous masses, the largest 15cm. in greatest diameter. Right Prescapular Gland—The right prescapular [Age about 14 weeks. ] [84 days after inoculation. } gland measured 4:2 by 1°8 by I-cm., and was normal on section. Prepectoral, Cervical, and Axillary Glands—Normal. Thorax. Pleura, Heart.—Normal. Lungs.—The lungs were perfectly normal in general appearance. One pinhead-sized reddish-grey tubercle was seen just under the pleura of the right caudal lobe : otherwise the lungs were normal. Thoracie Glands—The bronchial and mediastinal glands were normal in size. The left bronchial gland showed on section one greyish-white focus which was not gritty. The other glands were normal. Abdomen. Omentum and Peritoneum; Spleen, and Suprarenal bodies—Normal. Portal, Coeliac, Renal, Lumbar, and Iliac Glands.— Normal. ; Liver, Kidneys Testicles—Normal. Alimentary Tract. Longue, Pharynx, Palate, Tonsils—Normal, Retro - pharyngeal, Submaxillary, and Parotideal Glands.—Normal. Intestines and Mesenteric Glands.—Normal. Various Lymphatic Glands. Precrural, Popliteal, Gluteal, Ischiatic, Pudie— Normal. Microscopical Examination. Emulsion of Left Prescapular Gland.—Tubercle bacilli numerous. CALF 1191. Virus H 49. strain). Dose—50:0 milligrammes, 61 engy Subcutaneous inoculation of culture derived directly from the human mesenteric glands (Blythwood Date of Inoculation—February 26, 1907. [Age about 44 months. | Killed when in good health—May 31, 1907. [94 days after inoculation.] Clinical Notes. A fortnight after the inoculation on the left side of the neck there was an elongated firm local swelling, thick above and tapered below, extending into the dewlap and measuring 20 by 8 cm. The left pre- scapular gland was moderately enlarged, measuring 10 cm. in length. The tumour and gland afterwards diminished con- siderably in size. On April 19, 52 days after inocula- tion, the former measured 14 by 7 cm. and the latter was 7 cm. in length. The tumour subsequently became soft and fluctuat- ing, and eventually opened and discharged caseo- purulent matter. The general condition and health of the calf re- mained good during the experiment. Temperature.—On the 11th day the temperature rose suddenly to 40°7° C. It returned to the normal on the 19th day, and remained normal until the close of the experiment. Tuberculin Test—May 9, 1907. [72 days after inoculation.] Reacted. Rise of temperature, 1°8° C. Weights. ewt. qrs. lbs. February 26,1907... see 1 3 8 May 31, 1907 ... ae a 2 1 21 Lotal gain of weight during experiment.—2 qrs. 13 lbs. Average rate of gain per week.—5r1 lbs. POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion.—In the subcutaneous tissues on the left side of the neck there was an oval tumour measur- ing 13 by 6°5 by 3 cm. ; on section it was found to be composed of cysts or cavities of various sizes filled with creamy caseo-pus and surrounded by dense white fibrous tissue ; the cysts were lined internally with granulation tissue in which numerous calcareous ‘grains could be detected. The skin over the tumour was much thickened, measuring 1°5 cm. in greatest thickness. Left Prescapular Gland.—The left prescapular gland measured 6°5 by 4 by 2 cm. On section it showed five caseo-calcareous nodules varying in diameter from 1 to 2cm.; some of the nodules were partially softened, others contained small islets of translucent fibroid tissue. Right Prescapular Gland.—Measured 45 by 23 by lcm. Normal. Prepectoral and Awillary Glands.—Normal. Thoraz. Pleura, Lungs, Heart, Thoracic Glands.—Normal. Abdomen. Peritoneum, Liver, Spleen, Kidneys, Suprarenals.— Normai. Portal, Lumbar, Iliac, and Renal G'lands.—Normal. Alimentary Tract. Tongue, Pharynx, Palate, Tonsils——Normal. Submaxillary, Hyoid, Retro-Pharyngeal, and Parotid Glands.—Normal. Intestines and Mesenteric Glands.—Normal. Genito-Urinary System. Testes.— Normal. Various Lymphatic Glands. Precrural, Popliteal, Gluteal, Ischiatic, Pudie.— Normal. Microscopical Examination. Emulsion of Prescapular Gland.—Tubercle bacilli fairly numerous. Animal Inoculated. Rabbit 1318 was inoculated with an emulsion made from the prescapular gland. It showed a local lesion only when killed after 129 days. CALF 1193. Virus H 49. “ TC.” Subcutaneous inoculation of culture derived directly from the human mesenteric glands (Blythwood -strain). Dose—100-0 milligrammes. Date of Inoculation—February 26,1907. [Age about 43 months.] Killed when in good health—June 5, 1907. [99 days after inoculation.] Clinical Notes. A prominent tumour developed at the seat of ‘inoculation on the left side of the neck. On the 16th ‘day it was firm, and measured 12 by 8cm. The left m prescapular gland was 9 cm. in length. The calf was well. On the 52nd day the tumour measured 10 by 6 cm. and showed a prominent rounded fluctuating swelling 5 cm. in diameter at the upper part. The prescapular gland was now 7 cm. in length. The calf was in good condition. : Subsequently the tumour opened and discharged, afterwards diminishing greatly in size. ee calf was killed on the 99th day, when quite well. Temperature—The temperature rose on the 11th day and reached 39°8°C. on the 13th day, and then quickly fell to normal. Subsequently it was approxi- mately normal, but somewhat irregular, the maximum range of variation during 12 weeks being 1-4° C. (37°9-39°3° C.). _ Luberculin Test.—May 9, 1907. vee Slight reaction. [72 days after Rise of temperature, Weights. cwt. qrs. lbs. February 26,1907... sale IS 9 June 5, 1907 ... a due 2 3 GE, Total gain of weight.—2 qrs. 26 Ibs. Average rate of gain per week.—5'8 lbs. POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion.—The lesion at the seat of inoculation on the left side of the neck consisted of thickened skin anda thin patch (in the subcutaneous tissues) of fibrous tissue, about 5 cm. in greatest diameter ; the latter was beset with soft caseo-caleareous nodules with translucent fibrous walls, the largest the size of a hemp seed ; the thickened skin towards the lower part showed a healed sinus. Left Prescapular Gland.—The left prescapular gland 62 measured 6 by 3 by 2 cm., and on section showed about two-thirds of its substance replaced by three large nodules with caseo-purulent centres and brownish translucent fibroid margins closely beset with cal- careous grains. Right Prescapular Gland.—The right prescapular gland measured 4 by 2 by 1 cm., and was normal on section. Prepectoral and Axillary Glands.—Normal. Thorax. Pleura, Lungs, Thoracic Glands, Heart.—Normal. Abdomen. Omentum and Peritoneum, Liver, Portal Glands, Spleen, Kidneys, Suprarenals; Renal, Lumbar, and Iliac Glands.—Normal. Alimentary Tract. Tongue, Pharynx, Tonsils; Glands of the Neck and Throat ; Intestines and Mesenteric Glands.—Normal. Testes.—Normal. Various Lymphatic Glands. Precrural and Popliteal, Gluteal and Ischiatic,. Pudic.—Normal. CALF 1218. Virus H 49. “ TO Subcutaneous inoculation of culture derived from the human mesenteric glands (Royalcot strain). Dose—50'0 milligrammes. Date of Inoculation—April 20, 1907. Killed when in fair health—July 9, 1907. Clinical Notes. A large tumour developed at the seat of inoculation on the left side of the neck; 24 days after inocu- lation the tumour measured 17 by 9 cm. It was firm and extended into the dewlap, and the skin over the centre showed a fissure discharging serum. The left prescapular gland was much enlarged, measuring 10 cm. in length. During the fourth week the calf was off its food, and lay on the ground more frequently than during the earlier part of the experiment. The respiration [Age about 16 weeks. ] [80 days after inoculation. } was not increased, and there was no apparent loss in flesh. Subsequently the respiration became definitely in- creased and the calf began to look thin. Eighty days after inoculation the tumour was soft and fluctuating, partly detached from the muscles, and very pendulous, hanging below the dewlap. The fissure noticed in the overlying skin had closed up. The calf was fairly well but somewhat thin; the respiration was much increased in frequency. On this day it was killed. Temperature. Tuberculin Test.—May 9,1907. [19 days after inocu- Jation.] Reacted. Rise of temperature, 1:2° C. Weights. cwt. qus. lbs. April 20, 1907... tee sin 1 1 16 July 9,1907 ... eee Tf 1 2 19 Total gain of weight.—1 qr. 3 lbs. Average rate of gain per week.—2°7 lbs. POST-MORTEM EXAMINATION. Carcass in fair condition. Local Lesion—At the seat of inoculation on the left side of the neck there was a very large pendulous fluctuating tumour, which measured 22 by 13 by 11 cm. and,weighed with attached skin and muscles 1,278 grms. ; on section the greater part of the tumour was a cyst filled with thin curdy caseo-pus and irregular caseo- necrotic masses, most of which were attached to the wall of the cavity; the cavity was surrounded by fibrous tissue lined internally with granulation tissue very gritty to the touch ; short thick fibrous trabeculae projected into the cavity. The skin was thickened and beset with caseo-calcareous tubercles ; the muscles under the cyst were very fibroid and infiltrated with calcareo-caseous tubercles. ‘The lower part of the tumour tapered, and was composed of dense caseous substance surrounded by fibroid tissue infiltrated with calcareo-caseous streaks and foci. Left Prescapular Gland.—The left prescapular gland weighed 135 grms. and measured 8 by 5°5 by 5cm.; on section it was composed nearly throughout of dense pinkish-yellow caseous substance gritty only around the peripheral parts ; the rest of the gland was beset with calcareo-caseous tubercles. Right Prescapular Gland.—The right prescapular gland measured 4°5 by 2:2 by 1:2 cm. and showed on section two calcareo-caseous tubercles. Left Prepectoral Glands.—The rounded gland, 1 cm. in diameter, showed three-quarters of its substance caseo-calcareous and in the rest discrete tubercles. The kidney - shaped gland contained numerous minute calcareous foci. Another deeper gland had three or four calcareous miliary tubercles in the cortex. Right Prepectoral Glands.—Normal. Cervical Glands.—On the left side one of the lower cervical glands was the size of a thrush’s egg, and was dense and caseous throughout and slightly gritty from calcification. Another near it was beset with minute yellow foci ; a gland near the articulation of the jaw contained a calcareo-caseous tubercle. Other cervical glands were normal. Agillary Glands.—The left axillary gland contained one, the right two, calcareous tubercles. Thoran. Pleura.—The lymphatic fringes along the margins of some of the ribs were slightly hypertrophied and contained here and there a small fibrous nodule. On the diaphragmatic pleura there were some pinkish villous growths around the margins of the tendon, and on the left side an eruption of grey granules; there were also two nodules, one small and loosely pedunculated, the other about 1 cm. in diameter and not pedunculated ; both were fibrous and contained calcareous grains. Lungs. — The lungs were voluminous, emphy- sematous, and did not collapse on opening the chest ; they weighed 4,601 grammes. The surfaces of the small anterior lobes and the ventral surface of each caudal were thickly studded with raised pinkish-grey translucent nodules, varying in shape and size, the largest being about 8 mm. in greatest diameter. They were most numerous on the anterior lobes, particularly where the lobes were in contact, and became gradually less numerous towards the tips of the caudal lobes ; on the dorsal surfaces of the caudal lobes there were similar nodules, sparsely scattered. : On section they were found to extend into the lung substance, the pleura passing over them, and to have calcareous centres. 63 The substance of the lung under the pleura showed numerous irregular greyish-red patches and grey tubercles some calcareous in the centre, and calcareous nodules up to a pea in size irregularly distributed throughout each lobule. On section of the lung similar appearances were seen ; the reddish grey patches occupied the whole or part of a lobule, and in the latter cases were diffuse in outline and were situated chiefly in the centre of the lobule around the bronchus; they contained scattered calcareous tubercles and sometimes a large calcareous nodule. In the anterior lobes, particularly on the right side, there were larger areas of consolidation than in the caudal lobes. The fringes around the margins of the lungs were hypertrophied and showed a nodule here and there. Thoracic Glands.—The thoracic glands weighed 341 grammes, and were much enlarged ; they were firm and on section beset with yellow calcareous tubercles which formed in places irregular calcareous patches ; the bronchial glands were more severely affected than the mediastinal and in them there was less normal looking gland tissue than in the latter. The ventral mediastinal glands each contained a caseo-calcareous nodule. Heart.—Normal. Abdomen. There was a slight excess of fluid in the peritoneal cavity. Omentum.—The lymphatic fringes on the ventral surface were slightly hypertrophied and there were about a dozen loosely pedunculated nodules with calcareous centres and fibrous margins, the largest the size of a barley grain. Parietal Peritoneum.—There was an eruption of flattened grey nodules, varying in size, on the right side of the diaphragm. Spleen—The spleen weighed 284 grammes; it showed on the surface several flattened deep red firm growths containing calcareous foci, and numerous rounded projections caused by nodules in the sub- stance, the capsule over which was deeply congested. On section the pulp contained numerous nodules, varying in diameter up to 5 mm., with grey trans- lucent margins and calcareo-caseous centres. Liver.—The liver showed on the surface a small number of flat grey growths, loosely attached to the capsule and lying in shallow depressions in the liver substance ; the largest measured 1 cm. in diameter and all contained yellow gritty foci ; in the substance of the liver under the capsule fairly numerous evenly distributed grey tubercles with calcareous centres, the largest 2 mm. in diameter, were seen; on section similar tubercles were evenly distributed in the depth. Gall-bladder.—Normal. Portal Glands—The portal glands were slightly enlarged and beset with yellow calcareo-caseous tubercles. Kidneys.—In the cortex of each kidney on the surface as well as on section were fairly numerous nodules varying in size from a pin’s head to that of a barley grain ; the smaller ones were grey and trans-, lucent, the larger ones had yellow calcareous centres ; many of the latter projected slightly from the surface and extended in a radial manner deeply into the cortex. There were besides many pale patches on the surface which on section were seen to be the bases of wedge-shaped areas which extended into the cortex. Suprarenal Bodies——Hach showed in the cortex a hempseed-sized nodule with a grey margin and a caleareo-caseous centre, and several miliary tubercles. Alimentary Tract. Tongue, Pharynx, Palate, Tonsils—Normal. Intestines, Small.—The mucous membrane of the ileum showed half-a-dozen ulcers with raised con- gested margins, slightly thickened base and caseous floor. Intestines, Large.—In the coecum and large intestine there were three or four similar ulcers. Mesenteric Glands,—The mesenteric glands contained scattered calcareous nodules and tubercles. Gastrie and Colic Glands.—These glands contained discrete calcareous tubercles. Genito-Urinary Tract. Testicles.—Normal. Eyes.—Normal. Various Lymphatic Glands. Each Submazillary, Parotideal, and Retro-pharyngeal Gland contained a few calcareo-caseous tubercles. Coeliac Glands——One was slightly enlarged and contained calcareous nodules with translucent fibrous margins confluent in places; others, not enlarged, showed discrete tubercles and nodules. The Lumbar and Renal Glands were rather closely beset with discrete calcareo-caseous miliary tubercles ; a were also a few larger nodules with calcareous centres. 64 The Precrural, Popliteal,and Iliac Glands contained scattered calcareo-caseous nodules, mostly about 2 mm., but several as much as 3mm. in diameter. The Gluteal and Ischiatic Glands were normal. A Haemo-lymph Gland near a gluteal gland con- tained a millet-seed sized calcareous tubercle. Microscopical Examination. Emulsion of Prescapular Gland.—Tubercle bacilli fairly numerous. Emulsion of Mediastinal Gland.—Tuberele bacilli scanty. Animals Inoculated. Calf 1263 was inoculated subcutaneously with an emulsion made from the left prescapular gland. Two Rabbits, 1378-1379, were inoculated subcutane- ously with the same emulsion. (Died of G.T. In 52 and 78 days.) Two, Nos. 1876-1877, were inoculated subcutaneously with an emulsion made from the mediastinal gland. (Died of G.T. in 122 and 86 days.) CALF 1263. Virus H. 49. 66 TC.” Subcutaneous inoculation of an emulsion of the left prescapular gland from Calf 1213. Dose—10°0 cc. of the emulsion, containing fairly numerous tubercle bacilli. Date of Inoculation—July 9th, 1907. [Age about 17 weeks.] Killed when in good health—October 8th, 1907. [91 days after inoculation. ] Clinical Notes. The Calf remained in good health during the whole period of the experiment. Temperature. RuqusT (Chart to Sept. 13.) From Sept. 13 to Oct. 8 the temperature was a little above the normal, and varied only within narrow limits (maximum 39°4° C., minimum 3$-0° C.). Tuberculin Test—The calf was not tested subse- quent to inoculation. Weights. cwt. qrs. lbs. July 9,1907 ‘ie oe 1 2 3 October 8, 1907... we wie 2 OO 8 Total gain of weight.—2 qrs. 0 lbs. Average rate of gain per week.—4°3 lbs. POST-MORTEM EXAMINATION, The carcass was in good condition. Local Lesion.—At the seat of inoculation on th left side of the ueck there was a firm, prominank, well-defined tumour, which when removed measured 11 by 8 by 5°5 cm., and weighed with skin and attached muscles 14 ozs. On section it was composed of three layers, a middle one of dense yellow homogeneous cnmeaed tissue 1°5 cm. in greatest thickness, an outer one of greatly thickened skin beset with caseo-calcareous tubercles, and an inner one of fibroid muscular tissue infiltrated with irregular caseo-calcareous nodules and masses. Left Prescapular Gland.— The left prescapular gland measured 10 by 5 by 45 cm., and weighed 60zs.; on section it was composed throughout of dense yellow caseous substance slightly gritty from calcification ; the capsule was thickened. Right Prescapular Gland.—The right prescapular gland measured 6 by 2 by 1 cm. and was normal on section. Prepectoral Glands.—On the left side one round gland measured 1°5 cm. in diameter and contained a caseo-calcareous nodule 8 mm. in greatest diameter ; another was normal. Those on the right side were normal. Cervical Glands.—One near the angle of the jaw on the left side showed part of the cortex firm and in- filtrated with caseo-calcareous points. A small gland in the middle of the neck on the right side contained a small calcareous tubercle. Other cervical glands were normal. Thorax. Pleura.—The fringes along the margins of the ribs were hypertrophied in places ; there were no nodules. On the pleural surface of the diaphragm there was a loosely attached haemorrhagic nodule, the size of a split pea, with a caseo-calcareous centre. Lungs.—The lungs were crepitant and collapsed normally ; they showed on the surface a number of tuberculous growths similar to those on the liver, but containing caseo-calcareous patches, and under the pleura a moderate number of nodules ; on section nodules, not in very large numbers, were found evenly distributed throughout the parenchyma ; they varied much in size, the largest measuring 1 cm., the smallest about 1 mm. in diameter; the smaller ones were fibrous with calcareous centres, the larger were caseo- calcareous with fibrous margins; there were no patches of consolidation and no adhesions to the chest wall. Thoracic Glands.—The bronchial and mediastinal glands were enlarged, weighing together 10 ozs.; on section they were dense and calcareo-caseous practically throughout, with the exception of one or two small glands which contained discrete nodules. Heart and Pericardium.—Normal. Abdomen. Omentum and Peritoneum.—On the ventral surface of the omentum there were two small fibrous nodules, one with a calcareous centre. On the peritoneal surface of the diaphragm there were about a dozen flattened translucent fibroid nodules, some calcareous in the centre. Spleen.—The spleen was not enlarged and was normal on the surface ; on section the pulp contained scattered nodules with yellow calcareous centres and fibrous margins, the largest 4 mm. in diameter. Twenty-seven were counted on one cut surface after longitudinal section. Liver—On the surface there were half a dozen slightly raised flattened fibroid growths, the largest 65 1 cm. in diameter, containing yellow calcareous foci ; they were lobular in outline and extended only a short distance into the substance of the liver ; under the capsule scattered nodules, some projecting, were seen; on section, nodules similar to those in the spleen were somewhat sparsely scattered throughout the substance. Portal Glands.—Three were enlarged and closely beset with irregular yellow caseo-calcareous nodules, in places confluent, the largest 5 mm. in diameter ; the rest were normal. Kidneys.—The right kidney showed on the surface five and the left three greyish yellow nodules, the largest 6 mm. in diameter, some of which projected ; on section they had grey fibrous margins and calcareo-caseous centres; several were lobular in outline as if composed of a number of smaller nodules, and some were much elongated radially, penetrating in a few cases the whole thickness of the cortex ; in the depth of the cortex a few scattered nodules were seen. Suprarenal Bodies.—Normal. Lumbar and Coeliac Glands—Two lumbar glands and one coeliac were enlarged, and on _ section resembled the portal glands. Iliac Glands.—Normal. Testicles—Normal. Alimentary Tract. Tongue, Pharyna, Palate, Tonsils—Normal. Retro-pharyngeal, Submazillary and Hyoid Glands.— Normal. Parotideal Glands. —Iu the cortex of the left parotideal gland there was a hempseed sized caseo- calcareous nodule. The right was normal. Intestines —Normal. Mesenteric Glands.—One at the posterior end of the mesentery contained a caseo-calcareous mass 1:7 cm. in greatest diameter ; another a little anterior to the latter contained a nodule 5 mm. in diameter. Lleo-colic Glands.—Normal. Various Lymphatic Glands. Precrural, Popliteal, Gluteal, Ischiatic, Pudic.— Normal. Microscopical Examination. Emulsion of Prescapular ) Gland. Emulsion of Portal Gland + A few tubercle bacilli were seen in each, the bacilli were all very Emulsion short. Gland. of Thoracic Animals Inoculated. One rabbit, No. 1536, was inoculated subcutaneously with the emulsion of the left prescapular gland. (Died in 232 days of chronic progressive tuberculosis. ) Another, No. 1537, was inoculated subcutaneously with the emulsion of the portal gland. (Died prematurely.) 27076 CALF 1347. Virus H 49. 66 en? Subcutaneous inoculation of culture derived from the mediastinal gland of Calf 1263. Dose—-50°0 milligrammes. Date of Inoculation—December 30, 1907. [Age about 11 weeks. ] Killed when ill—February 20, 1908. [52 days after inoculation. ] Clinical Notes. A very large tumour developed at the seat of inocu- lation, which eventually ulcerated. . The calf gradually became ill and thin, and the respiration quickened ; weakness and emaciation con- tinued, and when killed on the 52nd day, the animal ‘could not have lived more than a few days longer. Temperuture.—The temperature rose to 40:0° C. three days after inoculation, and reached a maximum (40°5°C.) on the ninth day. Subsequently the tem- perature was raised and irregular (maximum 40-0° C., minimum 38°2° C.) until the calf was killed. Tuberculin Test.—The calf was not tested subsequent to inoculation. Weights. qrs. lbs. December 30, 1907... ste nae Oe 9 February 20, 1908 ... ave is! OS Y POST-MORTEM EXAMINATION. The carcass was emaciated. Local Lesion.—At the seat of inoculation there was a very large firm prominent tumour extending down in front of the shoulder into the dewlap ; it measured 15 by 12 by 7 cm. and weighed 2 Ibs. 6 ozs. On section it was composed of an outer layer of pinkish yellow caseo-necrotic substance 2°5 cm. in ‘greatest thickness and an internal layer (3 cm.) of fibroid muscular tissue very closely beset with caseous coalescing nodules. ‘The skin over the greater part of the tumour was necrosed and ulcerating. Left Prescapular Gland.—The left prescapular gland measured 9 by 5:5 by 5 c.m. and weighed 8 ozs. [t was adherent to the local lesion which partly sur- rounded it. On section, it was dense and caseous throughout. Right Prescapular Gland.—The right prescapular gland measured 3:5 by 1°8 by lcm. and contained several firm caseous nodules. Prepectoral Glands.—On the left side one was very large, measuring 3°5 cm. in greatest diameter, and ‘showed the major part of the cortex grey and beset with caseous foci: two other small glands showed ‘small caseous tubercles. Cervical Glands.—All the cervical glands were enlarged ; some contained discrete caseous nodules up to 3 mm. in diameter, others showed part of the cortex grey and infiltrated with caseous foci or a caseous network. Left Axillary Gland.—The left axillary gland was very large and oedematous, and showed numerous minute greyish-white points in the cortex. Thorax. Piewra.—On the costal pleura there were several lenticular caseous nodules, varying up to 8 mm.; in several places the lymphatic fringes were hyper- trophied and beset with small grey tubercles. There were similar nodules and tubercles on the pleural surface of the diaphragm. Lungs.—On the left side there were several recent adbesions to the chest wall, and the lobes on each side showed some slender adhesions to each other ; the surfaces of the lungs (except where covered with thick pleura) were studded with raised grey nodules varying in size up to that of a hempseed ; there were besides a few flattened caseating nodules similar to those on the parietal pleura ; on section the lung was found to be closely beset with nodules similar to those on the surface ; they were grey and slightly caseous centrally ; many lobules or parts of lobules were solid and caseating, especially in the caudal lobes. : The right anterior lobe was completely hepatised (old), and there were irregular patches in the other small lobes and in the ventral portions of the caudal lobes. Thoracic Glands. —The dorsal mediastinal and bronchial glands were enlarged, and weighed 7 ozs. ; their cortices were composed of translucent grey tissue infiltrated with a yellow caseous network. Heart.—On the pleural surface of the pericardium there were several lenticular caseating nodules ; in the auriculo-ventricular sulcus there was a large caseating nodule. The endocardium of the right auricle showed a greyish-white miliary tubercle, and under the endo- cardium of the left ventricle there was a minute grey tubercle. Trachea.—There was a quantity of mucus and muco- pus in the trachea. The tracheal mucous membrane and the larynx were normal. Abdomen. Omentum and Peritoneum.—On the ventral surface of the omentum there were about half-a-dozen len- ticular caseating nodules, the largest 5 mm. in diameter. The peritoneum was normal. Spleen.—There were no tubercles on the surface ; in the pulp there were fairly numerous completely caseous nodules ranging up to 2°5 mm. in diameter. Liver.—On the surface there was a moderate number of flattened slightly raised nodules, with yellow caseous centres and grey margins, the largest 5 mm. in diameter ; small yellow nodules were seen under the capsule in the substance. On section the substance contained evenly distributed caseous nodules, the largest the size of a hempseed ; they were not very numerous, 12 only being counted in an area 5 cm. square. Portal Glands.—The portal glands were enlarged, and their cortices composed of dense caseating nodular tuberculous masses. Kidneys.—In the cortex of each kidney there was a moderate number of caseous nodules varying in size from a pin’s head to that of a hempseed ; the larger ones extended into the cortex in a wedge-shaped manner. Suprarenal Bodies—Each showed in the cortex several miliary caseous tubercles. Lumbar Glands.—The lumbar glands were enlarged and contained more numerous nodules than the peri- pheral glands (se below). Coeliac Glands.—The coeliac glands were enlarged, their cortices were grey and caseating. Alimentary Tract. Tongue, Pharynx, Palate—Normal. _ Tonsils—One contained a caseous nodule 2 mm. in diameter. Intestines.— All the Peyer's patches contained one or two yellow caseous nodules up to 2 mm., some ulcerated, and there were a few under the mucous membrane. There were a few small nodules under the mucous membrane of the large intestine. Mesenteric Glands-—The mesenteric glands all con- tained nodules and some grey caseating patches. Lleo-colic Glands.—The ileo-colic glands showed the cortices extensively replaced by caseating patches. Gastric and Colic Glands.—These glands contained small caseous tubercles. CALF 138%. Virus H 49. 67 Testicles.—Normal. Peripheral Lymphatic Glands, All the peripheral lymphatic glands contained moderately numerous yellow caseous nodules, varying in size up to three and sometimes rather more millimetres in diameter, Eyes.—Normal. eng Subcutaneous inoculation of culture derived from a single colony (No. 3) isolated from a glycerin- serum plate sown from the sternal gland of Guinea-pig 2918, which received a culture of the original material. Dose—50-0 milligrammes. Date of Inoculation—May 13, 1908 Died—July 10, 1908. Clinical Notes. A large tumour developed at the seat of inoculation which afterwards ulcerated: the left prescapular gland became greatly enlarged. The calf gradually (58 days after inoculation. ] [Age about 11 weeks.] became ill and thio and the respiration embarrassed ; weakness and emaciation followed and the calf died on the 58th day after inoculation. Temperature. " MAY JUNE JOLX Weights. ewt. qr. Ibs. May 13, 1908 sea ss 1 O 14 July 10, 1908 sa 0 3 = «12 Total loss of weight.—1 qr. 2 lbs. Average rate of loss per week.—3'8 lbs. POST-MORTEM EXAMINATION. The carcass was emaciated. Local Lesion.—At the seat of inoculation there was a prominent swelling measuring 16 by 11 by 5 cm., the skin over which was necrosed and represented by a dry dark brown scab; the latter was fissured and partially detached, exposing foul smelling caseo-pus. On section the deeper parts of the tumour were com- posed of congested fibroid muscular tissue closely beset with caseating nodules and tracts some gritty from calcification. Left Prescapular Gland.—The left prescapular gland weighed 7 ozs. and measured 10 by 5°5 by 5-4 em. ; on section it was composed practically through- out of dense pinkish-yellow caseous substance slightly gritty from calcification and beginning to break down in places under the capsule. 27676 Right Prescapular Gland.—The right prescapular gland measured 4 by 2 by 0°8 cm. and contained three caseating nodules the largest the size of a hemp seed. Prepectoral Glands.—Two on the left side showed early caseation of the cortex: two others a caseous tubercle or two. Those on the right side each contained a few caseat- ing nodules. Cervical Glands.—-One on the left side near the first rib was enlarged, the cortex was grey and showed discrete caseous tubercles ; most of the other cervical glands contained one or two discrete caseous nodules. Axillary Glands.—The cortex of the left axillary gland was firm and showed minute caseous foci. The right axillary gland contained one caseous nodule. Thorax. There was a slight excess of fluid in the pleural cavities. Pleura.—The costal and diaphragmatic pleurae showed a few congested flattened nodules and patches of congested vegetations. Lungs.—The lungs filled the chest and weighed M2 5 lbs. 2 ozs. On the surface there were scattered congested caseating nodules up to 5 mm. in diameter ; the anterior lobes, a small portion at the antero- ventral end of the left caudal lobe and almost the whole of the right caudal lobe were red and _consolli- dated ; both caudal lobes on section showed mode- rately numerous caseating nodules varying in size up to about 3 mm. congested around the margins and in places confluent; there were similar nodules in the anterior lobes but much less numerous; the con- solidated tissue on section showed a greyish mottling (? early tuberculous infiltration); the trachea and many of the bronchi contained muco-pus. The mucous membrane of the trachea and larynx was normal, Thoracie Glands.—The bronchial and mediastinal glands were much enlarged, especially the long media- stinal gland, which measured 15 cm. in length; on section the cortices were composed of greyish fibroid tissue containing gritty caseous tubercles and irregular -caseous patches. Heart.—Normal. Abdomen. Omentum and Peritonewm.—On the ventral surface of the omentum there were sparsely scattered flattened congested nodules. The parietal peritoneum was normal. Liver.—The liver showed on the surface scattered flattened caseating growths, some loosely attached, others attached by a broad base, the largest about 1 cm. in diameter. On section the substance contained sparsely scattered grey tubercles with caseous centres the largest the size of a millet seed. Portal Glands.—The portal glands were enlarged and the cortices filled with nodular caseating masses. Spleen.—The spleen was vot enlarged and contained moderately numerous grey nodules with caseous centres varying up to 3 mm. in diameter. Kidneys.—On the surface of the left kidney there were twenty-eight caseous nodules varying in size up to that of a hemp-seed; some projected from the 68 surface, a few were elongated in a radial direction ; a few caseous nodules were seen on section. On the surface of the right kidney there were twenty-two caseous nodules and several were seen in the depth of the cortex. Suprarenal Bodies.—Normal. Coeliac Glands.—The coeliac glands were enlarged and composed practically throughout of greyish tissue surrounding yellow caseous nodules. Lumbar and Renal Glands.—The lumbar and renal glands were beset with caseating nodules. Alimentary Tract. Tongue, Tonsils, and Pharynz.—Normal. Small Intestines—On the mucous surface three small ulcerated caseous nodules were seen. Large Intestine. —Normal. Mesenteric Glands.—All the mesenteric glands were affected, containing caseating nodules, the largest 1°5 cm. in diameter. Ileo-colic and Colic Glands.—The ileo-colic ‘and several of the colic glands contained similar nodules. Testicles.—Normal. Various Lymphatic Glands. The peripheral lymphatic glands contained a vary- ing number of caseating nodules (up to 3 mm. in diameter), most numerous in the retro-pharyngeal and precrural glands. Microscopical Examination. Emulsion of Prescapular Gland.—A few tubercle bacilli seen. : ; Emulsion of Mediastinal Gland.—A few tubercle bacilli seen. Emulsion of Popliteal bacilli seen. Gland.—A few tubercle CALF 1415. Virus H 49. “ T.C.” Subcutaneous inoculation of culture derived from a single colony (No. 7) isolated from a glycerin- ‘serum plate sown from Guinea-pig 2918. This guinea-pig was inoculated with the original strain of culture. Dose—50-0 milligrammes. Date of Inoculation—May 18, 1908. Clinical Notes. The calf remained in good health during the experiment. Temperature. There was a slight rise of temperature commencing on the tenth day and lasting ten days (maximum 40:2° C.). With this exception the temperature was perfectly normal] during the experiment. Tuberculin Test. August 18, 1908. [92 days after inoculation.] Dose, 20 cc. Reacted. Rise of temperature, 1°5° C. Weights. ewt. qr. Ibs. May 18, 1908... ‘iis abe 1 O 18 August 28, 1908 ar ‘es 1 1 25 Total gain of weight.—1 qr. 7 lbs. Average rate of gain per week.—2'4 lbs. POST-MORTEM EXAMINATION. The carcass was in good condition. [Age about 13 weeks.] Killed when in good health—August 28, 1908. [102 days after inoculation. ] Local Lesion.—On the left side of the neck there was a prominent circumscribed fluctuating tumour measuring 9 by 5°5 by 4 cm. ; on section it contained a loose dense caseous sequestrum separated from a thick fibrous capsule by yellow pus. Left Prescapular Gland.—The left prescapular gland measured 6 by 3 by 2°56 cm. and showed more than three quarters of its substance composed of dense caseous material slightly gritty from calcification. Prepectoral Glands.—On the left side one contained a fibro-calcareous nodule 5 mm. in diameter. The rest were normal. Right Prescapular Gland.—The right prescapular gland measured 5 by 2:2 by 1 cm. and was normal on section. Other glands and all the organs were examined and found normal. Microscopical Examination. Emulsion of Prescapular Gland.—A few beaded tubercle bacilli seen. 69 CALF 1413. Virus H 49. “T.C.” Subcutaneous inoculation of culture derived from a single colony (No. 17) isolated from a glycerin- serum plate sown from Guinea-pig 2918. This guinea-pig was inoculated with the original strain of culture. Dose—50:0 milligrammes. Date of Inoculation—May 18, 1908. Clinical Notes. The calf remained well during the experiment. Temperature. On the 10th day after inoculation the temperature rose from the normal sharply to 40°3° C. and then slowly fell; on the 17th day it was again normal (38°9° C.), and remained normal until the animal was killed. Tuberculin Test. August 18, 1908. [92 days after inoculation.] Dose, 2:0 ce. Reacted. Rise of temperature, 2°1° C. Weights. ewt. qr. Ibs. May 18,1908 ... oie ana 1 0 5 September 2,1908 ... aoe 1 1 18 Total gain of weight.—1 qr. 13 lbs. Average rate of gain per week.—2°7 lbs. POST-MORTEM EXAMINATION. The carcass was in good condition. Local Lesion.—The lesion at the seat of inoculation on the left side of the neck consisted of a small subcu- taneous mass of fibrous tissue containing scattered calcareous nodules, and a cyst 1 cm. in diameter filled with caseo-pus; the skin showed a funnel-shaped depression, the apical part of which was formed by scar tissue. Left Prescapular Gland.—The left prescapular gland measured 7 by 4 by 3°5 cm. and was composed through- out of very dense pinkish yellow caseous substance, gritty from calcification; the capsule was much thickened, and was in places separated from the denss caseous substance by caseo-pus. Right Prescapular Gland—The right prescapular gland measured 4°8 by 1°5 by 0°8 cm. and was normal on section. Prepectoral Glands.—One prepectoral gland on the left side contained a hempseed-sized caseo-calcareous nodule. Other prepectoral glands were normal. Cervical Glands——On the left side two contained each one caseous miliary tubercle; in another there were two pinhead-sized caseous tubercles, one gritty. Thorax. Pleura and Heart.—Normal. Lungs.—The lungs were crepitant throughout ; they showed under the pleura evenly distributed not very numerous nodules varying in size from a pin’s head up to a little larger than that of a hemp seed; 78 were counted on the surface of the left lung. They were calcareous and had grey fibrous margins. Similar nodules were sparsely scattered throughout the substance of the lung. [Age about 11 weeks.] Killed when in good health—September 2, 1908. {107 days after inoculation. ] Thoracie Glands.—The bronchial and mediastinal glands were a little enlarged; their cortices were closely beset with highly calcareous tubercles mainly discrete, but here and there aggregated together. Abdomen. Omentum and Peritoneum.—Normal. Spleen.—The spleen was normal in size; the pulp contained scattered yellow calcareous tubercles ranging from 1:5 to 2°5 mm. in diameter ; each had a grey fibrous capsule from which the calcareous centre easily shelled ; three dozen were counted on the two cut surfaces after longitudinal section. Liver.—In the substance. of the liver under the capsule there were eight grey tubercles up to a milli- metre in diameter all with calcareous centres; two similar tubercles were seen in the depth. Portal Glands.—The portal glands were not enlarged. Each contained a few calcareous tubercles up te a millet seed in size. Kidneys and Suprarenal Bodies.—Normal. Renal Glands.—Between the kidneys there were four pigmented glands. each of which contained a few yellow calcareous foci. Coeliac Glands.—One coeliac gland—not enlarged— contained discrete yellow calcareous miliary tubercles. Lumbar and Iliac Glands.—Normal. Alimentary Tract. Tongue, Pharynz, Tonsils——Normal. Retro-pharyngeal Glands.—The right contained two nodules each 2 mm. in diameter, one caseous the other calcareous. Submaaillary Glands.—Normal. Parotideal Glands.—The left showed a cavity rather more than 1 cm. in diameter filled with tenacious yellow pus. Intestines —Normal. Mesenteric Glands.—The mesenteric glands contained two calcareous tubercles. Lleo-colic Glands——Two ileo-colic glands showed scattered calcareous tubercles. Mammary Gland.—Normal. Various Lymphatic Glands. Precrural, Popliteal, Gluteal, Ischiatic, Pudie.— Normal. Microscopical Examination. Emulsion of Lung Tubercle—Few tubercle bacilli. Emulsion of Bronchial Gland.—Few tubercle bacilli. Pus from Left Parotideal Gland—No tubercle bacilli. CALF 1501. Virus H 49. « TC.” Subcutaneous inoculation of culture derived from Colony No. 17 isolated from a glycerin-serum plate sown from the sternal gland of Guinea-pig 2918. Dose—48°0 milligrammes. Date of Inoculation—November 12, 1908. Died—December 27, 1908. Clinical Notes. The course of the disease was similar to that usually [Age 7 weeks. ] [45 days after inoculation. ] seen after the inoculation of 50:0 mg. of a virulent virus. 70 Temperature. On the day following the inoculation the tempera- ture was 39°5° C.; it rose slowly and reached 41-0° C. on the 9th day. The temperature was raised during the remaining period of the experiment, averaging about 401° O.; on the day of death it dropped sharply from 39°8° C. to 381 C. Weights. qrs. lbs. November 12, 1908 na ire 3 22 December 27, 1908 aoe ee 3 5 Total loss of weight.—17 lbs. Average rate of loss per week.—2°6 lbs. POST-MORTEM EXAMINATION. The carcass was thin. Local Lesion.—At the seat of inoculation on the left side of the neck there was a prominent firm tumour measuring 11°5 by 8 by 4 cm. composed of pinkish caseo-necrotic substance infiltrating both the skin and the muscles. Left Prescapular Gland.—The left prescapular gland measured 7:5 by 4:5 by 3°5 cm. ; the cortex was com- posed of dense pinkish-yellow caseo-necrotic substance, in which opaque yellow calcareous foci and fine streaks were seen ; a small portion of the medulla was com- posed of yellowish-white confluent caseous tubercles. Right Prescapular Gland.—The right prescapular gland measured 4:5 by 2 by 0°6 cm. and contained in the cortex a moderate number of caseous tubercles varying from 1 to 2 mm. in diameter. Left Prepectoral Glands.—The round gland measured 1-9 cm. in diameter and was composed throughout of firm greyish-red tissue beset with caseous foci; a second prepectoral gland showed confluent tubercles in its cortex and a few small discrete tubercles in the reddish medulla. A third showed in the cortex small patches of caseous tubercles and a few small yellowish- white discrete tubercles in the medulla. Right Prepectoral Glands—The right prepectoral glands contained a few small yellowish caseous tubercles and larger caseating nodules. Left Cervical Glands.—The lower cervical gland on the left side contained numerous small discrete caseous tubercles some of which were becoming confluent. A mid-cervical gland was entirely composed of greyish- yellow necrotic substance. The left upper cervical gland showed in its cortex a moderate number of yellowish-white caseous tubercles varying from 1 to 3 mm. in diameter ; each of the other cervical glands contained a few caseous tubercles. Right Cervical Glands—In each of the cervical glands on the right side there were a few tubercles the majority of which were a little more than 2 mm. in diameter. Asillary Glands—In the left axillary gland there were three yellowish nodules measuring rather more than 2 mm. in diameter and a moderate number of small discrete yellowish-white caseous tubercles in the cortex. The right contained four similar nodules. Thora. Lungs—The lungs were heavy weighing 4 lbs. 12 ozs. ; the cephalic, right middle, and antero-ventral portions of the caudal lobes were dark-red firm and almost completely airless ; the rest of the caudal lobes was congested and mottled with irregular patches of red hepatization ; the lung parenchyma was closely and evenly beset with firm caseous nodules varying up to about 2°5 mm. in diameter ; here and there through- out the lung they were aggregated together into groups in places forming irregular nodules. Thoracic Glands.—The bronchial and dorsal media. stinal glands were enlarged, the long mediastinal measuring 13°5 by 3:5 by 2°5 cm. ; they were firm and composed practically throughout of dense congested greyish-yellow caseating tissue showing here and there opaque yellow points probably of beginning calcifica- tion ; in the medulla of some of the glands there were discrete caseous tubercles. A ventral mediastinal gland on the left side measured 3 by 25 by 1 cm.and was composed practically throughout of firm reddish tissue infiltrated with a caseous network. Heart.—Under the endocardium of the right auricle four small greyish-white tubercles were seen: the heart was otherwise normal. Pleura.—The villous fringes on the pleural surface of the diaphragm were hypertrophied ; there was one lenticular growth measuring 4 mm. in diameter, yellowish-white and caseous in the centre. The fringes along the intercostal spaces were hyper- trophied but no tubercles were seen. Larynx.—The mucous membrane of the larynx showed four caseous tubercles with congested margins and ulcerated surface. Trachea.—There were a few congested spots with caseous centres in the mucous membrane of the trachea. Abdomen. Omentum and Peritoneum.—On the surface of the omentum there were counted seventeen loosely attached flattened caseating growths, the largest measuring a little more than 6 mm. in diameter. The peritoneum was normal. Spleen.—Thbe spleen was moderately closely beset with firm greyish-yellow nodules varying from 2 to nearly 3 mm. in diameter. Liver.—The liver showed on the surface just beneath the capsule numerous evenly distributed yellowish tubercles varying from 1 to 2mm. in diameter. On section similar tubercles were seen evenly distributed throughout its substance ; besides the caseous tubercles there were many minute grey tubercles. + Portal Glands.—The portal glands were enlarged and slightly oedematous ; their cortices were moderately closely beset with coalescing caseous tubercles, and there were a few discrete caseous tubercles in the medulla. Coeliac Glands.—The coeliac glands were enlarged, their cortices contained caseous tubercles becoming confluent, and the medullae a few discrete tubercles measuring 1 mm. in diameter. Kidneys.—On the surface of the right kidney twenty-one yellowish tubercles varying from 1 to 2mm. in diameter were counted. Some of the tubercles had grey margins. Two caseous tubercles with grey margins were seen in the cortex on section. Or the surface of the left kidney nine tubercles similar to those in the right were counted and four were seen in the cortex on sectior. Suprarenal Bodies——Hach suprarenal contained in the cortex one yellowish-white caseous tubercle measuring a little more than 1 mm. in diameter. Renal Glands.—The right was moderately closely beset with yellowish-white coalescing caseous tubercles, the left showed caseous tubercles becoming confluent. Lumbar Glands——All the lumbar glands showed discrete whitish caseous nodules, in places aggregated together, measuring from 2 to 3 mm. in diameter. Lliae G'lands.—Each iliac gland contained about ten caseous nodules. Alimentary Tract. Tongue.—Under the mucous membrane at .the base of the tongue there were a few small tubercles. Pharynax.—In the mucous membrane at the vault of the pharynx there were a few caseous tubercles. Tonsils—The left tonsil contained four, the right five firm caseous nodules, the largest almost the size of a hemp-seed. Parotideal Glands.—Each parotideal gland contained about twenty yellowish caseous nodules varying in size from 1 to 3 mm. in diameter, The Submazillary and Retro-pharyngeal Glands con- tained similar tubercles. Intestines.—The walls of the small intestine were thin ; every Peyer’s patch contained soft yellowish- white tubercles varying from 1 to 2 mm. in diameter ; they were moderately numerous in the lone Peyer’s patch in the ileum, and varied in number from two to eight or nine in the smaller patches. A few similar tubercles were seen scattered about in the submucosa of the duodenum and the jejunum. The lymphoid follicles in the large intestine were enlarged, and each showed a central depression but no sign of caseation. Gastric Glands.— All the gastric glands were enlarged and contained confluent caseous tubercles. Mesenteric Glands.—Some of the mesenteric glands were slightly enlarged; each showed in the cortex discrete caseous tubercles or caseating patches up to 5 mm. in diameter. Ileo-colic G'lands.—The ileo-colic glands were en- larged and showed in the cortex moderately numerous yellowish white tubercles becoming confluent. Various Lymphatic Glands. Precrural Glands.—Both contained a moderate number of yellow caseous nodules, the majority el measuring 3 mm. in diameter and in places becoming confluent. Popliteal Glands.—The popliteal glands were similar to the precrural. The Supramammary Gland contained three caseous nodules 3 mm. in diameter. Gluteal Glands.—In the left there were five yellowish- white tubercles measuring rather more than 1 mm. in diameter ; the right contained nine similar tubercles. Ischiatic Glands.—The left contained one similar tubercle ; the right was normal. Brain.— Normal. Microscopical Examination Smear from a Tubercle in the Small Intestine.— Tubercle bacilli numerous. CALF 1203. Virus H 60. “WB.” Subcutaneous Inoculation of culture derived from the human bronchial gland, through G.P. 1825 (The strain had been nearly a year in artificial cultivation and was tested after growth on glycerin-media). Dose—48°0 milligrammes. Date of Inoculation —March 26, 1907. Killed when in good health—July 1, 1907. Clinical Notes. A large tumour developed at the seat of inoculation on the left side of the neck ; three weeks after inocu- lation it was firm and prominent, measuring 15 by 8cem., and between 3 and 4cm. thick. The left prescapular gland was 10cm. in length; the left prepectoral gland was the size of a small walnut. The calf was in good condition. The tumour subsequently became pendulous and cystic, and eventually (on June 30th) opened and -discharged thick caseo-pus. The calf remained in good health during the whole period of the experiment. Temperature. During the first three weeks after the inoculation the temperature was a little raised (maximum 39°8° C.). ‘Subsequently it was approximately normal. Tuberculin Test. The Calf was not tested subsequent to inoculation. Weights. ewt. qrs. lbs. March 26, 1907 ... one - Ll 0 26 July 1,1907 ... sie - 2 0 15 Total gain of weight.—3 qrs. 17 lbs. Average rate of gain per week.—T 2 lbs. POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion At the seat of inoculation on the ‘left side of the neck there was a baggy partially collapsed oval cystic swelling, measuring 12 by 8 cm. ‘in superficial area; on section the cyst was partially filled with thick tenacious caseo-pus, and was sur- rounded by a thick wall of fibrous tissue lined internally with granulation tissue beset with caseo- -caleareous tubercles; the skin was adherent to the anterior wall of the cyst and was greatly thickened ; it was pierced towards the upper extremity by a sinus ‘the diameter of a crowquill, which communicated with the interior of the cavity and through which caseo- jpus could be expressed. Left Prescapular Gland.—The left prescapular gland [Age 13 weeks. ] [97 days after inoculation. ] measured 8 by 4°5 by 3cm., and showed on section two dense irregular caseous masses calcareous around the margins occupying rather more than half of the gland substance. Right Prescapular Gland.—(4:3 by 2:2 by 1cm.). Normal. Left Prepectoral Glands—The rounded gland was about the size of a thrush’s egg and had about a third of its substance replaced by dense caseo-cal- careous substance. The reniform gland was much enlarged and showed on section scattered calcareous foci. Right Prepectoral Glands.—Normal. Cervical Glands.—A lower cervical gland on the left side of the neck contained two or three calcareous tubercles. Other cervical glands were normal. Thorax. Pleura.—The parietal pleura was normal; on the diaphragmatic pleura there was one pin-head sized fibrous tubercle. Lungs.—The lungs were perfectly normal in general appearance; on close inspection sparsely scattered grey tubercles, ranging up to 1 mm. in diameter, were seen under the pleura ; twenty-four were counted on the surface of the left and there was about the same number on that of the right ; similar tubercles were sparsely scattered throughout the substance of the lung ; the larger tubercles had calcareous centres, the smaller ones were quite grey and translucent. Larynx and Trachea.—Normal. Thoracic Glands.—The bronchial aud dorsal medias- tinal glands were not enlarged ; on section they were beset, though not very closely, with discrete cal- careous tubercles which could be readily shelled out from normal-looking gland substance. The ventrai mediastinal glands were normal. Heart and Pericardiwm.—Normal. Abdomen. Omentum and Peritoneum.—Normal. Spleen.—The spleen, normal on the surface, showed on section altogether about fifty grey nodules with calcareous centres, ranging in size from a millet to a hemp seed. Liver.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Portal Glands.—The portal glands, not enlarged, contained discrete calcareous tubercles similar to those in the thoracic glands, but not so numerous. Coeliac Glands.—The coeliac glands were not enlarged ; one showed a small group of calcareous tubercles ; another contained one calcareous tubercle. Renal, Lumbar, and Iliac Glands—One lumbar gland contained two minute calcareous tubercles. The renal and iliac glands were normal. Ilio-sacral Glands.—Normal. Alimentary Tract. Tongue, Pharynx, Tonsils— Normal. Intestines —Normal. Mesenteric Glands.—In the cortex of one there was a minute calcareous tubercle. Gastric, Ileo-colic and Colic Glands.—Normal. Testicles.—Normal. 72 Various Lymphatic Glands. Retro-pharyngeal Glands.—One contained a small group of calcareous grains ; in the other there was a small calcareous tubercle. Submasillary, Pavrotideal, Mastoid, Awillary, Pre- crural, Popliteal, and Pudic Glands—Normal. Microscopical Examination. Emulsion of Prescapular Gland.—Tubercle bacilli fairly numerous. Emuision of Bronchial Gland.—One tubercle bacillus seen. Animals Inoculated. One rabbit was inoculated with the bronchial gland emulsion, No. 1363, subcutaneously (died 111 days, G.T.). Teo rabbits were inoculated with the prescapular gland emulsion, Nos. 1364, 1365, both subcutaneously (the former died prematurely, the latter died of chronic G.T. in 138 days). CALF 1481. Virus H 60. “WB.” Subcutaneous inoculation of culture derived from a single colony (No. 1) isolated from a glycerin- serum plate sown from Guinea-pig 2489. Dose—50°0 milligrammes. Date of Inoculation—July 30, 1908. Clinical Notes. The calf remained well during the experiment. Temperature. On the 9th day after inoculation the temperature rose to 40°3° C. and continued high (about 40:0° C.) for 17 days. It then fell to the normal and remained normal subsequently. Weights. ewt. qrs. lbs. July 30, 1908 ... ans si 1 2 1 November 3,1908 .... Be 1 38 21 Total gain of weight.—1 qr. 20 Ibs. Average rate of gain per week.—2-4 lbs. POST MORTEM EXAMINATION. The carcass was in good condition. Local Lesion.—At the seat of inoculation on the left side of the neck there was a tumour measuring 9 by 6 by 5 cm. which on section was composed of fibrous tissue with a cavity in the centre filled with caseo-pus. Left Prescapular Giland.—The left prescapular gland measured 6 by 3 by 25 cm. and contained one large fibro-caseo-calcareous mass and several smaller ones; they replaced about two-thirds of the gland tissue. . Right Prescapular Gland.—The right prescapular gland measured 5 by 2 by 1 cm. and was normal on section. Prepectoral, Axillary, and Cervical Glands.—Normal. [Age about 18 weeks.] Killed when in good health—November 3, 1908. [96 days after inoculation. | Thorax. Lungs.—Under the pleura seven submiliary grey tubercles with minute calcareous centres were seen. A few tubercles were seen on section. Thoracic Glands, Plewra, Heart.—Normal. Abdomen. Omentum and Peritoneum.—Normal. Liver.— Under the capsule there were two grey tubercles each with a minute yellow centre; no tubercles were seen on section. Spleen, Kidneys, and Suprarenal Bodies; Renal, Lumbar, Iliac, and Portal Glands.—Normal. Alimentary Tract. Tongue, Pharynx, Palate, Tonsils ; Submazillary, Retro-pharyngeal, and Parotideal Glands; Intestines and Mesenteric Glands.—Normal. Lleo-colic Glands.—One ileo-colic gland contained two or three calcareous grains. Mammary Gland.—Normal. Various Lymphatic Glands. Precrural, Popliteal, Gluteal, Ischiatic, and Pudic— Normal. Microscopical Examination. Focus from Liver.—No tubercle bacilli. A few thick bacilli and a few segmented bacilli stained blue. CALF 1425. Virus H 60. 7 3 “ W.B.” Subcutaneous inoculation of, culture dérived from a single colony (No. 5) isolated from a glycerin- serum plate sown from Guinea-pig 2489. Dose—50°0 milligrammes. Date of Inoculation—July 30, 1908. [Age about 19 weeks.] Killed when dying—September 14,1908. [46 days after inoculation. | Clinical Notes. The course of the disease was similar to that usually seen in calves inoculated with virulent viruses. Temperature. SULT usT SEpTEMBER ~ Weights. cwt. qrs. lbs. July 30, 1908 at 1 2 17 September 14, 1908 ... 1 0 22 Total loss af weight.—1 qr. 23 lbs. Average rate of loss per week.—T‘7 lbs. POST-MORTEM EXAMINATION. The carcass was thin. Local Lesion—The tumour at the seat of inocula- tion on the left side of the neck was very large : the left prescapular gland was incorporated with it: the mass weighed 3 lbs. 15 ozs. ; the tissues surrounding it were very oedematous. The tumour alone was pear-shaped measuring 20'5 cm., by 14 cm. in its widest part, by 7cm. On section the central part was composed of firm pink caseo-necrotic substance in two places commencing to break down; above it the skin was thickened and infiltrated right up to the surface with caseating areas ; below it there was a mass of infiltrated muscle. Left Prescapular Gland.—The left prescapular gland was very large measuring 10 by 6°5 by 5°5 cm. On section it was composed throughout of firm caseo- necrotic substance and showed two small crescent- shaped cavities containing fluid. Right Prescapular Gland.—The right prescapular gland measured 4 by 2 by 1 cm. and contained four congested caseous tubercles up to 2 mm. in diameter. Left Prepectoral Glands.—The rounded prepectoral gland was the size of a thrush’s egg, and was caseating throughout. The kidney-shaped gland measured 4 by 38cm. It was composed of glandular tissue thickly peppered with irregular whitish tubercles often con- fluent.’ A’smaller gland adjacent to it was similar. Right Prepectoral Glands.—One contained a single caseous tubercle, the others appeared normal. 27676 Left Cervical Glands.—One mid-cervical gland was enlarged and in an early stage of caseation. Auother contained caseous tubercles. An upper cervical gland was enlarged and oedema- tous, and showed caseous tubercles in the cortex. Right Cervical Glands—A mid-cervical gland con- tained two millet-seed sized yellow tubercles with congested marfins. An enlarged upper cervical gland contained two similar tubercles. Agillary Glands.—Each contained one hemp-seed sized congested caseous nodule. Thorax. Plewra.—Between the 7th and 8th ribs on the left side, close to the vertebra, there was a flattened loosely-attached fibro-caseous nodule 7 mm. in diameter. Two other smaller nodules were seen on this side, and there was one situated between the 5th and 6th ribs on the right side. Four were seen on the pleural surface of the pericardium, the largest 7 mm. in diameter. | Diaphragm.—On the pleural surface the villous tufts were hypertrophied aud a few opaque foci could be seen in them. On the peritoneal surface there were a few small scattered yellow tubercles. Lungs.—The lungs were heavy and voluminous, and weighed 5lbs.10z. The surtaces were thickly covered with irregular tubercles with grey translucent margins and large irregular yellow centres; these were fre- quently confluent forming patches up to lcm. in diameter. The tubercles varied in size from a pin- head upwards to a hemp seed. On the posterior and dorsal portions of the caudal lobes they were less numerous than elsewhere. On the surface of the anterior and middle lobes there were large and small dark red areas; the right middle lobe was almost entirely dark red. The posterior lobes showed nume- rous little irregular reddish patches frequently surrounding a tubercle or a group of tubercles. N On section the tubercles were equally numerous ; in the consolidated portions of the anterior and middle lobes they were especially thickly packed. The right middle lobe was consolidated almost throughout and sank in water; the other anterior lobes still contained a good deal of air-containing tissue. The right posterior lobe showed small irregular consolidated areas similar to those on the surface, but there was a large amount of air-containing tissue, and the tubercles were less numerous than elsewhere. The left posterior lobe was similar on section to the anterior lobes. The tubercles were almost entirely caseating. They were frequently confluent, but no large patches were seen. Thoracie Glands.—The bronchial and mediastinal glands were much enlarged, and weighed altogether 7 ozs. The surrounding tissues were very oedematous. The long mediastinal gland measured 12 by 4:5 by 15 cm. On section they were caseating almost throughout, very little gland tissue remaining ; the right bronchial was entirely caseated, and similar to the left prescapular. Heart.—Normal. Abdomen. Omentum.—The omentum contained several nodules, some loosely attached; the largest measured 0°6 cm. in diameter, and was composed of caseo-calcareous substance in a fibrous capsule ; the others were fibro- caseous. Parietal Peritoneum.—Normal. Spleen.—The spleen was slightly enlarged and packed with yellow caseating tubercles from 1 to 2 mm. in diameter. Liver—On the surface of the liver there were numerous whitish- yellow tubercles, the majority dimly seen through the capsule. A few of the largest were flattened on the surface and had slightly overhanging margins ; one measured 9 mm. by 4 mm., another 4 mm. in greatest diameter. Section showed the tubercles to be evenly and moderately thickly dis- tributed throughout the substance of the liver. They were firm and caseous, and varied from 1 to 2 mm. in diameter. Portal Glands.—The portal glands were enlarged and oedematous, the largest measuring 5 cm. in long diameter. On section they were packed with con- fluent caseating nodules, indefinite in outline. Coeliac Glands—The coeliac glands were similar to the portal. Kidneys.— Twenty - five yellowish - white tubercles were counted on the surface of the right kidney, some slightly raised, others dimly seen through the capsule. They measured from 1 to 2 mm. in diameter. Similar tubercles were seen here and there in the 74 substance of the kidney. The left kidney was simi- larly affected, 21 tubercles being counted on the a surface. a Suprarenal Bodies—The left showed four yellow tubercles up to 2 mm. in diameter ; the right con- tained one. Alimentary Tract. Tongue.—Normal. Tonsils.—The left contained one millet-seed sized caseating tubercle. The right was normal. Pharynx.—The corrugated mucous membrane at the back of the pharynx showed six congested caseous tubercles up to a millet seed in size. Larynz.—There were four yellow congested tuber- cles in the larynx. Trachea.—The trachea appeared normal. Submasillary Glands.--Each contained two or three yellow caseous tubercles with congested margins. Retro - pharyngeal Glands——Each contained a moderate number of congested caseous tubercles in the cortex. Intestines—The small intestine was normal. In the caecum there were two small opaque submucous tubercles with congested margins ; when picked out they were found to be caseous. Mesenteric and Ileo colic Glands.—Most of the mesenteric and ileo-colic glands contained scattered caseous foci. Testicles.—Normal. Various Lymphatic Glands. Lumbar and Renal.—These glands were packed with caseating tubercles. Llen-Sacral.—The ileo-sacral gland contained several caseating nodules. Lliac.—Two iliac glands contained each about a dozen caseous nodules up to a hemp seed in size. Precrural and Popliteal—Each contained about half a dozen similar caseous nodules. Pudic.—The one on the left side contained three, that on the might four, caseous nodules. Ischiatic.—The left contained a caseous tubercle, the right was normal. Gluteal—The right contained half a dozen small congested caseous tubercles. The left contained two caseous nodules. CALF 1489. Virus H 90. erp Subcutaneous inoculation of culture derived from the pudic gland of Calf 1383. Dose—50°0 milligrammes. Date of Inoculation—November 25, 1908. Died—January 6, 1909. Clinical Notes. The course of the disease was like that usually seen after the inoculation of a fatal dese of a virus virulent for bovines, being characterised by weakness, emaciation, and increasing difficulty of respiration. Temperature. The temperature was raised and irregular during the week following the inoculation ; 40°8°C. was [Age about 6 months.] [42 days after inoculation. ] reached on the 10th day, and the temperature was very high during the following 18 days (maximum 41°3°C., minimum 40°6°C.) ; it then slowly fell, and 386°C. was recorded on the day the animal died. : Weights. t. . lbs. November 25,1908 _.., eae ie o i: January 6, 1909... a - 1 2 16 Total loss of weight.—1 qr 27 Ibs. Average rate of loss per week.—8 lbs. POST-MORTEM EXAMINATION. The carcass was emaciated. Local Lesion—At the seat of inoculation on the left side of the neck there was a slightly raised tumour fluctuating in the centre measuring 15 by 7 by 3:°5cm.; on section it was composed of a'subcu- taneous mass of pinkish-yellow caseo-necrotic sub- stance infiltrating the skin and the muscles, and ‘containing in the centre a ragged cavity filled with yellow serous fluid. The tumour was attached to the nearest glands by thick caseous cords. Left Prescapular Gland.—The left prescapular gland measured 8:5 by 6 by 45cm. and was com- posed throughout of dense yellow caseo-necrotic substance ; the capsule was thickened and in places (i.e. where joined by the caseous cords) infiltrated with caseous substance. Right Prescapular Gland.—The right prescapular gland measured 4 by 1°7 by 1cm., and showed in the cortex four caseous nodules, the largest 3mm. in diameter. Prepectoral Glands.—On the left side one 1:3cm. in greatest diameter was indurated and yellow and ‘caseous almost throughout. Another, disc-shaped and 4cm. in greatest diameter, was firm grey and showed early foci of caseation. Another small one was in a similar condition. On the right side two contained altogether five -caseous nodules up to 2 mm. in diameter. Cervical Glands.—On the left side at the root of the neck there was a large gland measuring 4°5 cm. in greatest diameter which was dense and caseating practically throughout. Other cervical glands were ‘congested and contained each two or three caseous nodules with congested margins, the largest 25mm. in diameter. Axillary Glands—In the left there was one easeous tubercle, in the right there were about half a dozen up to 2mm. in diameter. Thoran. Lungs.—The lungs filled the chest and were very heavy weighing 11 lbs. 4 ozs. and all the lobes were ex- tensively hepatized, dark red in colour and engorged with blood ; in the posterior parts of the caudal lobes and around the margins of the cephalic there was some air-containing tissue, but this was firmer than normal and deeply congested ; the parenchyma of the lungs was closely beset with yellowish-white caseous tubercles the majority about 1mm. in diameter; a few were larger and here and there some had coalesced; the tubercles under the pleura of the anterior lobes and parts of the caudal projected slightly and were flattened out to a larger diameter than those in the -depths. Thoracic Glands.—The bronchial and mediastinal glands were greatly enlarged weighing together 12 ozs. ; on section their cortices were composed of firm pinkish- grey tissue mottled with irregular yellow caseous patches and foci. Pleura.—The fringes along the margins of the ribs -were slightly hypertrophied and deeply congested ; they -contained sparsely scattered flattened caseating nodules 75 varying up to 3mm. in diameter. The diaphragmatic pleura showed a few small flattened nodules and con- gested villous patches. Heart and Pericardium.—The heart muscles and valves were normal. There was a hemp seed-sized caseous pedunculated nodule on the surface of an auricle. Abdomen. Omentum and Peritoneum.—On the ventral surface of the omentum there were about half a dozen lenti- cular caseating nodules with congested margins the largest 5 mm. in diameter. The parietal peritoneum was normal. Spleen.—The pulp was soft and moderately closely beset with yellow caseous nodules the largest 3mm. in diameter. Liver.—The liver was much enlarged yellowish in colour and very friable; the substance contained moderately numerous evenly distributed tubercles ranging trom about 0°5 to 2mm. in diameter, the larger ones were yellow and caseous with grey margins, the smaller ones grey with opaque centres; on the sur- face there were a few flattened mushroomed nodules. Portal Glands.—The portal glands were enlarged ; their cortices were composed of grey caseating nodules closely aggregated together; in the cential parts of the glands the nodules were more discrete. Kidneys.—In the cortex of each kidney on the sur- face as well as on section there were scattered caseous nodules ranging from 1: to 2°5 mm. in diameter. Suprarenal Bodies.—In the cortex of the right there were two greyish-white miliary tubercles. The left was normal. Coeliac Glands.—The coeliac glands were enlarged and composed throughout of confluent grey nodules in a state of early caseation. Lumbar and Renal Glands——The lumbar and renal glands were enlarged and closely beset with caseating nodules. ' Alimentary Tract. The Tongue, Pharynx, Larynx, and Trachea were congested, but no tubercles were seen. Tonsils—One tonsil contained a caseous nodule 3 mm. in diameter. Intestines.—Some of the Peyer’s patches in the small intestine contained a few miliary caseous tubercles. The Mesenteric and Ileo-colic Glands showed in the cortex each a few grey caseating patches some 1 cm. and more in diameter. Testicles.—Normal. Various Lymphatic Glands. The Precrural, Popliteal, Mliac, Submazillary, Paro- tideal, and Retro-pharyngeal Glands contained a varying number of discrete caseating nodules in the cortex the largest 3 mm. in diameter, the iliac and submaxillary containing a moderate number. CALF 1493. Virus H 90. 21 pe Subcutaneous inoculation of culture derived from the bronchial gland of Calf 1471. Dose—45-0 milligrammes. Date of Inoculation—November 23, 1908. Died—December 14, 1908. [Age about 6 montas. j [21 days after inoculation.] Clinical Notes. ‘The illness was of the acute type frequently seen after the inoculation of virulent viruses. a 76 ‘Temperature. NovEMBeER, Tuberculin Test. The calf was not tested subsequent to inoculation. Weights. ewt. qrs. lbs. November 23, 1908 ... 1 38 «iil December 14, 1908 ... 1 2 3 Total loss of weight.—1 qr. 8 lbs. Average rate of loss per week.—12 lbs. POST-MORTEM EXAMINATION. The carcass was emaciated. Local Lesion.—In the subcutaneous tissues on the left side of the neck there was a lenticular mass of yellow caseo-necrotic substance measuring 10 by 8 by 2 cm., infiltrating the skin to a slight extent and the muscles in one place rather deeply. Left Prescapular Gland.—The left prescapular gland measured 9 by 5 by 3cm., and was dense yellow and caseous practically throughout. Right Prescapular Gland.—The right prescapular gland measured 5 by 2 by 1 cm., and showed scattered minute tubercles on section. Prepectoral Glands.—On the left side one 2 cm. in diameter was caseous throughout, another larger flat gland showed the cortex grey and in a state of early cageation ; another gland contained discrete tubercles. The glands on the right side contained scattered minute tubercles. : Cervical Glands.—One in the middle of the neck on the left side, the size of a thrush’s egg, was firm and in a state of early caseation ; the other cervical glands were deeply congested and showed scattered tubercles, as in the peripheral glands. Thorac. Pleura.—The fringes along the margins of the ribs were slightly hypertrophied and deeply congested ; no tubercles were seen. Lungs.—The lungs did not collapse and filled the chest ; they were heavy, weighing 9 lbs. 14 ozs., and with the exception of the postero-dorsal parts of the caudal ‘lobes, which were still air-containing, were dark-red and solid, portions sinking in water ; the parenchyma of the lungs was everywhere very closely beset with minute grey tubercles. Thoracic Glands—The bronchial. and mediastinal glands were enlarged, deeply congested, and showed in the cortex grey patches composed of aggregated grey tubercles and discrete grey miliary tubercles. DECEMBER. Heart.—The cavities of the heart were filled with red blood-clot ; the endocardium of the right auricle and right ventricle showed a few small caseous tuber- cles. The pericardium was normal. Larynz and Trachea.—The mucous membrane of the larynx and trachea was congested. Abdomen. Omentum.—There were four small tubercles on the omentum. Peritoneum.—The peritoneum appeared normal. Spleen.—The spleen was not enlarged ; the capsule. was wrinkled and the pulp was soft and seemed a little atropbied ; it showed numerous not very con- spicuous grey bodies, probably enlarged Malpighian ‘bodies, which showed no sign of caseation. Liver.—The liver was enlarged, pale and soft, and contained fairly numerous evenly distributed grey miliary tubercles. Portal Glands.—The portal glands were not obviously enlarged. The tissue was oedematous and aod cortex contained grey patches with early caseous oci. Kidneys.—In the cortex of each kidney there were a few minute grey tubercles, Suprarenal Bodies.—Normal. Alimentary Tract. Pharync.—The mucous membrane of the pharynx was congested. Tonsils.—The tonsils were congested, no tubercles. were seen. Intestines—Some of the Peyer's patches of the small intestine showed a few greyish-white points.. The large intestine was normal. Mesenteric Glands.—No tubercles were seen. Lleo-colic Glands.—A few tubercles were seen in one- gland. Testicles.—Normal. Various Lymphatic Glands. All the peripheral lymphatic glands showed in the cortex scattered opaque tubercles less than 1 mm. in diameter. There were similar but more numerous tubercles in- the coeliac, lumbar, and renal glands. Microscopical Examination. Smear from a focus in a Peyer’s patch.—A few- tubercle bacilli. & N pure J sorqey, 29g ((quarnata pax orm0ssp) aangqno perm ‘A IRL sag -(quafnna AYYSYs pure otuoSne) oyejd rede wnszos e woaz Luojoo e Lq paonpoad yyaors oy, *9 ‘ON OU} YUM pozv[nooar spemiuvy woIz poyepost sainyrno Aq paonpord sqyaory “OT put G ‘SON, Pe, 999 «*(quepnaia pure oruossXp) oyefd umnses e mors (F “ON') AuOJoo v Aq poonpoad qyMo013 oT, “FON ‘has 2 =. 0 IVL 229 *(quapnata ‘d MRL 9g “(quernsra ApqST[s pur ormo0Sna) aye [d wnaes & wosz (¢ on) AdoToo v kq poonpoad yyMo1s oy, “Eg “ON [HPSS puv otaosna) o4vyd mies © woz Auojoo afsuts v £q poonpoid yyaois oy, “9 ‘ON ‘CNL ag “(queynita pur ormosna) oyejd wnzas & U0Iz (QT pur Z SON) SetMOTOD Aq peonpoad sqyMoiy “G pur Z ‘SON ‘(qugynata pur otmoSna) aanqno jemtat eq} Aq peonpoad qaors ayy, *2 ‘ON “(quo[nata pUue oTUOSNs) san4qTNo [eIyIUL ey} Aq poonpoid yyAOoIs ONT, “[ “ON ‘OT 6 8 Bs) 9 g t € G I | Sey shel ||| Sesone S| Poe aquanasah rio swe 0 sua oe Gossaoy | I | Goswaye Gosia Op | 7 sande | | Ligtab | i sgaieb oy | Leearcatclir O\ ANOT0) : ty ANoI0) g snore? || F J sor oarvana6 408 | ZOs1 3D | (Breese = rrr i S09T th 164 ab 1645 db 4G a> ; \6ng ab — rem =a | rim at Ape NOEs PSH EMOGH} | |/awoon)|| | / No cJCUCIH ‘van 7 i Ue) “(umes pun6 ypryou0sg) IAL 09 (HH Shee “(ums 108 JD) OV 9 STH snszg fo saunyno pammandas ayy fo auos pun aanygno qoryiwe ay? JO (e) fo sasngna paypundas ay) Jo auos pun aunzyna qouiue ay JQ (q) —O1LVLOd GHLVNIYHOATS NO SHUNLTNANAAS 78 *(qQuepnata pue o1mosne) ain ~Tao yertur egy Aq pesonpoid = qyAors eqL ‘OT “ON ‘Or mo oat Te wen yST Lexar eb pre ee a {a lOOh] W pur yy soquy, aag-*(quopnata ¢ ‘on ‘otuossdp) @IN4[ND [VI4TUT 94} YIM paye[Nooul s[emtae wWoAZ peyvjost saingguo Aq peonpoid qyao1xy “g put g ‘SON *‘(qu9[nita pur o1u08ne) amypno yeytar oq} Aq peonpord yyaors oy, “2 ‘ON 6 8 Y Kg ah AY bi Ni - ad SESS Go ened 87 ne 2 i 5 ok eG aot tt (ee ms fe 5¢6 jee dows Wo SN CoH) [ [SMOqH : sf “V PAE L 298" (quoyn.11a puv o1uodsXp) spemrue oBessed wosz poqzepost soanqyjno Aq poonpoad syjmory, 9 pue ‘Gg ‘p ‘SON “dl IRL, 998" ‘(quafnita put ormosno) oyeyd uintes v worz (p put Z ‘SONT) SoruOjoo Aq poonposd syyModx) “gE pue Z ‘SON, ‘(qU9[NATA pUe dIdOSsNa) eAngjno [erytut oq} Aq poonpoad qymo13 oT, “T ‘ON 9 G T ‘€ 6 ‘lt ae sw OM 1 weed | cho sava 64° Bl) cio siwaoy) es sy] marr 45 | rents | ges — (2 No) | | (GANMOD, is 14 64ST abd | | 169e ‘ay | tos’) ay Gl 4 log: Ne “(ays punjb yoauojad -0njat) AT 99 06 (HH Snied ayy fo ainjgnd pour ayn JO (2) ae “(ums puoj6 7D2YIU0LG) I'M O9 “EL snl2gq 947 £0 sasngyna parpindas ayy fo omg pun aunqyno pore aya fo (q) ‘(apes 108 JM) OV SIO Shi yp JO saangyna pagnandss ay} {6 awos pun aingjna qoyae ayn {oO (®) —AVNV NIVMOATO NO SHHNALTINOGNS » e? * ’ “ ‘ < _ahit : . i , f * : ® \ is & . if ¢ . : : ‘ ‘ | | ‘ | | = qi ‘ oe, y See 8) ; 4 i ey de sys Sb on 79 REPORTS ON THE EXCRETION INTO THE MILK OF 27676 COWS AND GOATS, AND INTO THE MILK SINUSES OF THE UNDEVELOPED UDDERS OF HEIFERS, OF TUBERCLE BACILLI WHICH HAD BEEN SUBCU- TANEOUSLY OR INTRAVENOUSLY INOCULATED. BY A. STANLEY GRIFFITH, M.D. THE EXCRETION OF TUBERCLE BACILLI IN MILK. 80 TABLE OF CONTENTS. PART I. THE EXCRETION OF TUBERCLE BACILLI INTO THE MILK OF COWS AND GOATS. Introduction Technique Short account of each experimeat Summary Tabular summary of the experiments THE EXCRETION OF TUBERCLE BACILLI INTO THE UNDEVELOPED UDDER. {ntroduction PART II. Short account of each case investigated ... Summary wes Tabular summary of the cases investigated Details of the Experiments. Details of the experiments dealt with in Part I of the Report :— Experiment A (Cow 300) ... As B (Cow 77) ... ‘3 C (Cow 61) ... . D (Cow 565) ... a E (Goat 79) . ‘5 F (Goat 55) ... ™ G (Goat 59) ... a H (Goat 69) ... 43 I (Goat 61) ... 3 J (Goat 81) . 35 K (Goat 75) ... Abstracts of the cases, and detailed results of the investigation summarised in Part II of the Report :— Case 1 (Heifér-calf 1479) ... » 2 (Heifer 1601) » 3 (Heifer-calf 1461) ... » 4 (Heifer-calf 1535) ... » 5 (Heifer 1571) » 6 (Heifer-calf 1505) ... » 7 (Heifer-calf 1549) ... » 8 (Heifer 1577) . 9 (Heifer 1583) » 10 (Heifer-calf 1533)... » 11 (Heifer 80)... » 12 (Heifer 1569) » 13 (Goat 77) PAGE 81 81 82 90 92 94 95 96 97 . 137 . 138 . 138 . 139 . 139 . 140 - 140 - 140 . 141 e. 141 «. 142 . 142 oe 143 81 Part I. THE EXCRETION OF TUBERCLE BACILLI INTO THE MILK OF COWS AND GOATS. INTRODUCTION. In spite of numerous observations on naturally tuberculous cows, it has not yet been definitely settled whether the normal udder will permit the passage of tubercle bacilli from the blood-stream into the milk. Positive results have been obtained by many investigators with milk from tuberculous cows in which no disease of the udder could be detected either macro- scopically or microscopically, but these cannot be accepted as proof that the bacilli were excreted in the milk directly from the blood-stream, since the bacilli may have been introduced accidentally while collecting the samples or have escaped into the milk from some early and undetected tuberculous lesion in the udder. Accidental contamination can be avoided, and has been in the more recent experiments in which positive results have been obtained, but it is doubtful whether any examination can be so exhaustive as definitely to exclude the presence of early lesions in the udder. It seemed to me, therefore, that the question whether tubercle bacilli can pass from the blood into the milk without producing changes in the mammary tissue could be solved only by direct experiment, 7.e. by introducing tubercle bacilli into the blood-stream of a healthy cow and testing her milk at short intervals subse- quently. If such experiment showed that tubercle bacilli so introduced were excreted in the milk within a short period after their inoculation into the blood- stream there would arise strong presumption that a similar excretion takes place during the progress of a natural infection. A preliminary experiment was therefore made at Walpole Farm with a culture of bovine tubercle bacilli. A cow was inoculated subcutaneously with 100°0 milli- grammes of culture, and her milk tested at intervals after the inoculation produced tuberculosis in guinea-pigs. The subcutaneous method of inoculating the cow was chosen instead of the intravenous, as it was anticipated that the latter method of inoculating bovine tubercle bacilli would cause the too speedy death of the animal, and as it had been shown that after subcutaneous inoculation there is an immediate and for several days a rapid escape of bacilli into the blood-stream. It was inferred from the results of the guinea-pig inoculations in this experiment that the milk of a tuberculous cow is liable to contain tubercle bacilli whenever tubercle bacilli gain access to the general circulation. This experiment (Experiment A) has already been fully reported (Appendix to the znd Interim Report, Vol. I, pp. 35 and 226) but for convenience of reference the details have been reproduced in this report. In order to ascertain whether a similar excretion takes place after the inoculation of tubercle bacilli possessed of slight virulence for the calf, a series of three cows and six goats has been inoculated and the milk of these animals tested fr. ‘quently at short intervals subsequently ; in addition a goat has been inoculated wth a culture of human origin but exhibiting bovine characters. The clinical histories and post-mortem notes of these animals, and tli: results of the inoculation of their milk, are fully detailed in this report. All the animals previous to their inoculation had been tested with tn!-rculin and their milk inoculated into guinea-pigs ; the results in all cases were ne»: ve. TECHNIQUE. The milk of the first two cows (300 and 77) was collected by hand the teats, udder and surrounding skin were carefully cleansed and then washed \ a | in 1,000 solution of mercury perchloride ; finally the teats and udders were washed with aleohol ; the first few drops of milk were rejected and in the case of le. first two 27678 P 2 82 cows the samples for inoculation were received into sterile glass bottles held at an angle to avoid as far as possible the entrance of chance particles of dust. ; The milk from the other animals, with the exception of Goat 81, was obtained by catheterisation, the same precautions as to cleanliness of the udder and surrounding skin being observed as with the first two cows. In all cases the milk was collected by me or by a skilled attendant under my direct supervision. In every case the udder was milked dry just before the animal was inoculated, and whenever samples were taken no milk had been withdrawn during the previous twenty-four hours. The amount of milk inoculated into the guinea-pigs varied. Sometimes the milk was centrifuged with a hand centrifuge and the deposit was suspended in uncentrifuged milk for the purposes of inoculation, but in the large proportion of cases the milk was uncentrifuged, the dose varying from 0°5 or 1-0 ec. up to 30:0 cc. In the first two experiments the dose in nearly every case was 5°0 cubic centimetres of uncentrifuged milk ; in the others the dose was generally 10-0 cubic centimetres. EXpERIMENT A. Cow 300 was inoculated subcutaneously in the neck on September 2, 1905, with 100°0 mg. of a bovine culture and her milk tested on guinea-pigs subsequently at intervals of 7, 16, 21, 28 and 30 days after her inoculation ; also two pigs were fed with her milk for a week. The experiments on the guinea-pigs showed that the milk was capable of setting up tuberculosis at the end of the first week and on every subsequent occasion. The number of tubercle bacilli excreted in the milk did not appear to be large since most of the guinea-pigs developed only a mild type of disease and the swine though fed for a week with the milk did not become tuberculous. The cow died of acute generalised tuberculosis in 30 days ; the udder was normal to the naked eye and no tuberculous lesions or tubercle bacilli were found on microscopical examination. . EXpERIMENT B. Cow 77 was inoculated subcutaneously on November 28, 1906, with 100°0 milli- grammes of culture derived from the original material of Virus H 68. “ R.B.” (a case of tuberculous osteitis) ; this culture grew luxuriantly on artificial media and was only slightiy virulent for the calf and rabbit. The general health of the cow was unaffected by the inoculation, and the local lesion which developed at the seat of inoculation diminished greatly in size and was represented finally by a small patch of thickening. The cow, which was killed in good health 156 days after inoculation, showed at the post-mortem examination a fibrous local lesion, a caseo-purulent nodule in the prescapular gland, but no sign of tuberculosis elsewhere. ach quarter of the udder was examined histologically by Dr. Eastwood but no tuberculous lesions or tubercle bacilli were found. Throughout the first week of the experiment a series of eight guinea-pigs was inoculated every twenty-four hours, then for twelve weeks a series of eight at intervals of about a week ; the last two series were inoculated after 112 and 155 days and each included twelve guinea-pigs. Altogether seventeen series, comprising 144 animals, were -inoculated. Each guinea-pig in the first fifteen series received 5:0 cc. of milk ; two were inoculated from each quarter, one subcutaneously and one intraperitoneally. In the sixteenth series eight of the guinea-pigs received 5:0 cc. of milk, the other four each got the centrifuged deposit of 100°0 cc. For the last series the whole of the milk yielded during twenty-four hours was centrifuged, and the deposit from each quarter inoculated into three guinea-pigs. In all but two of the series (¢.e. the fifth, in which none of the animals became tuberculous, and the tenth, in which the one animal which was affected had spontaneous tuberculosis) one or more of the guinea-pigs inoculated developed definite inoculation tuberculosis. Of the first day’s milk, 7.e. that secreted durine the 24 hours after inoculation of the cow, that from two of the quarters only proved infective ; on the second and third days milk from all the quarters picoice tuberculosis. From the end of the third week to that of the thirteenth, one. or at most three, of the guinea-pigs in each series became tuberculous. In the last series of twelve guinea-pigs, which were inoculated with centrifuged deposit, one from each of three quarters and two from the other became tuberculous. 83 ‘Two rabbits were inoculated, one subcutaneously and one intraperitoneally, with mixed milk taken from all the quarters just before the cow was killed. They were killed 92 days later, the subcutaneously inoculated rabbit showing a local lesion and one tubercle in the lung, the intraperitoneally inoculated rabbit three or four tubercles in the omentum, one in a kidney, and one in the lung. In order to ascertain whether the tuberculosis in the guinea-pigs in the last three series was produced by bacilli of like kind with those inoculated into the cow, ten rabbits were inoculated with emulsions of the organs of eight guinea-pigs and in addition a culture was isolated from each of these guinea-pigs. Three of the rabbits died prematurely, the others showed slight retrogressive tuberculosis All the cultures isolated grew luxuriantly on the test media. The properties of the bacilli excreted in the milk were therefore precisely those of the bacilli inoculated under the skin of the cow. EXPERIMENT C. Cow 61 was inoculated subcutaneously on August 6, 1907, with 100°0 milli- grammes of a slightly virulent culture obtained from a tuberculous lung (Virus H 81. ‘“P.W.”). She was an old cow and was giving only a very small amount of milk ; her last calf had been born on October 15,1905. One of the quarters of her udder, the right fore, was atrophied, and had for some time yielded only a small quantity of serous fluid and yellow clots in which numerous streptococci were demonstrated. Nine series of guinea-pigs were inoculated with the milk, three series in the first week after her inoculation, the last series after eighty-four days. Only one of the guinea-pigs became tuberculous. This guinea-pig was one of the series inoculated on the seventh day and had received intraperitoneally 10°0 cc. of fluid obtained by filtering clots which had been withdrawn by hand from the right fore quarter. A culture isolated from the guinea-pig grew like the culture inoculated into the cow. Ninety days after the subcutaneous inoculation the cow was inoculated intravenously with 150°0 milligrammes of culture from Virus H 83. “G.C.,” a slightly virulent culture obtained from the spleen of a child who died of general tuberculosis. A series of guinea-pigs was inoculated with her milk every day (except on the fourth) during the first eight days, then on the tenth, twelfth, fifteenth and forty-fourth days, and one guinea-pig on the eighty-fifth day. All the guinea-pigs, fourteen in number, inoculated one and two days after the inoculation of the cow, became tuberculous ; in the third series four out of the six developed tuberculosis ; and the milk continued to be infective up to the forty-fourth day, the single guinea-pig inoculated on the eighty-fifth day remaining healthy. Cultures were isolated from seven of the tuberculous guinea-pigs, and these all exhibited the cultural characters of the bacillus intravenously inoculated. As in Cow 77, an irregularity in the infectivity of the milk was observed ; of a pair of guinea-pigs inoculated from the same quarter one would be found tuberculous, the other healthy, and the milk from a quarter which produced tuberculosis on one occasion would on the next fail to do so. By the 85th day, that is on January 28, 1908, the secretion of milk had practically ceased and the routine inoculations of guinea-pigs was discontinued. The milk had been examined microscopically for tubercle bacilli on several occa- sions during the first ten days after the cow was intravenously inoculated, but none was demonstrated in it. From January 28 to April 14, 1908, the udder of the cow was not meddled with. On the latter date (162 days after the intravenous inoculation) the two fore- quarters were milked and some pus was obtained from each ; smear preparations from the pus showed in each case very numerous tubercle bacilli, the majority being long and beaded; there were numerous cells, mainly polymorphonuclears with scattered lymphocytes and alveolar cells. On the following day 50:0 cc. of sterile salt solution were injected into each of the two fore-quarters, and after manipulating the quarters with the object of forcing the fluid into the smaller ducts, as much as possible was recovered by means of the apparatus which had been used for injecting the salt solution ; only a small quantity of the fluid was recovered, and this was slightly milky and contained small flakes and flocculi ; smear preparations of this fluid showed numerous tubercle bacilli. Subsequent microscopical examinations of the fluids which collected in the milk sinuses were made at intervals. The following table gives April 14, 1908 :— 84 the results of these examinations on and after Date. orn Description of Material. Result of Examination. ; Numerous T.B. Cells mainly April 14, 1908 LF... | Dus polymorpho-nuclears ; scattered RF... j lymphocytes and alveolar cells. . { Fluid recovered after | Numerous tubercle bacilli. April 15, 1908 a or injecting salt solu- ee tion. April 21, 1908 L.H. Fairly numerous T.B., definitely R.H. Pus | less numerous than iri the fore (quarters. June 2, 1908 S a A few drops of pus... | T.B. numerous. July 1, 1908 Mixed fluids | Yellow muco-purulent | T.B. numerous. from all the fluid. quarters. July 17, 1908 LF... 7 RB vss | A te dope. oF ‘pam T.B. numerous. LH. { lent fluid. R.H. 5 5 7 A few clumps of very short T.B., quite different from those in the : other quarters. August 6, 1908 ... | LF... ('T.B. fairly numerous, but dis- BR. Pin Pus tinctly less numerous than L.H. when first examined in April Sept. 15, 1908 LB. ses Yellow muco-pus with | A moderate number of tubercle small tlakes. bacilli. R.F. . Yellow muco-pus : LH. Thirketenaciouwyallow | Scattered beaded T.B. and a few pus \ clumps. R.A. Milky watery fluid ... | A few small groups of very short acid-fast bacilli and granules; the cells were chiefly polymor- pho-nuclears. October 6, 1908 ... | L.F... Yellow tenacious l muco-pus. : RF. ... Serous fluid containing T.B. moderately numerous. small whitish flakes L.H. Thick yellowtenacious | T.B. rather more numerous than pus. in the fore quarters. R.H. Muco - purulent sub- | One T.B.and a few groups of acid- stance and serous fast granules. fluid. October 15, 1908... | L.F... Yellow pus ... . | T.B. moderately numerous. R.F.... Yellow pus in which sane gram |’ T.B. in moderate numbers, L.H. Yellow pus... R.H. Milky fluid on which | A few small groups of short acid- on standing a layer fast bacilli; many short chains of cream formed. of cocci. January 14, 1909... | L.F.... Yellow pus, thick and | T.B. moderately numerous. tenacious. R.F.... Watery fluid with | Scattered T.B. seen. whitish flakes. L.H. Yellow pus, thick and | A moderate number of T.B. seen. tenacious. R.H. Turbid watery fluid | No T.B. seen. with flakes of yellow pus. April 15, 1909 ioe Yellow pus Numerous long and beaded TB. L.H. ” ‘8 ‘ moderate number of T.B. R.H. a 2 A tew acid-fast granules only were seen, 85 : Towards the end of June, 1908, that is nearly eight months after her intravenous inoculation, it was observed that three of the quarters of the cow had become enlarged and somewhat indurated, and that there was a sero-purulent discharge from the two fore teats. A fortnight later there was a distinct increase in the size of the three quarters ; the left fore and the left hind were uniformly indurated, the right fore was indurated and nodular ; the right hind showed no alteration in size. On July 1, 1908, each of the quarters was milked dry because it was noticed that three of the sinuses were becoming distended, Altogether 450-0 cc. of yellow tenacious muco-purulent fluid was withdrawn ; there was practically none in the right hind quarter. With the exception of the few drops required for microscopical examination nothing had been withdrawn from the udder for six months. On the next day 250°0 cc. were obtained, and on the next 100-0 ce. ; the latter was more serous than the first, but still very turbid. Subsequently the amount of discharge rapidly became less and after July 17 small quantities only of pus were obtained. After attaining its maximum size on or about July 1, the udder underwent no appreciable alteration for some time ; on October 15, 1908, it was thought to be on the whole a little smaller ; the quarters on the left side were each larger than the right tore and were uniformly very knotty and indurated ; the right fore was less uniformly indurated and could be resolved more easily into separate nodular or knotty areas ; the right hind was small and atrophic and no nodules were felt in it. Subsequent to this date there was a gradual diminution in the size of the organ, and it was found on April 15, 1909, the day before the cow was killed, when an attempt was made to wash out the quarters, that the milk sinuses of twu of the quarters were much contracted and the ducts apparently obliterated, and in the case of one of the quarters it was impossible to insert the cannula on account of the teat canal being filled with a firm cheesy substance. The cow was killed on April 16, 1909, 529 days after the intravenous inoculation ; the post-mortem examination showed in the caudal lodes of the lungs a few dense or softened caseous and calcareous nodules of various sizes, a large fibro-caseous mass with two small nodules in the long mediastinal gland, gritty caseous nodules in the suprarenal bodies, caseo-purulent nodules in the pancreas and tuberculosis of the udder which was atrophied and fibroid (see page 109). There was no lesion at the site of the subcutaneous inoculation and the adjacent prescapular gland was normal. The virulence of the tubercle bacilli in the milk sinuses has been tested by the inoculation into rabbits both of the fluids recovered from the quarters after injecting saline and of cultures obtained direct from the purulent discharge. Thus :—Washings from the quarters have been inoculated into rabbits on three separate occasions, on April 15, and October 15, 1908, and on April 15, 1909, and in each series of rabbits slight tuberculosis only was produced. And cultures of tubercle bacilli have been obtained direct from the pus of the mamma of the cow on four separate occasions, on April 15, August 6, and October 7, 1908, and on April 15, 1909. All these strains have grown on the test media exactly like the culture intravenously inoculated, and three, including one isolated on April 145, 1909, have been shown to possess for the rabbit the low virulence of the original culture. The bacilli from the milk sinuses had not, therefore, undergone any alteration in cultural characters and in virulence for the rabbit after a residence in the body of the cow of 528 days. In the early part of the experiment streptococci were often demonstrated in the milk and were particularly numerous in the clots from the right fore quarter. On October 7, 1908, a culture tube sown with fluid from the right hind quarter became covered in 48 hours with streptococcus colonies. The cultures of tubercle bacilli obtained from the other quarters did not show any streptococcus colonies and vo organisms other than tubercle bacilli had been demonstrated in smear preparations from these quarters during the period from April 14, 1908. The streptococci there- fore had disappeared from those quarters in which there had been a multiplication of tubercle bacilli and had persisted in the single quarter which had not become affected with tuberculosis. The positive results with the milk of this cow after tubercle bacilli had been inoculated directly into the blood-stream supplies a possible explanation of the 86 negative results after the subcutaneous inoculation, namely that the subcutaneous inoculation was followed by only a slight dissemination of tubercle bacilli. This slight dissemination of bacilli may have been due to the condition of the culture. It not infrequently happens that bacilli of Group IL grow on serum very quickly and luxuriantly and then apparently rapidly lose their vitality ; this loss in vitality is shewn in the difficulty sometimes experienced in obtaining subcultures (on serum) from such cultures when they are not more than three weeks or a month old. The faeces of the cow were inoculated into sixteen guinea-pigs, six after the subcutaneous inoculation and ten after the intravenous. Seven guinea-pigs died shortly after inoculation, eight were killed and found to be healthy, one only became tuberculous. The guinea-pig that became tuberculous was inoculated ten days after the intravenous inoculation of the cow. Experiment D. Cow 565 was inoculated intravenously on February 18, 1909 with 10:0 milli- grammes of a Group II. culture obtained from a tuberculous human lung (Virus H 104. “ E.R.”) The cow, 44 years old, was about 4 months pregnant; one of her quarters was “dry.” The milk from the three functional quarters was inoculated into a series of six guinea-pigs, two from each quarter, every day during the first seven days (the first series being inoculated twenty-four hours after the inoculation of the cow), and again at the end of a fortnight ; between the 14th and the 168th day seven series of guinea-pigs were inoculated, the intervals ranging from two to four weeks ; in the last fortnight four series of guinea-pigs were inoculated. In the first series of guinea-pigs, ¢.e. those inoculated with milk secreted during the first twenty-four hours of the experiment, only one animal became tuberculous. All the animals in the second, third, and fourth series remained healthy. In the fifth series there were two, in the sixth one and in the seventh four tuberculous animals. The milk from each of the three quarters withdrawn on the 14th day produced tuberculosis in one of each pair of guinea-pigs inoculated, the other remaining free from tuberculosis. Tuberculosis was not produced in any of the guinea-pigs inoculated subsequently. Two pigs were fed for seven days with milk withdrawn during the period included between the second and the eighth days after the inoculation of the cow; they together received all the milk yielded with the exception of that used for the guinea- pigs. They were killed 97 days after the experiment began ; one was healthy, the other showed a calcareous tubercle in one submaxillary gland. In this case therefore persistence of tubercle bacilli in the milk sinuses was not demonstrated. The negative results do not however prove that the milk sinuses were free from bacilli since only a small proportion of the total amount of milk secreted was inoculated ; tubercle bacilli were no doubt present in the milk secreted on the second, third, and fourth days but in such small numbers that the samples inoculated, 5 to giz of the total yield, did not happen to include any. It is to paucity of bacilli and the inoculation of only a small proportion of the milk yielded that the irregularity of the results in those series in which some of the guinea-pigs became tuberculous is to be attributed rather than to intermittent discharge of bacilli into the milk. An inspection of the table on page 118 shows that frequently only one of the guinea-pigs inoculated with milk from the same quarter became tuberculous and that milk from a quarter which on one occasion produced tuberculosis was not on the next infective. The cow 153 days after her inoculation gave birth to a dead foetus (eight months old) ; during parturition a large amount of thin purulent fluid was discharged from the uterus ; microscopical examination showed this fluid to contain very numerous tubercle bacilli. During the next two or three days there was a slight discharge and on the sixth day some membranes and purulent fluid came away ; numerous tubercle bacilli were again demonstrated in the fluid. The foetus showed no sign of tuberculosis and emulsions made from its spleen and a portal gland did not produce tuberculosis in guinea-pigs. The cow was killed 182 days after inoculation ; there was no sign of tuberculosis in any of her organs or glands but a few tubercle bacilli were found in smear preparations of scrapings from the mucous membrane of one cornu of the uterus. 87 EXxprErimMent E. Goat 79 was inoculated subcutaneously on November 12, 1908, with 50-0 milli- grammes of a lupus culture (H 110. “J.B.” (a)) possessing all the properties of a culture of bovine tubercle bacilli, and was killed 9 days later. The post-mortem examination revealed a local lesion, early caseation of the pre- scapular gland and moderately numerous small tubercles in the lungs ; there was no sign of disease elsewhere. When inoculated the goat was giving only a small quantity of milk and after inoculation she soon became “ dry,” no milk being obtained from the right half of her udder after the fourth day or from the left after the sixth day. The milk withdrawn at intervals of twenty-four hours, during four days in the case of the right half of her udder and six days in that of the left, was inoculated into guinea-pigs. On the first and also on the second day four guinea-pigs were inoculated, two from each half of the udder, the doses varying from 10 to 4°5 cubic centimetres, but subsequently sufficient milk was yielded to inoculate only one guinea-pig from each half; the amount obtained from the left half of the udder on the fifth as well as on the sixth days was only 0°5 cubic centimetre. Only one of the eight guinea-pigs inoculated with milk from the left side became tuberculous. Milk withdrawn from the right side twenty-four hours after inoculation did not produce tuberculosis, while that withdrawn on the second, third, and fourth days did. Just before the goat was killed the milk sinuses of the udder were washed out with sterile salt solution, and a milky fluid was recovered from each side which was inoculated into two guinea-pigs. The two guinea-pigs inoculated with the fluid from the left half remained healthy while those from the right half became tuberculous. Microscopical examinations of the milk on several occasions and of the recovered saline solution were negative as regards presence of tubercle bacilli. In this experiment tubercle bacilli had passed out into the milk mainly from one half of the mamma and from that half which was giving the least amount of milk. EXPERIMENT F, Goat 55 was inoculated subcutaneously on November 14, 1907, with 50-0 milli- grammes of culture* obtained from the mesenteric glands of an old man (Virus H 90. “I.P.”), and was killed 103 days after. At the post-mortem examination there was a scar at the seat of inoculation with caseous nodules in the subcutaneous tissues around ; both prescapular glands were caseous the left more than the right ; one kidney contained a nodule ; the mesenteric glands were tuberculous ; and there were several miliary tubercles in the left quarter of the udder, and a caseating patch in the right. “Danna the first eight days guinea-pigs were inoculated with the milk at intervals of twenty-four hours ; during the remaining period guinea-pigs were inoculated on the fourteenth, twenty-first, and seventy-fifth days only. In each series some or all the guinea-pigs developed tuberculosis ; the milk from the left half of the udder was invariably infective, that from the right occasionally failed to infect. It is to be noted that the left quarter during life generally yielded less milk than the right, and the post-mortem examination showed in it a greater amount of tuberculosis. Cultures isolated from two of the tuberculous guinea-pigs, Nos. 2736 and 2791, grew luxuriantly on the test media, and one (Guinea-pig 2791 strain) had low virulence for the rabbit. Eight guinea-pigs were inoculated intraperitoneally each with 1:0 cc. of an emulsion of faeces prepared by diluting 1°0 cc. of thick filtered suspension to 20-0 ce. Four of the guinea-pigs died soon after inoculation ; the otier four were killed after several months and found to be free from tuberculosis. EXPERIMENT G. Goat 59 was inoculated subcutaneously on November 28, 1907, with 100°0 milli- * When used for inoculation this culture was believed to be an ordinary slightly virulent human tubercle bacillus; there'is now evidence to show that it contained a small proportion of virulent tubercle bacilli. 27676 Q 88 grammes of culture derived from the lung of aman (H 79. “J.N.”) ; the culture was slightly virulent for the calf and rabbit, and grew luxuriantly on culture media. The goat was killed in good health 106 days after the inoculation, and showed a scar at the seat of inoculation with caseo-calcareous tubercles in the subcutaneous tissues around, caseation of the prescapular gland, some doubtful tubercles in the lung, a few tubercles in the spleen, and tuberculosis of the mesenteric and ileo-colic glands ; the udder appeared normal. ; ; Guinea-pigs were inoculated with the milk every day except the third during the first six days. All these guinea-pigs, twenty in number, became tuberculous. After the sixth day no further observations were made on the infectivity of the milk. At the time of inoculation the goat was giving a very small quantity of milk, and after the sixth day the amount rapidly diminished, and in a short time the secretion ceased altogether. A culture isolated from one of the tuberculous guinea-pigs, No. 2797, grew like the culture inoculated into the goat. ‘ EXPERIMENT H. Goat 69 was inoculated subcutaneously on June 3, 1908, with 10:0 milligrammes of culture obtained from cerebro-spinal fluid (H 98. “B.R.”). This culture grew luxuriantly on artificial media and had low virulence for the rabbit. ; The milk of the goat was subsequently inoculated into a series of four guinea-pigs on the first, second, third, fifth, and seventh days during the first week, and then once a week for three weeks ; a final series was inoculated on the 44th day after the inoculation of the goat. Altogether 36 guinea-pigs were used. All the guinea-pigs inoculated with the milk withdrawn during the first week remained healthy ; all those inoculated on the 14th day (with one exception) and subsequently, developed tuberculosis. In this case, therefore, the bacilli did not appear in the milk so quickly as in the other like experiments, probably because the dose was so much smaller. The general health of the goat was good throughout the experiment. The amount of milk secreted in a day was never very large; during the first month the daily yield varied from 100-U to 300-0 ce. of which usually only 40-0 cc. were inoculated into guinea-pigs ; after the first month the secretion of milk gradually diminished and towards the end of the second month ceased altogether. On July 15, 42 days after inoculation, the mamma of the goat was found to be enlarged ; on palpation the enlargement affected mainly the right half which was uniformly indurated and knotty ; the left half was not obviously enlarged but contained some hard nodules just above the teat ; the milk at this time was much diminished in amount but showed no alteration in quality ; later on the left halt became more uniformly indurated ; during the latter part of the experiment the mamma remained indurated but decreased a little in size. Microscopical examinations of the milk were made at intervals for the presence of tubercle bacilli. One (?) tubercle bacillus was found in the centrifuged deposit of milk withdrawn on the second day from the right half of the mamma ; in centrifuged deposit of milk taken from the same side on the 14th day five tubercle bacilli were found ; other examinations were negative. On the 42nd day a loopful or two of the milk from each haif of the mamma was sown on to egg tubes ; pure cultures of the tubercle bacillus were obtained in each case, the colonies being more numerous on the tube sown with milk from the right quarter than on that sown with milk from the left. The cultures grew on glycerin media like the original, The goat was killed 114 days after inoculation, and showed at the post-mortem examination an encapsuled caseous mass at the seat of inoculation and enlargement and induration of the mamma which was beset with miliary caseous tubercles, but no sign of tuberculosis elsewhere. No tubercle bacilli were found in the milky fluid in the sinuses nor in some of the distended alveoli (milk cysts) ; a few tubercle bacilli were found in a smear made from the miliary tubercles. Cultures were sown with some of the fluid withdrawn with a pipette from the milk sinuses after death and also with fluid from two milk cysts. All the tubes sown 89 with milk from the sinuses grew numerous colonies of tubercle bacilli in pure culture ; numerous colonies were also obtained from one of the milk cysts on the left side ; the tubes sown with milk from a cyst on the right side remained sterile. The different strains grew on glycerin media like the culture inoculated. Experiment I, Goat 61 was inoculated subcutaneously on July 27, 1908, with 1:0 milligramme of one of the easy growing slightly virulent lupus cultures (H 109. “M.W.”). The milk of the goat was inoculated into guinea-pigs on thirteen separate occasions, at frequent intervals during the first month and at longer ones subsequently, the last series being inoculated 197 days after the inoculation of the goat. Altogether fifty guinea-pigs were inoculated, the majority receiving 10 cubic centimetres each. Not one of the guinea-pigs developed tuberculosis. The goat when inoculated was suckling a kid ten weeks old. During the first seventeen days after her inoculation the kid was not allowed to suck, but from the 17th to the 171st days the kid took all the milk with the exception of that which had been withdrawn for the purpose of inoculating guinea-pigs. On the latter date, that is when the feeding experiment had lasted 154 days, the kid was killed; there was no sign of tuberculosis in any of its organs or glands. The mother goat was killed 197 days after inoculation and showed slight tuberculosis at the seat of inoculation only. The experiment with this goat was negative probably because on account of the smallness of the dose subcutaneously inoculated there had been little or no dis- semination. EXPERIMENT J. Goat 81 was inoculated intravenously on December 18, 1908, with 2°0 milli- grammes of a slightly virulent human tubercle bacillus (H 115. “ N.G.”). The milk of the goat was inoculated into five series of guinea-pigs at intervals of twenty-four hours during the first three days after inoculation, then on the 8th and 15th days, four guinea-pigs each receiving 10 cc. being inoculated on each occasion. All the guinea-pigs in the first two series became tuberculous, while in the third only one developed tuberculosis ; in the fourth and fifth series three of the four guinea-pigs became tuberculous, the fourth remaining healthy. A few days after the last series of guinea-pigs was inoculated, the secretion of milk ceased and the udder became dry. At this time the goat was obviously © ill; she gradually became worse and died on January 28, 1909, 41 days after inoculation. The post-mortem examination showed tuberculous pneumonia, enlarge- ment and congestion of the thoracic glands, but no sign of disease elsewhere. Before the sectio cadaveris was begun the milk sinuses of each half of the udder were washed out with salt solution ; 4:0 cc. of milky fluid was recovered from the right and 5:0 cc. from the left half, and each of these quantities was divided and inoculated into two guinea-pigs; all four guinea-pigs developed general tuberculosis. In this case therefore tubercle bacilli which had been injected intravenously were demonstrated in the milk secreted during the twenty-four hours following inoculation and on every subsequent occasion up to the 15th day. At the end of the third week the secretion of milk had ceased, but tubercle bacilli were shown to be present in the small quantity of milk which existed in the milk sinuses when the animal died on the 41st day. EXPERIMENT K. Goat 75 was fed, on a single occasion, September 22, 1908, through a stomach tube with 74:0 milligrammes of culture of a slightly virulent human tubercle bacillus (H 76. “G.M.”). Guinea-pigs were inoculated with her milk at frequent intervals during the first three weeks and at longer ones subsequently, the last series being inoculated 258 days after the goat was fed. Altogether eleven series comprising 42 guinea-pigs were used. None of the guinea-pigs became tuberculous. Eight months after the experiment began the gcat gave birth to two kids which she suckled for nearly two and a-half months ; when killed at the end of this period they were found to be perfectly healthy. 27676 Q2 90 The mother goat was killed 328 days after the experiment began and showed caleareous tubercles in some of the neck and mesenteric and ileocolic glands and in one portal gland but no sign of tuberculosis elsewhere. SUMMARY. With the object: of ascertaining whether tubercle bacilli are excreted in the milk from a normal udder, four cows and seven goats which had not reacted to the tuberculin test were inoculated with culture of such bacilli and their milk tested subsequently on guinea-pigs. Three cows and five goats were inoculated subcu- taneously (one of the cows being subsequently inoculated intravenously), one cow and one goat intravenously, and one goat was fed. The first cow, Cow 300, was inoculated subcutaneously with 100-0 mg. of tubercle bacilli of bovine origin ; the milk withdrawn by hand on the seventh, sixteenth, twenty-first, twenty-eighth, and thirtieth days after the inoculation of the cow pro- duced tuberculosis in guinea-pigs. ; The second cow, Cow 77, was inoculated subcutaneously with 100°0 mg. of a culture of human origin which was slightly virulent for the calf and rabbit ; the milk withdrawn by hand twenty-four hours after the inoculation of the cow produced tuberculosis in four of the eight guinea-pigs inoculated, and tuberculosis was produced with two exceptions on every subsequent occasion, including the last, on which the milk was tested ; the last series of guinea-pigs was inoculated after the experiment had lasted 155 days. The udders of these two cows were normal to the naked eye at the post-mortem examination, and showed no tuberculous lesions or tubercle bacilli on microscopical examination. The third cow, Cow 61, was also inoculated subcutaneously with 100-0 mg. of a culture of human tubercle bacilli ; the samples of milk were collected by catheterisa- tion ; guinea-pigs were inoculated at intervals of twenty-four ‘hours during the first week and at gradually lengthening intervals subsequently, the last series 84 days after the cow was inoculated. Altogether 76 guinea-pigs were inoculated, and out of these one only developed tuberculosis. Ninety days after the subcutaneous inoculation this cow was inoculated intra venously with 150°0 mg. of a slightly virulent culture (Virus H 83. “G.C.”). During the first fifteen days of the renewed experiment nine series of guinea-pigs were inoculated at intervals of one or two days; in the first two series (1st and 2nd day’s milk) all the guinea-pigs, fourteen in number, developed tuberculosis ; and the milk continued to be infective up to the forty-fourth day when the routine poe of guinea-pigs was discontinued on account of the cow’s udder becoming ‘dry. About five and a half months after the intravenous inoculation of the cow some pus was obtained from the two fore quarters, and this on microscopical examination was found to contain very numerous tubercle bacilli ; subsequently tubercle bacilli were demonstrated from the other quarters also, Two and a half months later it was noticed that three of the quarters had become enlarged and indurated and that a large amount of purulent substance which microscopically contained numerous tubercle bacilli had collected in the sinuses. The udder decreased in size and the discharge of pus rapidly diminished, a few drops only being obtained at intervals during the remaining period of the experi- ment. Tubercle bacilli were demonstrated on every occasion. Cultures were obtained direct from the pus on four different occasions, the last one day before the cow was killed ; these cultures were identical in their cultural characters and in their virulence for the rabbit with the culture intravenously inoculated. The cow was killed 529 days after the intravenous inoculation, and 619 days after the subcutaneous inoculation ; she showed slight generalised tuberculosis and tuberculosis of the udder. The faeces of this cow were inoculated into 16 guinea-pigs, six after her sub- cutaneous inoculation and ten after her intravenous; 7 died shortly after inoculation the others survived and when killed one alone was found to be tuberculous : this guinea-pig had been inoculated 10 days after the intravenous inoculation of the cow. The fourth cow, Cow 565, was inoculated intravenously with 10-0 milligrammes of a culture of human tubercle bacilli (H 104.“ E.R.”). Her milk was tested on guinea- 91 pigs eight times during the first fortnight and 11 times during the remaining period of the experiment (168 days). The milk collected 24 hours, 5, 6,7, and 14 days after inoculation produced tuberculosis in from one to four of the six guinea-pigs inoculated on each occasion ; milk collected subsequent to the 14th day did not produce tuberculosis in any of the guinea-pigs inoculated. 153 days after inocula- tion the cow gave birth to a dead 8 months old calf; the fluid which came away with it was purulent and contained numerous tubercle bacilli; the spleen and a portal gland of the foetus did not produce tuberculosis in guinea-pigs. The cow was killed after 182 days and showed no tubercvlous lesions. One of the goats, Goat 79, was inoculated subcutaneously with 50:0 milligrammes of a bovine culture from a case of lupus and was killed 9 days later. The milk of the goat was inoculated into guinea-pigs at frequent intervals ; that withdrawn from the right half of the udder on the 2nd, 3rd, 4th, and 9th days was infective for guinea-pigs, that from the left side produced tuberculosis in only one of the guinea- pigs inoculated. Another goat, No. 55, was subcutaneously inoculated with 50-0 mg. of a eugonic culture which was shown subsequently to contain a small proportion of virulent tubercle bacilli. Milk collected at intervals of 24 hours during the first eight days, and on the 14th, 21st, and 75th day, produced tuberculosis in guinea-pigs. At the post- mortem examination of the goat several miliary tubercles were found in the left half of the udder and a caseating patch in the right. Two cultures isolated from the milk of this goat were eugonic and one has been proved to have low virulence for the rabbit. . The third goat, No. 59, was inoculated subcutaneously with 100-0 mg. of a slightly virulent culture. All the guinea-pigs, twenty in number, inoculated with her milk during the first six days became tuberculous. At the post-mortem examination the udder of the goat was normal to the naked eye. The fourth goat, No. 69, was inoculated subcutaneously with 10-0 mg. of a human tubercle bacillus. Her milk was not infective for the guinea-pigs inoculated durin the first week but produced tuberculosis in every guinea-pig inoculated on the 14th day and subsequently. Tuberculosis of her mamma developed on the 42nd day after inoculation. Cultures isolated at various times from her milk resembled the culture inoculated. . _ The 5th goat, Goat 81, was inoculated intravenously with 2°0 milligrammes of a culture of human tubercle bacilli ; she died in 41 days of tuberculosis of the lungs and thoracic glands, no disease being found elsewhere in the body. Milk with- drawn from her on the Ist, 2nd, 8rd, 8th and 15th days, and the fluid recovered from the mammary sinuses on the 4tst day after injecting saline solution produced tuberculosis in guinea-pigs.. A sixth goat, Goat 61, was inoculated subcutaneously with 1:0 milligramme, and a seventh (No. 75) was fed with 74:0 milligrammes, of culture, the culture in each case being slightly virulent and easy-growing. The milk of each of these goats was inoculated at intervals into a series of guinea-pigs, 13 series in one case and 11] in the other being used. Not one of the guinea-pigs became tuberculous. These are the only two out of eleven experiments in which the results were entirely negative. The other nine experiments have shown clearly that the normal mammary gland will permit the escape of tubercle bacilli whether of bovine or of human type from the blood-stream into the milk. | There arises, therefore, strong presumption that the milk of naturally tuberculous cows without udder tuberculosis will contain tubercle bacilli whenever in the course of the disease tubercle bacilli circulate in the blood stream. A. 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THE EXCRETION OF TUBERCLE BACILLI INTO THE UNDEVELOPED UDDER. INTRODUCTION. Tur foregoing experiments demonstrate that when lactating animals are inoculated subcutaneously or intravenously with tubercle bacilli of relatively slight virulence such bacilli appear in their milk within 24 hours of their inoculation and may continue to be eliminated therein for long periods subsequently. This immediate discharge into and persistence in the milk sinuses of tubercle bacilli which have been introduced into the tissues of an animal renders the vaccination of milch cows with living human tubercle bacilli inadvisable ; cows treated in this way might prove even more dangerous to human health than naturally tuberculous cows since, as the above experiments have shown, the bacilli may exist in the milk sinuses for a long time without altering the quality of the milk or affecting the udder or the general health of the animal. The vaccination of the bovine animal with living human tubercle bacilliis practised however only on calves and not on animals in use for dairy purposes. It becomes important therefore to ascertain whether the vaccination of calves intended eventually to supply milk for human consumption is free from the danger which would attend the vaccination of milch cows, whether in fact tubercle bacilli can gain access to the sinuses of the immature mamma, live and multiply therein, and be present in the milk when the animal begins to lactate. With a view to throwing some light on these questions I have examined the contents of the mammary sinuses of several heifer calves which had been inoculated to test the virulence of various sorts of cultures, and the milk of a goat which was inoculated when pregnant has also been tested. Observations have been made on eleven heifer calves, all of which had been inoculated subcutaneously with culture. Two of these cultures had the properties of the human tubercle bacillus, one those of the bovine tubercle bacillus, the others resembled bovine tubercle bacilli in their manner of growth, but exhibited lower virulence for the calf and rabbit. All the eleven’ calves were in good condition when killed and had increased normally in weight ; the disease produced ranged from tuberculosis limited practically to the site of inoculation and the nearest glands to generalised tuberculosis not severe and only apparently progressive in one case. The material investigated was obtained after the death of the animal in the following way: the skin and teats were thoroughly cleansed and a few cubic centimetres of sterilised salt solution (up to 10°0 cc.) were injected into each of the mammary sinuses, and after massaging the quarter as much of the fluid as possible was withdrawn by means of the pipette used for the injection; the fluid recovered was usually only slightly milky or turbid, and was always less in amount than that injected. The following summaries give in the case of each calf the details of the experimental inoculation, an abstract of the post-mortem notes and the results of the investigation of the fluids from the mammary sinuses. I include an account of a case, a heifer (No. 80) inoculated with bovine tubercle bacilli, the details of which have already been reported in the Appendix to the Second Interim Report, Vol. L., pp. 198, 199. The ages of the calves when killed varied from six to ten months, and the duration 95 of the experiments from 91 to 127 days; the heifer inoculated with bovine tubercle bacilli was 18 months old, and when killed was five months pregnant. Case 1. A heifer-calf (Calf 1479) inoculated subcutaneously when six months old with 100-0 milligrammes of culture from Virus H 79. “J.N.” (Calf 1325, 3rd passage). The cultures of this virus grow luxuriantly on artificial media and have low virulence for the calf and rabbit. The calf was killed 110 days after inoculation and showed a caseous abscess at the seat of inoculation, slight tuberculosis of the nearest gland, and calcareous tubercles in the suprarenal bodies, the thoracic glands, the mesenteric, ileo-colic and a few other abdominal lymphatic glands. The mamma was small and free from visible tuberculous lesions ; the fluid recovered from the sinuses after injecting saline was small in amount and opalescent. No tubercle bacilli were demonstrated in smear preparations of the fluid, but four guinea-pigs inoculated two with the mixed fluids from the fore quarters and two with the fluids from the hind quarters. became tuberculous. Cultures of the tubercle bacillus were obtained direct from the fluids, numerous colonies. developing on tubes sown with a loopful or two; the cultures exhibited the cultural characters of the original cultures of the virus. Case 2. A heifer-calf (Calf 1601) inoculated sudcutaneously when 4 months old with 68-0 milligrammes of culture from the original material of Virus H 143. “ L.L.” The calf was killed 91 days after inoculation and showed a caseous abscess at the seat of inoculation, tuberculosis of the nearest glands, a few minute tubercles in the lungs, some caseous foci in the ileo-colic glands and about fifteen miliary caseous tubercles in the suprarenal bodies. The mamma was free from macroscopic tuberculous lesions. Slightly turbid fluid was recovered from each of the sinuses after injecting saline solution. One or two tubercle bacilli were seen in smear preparations of fluid from each of three of the quarters, none in that from the fourth ; the fluid from each of the four quarters produced numerous colonies on culture tubes, and tuberculosis in guinea-pigs. Case 8. A heifer-calf (Calf 1461) inoculated subcutaneously when six months old with 50:0 milligrammes. of culture from Virus H 53. “D.H.” (a) (Calf 1367, 3rd passage). The calf was killed 111 days after inoculation and showed a caseous local lesion, tuberculosis of the nearest glands, a few scattered calcareous foci in the thoracic glands, and three softened caseous tubercles in a mammary lymphatic gland. The mamma was well-developed and no tuberculous lesions were found in it, There was a small quantity of opalescent fluid in each of the sinuses which in one case showed microscopically moderately numerous tubercle bacilli. A culture of tubercle bacilli was obtained direct from the fluid, and one guinea-pig inoculated with a few drops developed tuberculosis. Case 4. A heifer-calf (Calf 1535) inoculated subcutaneously when about three months old with 50-0 milligrammes of culture from the original material of Virus H 53. “ D.H.” (6). The calf was killed when in good health 103 days after inoculation and showed a local abscess, tuberculosis of the adjacent gland, scattered calcareous tubercles in the lungs and the spleen, tuberculous foci and ulcers in the intestines and caseous or calcareous tubercles in nearly every lymphatic gland. No tubercles were found in the tissue of the mammary gland. In the sinuses and ducts of the right fore quarter there was a small quantity of purulent fluid which microscopically showed tubercle bacilli: from the other quarters slightly milky fluid was obtained after injecting saline ; no tubercle bacilli were found in smear preparations of this fluid. A guinea-pig inoculated with the pus from the right fore quarter died of severe general tuberculosis ; those inoculated with the milky fluid recovered from the left fore and right. hind quarters remained healthy. A pure culture of the tubercle bacillus was obtained from the pus; tubes sown with the milky fluids remained sterile. The culture grew on the differential media like the culture inoculated into the calf. Case &. A heifer-calf (Calf 1571) inoculated subcutaneously when 3} months old with 50-0 milligrammes. of culture from Virus H 53. “ D.H.” (6) (Calf 1561, 1st passage). The calf was killed 111 days after inoculation and showed a small caseous abscess at the seat of inoculation, tuberculosis of the nearest glands, three tubercles'in the spleen, a caseous nodule in a suprarenal body and one or more minute foci in each of seven lymphatic glands. The mamma was normal to the naked eye; the sinuses were washed out with saline and a small quantity of very slightly turbid fluid recovered from each. Microscopical examina- tions and guinea-pig inoculations of these fluids gave negative results, and culture tubes sown with them remained sterile. Case 6. A heifer-calf (Calf 1505) inoculated subcutaneously when five months old with 100-0 milli grammes of culture from Virus H 107. “ H.H.” (Calf 1453). The calf was in good health when killed 101 days after inoculation. The post-mortem examination showed a small local lesion, caseo-calcareous changes in the nearest glands and some ulcerated fuberculous nodules in the intestines; there was no sign of tuberculosis. elsewhere, and the mamma was normal to the naked eye. The mammary sinuses were washed out with saline, and the slightly turbid fluid recovered was used to sow cultures and inoculate guinea-pigs. The results were negative. No tubercle bacilli were found in smear preparations of the fluid. Case 7. A heifer-calf (Calf 1549) inoculated subcutaneously when 23 months old with 50-0 milli- grammes of culture from Virus H 107. “ H.H.” (Calf 1497). The calf was killed in good health 97 days after inoculation, and was found to have slight general tuberculosis, apparently progressive in the thoracic glands. The mamma was well- developed, and was free from macroscopic tuberculosis ; many of the lactiferous ducts contained plugs of purulent substance. . : The mammary sinuses were washed out with saline, the recovered fluids depositing pus in the form of casts. Tubercle bacilli were found in smear preparations, and numerous colonies grew on culture media sown from the mixed fluids from the fore quarters and the mixed fluids from the hind quarters. No guinea-pigs were inoculated. Case 8. A heifer-calf (Calf 1577) inoculated subcutaneously when 33 months old with 50-0 milligrammes of culture from Virus H 107. “H.H.” (Calf 1551, 2nd Passage). The calf was killed 103 days after inoculation and showed a caseo-purulent abscess at the seat of inoculation, caseation of the adjacent glands, one tubercle in each suprarenal body and one or more minute foci in the thoracic and three abdominal glands. 27676 R 96 The mamma was normal to the naked-eye ; the sinuses were washed out with saline ; the fluid recovered was in each case very ‘slightly turbid. Microscopical examinations and guinea- pig inoculations of these fluids gave negative results, and culture tubes sown with them remained sterile. Case 9. A heifer-calf (Calf 1583) inoculated subcutaneously when 11 weeks old with 50-0 milligrammes of culture from Virus H 107.*H.H.” (Calf 1549, 1st Passage.) The calf was killed 127 days after inoculation and showed a small abscess at the seat of inoculation, caseous and calcareous nodules in the adjacent prescapular gland, a minute focus in one suprarenal body, and one or more foci, mostly minute, in the thoracic and two or three abdominal glands. ' The udder was small and showed very little glandular tissue ; the sinuses were washed out with saline. Microscopical examination of the fluids recovered from the sinuses showed no tubercle bacilli, guinea-pigs inoculated with the fluids remained healthy, and culture tubes sown with them sterile. Case 10. A heifer-calf (Calf 1533) inoculated subcutaneously when four months old with 50-0 milli- grammes of culture from Virus H 108. “ H.R.” (Calf 1417). The calf was killed in good health 98 days after the inoculation and was found to have widespread generalised tuberculosis ; all the lymphatic glands were affected some rather severely, but in the chief organs the lesions were not very numerous, and all had retro- gressive characters. The mamma was free from macroscopic tuberculous lesions. A small quantity of slightly turbid fluid was recovered from each of the sinuses after injecting saline. The mixed fluids from the two fore quarters and the mixed fluids from the two hind quarters were each used to inoculate a guinea-pig and to sow cultures. The guinea-pig inoculated with the fluid from the fore quarters became tuberculous, the other remained healthy. Two egg tubes sown with the fluid from the fore quarters produced altogether eight colonies of tubercle bacilli ; those sown from the hind quarters remained sterile. Case 11. A heifer (Heifer 80) inoculated subcutaneously when about 18 months old with an emulsion made from a bovine tuberculous gland (Virus B. V) the dose containing 44 million tubercle bacilli. The fuil details of this case are given in the Appendix to the 2nd Interim Report, Vol. I., p. 198. The heifer was killed 62 days after inoculation and showed slight generalised tuberculosis ; there was a five months old foetus in the uterus which was not tuberculous; the mammary tissues showed no naked-eye evidence of tuberculosis and the milk sinuses contained a small quantity of thick yellowish fluid ; no tubercle bacilli were demonstrated in a smear preparation made from the fluid, but a guinea-pig inoculated with it died of general tuberculosis. ‘Case 12, A heifer-calf (Calf 1569) inoculated subcutaneously when 3 months old with 50:0 milligrammes of culture from the original material of Virus H 127. “ R.R.” (a). The calf was killed when in good health 105 days after inoculation and showed slight general tuberculosis apparently retrogressive. The mamma appeared normal (the supramammary lymphatic glands were normal). Thin grey purulent fluid was expressed from the teats of three of the quarters ; these fluids showed tubercle bacilli on microscopical examination, and produced numerous colonies on culture tubes sown with them. From the remaining quarter (L.H.) no pus could be expressed ; the sinus was washed out with salt solution ; microscopical examination of the recovered fluid showed no tubercle bacilli and culture tubes sown with it remained sterile. Case 13. Goat 77,a young pregnant primipara, was inoculated subcutaneously with 25-0 milligrammes _ of a human Group II culture obtained from sputum (Virus H 118, “ F.C.”), This experiment was made for the purpose of ascertaining whether tubercle bacilli can pass from the blood into the milk sinuses of an immature mamma and remain alive therein until the animal begins to lactate. The experiment had a negative result. The goat gave birth to two dead full-time foetuses 52 days after inoculation. The udder was well-developed and yielded a good supply of milk. Guinea-pigs were inoculated with milk withdrawn 8 hours, 1, 2, 8, 14, 28, and 59 days after the birth of the kids, and all remained free from tuberculosis. Microscopical examinations and culture experiments were also negative. SUMMARY. In seven out of eleven heifers tubercle bacilli of various types which had been inoculated in large dose into the subcutaneous tissues had found their way into the milk sinuses of the undeveloped mamma, and in four cases at least (Calves 1461 1535, 1569, and 1601) were present in such numbers as to suggest that since thei arrival there they had undergone multiplication. s These observations clearly establish the important fact that tubercle bacilli which have been introduced into the subcutaneous tissues of a calf can find their way into the ducts and milk sinuses of the immature mamma, and suggest that the ie live and multiply therein for considerable periods. . oe Whether the bacilli in these cases would have remained alive until the eriod of lactation is a matter of conjecture, but judging from the lenoth of time bacilli ae ee e ee of living in the milk sinuses of cows there is reasonable probability that the first milk of so i 7 contained ine Ghent bacilli. pier cee ean Though there is not complete evidence that bacilli excreted into the milk sinuses of the immature udder do remain alive therein until the period of lactation becins the facts established point strongly to the desirability of testing, before Dannie its 5 use as human food, the milk of all cows which have been vacc} i . . . . : "ACC 5 living human tubercle bacilli. . BEDS cane mu reqyrenb pury yoy be eas: oe Geis Jayunb pury FYSTY (HH »s ee a PANOON dL ‘ON Jayzenb a10F 1JO'T *({VULIOU BUUULRTH) SUOTS9T ‘sfkUp COT | ‘LOT H) eutaoq *(syjuout 4) Iaqyrenb o10F FYSTY | payVUrulesstp Moy V YA |, [BOOT | 109Je polly | poyenusjy “SUI ().0C | ZLZGT 10710 8 "S9TTO]09 ShuLetunu A TayBLIpoO — ‘EL AO} VW | sdozVnb pury omy *(jew10U (HH » *SaTUO]OO BUUUUVUL) SpUL[S O1OVLOY} UL OATS ‘sfep 16 | "LOT EH) outa0g “(SOOM QT) suodauluu ATa}BIIPOTFL —_— ‘EY, Mop VW | Sdoyrenh arog omy, | -seasoad Apyuesedde *y, 9 YSIS | 1eyye pel[Ly | peyenueiy ‘SUL (JC | PST IeFOTT 4 C(CH'H » ‘ATLMaIS PIULBULAL SIqu, AL]VSAN ‘TL ON siaqyrenb pury OA, *([VULIOU VUIUIVUL) STO1SE] ‘skep TOL | "LOT H) eutaog ‘(syjuoUt G) ‘OTL194S PoulVUlel saq Uy, OATPRBON ‘TT ON ——! aLOJ OAT, | POYCUITMIOSSIP MOT V TIA “Y TRIO'T | 109Fe paT[Iy | veyenueyy | ‘Sul Y.OOLT | GOGT JeJtOH 9 qey1enb pury eT S pe Joqenb pury wy sry @) No. of Mode Quarter | Dose | Killed : Date. re - Guinea- of of ‘cn ao ae of Result. i jecti r. ilk. ied. ; 24 hours.| PIs Injection.| Udder i Nov. 28, 1906 2021 Intrap. L.F. 5 cc. K. 36 days Healthy, [Before inoculation]. 2022 3 L.H. 5 3 Healthy. 2023 3 RF. 3 : ” Healthy. 2024 3 R.H. # si i Healthy, 1 Nov. 29, 1906 2350 ec. 2027 Intrap. LF. 5 ce. K. 46 days No tuberculosis. [24 hours after inoculation ]. 2028 Subcut. Fi $i Dz. 17 days Carcass eaten by other guinea-pigs. 2031 Intrap. L.H. i K. 46 days No tuberculosis. 2032 Subcut. us Fr 5 e No tuberculosis. 2025 Intrap. RF * 5 “3 (2) tuberculosis of omen-- tum. 2026 Subcut. sj 5 5 sf Slight tuberculosis. 2029 Intrap. R.H. i +6 $3 Slight tuberculosis. 2030 Subcut. 9% 3 5 ze Slight tuberculosis. Nov. 30, 1906 1450 ce. 2049 Intrap. LF. 5 ec. K. 45 days Slight general tubercu- [2 days after losis. . inoculation ]. 2050 Subcut. 33 5 i <5 No tuberculosis, 2051 Intrap. L.H. 3 ‘i 9 Slight tuberculosis. 2052 Subcut. rr Sy 55 +5 Slight general tubercu- losis. 2047 Intrap. RF +5 * 6 Slight tuberculosis. 2048 Subcut, 3 9 7 %y Slight tuberculosis. 2045 Intrap. R.H. 4 es eA Slight tuberculosis. 2046 <5 “ ‘5 5 * General tuberculosis. Dec. 1, 1906 1600 ce. 2057 Intrap. LF, 5 ce. K. 46 days Slight general tubercu-. [3 days after losis. inoculation ]. 2058 Subcut. 5, ; es 3 Slight tuberculosis. 2059 Intrap. L.H. 3 “ Slight general tubercu-- losis. 2060 Subcut. 4) i “ No tuberculosis. 2053 Intrap. RF. 3 5 45 Slight tuberculosis, 2054 Subcut. Pe 5 “ ‘3 Slight tuberculosis. 2055 Intrap. R.H, iy 4 45 Slight tuberculosis. 2056 Subcut. y + 5 * Slight tuberculosis. Dec. 2, 1906 2150 ce. 2065 Intrap. LF 5 ee. K. 46 days Very slight tuberculosis. [4 days after inoculation ]. 2066 Subcut. 5 3 6 4 Slight general tubercu- losis, 2067 Intrap. L.H. ‘5 " 5 Slight tuberculosis, 2068 | Sukcut. » " ” 7 No tuberculosis. 2069 Intrap. RF. 5 ai ia One (?) tubercle in és omentum, 2070 | Subeut. ” " » si No tuberculosis, 2071 Intrap. R.H. . ” ” ee general tubercu- e osis, 2072 Subcut. 3 si a No tuberculosis, 103 GUINEA-PIGS INOCULATED WITH THE MILK or Cow 77—continued. Total : No. of Mode Quarter | Dose | Killed : Date. Lie - Guinea- of of of or eee ee . Result. 24 hours,| Pig: | Injection.| Udder. | Milk. | Died. re Dec. 3, 1906 1900 cc, | 2073 Intrap. LF, 5 ee. K. 53 days No tuberculosis. [5 days after inoculation]. 2074 Subcut. i ‘5 ‘j 7. (2) Slight tuberculosis of inguinal and left iliac glands. 2075 Intrap. L.H. 5 3 * No tuberculosis. 2076 Subcut. a i 3 4 No tuberculosis. 2077 Intrap. RF. ‘3 is " No tuberculosis. 2078 Subcut. a ' 3 +5 No tuberculosis. 2079 Intrap. R.H. 43 is ‘3 No tuberculosis. 2080 Subcut. iy i i 4 No tuberculosis. Dec. 4, 1906 1850 ce. 2081 Intrap. LF... | 5 ce. K. 56 days General tuberculosis. [6 days after inoculation]. 2082 Subcut. . 4 a y No tuberculosis. 2083 Intrap. L.H. i a i No tuberculosis. 2084 Subcut. “s ip +3 3 General tuberculosis. 2085 Intrap. RF or 5 sy General tuberculosis. 2086 Subcut. ‘i 7 D. 13 days Locallesion only, (?) cause of death. 2087 Intrap. R.H. 54 56 days General tuberculosis, 2088 Subcut. sj 9 ee $5 No tuberculosis. Dec. 5, 1906 1800 ec. 2089 Intrap. LF. 5 ec. K. 57 days Slight general tuber- [7 days after culosis. inoculation], 2090 Subcut. “3 $i % 5 No tuberculosis, 2091 Intrap. L.H. a D. 9 days Pseudo-tuberculosis. 2092 Subcut. r Fs K. 57 days No tuberculosis, 2093 Intrap. R.F 5 ‘i 4 Slight general tuber-. culosis. 2094 Subcut. as ‘i a 4 Slight general tuber- culosis, 2095 Intrap. R.H. ” 3 3 General tuberculosis. moderately severe. 2096 Subcut. a e i‘ % Slight general tuber- culosis. Dec. 12, 1906 _ 2109 Intrap. LF. 5 ce. K. 55 days General tuberculosis. [14 days after inoculation]. 2110 Subcut. 3 9 53 s No tuberculosis. 2111 Intrap. L.H. #5 D. 1 day No sign of disease, (2?) cause of deaih. 2112 Subcut. 43 $3 K. 55 days No tuberculosis. 2113 Intrap. RF. ” ” 3 General tuberculosis. 2114 Subcut. 3 sh Pe a5 No tuberculosis. 2115 Intrap. R.H. 4 Ls % General tuberculosis. 2116 Subcut. 3 55 ‘5 5 No tuberculosis. Dec. 19, 1906 2600 ce, 2128 Intrap. LF. 5 ec. D. 16 days Pseudo-tuberculosis. (21 days after : inoculation]. 2129 Subcut. 5 $3 K. 54 days No tuberculosis. 2130 Tntrap. L.H. $3 3 +3 No tuberculosis. 213] Subcut. : 54 3) No tuberculosis, 2132 Intrap. R.F ‘5 ‘3 3 No tuberculosis. 2133 Subcut. is Pe i + No tuberculosis. 2134 Intrap. R.H. 5 43 +3 Slight tuberculosis. 2135 Subcut. as 7 ” 55 No tuberculosis, 27676 104 GUINEA-PIGS INOCULATED WITH THE MILK oF Cow 77—continued. Total : : No. of Mode Quarter | Dose | Killed ‘ Date. ea i QGuinea- of of of eA mae of Result. 24 hours.| Dis Injection.) Udder. | Milk. ied. Dec. 26, 1906 _— 2141 Intrap. LF. 5 ec. K. 58 days No tuberculosis, (28 days after 5 inoculation]. 2142 | Subeut. is ” ” ” No tuberculosis. 2143 Intrap. LH. o 34 No tuberculosis. 2144 Subcut. 3 a 9 53 No tuberculosis. 2145 Intrap. R.F. i 3 No tuberculosis. 2146 Subcut. Pr #3 i 5 No tuberculosis. 2147 Intrap. R.H. % a ss No tuberculosis. 2148 Subcut. is 43 # 33 General tuberculosis (spontaneous). Jan. 9, 1907 1457 ce. 2157 Intrap. LF. 5 cc K 57 days No tuberculosis, [42 days after : inoculation }. 2158 Subeut. ‘ a “5 sf No tuberculosis. 2159 Intrap. L.H. ‘3 re 4} No tuberculosis. 2160 Subcut. ; é ‘3 si No tuberculosis. 2161 Intrap. R.F. j ‘6 i No tuberculosis. | 2162 Subcus. ee | ry 45 4 Slight general tuber- culosis. 2163 Intrap. R.H. i 3 a; No tuberculosis. 2164 Subcut. 4 “ 45 re No tuberculosis. Jan. 23, 1907 _— 2195 Intrap. L.F. 5 cc. K. 61 days No tuberculosis, [56 days after inoculation]. 2196 Subcut. i i %s 4 No tuberculosis, 2197 Intrap. L.H. *5 53 : No tuberculosis, 2198 Subcut. 8 “ i a No tuberculosis, 2199 Intrap. R.F. 3 a % Slight general tuber- culosis. 2200 Subcut. ¥5 5 *s 55 No tuberculosis. 2201 Intrap. R.H. 3 D. 41 days | General tuberculosis. 2202 Subcut. 3 i K. 61 days No tuberculosis. Feb. 1, 1907 1800 cc. 2216 Intrap. LF. 5 ce. K. 76 days General tuberculosis, [65 days after inoculation ]. 2217 Subcut. 53 is a General tuberculosis, 2218 Intrap. L.H. i ss 3 No tuberculosis. 2219 Subcut. af “3 3 No tuberculosis. 2220 Intrap. R.F, 6 Ps General tuberculosis. 2221 | Subcut. s ” ” 4 No tuberculosis, 2222 Intrap. R.H. ” "9 ” No tuberculosis. 2223 Subcut. 4 ay s ‘ No tuberculosis. Feb. 14, 1907 = 2251 Intrap. LF. 5 ce. K. 69 days No tuberculosis. {78 days after inoculation ]. 2252 | Subcut. ‘5 te 55 be No tuberculosis, 2 2253 Intrap. LH, ” ” i No tuberculosis. 2254 Subcut. 3 9 i No tuberculosis. 2255 Intrap. R.F. 3 $5 43 General tuberculosis. 2256 Subcut. 5 5 49 a General tuberculosis. 2257 Intrap. R.H. ” ” fs No tuberculosis. 2 2258 | Subcut. ” » * as No tuberculosis, 105 GUINEA-PIGS INOCULATED WITH THE MILK or Cow 77—continwed. Total ‘ No. of Mode Quarter | Dose | Killed : 2 Date. wee = Guinea- of of of or ma of Result. 24 hours, | Dis. | Injection.) Udder. | Milk. | Died. ; Feb. 27,1907 | 1150cc, | 2280 Intrap. LF. 5 cc. K 68 days No tuberculosis. {91 days after inoculation ]. 2281 Subcut. 3 ‘ 4% 3 No tuberculosis. 2282 Intrap. LH. 3 6% 3 No tuberculosis. 2283 Subcut. i 55 i is No tuberculosis. 2284 Intrap. RF a i; No tuberculosis. 2285 Subcut. ‘5 5 i ” General tuberculosis. 2286 Intrap. RH, ‘5 ‘ i General tuberculosis. 2287 Subcut. ‘ i 3 ¥i General tuberculosis. March 20, 1907 | 1360 cc. 2309 Intrap. LF. ) rs 3 g Ry K. 63 days No tuberculosis. (112 days after E 234 inoculation]. 2310 Intrap. LH. | | be a i 3 General tuberculosis. S629 2311 Intrap. RF. 4 8 28 | 6 i General tuberculosis. sooo 2312 Intrap. RH. JRS7SIL ,, 3 General tuberculosis. 2313 Intrap. LF. 5 ec, + 65 days No tuberculosis. 2314 Subcut. i i D. 9 days No tuberculosis, 2315 Intrap. L.H. 44 PP 10 days Slight T. of omentum. Death from pseudo-. tuberculosis. 2316 Subcut. rr 8 K. 65 days No tuberculosis. 2317 Intrap. R.F ‘ji 55 4s General tuberculosis. 2318 Subeut. FF 3 Pe 66 days General tuberculasis. 2319 Intrap. R.H. iy 5 65 days General tuberculosis. 2320 Subcut. 3 D. 41 days Slight tuberculosis. May 2, 1907 975 cc. 2402 Intrap. LF. ( 63 days General tuberculosis. [155 days after 8 inoculation]. 2403 Intrap. 5 E, D. 23 days Pseudo-tuberculosis. a3 2404 | Subcut. i 28 K, 63 days | No tuberculosis. an 2405 Intrap. L.H. a4 i si No tuberculosis. qe 2406 Intrap. +3 ag 55 “ Slight general tubercu- BS losis. 2407 Subcut, 53 8 fe 43 5 No tuberculosis. a 2408 Intrap. RF. oe Dz 6 days No tuberculosis. Ba 2409 | Intrap. 7 oo K. 53 days | General tuberculosis. ae) 2410 Subcut, - = 2 - ay Slight general tubercu- aCe losis. 2411 Intrap. R.H. a a 5 ss No tuberculosis. 2412 Intrap. 5 = e 3 a No tuberculosis. 2413 Subcut. ae ers is ne general tubercu- osis. * The deposit from the milk of each of the four quarters was divided equally among three guinea-pigs. 27676 vy tw 106 RABBITS INOCULATED WITH THE MILK OF COW 77. Mode uarter | Dose | Killed 3 Date. No. of of 8 of : of or Poween of Result. Rabbit. | tniection.| Udder. | Milk. | Died. aE May 3, 1907 1286 Subcut. Milk | 10 ce. K. 92 days Local lesion, and one tubercle [156 days after | from all | in lung. inoculation ]. ‘ | quarters, | } 1287 Intrap. mixed. | , zs af Three or four tubercles in | | | omentum, one in a kidney, and one in the lung. RABBITS INOCULATED WITH EMULSIONS OF TISSUES OF GUINEA-PIGS INJECTED WITH THE MILK OF COW 77. Source of Mode Killed F coe . Tissue Emulsion a a of or ee PE Result. J : injected. * | Injection.| Died. . May 7, 1907 G.P. 2287 1288 Subcut. K, 86 days Local tuberculosis only. Cnguinal Gland). May 22, 1907 G.P. 2310 1295 Subcut. Dz. 9 days Death due apparently to local (Omentum). suppuration, G.P. 2311 1296 Subcut. K. 119 days | Local lesion and slight tuber- (Spleen). culosis of lungs, 7 i 1297 Subcut. D. 15 days Local tuberculosis only. Cause of death not determined, 3 G.P. 2312 1298 Subcut, K, 91 days Local tuberculosis and tuber- (Omentum). culosis of the lungs, May 24, 1907 G.P. 2317 1299 Intrap. K. 62days | Very slight tuberculosis of (Spleen). omentum. G.P. 2319 1300 Intrap. D. 22 days Local tub i "3 cal tuberculosis. Death from (Spleen). pseudo-tuberculosis, July 4, 1907 G.P. 2413 1373 Intrav. K. i iosi y 4, oa rav 127 days | Slight tuberculosis of lungs, Gland). G.P. 2409 1374 Intrav. K. 127 days | T Osi ” ree ae y' uberculosis of lungs. Spleen). o 1375 Subcut. K, 127 days | Local lesion only. 107 EXPERIMENT C. cow 61. Cow 61 was an old cow, and had lived at Blythwood nearly five years; she had been tested with tuberculin seven times in all, the last test having been made on July 2, 1907. Her last calf was born on October 15, 1905, and for several months the amount of milk yielded from all the quarters had rarely exceeded one-and-a-half pints. The R.F. quarter had for some time been practically ‘dry, and it had been possible to obtain from it only a small quantity of serous fluid and yellow clots. First Inoculation August 6, 1907. Subcutaneous inoculation of culture derived from the ischiatic gland of Calf 1145 (Virus H 81. “P.W.”). Dose—100°0 milligrammes. Second Inoculation—November 4, 1907. Intravenous Virus H 83. “G.C Dose—150-0 milligrammes. Killed when in good health—April 16, 1909. the 2nd inoculation.] 42 41 34 Tuberculin Tests. I. October 24, 1907. [79 days after 1st inoculation.] Dose, 3:0 cc. of tuberculin. No reaction. Rise of temperature, 0°:2° C. II. October 30, 1907. [85 days after Ist inocula- tion.] Dose, 6:0 cc. of tuberculin. Ill. February 21, 1908. [109 days after 2nd [90 days after ist inoculation. ] inoculation of culture derived from a human spleen through Guinea-pig 2274, [619 days after the first inoculation and 529 days after inoculation.] Dose, 1:0 cc. of tuberculin. Reacted Rise of temperature, 1°5° C. IV. May 20,1908. [198 days after 2nd inoculation. } Dose, 3:0 cc. of tuberculin. No reaction. Rise of temperature, 0°4° C. V. November 4, 1908. tion.] Dose, 3:0 cc. of tuberculin. No reaction. of temperature, 0°1°C. [366 days after 2nd inocula- Rise Temperature. The temperature remained normal after the subcutaneous inoculation on August 6, 1907. Four days after the intravenous inoculation on November 4, 1907, the temperature rose to 40:7° C.; it reached a maximum of 42-0° C. on the 10th day, and then fell rapidly to the normal ; the period of pyrexia lasted 12 days. OCTOBER '90). BER Clinical Notes. Ist Lnoculation—A small hemispherical tumour developed at the site of the subcutaneous inocula- tion ; on October 18 it measured 7 cm. in diameter ; the adjacent prescapular gland was not appreciably enlarged. The cow remained well. 2nd Inoculation—On November 8, four days after the intravenous inoculation, the cow first appeared to ‘be unwell, and refused her food. On November 11 and 12 she was very ill, the respirations were quick, and the temperature rising (40:3° C.). On November 15 the temperature was very high (41°7° C.), but the cow was much better, and the respirations appeared to be normal. The cow remained well subsequently. Microscopical Examinations of Milk and Faeces Sub- sequent to the Intravenous Inoculation ; and Notes on the Condition of the Udder. On November 5, 24 hours after inoculation, smears from thick mucoid deposit obtained from the R.F. Subsequently (for 17 months) the temperature was normal. DECEMSER quarter, and deposit of centrifuged milk from the L.A. quarter, were examined; no tubercle bacilli were seen in either. On November 6, 48 hours after inoculation, the centrifuged deposit of milk from the L.H. quarter was examined ; two or three groups of bodies stained red which resembled tubercle bacilli were seen ; another smear showed nothing of a doubtful nature. No acid-fast bacilli were seen in an emulsion of faeces examined on the same day. On November 7, 3 days after inoculation, one of the coagula from the R.F. quarter, and one from the L.F. quarter, were examined ; chains of streptococci were seen, but no tubercle bacilli. The centrifuged deposit of milk from the L.H. quarter was also examined ; no tubercle bacilli were seen. No acid- fast bacilli were seen in an emulsion of the faeces examined on the same day. On November 9, 5 days after inoculation, the centrifuged deposit (stringy and muco-purulent in character) of mixed milk from the L.F., L.H., and R.H. quarters was examined ; no tubercle bacilli were seen. The centrifuged deposit of mixed milk from the 108 same quarters was examined two days later with the same result. On November 12, 8 days after inoculation, some muco-pus from the mixed milk was examined; no tubercle bacilli were seen. On November 14, 10 days after inoculation, centri- fuged deposit of mixed milk from the R.H. and L.H. quarters was examined ; no tubercle bacilli were seen. Coaguium from the R.H. quarter was examined with the same result. Two days later a smear from au emulsion of faeces was examined ; no tubercle bacilli were seen. | No further microscopical examinations were made during November or December, and in January, 1908, the secretion of milk practically ceased. On April 14, 1908, 162 days after inoculation, some pus was expressed from the two forequarters of the udder and examined microscopically. Itwas found to contain very numerous tubercle bacilli and the cells were composed largely of polymorphonuclears with scattered lymphocytes and alveolar cells. On the following day 50-0 cc. of sterile salt solution were injected into the L.F. quarter and 50:0 cc. into the R.F., and then after manipulating the quarters with the object of flushing out the smaller ducts the fluid was withdrawn into the flasks ; not all the fluid was recovered and that which was obtained was slightly milky and contained small flakes and flocculi. Tubercle bacilli were easily found in the fluid. Guinea- pigs and rabbits were inoculated with the fluid (see table). On April 21, 1908, some pus was expressed from the two hind quarters. Fairly numerous tubercle bacilli were seen in smears, definitely less numerous than in the pus from the fore quarters. On June 2 drops of pus from the L.F. and the R.F. quarters were again examined ; tubercle bacilli were numerous in both. Towards the end of June it was observed that three of the quarters of the udder had become enlarged and somewhat indurated and that there wasa sero-purulent discharge from the two fore teats. A fortnight later there was a distinct increase in the size of the three quarters ; the L.F. and L.H. were uniformly indu- rated, the R.F. was indurated and nodular ; the R.H. showed no alteration in size. On July 1 each of the quarters was milked and altogether 450-0 cc. of yellow tenacious muco-purulent fluid were withdrawn ; there was practically none in the R.H. quarter. The fluid contained very numerous tubercle bacilli. [With the exception of a few drops for microscopical examination nothing had been with- drawn from the udder for about six months. ] On July 2 250:0 cc. were withdrawn from the four quarters ; and on July 3 only 110-0 cc. were obtained, and this was more serous but very turbid. On July 17 a few drops of muco-purulent fluid were expressed from each quarter. Tubercle bacilli were numerous in the fluid from three quarters; in that from the R.H. quarter a few clumps of very short tubercle bacilli were seen, quite different from those in the other quarters. On August 6 pus from the L.F., L.H., and R.F. quarters was microscopically examined. Tubercle bacilli were moderately numerous in the pus from each quarter, but distinctly less numerous than when first examined. On September 15 pus or fluid was expressed from each of the four quarters and microscopically examined :— From the L.F. quarter yellowish muco-pus with some small opaque flakes in it was obtained ; a smear preparation from it showed a moderate number of tubercle bacilli, moderately numerous in places, and occasionally in clumps. From the L.H. quarter thick tenacious yellow pus was obtained ; a smear preparation showed scattered beaded tubercle bacilli with a few clumps. From the RK.F. quarter yellow muco-pus was obtained ; a smear preparation showed scattered beaded tubercle bacilli, mostly single, and a few clumps. From the R.H. quarter milky watery fluid was obtained ; a smear preparation showed a few small groups of verv short acid-fast bacilli, some like granules ; the cells were chiefly polymorphonuclears. On October 6 pus or fluid expressed from each of the quarters was microscopically examined :— In yellow muco-pus from the U.F. quarter a mode- rate number of tubercle bacilli was seen. In tenacious yellow pus from the L.H. quarter tubercle bacilli were moderately numerous. In serous fluid containing small white flakes from the R.F. quarter a moderate number of tubercle bacilli was seen. In serous fluid from the R.H. quarter one tubercle bacillus was seen. On October 15, nine days later, milk was obtained from the R.H. quarter ; from the other quarters pus was obtained, and this was diluted with saline for microscopical examination and animal inoculation. The fluid withdrawn from the R.H. quarter was like milk, and on standing a layer of cream separated. In a smear preparation two small groups of acid-fast granules or short bacilli, the majority oval, some almost spherical, were seen, also a good many short chains of cocci. Cells were not numerous and mainly polymorphonuclears ; a few alveolar cells and mono- nuclears were seen. In the pus from the L.F. quarter tubercle bacilli were moderately numerous ; in that from the L.H.a moderate number was seen ; in the pus from the R.F. quarter also a moderate number of tubercle bacilli was seen, chiefly in small groups (in the pus from this quarter a calcareous grain was found). On January 14, 1909, pus or fluid expressed from each of the quarters was microscopically examined :— In yellow pus from the L.F. quarter moderately numerous tubercle bacilli were seen the majority long and beaded; only about 5:0 cc. of the pus were obtained, it was thick and tenacious and issued from the teat like paint from a tube. From the L.H. quarter rather less than 1-0cc. of pus of similar appearance and consistency was obtained ; a moderate number of tubercle bacilli was seen in the smear preparation made from it. In watery fluid with whitish flakes from the R.F. quarter (a few drops only were expressed) scattered tubercle bacilli were seen, much less numerous than in the pus from the L.H. quarter. In turbid watery fluid containing a few flakes of yellow pus from the- R.H. quarter (only a few drops were obtained) no tubercle bacilli were seen. No organisms other than tubercle bacilli were seen in any of the smear preparations. On April 15, 1909, the milk sinuses of three of the quarters (L.F., R.F.,and R.H.) were washed out with normal salt solution. The apparatus used for each quarter was a flask stoppered with a rubber cork con- taining two glass tubes ; one a short one was for the passage of air and was plugged with wool, the other was along one and extended io the bottom of the flask ; to the outer extremity of the long tube was attached by means of semi-pressure tubing a metal catheter which was passed through the teat canal; each flask contained 100-0 cc. of fluid and this was forced into the sinuses by means of an air pump which was fixed on to the air tube. In the case of the two fore quarters the milk sinuses appeared to be much contracted and it was possible to inject only a small quantity of the fluid at one time, and the fluid recovered was only slightly milky. The right hind quarter took all the saline and the fluid recovered was opaque milky and deposited on standing yellow pus. The teat canal of the left hind quarter was plugged with dry cheesy substance which blocked up the eye of the cannula and on this account the attempt to wash out this quarter was abandoned. The teat canals of each of the other quarters contained yellow pus which was squeezed out before inserting the cannula. The amount of fluid recovered from each of the three quarters was about 95 cubic centimetres and this was used to inoculate six rabbits intravenously and three guinea-pigs intraperitoneally, two rabbits and one guinea-pig from each quarter ; ‘the dose in each case was 4:0 cc. of the fluid. Microscopical Examinations. (L.F. Numerous long and beaded tubercle bacilli. L.H. A moderate num- Pus from the teat canals4 ber of tubercle bacilli. | R.H. No tubercle bacilli. A few acid-fast. lL granules. ; 7 | RF. A few tubercle Watery fluid recovered bacilli. From the sinuses after the- RE Bae : -H. A few _ tubercle . \ anjection of saline. | bacilli, long and L beaded. Since the last note the mamma had undergone appreciable diminution in size ; the left fore quarter was still very knotty and indurated, the hind quarters felt uniformly indurated. The general health of the cow was good. She was killed the following day, April 16, 1909. [ L.F. No tubercle bacilli. Weights. cwt. qrs. lbs. August 6,1907 ... wee we 8 2B 2 April 16,1909... wis se f 1 3 Loss of weight.—1 cwt. 0 qrs. 27 Ibs. POST-MORTEM EXAMINATION. The carcass was fat. Seat of Inoculation—There was no sign of tuber- culosis at the seat of the subcutaneous inoculation on the left side of the neck. Prescapular Glands.—The left prescapular gland measured 6 by 2°5 by 2 cm. and was normal on section. The right measured 6 by 2°5 by 1:5 cm. and was also normal on section. Thorax. Lungs —The lungs were crepitant and showed no areas of collapse or consolidation ; nothing abnormal was seen from the surface. In the depth of the right caudal lobe towards the posterior extremity there was a dense yellow gritty caseous nodule about 1:2 cm. in diameter, irregular in outline, and formed apparently by two or three nodules aggregated together ; in the lung tissue immediately around it were three or four discrete firm caseous nodules the largest the size of a hempseed ; adjacent to the above group was a nodule 2 cm. in diameter composed of a number of fibrous- walled cysts filled with tenacious muco-pus. There was also a pea-sized calcareous nodule with fibrous walls. Ia the posterior part of the left caudal lobe three hard nodules were found after cutting the lung into thin transverse strips; the largest of these nodules was perhaps 1 cm. in diameter and all showed an irregular calcareous centre and a thick fibrous wall; the cal- careous centre (the largest about 5 mm. in greatest diameter) readily shelled out and under a hand-lens showed its surface covered with little excrescences causing it to resemble a piece of coral. Within the fibrous wall of one of the nodules there was a small quantity of yellow pus. The rest of the lung was normal. Thoracic Glands.—The long mediastinal gland con- tained a mass measuring 3°5 by 3 by 1°5 cm. composed of dense canary-yellow caseous areas set in a scanty matrix of fibrous tissue. Elsewhere in the cortex there were two nodules one the size of a pea the other that of a hempseed with caseous centres and fibrous margins. - Other thoracic glands were normal. Pleura and Heart.—Normal., Abdomen. Omentum and Peritoneum, Kidneys.—Normal. Pancreas.—In the head of the pancreas there was a mass composed of yellow caseo-purulent nodules set in white fibrous tissue ; the largest was rather more than 1 cm. in diameter ; towards the middle of the pancreas there were two similar encapsuled caseous and softened nodules ; the purulent substance contained calcareous grains. Suprarenal Bodies.—The left suprarenal body con- tained altogether about twenty dense yellow gritty caseous nodules the largest 5 mm. in diameter. The right contained ten similar nodules. Portal, Renal, Lumbar, and Wiae Glands.—Normal. Spleen, Liver, and 109 Alimentary Tract. Longue, Pharynx, Palate, Tonsils, Intestines and Mesenteric Glands.—Normal. Genito-Urinary System. Uterus.—Normal. Udder.--The udder on the whole was small. The right fore-quarter was much atrophied and was smaller than any of the others ; the milk sinuses were almost completely, and the milk ducts completely, obliterated ; all that remained of the milk reservoir was a small smooth-walled cavity with short diverticulum, above the root of the teat, and this contained a small quantity of pus; the tissue of the quarter was very dense, and nodules could be felt in various parts of it. On section it was composed of a network of fibrous tissue enclosing light brownish islands of atrophied glandular tissue; in the peripheral parts of the quarter chiefly, i.¢., around the margins of the base and the root of the teat, was a number of encapsuled caseo-calcareous nodules, the largest the size of a kidney bean ; in the middle of the quarter there were sparsely-scattered caseous gritty nodules the largest the size of a hempseed. The left fore-quarter was larger than the right but smaller than the two hind quarters, and was very knotty and indurated, the milk sinus except for a small cavity about 2cm. in length was obliterated ; the tissue of the quarter was even more fibrous than that of the right fore, and was closely beset with encapsuled caseo-calcareous nodules ranging in size from a millet seed to a large pea. Around the root of the teat there were some larger nodules up to 1°5 cm. in diameter, some of which were brownish translucent and beset with calcareous foci, others caseo-calcareous and encapsuled. The two hind-quarters were of about equal size ; the tissue was very fibroid, and showed on section a white fibrous network and brownish translucent tissue representing the glandular tissue; there was no definite lobulation. Scattered evenly throughout both of the quarters were caseous gritty nodules with fivrous walls varying up to 5 mm. in diameter. The milk sinuses and ducts of the left bind quarter were completely obliterated, the former being repre- sented by a small cavity lined with granulation tissue and filled with yellow pus, which did not communicate with the teat canal; the teat canal was filled with cheesy substance. In the right hind quarter there were a good many patent milk ducts, and the milk sinuses were present, though apparently a good deal contracted ; the walls of the sinuses and ducts were smooth, and the sinuses contained a small quantity of turbid fluid. Supramammary Glands.—These glands showed no naked-eye evidence of tuberculosis. Peripheral Lymphatic Glands.—All the peripheral glands were examined aad found normal. Microscopical Examination, Emulsion of Left Prescapular Gland.—No tubercle bacilli. Emulsion of Long Mediastinal Gland.—A moderate number of tubercle bacilli. Emulsion of a Nodule from a Suprarenal Body.— Numerous tubercle bacilli. Emulsion of a Nodule from the Pancreas.— A moderate number of tubercle bacilli, short and uniformly stained. Smear of Muco-pus from a Nodule in the Lung.— Numerous tubercle bacilli. Emulsion of a Supramammary Gland.—A moderate number of tubercle bacilli. Animals Inoculated. Guinea-pigs.—T wo guinea-pigs, Nos. 3716 and 3717, were inoculated intraperitoneally with an emulsioz made from the left prescapular gland; they were killed after 77 days, and showed slight general tuberculosis. 110 Two, Nos. 3718 and 3719, were inoculated initra- Two, Nos. 3714 and 3715, were inoculated intra- peritoneally with an emulsion of a nodule from the peritoneally with an emulsion of a supramammary long mediastinal gland; they died of general tuber- gland. They died of general tuberculosis in 34 and culosis in 26 and 28 days. 21 days. PIG 99. (Age—5 months 10 days.) Feeding with milk of Cow 61, after her subcutaneous inoculation with the culture derived from Calf 1145, Virus H 81. “ P.W.” Dose—T he pig was fed daily for 21 days, receiving in all 6535 cc. of milk. The feeding commenced on August 10, 1907 (4 days after the inoculation of the cow) and ended on August 30, 1907. Killed when in good health—October 28, 1907. [79 days after the commencement of the experiment. ] Clinical Notes. Total gain of weight.—2 qrs. 3 Ibs. The pig showed no sign of ill-health during the Average rate of gain per week—5‘0 lbs, experiment, and grew normally. Telhercatin Pack POST-MORTEM EXAMINATION. 2 : ion. No ri October 24, 1907 (78 days). No reaction. No rise Ph dcamanes GN TAR. of t ture. ca Weights. There was no sign of tuberculosis. 2 ewt. qrs. Ibs, All the organs and lymphatic glands were examined August 10, 1907 a ‘chad Q 1 and found to be perfectly healthy. October 28,1907 ... =... 2) 0 24 111 GUINEA-PIGS INOCULATED WITH MILK OF COW 61. Mode Quarter Killed . Date. Given ¢ of of Dose of Milk. or |? aes e Result. ‘| Injection,} Udder, Died, tae T Aug. 2, 1907 2512 Tntrap. LF 50 ec, K, 45 days Normal, [before inoculation]. 2513 “ 5 5'0 cc, 5 ‘i Normal, 2514 ” LH. 5'0 ce. 5 Normal. 2515 ‘5 is 5:0 ce. 5s 7 Normal. 2516 ‘5 RE. 4:0 ce. % * Normal. 2517 5 a 4:0 ce. ‘i is Normal. 2518 + R.H. 50 ce. ‘ wi Normal, 2519 ‘j 33 5'0 ec. +s i Normal, Aug, 7, 1907 2524 Intrap. LF. 10°0 ec, K. 54 days Normal. [24 hours after subcutaneous 2525 ” 3 10°0 ec, ‘5 4 Normal. inoculation with virus 2526 < LH. 10-0 ce. fs a Normal. H 81, P.W.]. 2527 ” i 10°0 cc. 7 4 Normal. 2528 a R.H. 10:0 cc. ss i) | Normal, 2529 + +3 10°0 ce, es rs Normal. 2530 ” RF 0'5 cc, ‘ % Normal. 2531 Intrap. L.F, ) | Each received the K. 41 days Normal. centrifuged deposit of 30°0 cc. plus 4 10°0 ce. of I 2532 5 - uncentrifuged milk.}l ,, 3 Normal. 2533 3 LH. | f 4 a Normal. 7 Do. do. 2534 a 2b lL, i Normal. 2535 ss R.H. ) { #5 9 Normal. - Do. do 2536 : = 4 cae s Normal. Aug. 9, 1907 2537 Intrap. LF. 10°0 ce. K, 53 days | Normal. {3 days after inoculation ]. 2538 7 is 10°0 ce. 3 S Normal. 2539 5 L.H. 10°0 ce. 5 * Normal. 2540 i 33 10°0 ce. 5 53 Normal. 2541 5 R.H. 10°0 ce. a ae Normal. 2542 5 # 10°0 ce. $5 + Normal. 2543 5 RF. About 0°5 cc. 4 i Normal, 2544 Intrap. LF, 1 Each received the [ K. 46 days Normal. centrifuged deposit t of 30:0 ce. plus ; 100 ce. of } 2545 9 5 uncentrifuged milk.| | ,, i Normal. 2546 3 LH.) ( 5s %) Normal. ; Do. do. ) 2547 5 3 J | L 43 sy Normal, 2548 o R.H. eG ¥ Normal. Do. do 2549 r 4 t 8 ss Normal. Aug. 13, 1907 2550 Intrap |) ( i. 58 days Normal. [7 days after | LF Each received | ! ras E, N 1. inoculation j. 2551 ” \ LE. 10°0 cc. of mixed i ” ” worma 2 and milk from ; ] 2552 ” | R.H. these quarters. | | 2 Boral. 2553 33 J Mey ‘ Normal. 2554 3 RF 10-0 cc, drawn by ji Chronic general hand and filtered. tuberculosis ta 27676 T - 112 GUINEA-PIGS INOCULATED WITH MILK oFf Cow 61—continued. Mode Quarter Killed : Date. aoe of of Dose of Milk. or es of Result. PIS: | Injection.| Udder. Died, , ie ee 2555 Intrap. | Mixed milk. Each ( K. 49 days Normal, z 5 LF. guinea-pig received | sr Bone ” L LH. the centrifuged de- |; ” oY Normal. : a and posit of 30°0 cc. plus | } = x7 2557 ” R.H. 10°0 cc. of uncentri- | an 58 days Normal. 2558 z J fuged milk. Lod . Normal. Aug. 20, 1907 2570 Intrap. LF. 10°0. ec. K. 62 days Normal, [14 days after jinoculation ]. 2571 ” ” 10°0 ce. ‘3 s Normal. 2572 i LH. 10°0 ce. v2 pS Normal, 2573 * 3 10:0 cc. + i Normal. 2568 4 R.H. 10°0 ec. $5 Fe Normal. 2569 ” ” 10°0 ce. 5 : Normal. 2566 ‘a R.F 15 ce. 3 55 Normal. 2567 53 5 155 ce. 3 és Normal. ae an ere aes 0 ———. Sept. 10, 1907 2585 Intrap. LF 10-0 ce. K. 5ldays | Normal. [35 days after inoculation J. 2586 ” ” 10:0 cc. 5 4 Normal. 2587 ” LH. 10:0 ce. sg i Normal, 2588 ” sa 10°0 cc. = PS Normal, 2583 3 R.H. 10°0 ce. 9% Normal. 2584 ” ” 10°0 ce. » 7 Normal. Sept. 17, 1907 2598 Intrap. LF. 10:0 ce. K, 59 days, | Normal, [42 days after inoculation ]. 2599 ” ” 20°0 ce. ” oy Normal. 2606 a L.H. || Each received the [ i 5 Normal, | | centrifuged deposit ‘ of 300 ce. plus} | 10°0 ce. of | 2601 si » }| uncentrifuged milk. | L ,, 3 Normal. 2602 » RH. ) ( ‘ Normal. he Do. do. “4 2603 ” » J Loss ” Normal. 2604 ” RF 1-0 ce, 5 a Normal. Oct. 8, 1907 2617 Intrap. LF. 16-0 ee. K. 48 days Normal [63 days after , inoculation ]. 2618 ” % 10:0 ce. + si Normal. 2618 5 L.H. 10°0 ce. ; ” Normal. 2614 ” si 20-0 ce. sf * Normal. 2615 ” ” 20°0 cc. 3 i Normal. 2616 ” % 30°0 ce. D. 9 days Peritonitis. 2619 * R.H. 20:0 cc. K. 48 days Normal. 2620 ” : 30°0 ce. ” ” Normal. Oct. 12, 1907 2622 Intrap. L.H. ) 10°0 ec. of milk ro, 54 days [67 days after \ withdrawn | o£ days | Normal. inoculation]. 2623 ” » | with catheter. L » ” Normal. 9694 7 2624 Intrap. L.H. \ 10°0 ec. of milk { 5 ‘3 Normal. Stor withdrawn by hand, anes a » J " \ ” Normal. 113 GuINEA-PIGs INOCULATED WITH MILK oF Cow 61—continued. Mode Quarter Killed F Date. Ge ioe of Dose of Milk. ge es Result. . * | Injection.| Udder. Died. nes _ Oct, 29,1907 2650 Intrap. LF. 10°0 ce. K. 65 days Normal. [84 days after inoculation j. 2651 . 8°0 cc. 5 3 Normal. 2652 53 L.H. 10:0 ec. i - Normal. 2653 5 of 10°0 ce. 3 5 Normal, 2654 a R.H. 10°U ce. ‘5 3 Normal. 2655 3 i |10°0 cc. 4s 5 Normal. 2656 a R.F. Small quantity with- si se Normal. drawn with catheter. 2657 % 45 About 5:0 cc. with- 5 3 Normal. drawn by hand. Total z : No. of | Mode of | Quarter Killed ‘ ie a - Guinea-| Inocu- of Dose of Milk. or ri Result. "| 94 hours pig. lation. Udder. Died. : Nov. 5, 1907 LF. 2677 Intrap. LF. 50 ca K. 59 days | Slight general tuber- hours 10°0 cc. culosis. after second 2678 i 3 5-0 ce. a Slight general tuber- Cintraven- culosis. ous) inocu- L.H. 2671 4 L.H. 10°0 ce. 4 es Slight general tuber- lation with | 65:0 cc. culosis, Virus H83. 2672 i * 20°0 cc. + 3 Slight general tuber- “G0.” ]. culosis, cn 2675 % R.H. 10°0 ce. ss on General tuberculosis. 0:0 ec. 2676 5 ss 10°0 cc. 5 5 Slight general tuber- culosis.: R.F. 2673 a RF. About 3:0 cc. of 5 a Slight general tuber- 6-0 ce. filtrate (uncentri- culosis. 2674 os " fuged) each. “ 5 Slight general tuber- culosis. Nov. 6, 1907 LF. 2681 Tntrap. LF. About 6°0 cc. K. 61 days | Slight general tuber- (48 hours 6:0 ce. culosis. afterreinocu-| L.H. 2684 45 LH. 10-0 ec. 5 oH Early general tuber- lation]. 62°0 ce. culosis. 2685 Fe ‘3 10°0 ce. 4 % General tuberculosis. R.H. 2682 “5 R.H. 10°0 ce. 8 <9 Early general tuber- 35°0 ce. culosis. 2683 Ps 5 20°0 ce. or ‘i Early general tuber- culosis, RF. 2686 94 RF About 2°0 cc. of A $5 General tuberculosis. 20 ce. filtrate (uncentri- fuged). Nov. 7, 1907 LF. 2693 Intrap. LF. 10°0 ce. K. 64 days | Early general tuber- [3 daysafter | 10°0 ce. culosis. reinocula- LH. 2689 es L.H. 10:0 ce. i ay Slight general tuber- tion]. 50°0 ce. culosis. 2690 ‘ s 10:0 ce. ‘3 5 Gencral tuberculosis. R.H. 2691 3 R.A. 10:0 cc. 3 <4 No tuberculosis. 40:0 ce. 2692 i i 10°0 ce. a5 oe Early general tuber- culosis. R.F. 2694 7 RF 1:0 cc, of filtrate. i 5 No tuberculosis. - 1:0 ce. Nov. 9, 1907 | 110°0 cc. | 2708 Intrap. LF. Each received { K. 62 days | Early general tuber- [5 days after L.H.& | 20°0 cc. of mixed culosis. : reinocula- 2709 45 R.H. milk from these D. 6 days | No tuberculosis. tion]. quarters. Cause of death not determined. Nov. 11,1907 2711 Intrap. L.F. Each received f K 60 days | Early general tuber- [7 daysafter | 65°0 ce. L.H. & | 20°0 ce. of mixed |{ culosis. “ veinocula- 2712 ey R.A. milk, ioe is Slight general tuber- tion}. culosis. ——— 27676 TS 114 GuINEA-PIGS INOCULATED WITH MILK oF Cow 61—continued. Total Killed Date of ield of No, of | Mode of | Quarter : i Duration Inoculation. Milk in | Suinea- ee U. ad Dose of Milk. it of Life. Result. 24 hours.| PS: ation. er, . f Nov.12,1907} 24:0 ec. 2713 Intrap. LF. 10:0 cc. of mixed K. 59 days | Early general tuber [8 days after } L.H. & milk, filtered. culosis. reinocula- 2714 4 R.H. 14:0 cc. of mixed rf 7 Early general tuber. tion]. milk, filtered. culosis. Nov.14,1907 LF. 2727 Intrap. LF. About 1:0 cc. K. 63 days | No tuberculosis. [10 days 1:0 cc. after rein- L.H. 2725 Fy LH. 10:0 ce. ” ” Early general tuber- oculation]. | 20-0 ec. culosis. R.H. 2726 4 Fa 10°0 ce. * ‘i General tubercu- 10°0 ce. losis of moderate severity. 2724 *) R.H. 10:0 ce. 7 i No tuberculosis, Nov. 16,1907 L.H. 2741 Intrap. L.H. 18°0 ce. D. 8 days | An acute infection. {12 days 18:0 ce. after rein- R.H. 2740 7 R.H. 18°0 ce. K. 69 days | Slight general tuber- oculation]. | 18°0 ce. culosis. Nov. 19,1907 L.H. 2751 Intrap. L.H. 10°0 ce K, 69 days | No tuberculosis. {15 days 20°0 cc. after rein- R.H. 2750 i R.H. 18-0 ce 95 A General tuberculosis, oculation]. | 18:0 cc. . Dec. 18, 1907 L.H. 2855 Intrap. L.H. About | bs K, 97 days | No tuberculosis. [44 days 22'0 ce. 15:0 ce. oe ae after rein- 2856 54 5 About | 8 g 3 Dz 86 days | Severe general tuber- oculation ]. 7T0ce, +} ESS culosis, R.H. 2857 43 R.H. 20:0 ec | ea 3 K. 97 days | Spontaneous tuber- 30°0 ce. “ao culosis. 2858 $5 3 10:0 cc. J Z 5 D. 8 days | No tuberculosis. Jan. 28,1908 | Less than |; 2919 Intrap. R.H. Less than 0°65 cc. K, 62 days | No tuberculosis. [85 days 0°5 ce. after rein- oculation]. Guinea-pigs inoculated with the fluid obtained after flushing the two forequarters with normal saline solution on April 15, 1908 (see clinical history). : Mode Quarter Killed Z No. of : Durat f Date. 2 of of Dose. ore Teton Guinea-pig- | Trjection.| Udder. Died. Life. sient April 15, 1908 3027 Intrap. LF. 1-0 cc. of fluid | D. 34 da E [163 days after oe pea ya reinoculation }. 3028 Intrap. RF, 1:0 ce. of fluid D, 36 days |General tuberculosis moderately severe. Guinea-piqs inoculated with the fluid obtained after flushing three of the quarters wath normal saline solution on April 15, 1909 (see clinical history). April 15, 1909 [528 days after reinoculation |. 3712 3711 3713 Intrap. Intrap. Intrap. L.F. RF, RH. 40 ce. of fluid, 4:0 cc, of fluid. 4:0 cc. of fluid. D. D. 25 days 34 days 21 days General tuberculosis. General tuberculosis. General tuberculosis. 115 RABBITS INOCULATED WITH MATERIAL OBTAINED FROM THE UDDER OF COW 61. (a) Inoculation of rabbits with the fluid obtained after flushing the two forequarters with normal saline solution on April 15, 1908 (see clinical history). Mode Quarter Killed No. of ee i Date. Rabbit. of of Dose, or aon af Result, Injection. | Udder. Died. : April 15, 1908 1825 Tntrav, LF. 1:0 ce. of fluid. K. 174 days | Very slight T. of [163 days after lungs and kidneys. reinoculation ]. 1826 Intrav. R.F. 30 ce. of fluid. D. 29 days. Fairly numerous dis- crete grey sub- miliary tubercles in the lungs ; scatter- ed minute foci in the liver. (b) Inoculation of rabbits with pus [ from LF, LH, and RF. quarter: s] and milk [from RH. quarter] on October 15, 1908, Oct. 15, 1908 [346 days after reinoculation ]}. 2045 2046 2044 2043 Intrav. Intrav. Intrav. Intrav. LF. LH. RF. R.H. Rather larger dose than 2044. Do. do. Small quantity of pus suspended in saline. About 3°0 ce. milk. K. 161 days K. 161 ,, K, 161 ,, of | K. 161 ,, Tuberculosis of lungs and kidneys (slight). Tuberculosis of lungs and kidneys. Tuberculosis of lungs and one kidney. Slight tuberculosis of lungs. (c) Rabbits inoculated with the fluid obtained after flushing three of the quarters with normal April 15, 1909 (see clinical history). . saline solution on April 15,1909 2305 Intrav. LF. 4:0 ce. of fluid D. 102 days | Slight general tuber- [528 days after culosis apparently reinoculation ]. insufficient to ac- count for death. 2306 Intrav. i Do. do, K. 138 —,, Slight T. of lungs and kidneys. 2303 Inirav. RF Do. do. K. 138, Slight T. of lungs, kidneys and mam- mary gland. 2304 Intrav. 43 Do. do. K. 138, Slight T. of lungs and kidneys. 2307 Intrav. B.H. Do. do. 138 ,, Slight T. of lungs only. 2308 Intrav. ay Do. do. D. 102 —C««, Slight T. of lungs and kidneys. ? cause of death. GUINEA-PIGS INOCULATED WITH THE FAECES OF COW 61. Killed . . No. of Mode of Duration Date. Guinea-pig, | Injection. Dose of Faeces. aa of Life. Result. Oct. 12, 1907 2627 Intrap. | 1:0 cc. K. 54 days | No tuberculosis. [67 days after Emulsion of subcutaneous 2628 3 10 ce. faeces—1 in i r No tuberculosis. inoculation], 20 dilution. 2629 5 0°5 ce, i 7 No tuberculosis. Nov. 4, 1907 2666 Intrap. | 1:0 ce. D. 1l-days | ? Tuberculosis of omentum. [90 days after Emul Pseudo - tuberculosis of inoculation]. oa a o spleen. ; 2667 3 1°0 cc, 90 dilution. K. 63 days | No tuberculosis. 2668 33 2°0 cc. f L| K. 63 days | No tuberculosis. 116 Guinna-Pics INOCULATED WITH THE FAECES OF Cow 61—continued. Mode Killed . Date. No. of | of Dose of Faeces. or ae Result. Guinea-pig. Injection. Died. ; Nov. 5, 1907 2679 Intrap. | 0°5 ce. | pes : of ;| K. 59 days | No tuberculosis. {24 hours after aeces—1 in : intravenous 2680 ‘5 0°5 cc. 10 dilution. D. 6 days | No tuberculosis. inoculation]. ‘ Nov. 6, 1907 2687 Intrap. | 1:0 cc. ) Emulsion of D. 8 days | The cause of death was not {48 hours after faeces—1 in apparent. — reinoculation ]. 2688 e 1:0 ce. 20 dilution. K. 61 days | No tuberculosis. Nov. 7, 1907 2695 Intrap. | 1:0 ce. ( D. 7 days | The cause of death was not [8 days after Do. do. apparent. reinoculation ]. 2696 i 1-0 ce. 64 days | No tuberculosis. Nov. 14, 1907 2728 Intrap. | 1:0 ce. | ( D. 5 days | Subacute infection. [10 days after Do do. . reinoculation ]. 2729 $s 1:0 ec. | K. 63 days | Early general tuberculosis. Nov. 16, 1907 2742 Intrap. | 1°0 ce. } D. 7 days | No macroscopic evidence of [12 days after > Do. do. { tuberculosis. reinoculation }. 2743 + 10 cc. ( 6 days | Subacute infection. GUINEA-PIG INOCULATED WITH THE EPITHELIAL SCALES FROM THE UDDER OF cow 61. Killed é No. of Mode of nas Duration Date. Guinea-pig. | Injection. Material inoculated. Boe of Life. Result. Oct. 12, 1907 2626 Intrap. | Emulsion of epithelial K. 54 days | Normal. [67 days after scales scraped from subcutaneous the udder. inoculation }. RABBITS INOCULATED WITH CULTURES DERIVED FROM THE MILK OF COW 61. i Weights. Date of Immediate Source a mm No. of arate - f Duration of Inoculation. of Culture. nee Rabbit. a a Life. Result. 8 , ton. | Initial.) Final. Aug. 5, 1908 | On April 15, 1908, 1-0 mg. 1957 Intrav. | 2,000 | 2,570] K. 83 days | Slight T. of lungs a little of the and kidneys. milky fluid with- a drawn from the O-l mg. 1958 Intrav. 1,500 | 2,300] K, 83 ,, |Slight T. of lungs. RF. quarter and kidneys. after flushing with saline was Ol mg. 1959 Intrav. 2,300 | 2,620) K. 83 ,, | Very slight T. of spread on a tube lungs and one and a culture kidney. raised from it. Oct. 5, 1908 | On Aug. 6, 1908, 1:0 mg. 2027 Intrav. 2,300 | 1,450] D. 33 days |Generalised T. not some of the pus severe. expressed from ‘ the L.F. quar- Ol mg, 2028 Intrav. | 2,270 | 2,200) K.164 ,, | Slight T. of lungs ter was sown and kidneys on a culture , ho and a 0-01 mg. 2029 Intrav. | 2,070 | 2,970) K.164 ,, |Slight T. of lungs ure was raised i eee and kidneys. July 23, 1909 | On April 15, 1909, O-1 mg. 2397 Intray. | 1,500 | 2,000) K. 75 days |Slight T. of lungs. the day before and kidneys. the cow was ue killed, a little of Ol mg. 2398 Intray. 1,859 | 2,350] K, 75 Sligh the fluid with. i a oe . a drawn from the tracheal gland; - LF. quarter after 0-01 mg. 2395 Intrav. 1,800 | 1,100; D. 28, Slight general flushing with tuberculosis. saline was spread on a tube and aj 0°01 mg. 2296 Intray 1,650 | 2,320) K. 75, Slight T. of lungs culture raised and kidne. 8 g froin it. os Le EXPERIMENT D. COW 565. (Age—4} years.) Intravenous inoculation of culture derived from a human lung direct (Virus H 104. “ E.R.”). Dose—10°0 milligrammes. Date of Inoculation—February 18, 1909. Killed when in good health—August 19, 1909. Clinical Notes. On February 21, 1909, 100 cc. of milk taken from each of 3 quarters (left fore, right fore, and right hind) were centrifuged; there was no deposit. The milk from each quarter was examined micro- scopically, and no tubercle bacilli were seen. The left hind quarter of the udder was dry when the cow was inoculated. The following day milk from the R.F. and R.H. quarters was centrifuged ; the deposit which was very small in amount was examined microscopically ; it consisted in each case of cells mainly leucocytes, a few lymphocytes and alveolar cells; no tubercle bacilli were seen. No colonies of the tubercle bacillus appeared on culture tubes sown from the deposit. Centrifuged deposits from the milk of the L.F., R.F., and R.H. quarters obtained on the 77th and 133rd days after the inoculation of the cow were examined microscopically, and culture tubes were sown ; no tubercle bacilli were seen in any of the smears, and of the culture tubes some remained sterile and others became contaminated. On July 21, 1909, 153 days after her inoculation, the cow gave birth to a dead foetus (8 months). The foetus showed no sign of tuberculosis. Emulsions were made from its spleen and a portal gland (no tubercle bacilli were seen in smears made from either emulsion) and inoculated intraperitoneally each into two guinea-pigs. The guinea-pigs were killed after 41 days and found healthy. A large amount of thin purulent fluid came away from the uterus at the time of the birth of the foetus. Microscopically this contained very numerous tubercle bacilli. During the next two or three days there was slight discharge from the vagina, and on July 27 some membranes and purulent fluid were passed. Numerous tubercle bacilli were again demonstrated in the fluid. On August 5, 1909, centrifuged deposits from the milk of the L.F., R.F., and R.H. quarters were examined microscopically. The smear preparation from the R.F. quarter showed one doubtful tubercle bacillus ; no tubercle bacilli were seen in the others. On August 16, centrifuged deposits from the milk of the three quarters were examined ; no tubercle bacilli were seen in any of the smear preparations. Culture tubes sown with the deposit grew other organisms but no tubercle bacilli. The cow remained well during the experiment, and -was in moderately good condition when killed. Temperature. There was a very slight rise of temperature com- [182 days after inoculation. ] mencing on the 11th day after inoculation and lasting twelve days (maximum 40°0° C.). With this ex- ception the temperature was normal daring the experiment. Luberculin Test. August 16, 1909. [179 days after inoculation.] Dose 3:0 cc. Reacted. Rise of temperature, 1:7° C. Weights. ewt. qrs. lbs. February 18, 1909 ... ae 8 2 16 August 19,1909 1. 2. 7 O 16 POST-MORTEM EXAMINATION. The carcass was fat. The Lungs and Thoracic Glands, the Liver, Spleen and Suprarenal Bodies were normal. . Kidneys.—In the medulla of one of the kidneys there were two grey foci of a doubtful nature (a smear showed no tubercle bacilli). The other kidney was normal. Iliac Glands.—One showed « soft greenish-yellow focus (in a smear preparation made from it no tubercle bacilli were seen). Mamma.—One quarter (the left hind) was atrophied and fibroid ; the others were normal. Uterus——The right cornua of the uterus was enlarged, being about twice the size of the left; the mucous membrane was slightly swollen and showed signs of old congestion ; it had a brownish-red colour, that of the other cornua being pinkish; the walls were a little thickened but were not otherwise abnormal. There was no naked-eye evidence of tuberculosis anywhere. All the remaining organs and glands were examined and found normal. Microscopical Examinations. Scrapings from the Mucous Membrune of the Right Horn of the Uterus——Three smear preparations from each of three separate scrapings were examined ; one, two, and three tubercle bacilli respectively were found in them after long search. Guinea-pigs Inoculated. Two guinea-pigs, Nos. 3870 and 3871 were inocu- lated intraperitoneally each with a separate emulsion of scrapings from the mucous membrane of the right horn of the uterus. They were killed after 50 days and showed general tuberculosis. 118 GUINEA-PIGS INOCULATED WITH THE MILK OF COW 565. Total i : No. of | Mode of |Quarter| Dose | Killed ti f Date. an e Guinea- Tnocu- oO tee ne mae a ° Result. oO hours: pig. lation. | Udder, ilk, led. Feb, 18, 1909 ae 3511 Intrap. LF. | 9°0 ce, K, 107 days | Healthy. {Before inocula- tion]. 3509 ss R.F. | 10:0 ce. K. 107, ,, » 3510 R.H, | 10°0 ce, K. 107 ” Feb, 19, 1909 LF. 3518 Intrap. L.F. | 10-0 ce. K. 43 days No tuberc ulosis. [24 hours after 1070°0 ce. { + the ‘inoculation ]. 3519 ” ” centri- K. 43 ” ” fuged | RF 3520 ‘4 R.F, | deposit} K. 43, ” ” a: . of 1064:0 at 3521 Intrap. ‘“ 30°0 ce. K, 43 or ” ” and partly each, Sub-perit. RH. 3516 Intrap. | R.H, K. 43, ” ” 1366°0 ce, 3517 i Ri 43 ,, Early slight general. tuberculosis. Feb. 20, 1909 LF. f 3546 Intrap. LF. | 10°0 ce. K. 45 days No tuberculosis. [48 hours after 1471-0 ec. inoculation]. ( 3547 3 ” ” K. 45 ” ” { 3544 x RF - K. 45 ,, a i ute a ce. 3545 : ‘ 3 K. 45 ,, ” ” { 3542 ” R.H, ” K. 45 ” ” ” ian 0. cc, 3543 a “ 5 K. 45 ,, ” ” Feb, 21, 1909 3552 Intrap L.F, | 10:0 cc K. 50 days No tuberculosis [3 days after a - 66: { ; inoculation ]. 3553 $i ee ” K. 50 4, ” ” { 3550 $3 RF, ” K. 50 ” ” ” 10130 eo, 3561 P 7 : K, 50 y i ‘i { 3548 3 RA, 3 K, 50 ” ” 18040, 3549 ‘ ‘i ‘ K [Oy fn Feb, 22, 1909 3558 Intrap LF, | 10-0 ce K. 5C days No tuberculosis [4 days after we. eek { inoculation ]. 3559 ” ” ” K. 50 ” ” ” { 3556 53 R.F % K. 50, 49 . 1006 e, 3567 i , le EGS Sitges, ge gy { 3554 a R.H. 6 K. 50, a - re 7 ce, x 3555 ”? ” ” K. 50 ” @ ” Feb. 23, 1909 3562 Intrap L.F. | 10:0 ce. K, 55 days | No tuberculosis. [5 days after ee f a6: rf inoculation]. 3563 “3 ‘s 54 K. 55, a Ss { 3564 3 RF, 4 K. 55, i a 980: 3 ‘i ar e cc 3565 i 3 ss K, 55, Slight general tubercu-- losis. { 3560 sf R.H. 3 K, 55, General tuberculosis. gene 0 ce, 3541 . ‘ 3 K, 55, No tuberculosis, Feb. 24, ae 3572 Intrap LF. | 100 ce. K. 55 days No tuberculosis. {6 days after { inoculation]. 1286 0 Ce 3573 ” ” ” k. 55 ” ” ” RE. { 3574 i RF j K. 55, Slight general tubercu- 900°0 ae 2 losis. ee 3575 si “ rn K, 55, No tuberculosis, RH. {| 3570 fs R.H. : K. 5B, ‘ . ae ec. \{ 3571 e K. 5B, 119 GUINHA-PIGS INOCULATED WITH THE MILK or Cow 565—continued. q ne No. of | Mode of | Quarter! Dose | Killed D ‘ Date. walk in |Guinea-| Inocu- of of or ve of Result. 24 hours, | Pls: lation. | Udder. |} Milk. | Died. Ee Feb. 25, 1909 3580 Intrap. LF. | 10:0 cc. K. 57 days General tuberculosis. [7 days after 1 on . Ae inoculation]. 3581 ‘5 7 ” K. 57 ,, 4 ‘5 { 3578 ” RF ” K. 57 ” ” » 1105 0 oe 3579 ” ” ” K, 57 ” ” ” { 3576 i R.H. ¥ K, 57 No tuberculosis 18000 377 | j so [A URSA etd, ch, gy March 4, 1909 3589 Intrap. L.F. | 10-0 ce. K, 50 days Slight tuberculosis, [14 days after 4a a és { inoculation]. 3590 “ re i K. 50 ,, No tuberculosis. { 3591 i R.F. 4) K. 50 ,, General tuberculosis, 1160 ° oes 3592 43 = a K. 50 ,, No tuberculosis. 3587 9 R.H. a5 K. 50 ,, General tuberculosis, not a 0.00: { i severe. . 3588 43 a5 9 D. 27 «yy No tuberculosis, March 18, 1909 | All three 3627 Intrap. L.F. | 100 ce. D. 11 days No tuberculosis, [28 days after quarters inoculation]. 4815°0 cc. | 3628 i 45 " K, 42 ,, 3 i 3625 9 RF. % K. 42 ,, % 9% 3626 ” ” ” K. 42, ” ” 3629 4s R.H. ‘5 K, 42, mE 5 3630 ” ed ” K. 42 ” ” ” April 1,1909 | Allthree | 3657 Intrap L.F. | 10:0 ce, D. 32 days No tuberculosis. [42 days after | quarters inoculation]. 4300-0 cc, | 3658 rn 5 a K, 56, 5 5 3659 i RF. ‘ K. 56 y 7 . 3660 ” ” ” K, 56 ” ” ” 3655 ” R.H, F K, 56 ss : 3656 ” ” ” K. 56 ” ” ” April 22, 1909 _ 3728 Intrap. | | L.F 10°0 cc K. 51 days No tuberculosis [63 days after inoculation ]. 3729 5 is 5 K, 51 ,, ” ” 3724 a R.F. ” K. 51 ,, " ” 3725 » ” ” K, 51 ” ” ” 3726 ” R.H, ” K, 51 » ” ” 3727 ” » ” K. 51 ” ” May 6, 1909 3755 Intrap. L.F. | 10:0 ce. K. 71 days No tuberculosis, (77 days after { - inoculation]. 200 0 ec, 3756 ” ” ” K. 7 ” ” 3757 a RF. i . 12 ,, No tuberculosis. (Death from pseudo-tubercu- ie 7 ce, losis.) 2 3758 oy i s K. TM os, No tuberculosis. 3753 n R.H. ” K, 71 ” ” ” ine 0 oat 3754 55 53 3s K. 71 ” ” ” June 3, 1909 3773 Intrap. L.F. | 10:0 ce K, 56 days No tuberculosis [105 days after { inoculation]. s80 ee. 3774 43 i ; K. 56, 5 + 3775 49 RE % K. 56s, ” ” ae " ce. { 3776 ; “ i K, 56 ” ” ” 3771 sf R.H, mi K, 56 yy ” ” pon: re cc. { 3772 Fr K 56 ” ” ” 27676 120 GUINEA-PIGS INOCULATED WITH THE MILK oF Cow 565—continued. Total : P : No. of | Mode of | Quarter} Dose -: Killed Duration of Date. ae of |@uinea-| Inocu- of of | or ete. Result. milkin |" pig. | lation. | Udder.| Milk. | Died. 24 hours. i July 1, 1909 LF 3791 Intrap. | L.F. | 10°0 ce. K, 58 days No tuberculosis. [133 days after “6 | : inoculation]. es 3792 ” ” ” K. 53 oy ” ” RF. ( 3795 W R.F. Pr K. 53, ” ” 2310 ae, | 3796 i 5 ip K, 53 oy ” ” RH. 3793 a R.H. si K, 53 4 ” ” 740:0 ce. 3794 i 33 4 K. 53 ” ” ” August 5, 1909 _ 3822 Intrap. | LF. | 100cce.) K 60 days | No tuberculosis {168 days after inoculation, and 3823 5 5 3 K. 60 ,, ” ” 15 days after . giving birth to 3826 = RF. ‘ K. 60 ” ” a dead foetus]. 3827 ‘i F a K, 60 ” 3824 yy R.H. ” K. 60 ” ” ” 3825 * rr 2 D. 4 ” » August 12, 1909 LF. 3842 Intrap. | LF. [10:0 ce, K. 53 days No tuberculosis (175 days after | 9950-9 do. { inoculation]. ¢ 3843 4 + 4) K, 53 Cy ” ” R.F. { 3840 2 R.F. ” K. 53, ” ” soe os 3841 9 . 7 ” K. 53 ” ” ” 3844 gt es RH. ” K. 53 OC, General tuberculosis ap- R.H. : parently of spontaneous 2440°0 ce. | origin. 3845 “5 4 5 K 53 OC, No tuberculosis. August 14, 1909 LF. 3850 Intrap. L.F. | 10°0cce.} K. 52 days No tuberculosis. {177 days after 1670°0 cc. { inoculation ]. 3851 r 45 a K. 52 4, ia ‘i { 3848 i RF. ‘ K. 52, i x 120° oe, 3849 4 8 ‘ KT) hie Bye og { 3846 r R.H. aera K. 52 ., bd r 1580" : cc. 3847 " ; r K. 52 y 7 : August 16, 1909 f ~ 3852 Intrap. | L.F. | 100 ce. K. | 52 days No tuberculosis [179 days after sie ie ce: ; inoculation]. | 3883 » 7% K | 52, ” ” { 3856 ‘ RE. ; kK. | 52, 7 9990" a ec. 3857 f Ps 7 K, \ ey, f { 3854 ‘5 R.H. 3 K 52, _ / aree. ce. 3855 . ‘ , K 52, ‘ August 19, 1909 3858 | Intrap. | LF. | 100cc.) K, 5 losis [182 days after 15000 {| y ee | 50days | No tuberculosis. inoculation}. on 3859 0 i ay K, 50, ’ ” ” Bd 3862 ts R.F, ‘a K, 50 y - ; ane 3863 ” ” ” K, 50 ” ” ” { 3860 in R.H, i K, 50, & 5 seo a ce. 3861 ” ” ” K, a 50 ” ” at 121 PIG 145 (Boar) and PIG 147 (Sow). (Hach 11 weeks old.) Fed daily for seven days from February 20 to 26, 1909, with the milk of Cow 565. Dose—All the milk drawn from the cow during the seven days commencing February 20, 1909, was given to the pigs. Killed when in good health—June 3, 1909. [97 days after the commencement of the experiment. ] Weights. Pig 145. Pig 147. ewts. qrs. Ibs. — qrs. Ibs. February 20, 1909 ... 1 12 1 7 June 3, 1909 ww 1 0 18 3 26 Total gain of weight— 3. «6 2 19 POST-MORTEM EXAMINATIONS. The carcases were fat. Pig 145 was healthy throughout. Pig 147 showed one pinhead-sized calcareous tubercle in the left submaxillary gland (smear, no tubercle bacilli) ; all the other glands and the organs were healthy. EXPERIMENT E. GOAT 79 [Adult Female]. Subcutaneous inoculation of culture derived from the original material of Virus H. 110. “J.B.” (a): through Guinea-pig 3007. Dose—50-0 milligrammes. Date of Inoculation—November 12, 1908. Killed when apparently in good health—November 21, 1908. Clinical Notes. ‘Centrifuged deposit of milk from the left quarter was examined microscopically 24 hours after inocula- tion ; no tubercle bacilli were seen. Centrifuged deposit of milk from the same quarter was examined 48 hours after inoculation with the same result. On the sixth day after inoculation milk from the left quarter was examined microscopically (only 0°5 cc. was obtained) ; and on the following day (November 19) adrop of milk from the left quarter was examined ; no tubercle bacilli were seen on either occasion. The goat was now “dry,” the left side of the udder having ceased to yield any milk; the right side had ceased to yield three days previously. She was therefore killed, on November 21, 1908, 9 days after inoculation. Twenty cubic centimetres of saline were injected into each milk sinus of the goat before she was killed, and then as much as possible was withdrawn, 16:0 ec. in the case of the right and 18:0 cc. in that of the left ; the fluid was in each case slightly milky. Smear preparations from the fluid showed no tubercle bacilli, Temperature. The day after inoculation the temperature was 40°0° C., and the pyrexia continued until the goat was killed on the 9th day; the highest temperature recorded was 40°6° C. Weights. qrs. lbs. November 12, 1908 aoe ace 3 26 November 21, 1908 ves ae 3 20 Less of weight.—6 lbs. POST-MORTEM EXAMINATION. The carcass was in fair condition. [9 days after inoculation. ] Local Lesion.—In. the subcutaneous tissues on the: left side of the neck there was a patch about the area of the palm of one’s hand and 1 cm. thick, of yellow necrotic breaking-down substance adherent to and infiltrating both skin and muscles. Left Prescapular Gland.—The left prescapular gland was much enlarged and showed the greater part of its cortex in a state of early caseation. Right Prescapular Gland.—Normal. Thorax. Lungs.—The lungs were crepitant and contained a moderate number of deeply congested tubercles up to 1 mm. in diameter which could be distinctly felt between the finger and the thumb ; a few had opaque yellowish centres. Thoracic Glands.—Normal. Mamma.—The left side of the mammary gland was larger than the right ; the gland tissue and the mucous membrane of the milk sinuses were normal to the naked eye ; the milk sinuses and ducts were practically empty. Other organs and glands were normal to the naked eye. There were two small foetuses in the uterus. Microscopical Examinations. Smears from emulsions of—- Lung.—No tubercle bacilli seen. Liver.—No tubercle bacilli seen. Spleen.—No tubercle bacilli seen. Supramammary Gland.—No tubercle bacilli seen. 27676 122 GUINEA-PIGS INOCULATED WITH THE BLOOD, AND WITH EMULSIONS OF VARIOUS TISSUES, FROM GOAT 79. . No. of Mode of Killed or Duration of Result. Material Inoculated. | quinea-pig. | Inoculation. Dose. Died. Life, 3376 Intrap. 5°5 cc. K. 68 days No tuberculosis, Blood a : { 3377 Intrap. 5°5 ce. K. 68 C,, No tuberculosis. Emulsion of lung ... 3378 Intrap. _— D. 27 days Moderately severe general tuberculosis. : Emulsion of supra- 3379 Iutrap. — K, 68 days General tuberculosis, mammary gland. 4 x Emulsion of liver ... 3380 Intrap. _- K. 68 General tuberculosis, Emulsion of spleen 3381 Tntrap. _ Dz. 56 days | Moderately severe general tuberculosis. GUINEA-PIGS INOCULATED WITH MILK OF GOAT 79. Total ‘ yield of No. of Mode of Half Dose Killed Duration of a Date. milk in Guinea- Injection - a oe Life. ent, of hours: pig. Udder. | Milk. Died. Nov. 12, 1908 3355 Intrap. Left | 10°0cc.| K. 205 days | Healthy. [ Before inocula- tion]. 3356 3% ‘5 10:0 ce. D, 81 ,, No tuberculosis. 3357 5 Right | 10:0 ce. 205 ,, Healthy. (and a small quantity from Left) Nov. 13, 1908 Left 3360 Intrap Left | 10:0 ce. K, 56 days No tuberculosis. [24 hours after half inoculation ]. 36°0 ce. 3361 5 ‘ 100ce.| K. 55g, 3 ‘ Right 3358 8 Right | 10°0cc.}| K. 55, om is half 16:0 ce. 3359 ” ” 6°0 ce, K. 55, rr ” Nov. 14, 1908 Left 3364 Intrap Left | 100cc.| K. 54 days No tuberculosis. {48 hours after half inoculation ]. 24-0 ce, 3365 a 100 cc.| K. b4 i, 5 . Right 3362 ” Right | 4°5 ce. K. 54 4, * 3 half 9:0 cc. 3363 “4 * 4°5 cc. K. 54, General tuberculosis. Nov, 15, 1908 Left half | 3367 Intrap. Left {| 10°0cc.) D, 26 days | T. of omentum and portal [3 days after 10°0 ce, gland. Death from inoculation]. : . : other causes. Right half | 3366 4 Right | 3:0 cc. K. a3 Sy Slight general tubercu- 3:0 ce. losis, Nov. 16, 1908 Left half | 3369 Intra Left 3:0 ce. i {4 days after 3:0 ec. ' 57 days No tuberculosis, inoculation]. | Right half | 3368 53 Right | About D. 24, Onetubercleinomentum. 3 drops. 3 drops Death from other causes. Nov. 17, 1908 : : ov. 17, a a 3370 Intrap. Left ol K. 71 days No tuberculosis. Noy. 18, 1 f : 5 = : ov. 18, 1908 ve : a 3371 Intrap. Left | 0°5 cc. K. 70 days | No tuberculosis. 123 Guinea-pigs inoculated with the fluid obtuined- after flushing the two sides of the udder with normal saline on November 21, 1908 (see clinical history). uinea-pig.| Injection. Udder, Noy. 21, 1908 3374 Intrap. Left {9 days after inocula- tion]. 3375 ‘i ss 3372 ¥ Right 3373 ” ” Dose, ao, i u Result. 10°0 ce, | K, 67 days {| No tuberculosis. aie ye 4 10°0 ce. K, 67 4, General tuberculosis. ae eT 4 : " EXPERIMENT F. GOAT 55 [Adult Female]. Subcutaneous inoculation of culture derived from Dose—50°0 milligrammes. Date of Inoculation—November 14, 1907. Killed when in good health—February 25, 1908. Clinical Notes. Three weeks after inoculation there was an elon- gated raised local tumour measuring 11 by about 6 cm., the margins being rather ill-defined, soft, and fluctuating. A small opening had appeared at the inferior margin of the tumour a few days previously, from which a small quantity of pus was discharged. The discharge continued intermittently and the tumour decreased considerably in size; finally the discharge ceased and the opening healed, leaving a linear scar. The condition of the goat at the close of the period of high temperature (see below) was fairly good ; it had not shown any symptoms of illness, and it quickly improved in condition and remained well subsequently. Centrifuged deposits of milk from the left and right halves of the udder were examined microscopi- cally twenty-four hours after inoculation. No tubercle bacilli were seen in that from the left half, some (?) tubercle bacilli were seen in that from the right. On the third and fourth days after inoculation centrifuged deposits from both halves were again examined, and no tubercle bacilli were found. On the second and fourth days after inoculation the faces were microscopically examined; no tubercle bacilli were seen, but a few short thick acid-fast bacilli were ‘seen on each occasion. (For milk inoculations see table.) Temperature. On the second day after inoculation the temperature rose to 40°1° C, and reached a maximum of 40°5° C. ‘on the 11th day ; the temperature was raised for a total peuiee of 29 days, after which it was quite normal, Tuberculin Test. February 21, 1908. [99 days after inoculation.] Reacted. Rise of temperature, 2°3° C. Weights. qrs. Ibs, November 14, 1907 ... aa way. Oe | February 25, 1908 .., waa ce 3. 2 Loss of weight—1-Ib;---- --— - a human mesenteric gland (Virus H 90. “I.P.”), [103 days after inoculation.] POST-MORTEM EXAMINATION. The goat was in good condition. Local Lesion.—The skin on the left side of the neck showed a linear scar 7 cm. in length ; on section of the skin, through the centre of the scar and at right-angles to it, the subcutaneous tissues were seen to be thickened for a good distance around the scar and beset with softened yellow caseous nodules slightly gritty from calcification, ranging in size from a millet seed to a large pea, the larger nodules being situated chiefly towards the periphery of the thickened patch. Left Prescapular Gland.— The left prescapular gland measured 5 by 2°5 by 1:7 cm. ; it contained two large nodules, one 2 cm. in diameter, the other rather less, composed of soft caseous substance gritty around the margins. Right Prescapular Gland.—The right prescapular gland measured 4:3 by 2°5 by 1:3 cm.; on section it showed a softened caseous nodule 1 cm. in diameter and about half-a-dozen miliary caseous tubercles. Thorax. Pleura.—Normal. Lungs.—The lungs were pink and crepitant and collapsed normally ; they showed under the pleura as well as in the depth fairly numerous evenly distributed grey fibrous tubercles, the largest 1 mm. in diameter ; some were slightly opaque in the centre. In the caudal lobes near the surface were many firm slightly raised patches, not so pink as the surrounding lung and composed apparently of aggregations of grey foci (pieces floated in water). They were caused by filarie, the embryos of which could be demonstrated in large numbers in a smear preparation. Bronchial and Mediastinal Glands.—Normal. Heart.—Normal. Abdomen. Omentum and Peritoneum.—Normal. Spleen.—Normal. Liver.—A few yellow and greenish-yellow foci of a doubtful nature were seen in the substance. Portal Glands.—Normal. Kidneys.—In the cortex of one there was a pale grey streak; in that of the other just under the capsule there was a grey nodule (2 mm. in diameter). This on section penetrated the whole thickness of the cortex and had a yellow caseous centre and grey margins. Suprarenal Bodies —Normal. Renal, Lumbar and Tliae Glands.—Normal. Genito-Urinary System. Mammary Gland.—The halves were cut up into thin longitudinal strips. On passing those of the left between the finger and thumb, a number of hard bodies were felt ; these on section were tubercles about the size of millet seeds with fibrous margins and yellow caseous gritty centres which were readily shelled out ; one of the tubercles projected into one of the ducts: besides these tubercles there were two hard nodules, the largest 1 cm.in diameter, which were composed of denser tissue than the surrounding gland tissue and contained yellow caseous foci ; no tubercles were seen in the right gland, but there was a small firm patch beset with caseous points. The milk sinuses and ducts did not contain any milk and the mucous membrane was normal. Supramammary Lymphatic Glands.—Normal. Alimentary Tract. Tongue, Pharynx, Tonsils.—Normal. Submavillary and Retro - pharyngeal Glands.— Normal. [ntestines.—Normal. 124 Mesenteric Glands.—Most of the mesenteric glands contained a number of soft yellow gritty tubercles arranged around the cortex, and in places aggregated together to form irregular caseo-calcareous patches. Peripheral Lymphatic Glands.—Normal. Microscopical Examination. Tubercle from Kidney.—One tubercle bacillus seen. Focus from Liver —No tubercle bacilli. Firm Patch from Lung.—No tubercle _ bacilli. Numerous embryos. Tubercle from Lung.—No tubercle bacilli seen. Emulsion of Prescapular Gland.—Tubercle bacilli in moderate numbers. Tubercle from Mesenteric Gland.—One tubercle bacillus seen after prolonged search. (The yellow tubercle. bacilli seen. Mammary Gland—4 Three foci from one of the nodules. One tubercle L bacillus seen. centre No of a tubercle Animals Inoculated. Rabbits 1741 and 1742 were inoculated subcu- taneously with an emulsion made from the left prescapular gland. Both died of general tuberculosis, the former in 145, the latter in 157 days. Guinea-pig 2980 was inoculated intraperitoneally with an emulsion made from the spleen. It was healthy when killed after 94 days. GUINEA-PIGS INOCULATED WITH THE MILK OF GOAT 55. | Total ass No. of Mode Half Killed z Date. a of Guinea- of of Dose of Milk. or Dees Result. 24 fae. pig. | Injection.| Udder. Died. , | Nov. 13, 1907 | 2721 Intrap. Right 10°0 cc, K, 57 days | Healthy. {before in- oculation }. 2722 5 Left 10°0 ec, % ” ” 2723 55 Right 10:0 ce. Pe ‘5 and Left (mixed). Nov. 15,1907} 130°0 cc. | 2730 Intrap Left 10°0 ce. K. 60 days | Early general tuber~ i hours both culosis, after inocu-} halves. 2731 ‘ ss 20°0 ec. K. 60 days | Early general tuber. lation]. culosis, 2732 45 Right 10°0 ce. D. 5 days | Pseudo-tuberculosis, 2733 ‘i “4 20:0 ce. K. 60 days | Very slight general tuberculosis, Nov. 16,1907] 28°0 ce. 2736 Tntrap. Left Centrifuged de- K, 53 days | General tuberculosis. [2 days after | left half. posit of 10:0 cc. inoculation]. 2737 a i Centrifuged de- K, 53 days | Harly general tuber-. posit of 18:0 cc. culosis, 65:0 ce. 2738 35 Right Centrifuged de- D. lday | Death from injury. right posit of 18:0 ce. half, plus 10:0 cc. of uncentrifuged milk, 2739 ie ‘5 Centrifuged de- K, 53 days | No tuberculosis, posit of 27:0 cc. plus 10:0 cc, of uncentrifuged milk, Nov.17,1907| 95:0 ec. |. 2744 Intrap Left Centrifuged de- K, 53 days | Early general tuber- [3 days after both posit of 30°0 ec, culosis. inoculation ].| halves, 2745 + Right Centrifuged de- K, 53 days | Slight general tuber- posit of 65°0 ce. culosis. GUINEA-PIGS INOCULATED WITH THE MILK or 125 Goat 55—cuntinuwed. Total Fi No, of Mode Half Killed j Date. le a Guinea- of of Dose of Milk. or mn ie Result, od4hours. | PIs: Injection, | Udder. Died. | °F *1%e. Nov. 18,1907} 38:0 cc. 2748 Intrap. Left Centrifuged de- K. 52 days | Barly slight general [4 days after | left half. posit of 38:0 ce. tuberculosis. inoculation}.}| 68°0 cc. 2749 % Right Centrifuged de- K. 52 days | No tuberculosis. right half, posit of 68:0 ec. Nov. 19,1907] 34:0 ce. 2752 Intrap. Left Centrifuged de- K. 69 days | General tuberculosis [5 daysafter | left half. posit of 30°0 cc. of moderate inoculation }. plus +0 ce, of severity. uncentrifuged milk. 74:0 cc, 2753 rf Right Centrifuged de- K, 69 days | Slight general tuber- right half. posit of 70:0 cc. culosis. plus 4:0 cc. of uncentrifuged milk, Nov. 20,1907] 47:0 ce. 2758 Intrap. Left 10°0 ce. D. 72 days | Severe general tuber- [6 days after | left half. culosis. inoculation]. 2759 is ‘i 10°0 ce. K, 73 days | General tuberculosis of moderate severity. 77-0 ce. 2760 3 Right 10°0 cc. K. 79 days | No tuberculosis. right half. 2761 i i 10:0 ce. K. 79 days | No tuberculosis, Nov. 21,1907] 28°0 cc. 2764 Intrap. Left 10°0 cc. Dz. 73 days | General tuberculosis (7 days after | left half. of moderate inoculation]. severity. 2765 rt r Centrifuged de- D. 86 days | General tuberculosis. posit of 18-0 cc. emulsified in saline. 45:0 cc, 2766 i Right 10°0 ce. D. 12 days | Slight pseudo-tuber- es right half. : culosis, No sign’ of tuberculosis. 2767 is ‘ Centrifuged de- K. 130 days | Slight general tuber- posit of 35°0 cc. culosis. emulsified in saline. Nov. 22,1907 | 20°0 cc. 2770 Intrap. Left 10°0 ce. K. 112 days | General tuberculosis, [8 days after | left half. not severe. inoculation]. 2771 5 ‘i 10°0 ee. D. 102 days | General tuberculosis. ~ 60°0 ce. | 2772 3 Right 10:0 ce. K. 112 days | General tuberculosis, * | right half, not severe. : | 2773 ‘3 55 100 ce. K. 112 days | No tuberculosis. Nov. 28,1907] 40°0 cc. | 2789 Intrap. Left 10°0 ce. D. 99 days | General tuberculosis. - [14 days | left half. after inocu- . ‘| 2790 5 43 10°0 cc. D. 104 days | General tuberculosis. -lation]. ae 30 os J 117°0 ce. | 2791 5 Right 10°0 ec, D. 88 days | Severe general tuber- right half. culosis. 2792 53 6 10°0 ce. K, 101 days |} Chronic general j tuberculosis. Dec. 5, 1907 | 67:0 cc. 2831 Intrap. Left 6-7:0 ce. Dz 95 days | Severe general tuber- i2l days left half, culosis. after inocu-| 40°0 cc. 2832 5 Right Centrifuged de- 116 days | General tuberculosis, lation]. | right half - posit of 40°0 cc, Jan, 28,1908| About 2920 Intrap. Left About 0°5 ce. K. 62 days | Early slight general [75 days |0°5 ce. left tuberculosis. after inocu- half. dp Dong lation ]. About 2921 55 Right About 1°5 ce. K, 58 days | Chronic general 15 cog tuberculosis. alf, 126 GUINEA-PIGS INOCULATED WITH THE FAECES OF GOAT 55. Mode Killed ‘ Date, No OE of Dose of Faeces. or Dune Result. Guinea-pig. Injection. Died. ; | Nov. 16, 1907 | 9734 Intrap. | 1:0 cc.) Emulsion of D. 6 days ‘| The sause of death was. [2 days after | + Faeces—1 in I not apparent. inoculation]. 2735 5 10 ce.) 20 dilution. \ D. 6 days Nov. 18, 1907 2746 Intrap. | 1-0 ce. | (| D. 7 days | The cause of death was not (4 days after Do, do 4 sah ; inoculation]. 2747 5 1:0 cc. { UD. 2 days | An acute infection, Nov. 21, 1907 2768 Intrap. | 1:0 cc.) f K. | 180 days | No tuberculosis. {7 days after tr Do. do. inoculation ]. 2769 i, 10 cc. } L| K. 130 days | ,, ” Nov. 23, 1907 - 2774 ° Intrap. | 1:0 ce.) (| K. | 111 days | No tuberculosis. [9 days after + Do. do ¢ inoculation ]. 2775 5 1:0 ce. J LK. 111 days | ,, ” RABBITS INOCULATED WITH CULTURE DERIVED FROM THE MILK OF GOAT 55. THROUGH GUINEA-PIG 2791. Dose in No. of Mode of | Killed Date of Immediate ee Duration Inoculation. Source of Culture. Po Rabbit. se a Died. of Life. Result. Aug. 14,1908. | The culture was; 1:0 mg. 1967 Intray. K, 167 days | Tuberculosis of lungs obtained from and kidneys. G.P. 2791, in- | 01 mg. 1968 Intrav. K. 167 =O, Very slight tubercu- oculated on losis of lungs and Nov. 28, 1907, kidneys. with 10-0 cc. of | 0°01 mg. 1969 milk from the right half of the udder. 50°0 mg. 1970 The7th generation 11 days old was used for inocu- lation. Intrav. K. 167_—O,, Very slight tubercu- losis of lungs and kidneys, Subcut. K. 167 SC, Local lesion, 2 modee rate number of case- ating nodules in lungs, and one mi- nute grey tubercle in one kidney, EXPERIMENT G. GOAT 59 [Adult Female]. Subcutaneous inoculation of culture derived from a tuberculous lung (Virus H 79. “J.N.”). Dose—100:0 milligrammes. Date of Inoculation—November 28, 1907 Killed when in good health—March 13, 1908. Clinical Notes, At the close of the first week after inoculation there was a large somewhat flattened and firm tumour measuring 9 by 5cm. at the seat of inoculation with rather ill-defined margins ; the skin over it was hot and slightly tender. The adjacent prescapular gland was enlarged, measuring 5cm. in diameter. [106 days after inoculation.] For the previous two or three days the appetite- had not been so good and the goat was losing flesh ; the milk was diminished in amount. The respiration was not increased. The tumour subsequently became soft and fluctua- ting, and then opened and discharged. The goat quickly improved in general health, and was quite. well during the last two months of the experiment. 127 ‘Temperature. The temperature rose to 40°6°C. on the 5th day after inoculation ; it was raised and irregular for a period of six weeks, the temperature varying for the most part between 39-0° and 40-:0°C. Subsequently it remained normal, _ Tuberculin Tests. ! February 21, 1908. [85 days after inoculation.] 10 cc. tuberculin (human). Reacted. Rise of temperature, 1°8° C. March 4, 1908. [97 daysafterinoculation.] 4:0 ce. ene (avian). Reacted. Rise of temperature, Weights. grs. lbs. November 28, 1907 ... ons we 38 0 March 13, 1908 aes esi wea ioe) a Total gain of weight.—2 \bs. POST-MORTEM EXAMINATION. The carcass was in good condition. ‘Local Lesion—At the seat of inoculation on the left side of the neck there was a scar measuring 5 by 1-5 cm. in area ; on section through it the sub- cutaneous tissues under and for a short distance around it were slightly thickened and beset with yellow caseo-calcareous tubercles varying in size from a pin’s head to a hemp seed. Left Prescapular Gland.—The left prescapular gland was enlarged, measuring 3 by 2 by 1-5cm.; the major part of its substance was occupied by an oval firm caseous mass slightly gritty from calcification. Right Prescapular Gland (3 by 1:7 by 0°8 cm.).— Normal on section. ' Prepectoral, Cervical, Normal. and Axillary Glands.— Thorax. . Pleura.—Normal. Lungs.—The lungs were pink and collapsed nor- mally ; on the dorsal surface of each caudal lobe there were several firm slightly projecting greyish patches which extended into the lung substance for some distance (similar to those in Goat 55); else- where the lung parenchyma showed fairly numerous more or less discrete translucent grey tubercles vary- ing up to about 1:5 mm. in diameter, several of which showed in the centre a minute soft whitish focus. The patches in the caudal lobes were undoubtedly parasitic in origin; the separate tubercles were robably also caused by parasites, though these were not detected in smear preparations. Bronchial and Mediastinal Glands.—Normal. ‘ Heart.—Normal. Abdomen. Omentum and Peritonewm.—Normal. Spleen.—Normal. Liver.—One miliary tubercle with a minute opaque white centre was seen in the substance of the liver just under the capsule. Portal Glands.—Normal. Kidneys.—The kidneys showed on the surface a number of pale grey spots of various sizes, these penetrated the cortex as radial streaks ; the majority were obviously not tuberculous, but a few showed a whitish focus or two, and without microscopical examination a decision as to the nature of these could not have been made. Suprarenal Bodies——Normal, Iliac Glands.—One of the left iliac glands showed in the cortex an irregular caseous gritty patch 5 mm. | in diameter. Renal and Lumbar Glands.—Normal. Alimentary Tract. Tongue, Pharynx, Palate, Tonsils.—Normal. Submawillary Glands.—The left contained a caseous gritty focus ; the right was normal. Intestines. —Normal. Mesenteric Glands.—All the mesenteric glands were tuberculous; the large one at the posterior end showed several patches composed of a coarse yellow caseous slightly gritty network ; the rest of the glands contained scattered discrete irregular caseous foci and streaks, some just perceptibly gritty. Lieo-Colic Glands.—All the ileo-colic glands con- tained discrete irregular caseous tubercles. The tubercles in these and the mesenteric glands were situated at the outer margin of the cortex just under the capsule. Genito-Urinary System. Mammary Gland.—The gland was cut up into thin longitudinal strips, but no tubercles were seen. There was no milk in the sinuses or ducts. Supramammary lymphatic Glands.——Normal. Peripheral Lymphatic Glands. Precrural, Popliteal, Gluteal, [schiatic—Normal. On the right side of the neck at the site of inocula- tion of tuberculin (see above) there was a small firm swelling ; on section the swelling was caused by a layer of fibrous tissue, mottled with hemorrhages, between the muscles, which were oedematous. Microscopical Examination. (Smears from :—-) Submazillary Gland (caseous focus).—No tubercle bacilli seen. Mesenteric Gland (caseous patch).—No tubercle bacilli seen. Kidney (whitish focus).—Tubercle bacilli, few seen. Lung (firm patch in caudal lobe). — Numerous embryos of the filiaric. Lung (soft central foci of small smears).—No tubercle bacilli seen. Tuberculin Swelling.—Tubercle bacilli moderately numerous. (These bacilli were no doubt inoculated with the tuberculin ; see result of guinea-pig inocula- tion with an emulsion of the swelling.) tubercles, 2 Animals Inoculated. Guinea-pig 3000 was inoculated intraperitoneally with an emulsion of the tuberculin swelling; and Guinea-pig 3001 intraperitoneally with an emulsion of spleen. The former died in 54 days, the latter in 49 days ; neither showed any sign of tuberculosis. 27676 128 GUINEA-PIGS INOCULATED WITH THE MILK OF GOAT 59. Total Killed | Duration : No. of Mode Half e u Date. La of lqninea-| of of Dose of Milk. or of Result, Muk In Fi aati : Died. Life. 24 hours. | Pis- | Injection. Udder. Nov. 28, 1907 2793 Intrap. Left 10°0 ce. K. 95 days | Healthy. [before in- . oculation }. 2794 5 Right 10°0 ec. 0 * Healthy. 2795 4 Leftand 10°0 ec. i is Healthy. Right (mixed). Nov.29,1907| Left 2796 | Intrap. Left 100 ec. K, 55 days | General tuberculosis, [24 hours half . ; after inocu- | 202-0 cc. | 2797 $5 4 Centrifuged de- ‘4 is General tuberculosis, lation]. : posit of 96°0 cc. : 2799 3 5 Do. do. a5 s General tuberculosis, Right 2798 5 Right 10°0 cc. i - Early general tuber- half culosis. ; 202°0 cc. | 2800 ' 53 Centrifuged de- * 5 General tuberculosis, posit of 96:0 cc. : 2801 ” ” Do. do. # ay General tuberculosis, Nov.30,1907| Left 2807 Intrap. Left 10°0 ce. D. 75 days | Severe general tuber- [2 days after half culosis, inocula- | 1140 cc. | 2808 Intrap. |- ,, 10:0 ce. D. 75 days | Severe general tuber- tion]. and partly culosis, Subcut. : Right 2809 Intrap. | Right 10°0 ce. 87 days | General tuberculosis half 1040 ce. | 2810 55 i 10'0 ce. D. 80 days | General tuberculosis Dec. 2, 1907 Left 2811 Intrap. Left Centrifuged de- D. 109 days | General tuberculosis [4 days after half posit of 35:0 ce. inocula- 70:0 ce. 2812 si 45 Do. do. K. 113 days | General tuberculosis, tion]. Right 2813 3 Right Centrifuged de- D. 89 days | General tuberculosis, half posit of 37-0 ce, 74:0 cc. 2814 5 ‘5 Do. do. Dz 90 days | General tuberculosis, Dec. 83,1907} Left 2821 Intrap. Left 10:0 ce, D. 58 days | General tuberculosis, [5 days after half inocula- 75:0 ce. tion]. Right 2822 re Right 10:0 ce. K. 112 days | General tuberculosis. half 50°0 ce. Dec. 4, 1907 Left 2827 Intrap. Left Centrifuged de- D. 109 days | General tuberculosis. (6 days after half posit of 19°0 ce, inocula- 38°0 ce, 2828 i 3 Do. do. D. 97 days | Severe general tuber- tion]. : . culosis, Right 2829 5 Right Centrifugal de- D. 73 days | General tuberculosis, half posit of 20°0 ec. 40:0 cc. 2830 5 35 Do. do. D. 72 days | General tuberculosis not very severe EXPERIMENT H. GOAT 69 [Adult Female]. Subcutaneous inoculation cf culture derived from cerebro-spinal fluid through Guinea-pig 2878 (Virus H 98. Dose—10-0 milligrammes. Date of Inoculation—June 3, 1908. “BLR.”) Killed when in good health—September 25, 1908. Clinical Notes. On June 5, two days after inoculation, the centri- fuged deposit of milk from the right and the left halves of the udder was examined microscopically. No tubercle bacilli were seen in that from the left half, one bacillus stained red w the right. On June 17, the centrifu again examined ; no tube from the left half, five (114 days after inoculation. ] seen in that from the right. as found in that from ged deposit of milk was rele bacilli were seen in that long tubercle bacilli were 129 On July 15 (42 days after inoculation) before withdrawing the milk for the inoculation of guinea- pigs, the udder was noticed to be enlarged : on palpa- tion the right side was found to be uniformly enlarged, knotty, and indurated ; the left side was not obviously enlarged but contained some hard nodules just above the teat. Anenlarved supramammary lymphatic gland was felt on the right side. The milk was much diminished in amount (only about 5 cc. being obtained from the right half and 14 cc. from the left half), but showed no alteration in quality. On July 15 culture tubes were sown from the milk of each half, and pure cultures of tubercle bacilli were obtained, the colonies being more numerous on the tube sown with milk from the right side than on that sown with milk from the left side. The general health of the animal was good. On July 31 a few drops of milk were obtained from the right side of the mamma, and about 1°0 cc. from the left. No tubercle bacilli were found in smear preparations made from either of the samples. On September 24 both sides of the udder were still indurated, but had decreased in size. The goat was in good health when killed on September 25. Temperature. On the 45th day after inoculation the temperature rose to 39°5°C. and reached a maximum of 39°8°C. on the 50th day : it was raised during 10 days in all (July 18-28). With this exception, the temperature remained approximately normal, though somewhat irregular, during the experiment. Tuberculin Test. August 18, 1908. [76 days after inoculation.] Dose, 1'0cc. Reacted. Rise, 1:8° C. Weights. qrs. Ibs. June 3, 1908... tes ae ve «82 22 September 25, 1908 ... eos we «2 21 POST-MORTEM EXAMINATION. The carcass was in good condition. Local Lesion.—In the subcutaneous tissues at the seat of inoculation on the left side of the neck there was a softened caseous encapsuled mass rather less than a pigeon’s egg in size ; the capsule was adherent to the skin but not to the muscles; it was about 1 mm. thick and lined internally with granulation tissue. Left Prescapular Gland.—The left prescapular gland was the same size as the right, and was normal on section. Right Prescapular Gland.—Normal. Prepectoral and Axillary Glands.—Normal. Thorax. Pleura.—Normal. Lungs.—On the dorsal surface of each caudal lobe there were many irregular firm reddish patches which on section were found to extend a short distance into the substance of the lung; the patches were due to parasites, numerous embryos of the filaria being found in a smear preparation ; no tubercles were seen in the lung. Bronchial and Mediastinal Gland.—Normal. Heart.—Normal. Abdomen. Omentum and Peritoneum.—Normal. Spleen, Liver, Kidneys, and Suprarenal Bodies.— Normal. Portal, Normal. Coeliac, Lumbar, and Iliac Glands.— Alimentary Tract. Tongue, Pharynx, Palate, Tonsils.—Normal. Submasillary and Retro - pharyngeal Glands.— Normal. Intestines and Mesenteric Glands.—Normal. Mammary Gland.—The mammary tissue felt indu- rated and nodular, particularly in the posterior parts ; the right half of the gland was larger than the left, which was on the whole probably not much larger than it should be. On removal the upper surfaces showed several irregular spherical nodules which were fibrous-walled cysts containing a substance resembling cream cheese ; at the outer margin of each half of the mamma there was a thin-walled cyst containing about a cubic centi- metre or more of milky-white fluid. On section the tissue of the right half of the mamma was very tough and fibrous and beset with shotty tubercles from 1: to 1°5 mm. in diameter which stood up from the cut surface; these tubercles had fibrous margins and yellow centres which in the majority of cases were calcareous or caseo-calcareous and in a few caseous and softened; towards the posterior part there was a yellow caseo-calcareous nodule the size of a wheat-grain. The left half of the mamma was similar to the right, but the tubercles were not so numerous. The milk sinuses contained a small quantity of turbid or pale milky watery fluid; some of this was pipetted from each sinus through the teat before the mamma was cut into and cultures sown; the mucous membrane of the sinuses was normal. Supramammary Lymphatic Glands. — Both supra- mammary lymphatic glands were enlarged, the left only slightly, the right was about twice the size of the left ; on section the tissue of both was oedematous, but there was no sign of tuberculosis. Various Lymphatic Glands. Precrural, Popliteal, Gluteal, Ischiatic, Cervical.— Normal. Microscopical Examination. Smear from Reddish Patch in Lung.—No tubercle bacilli. Numerous embryos of the filaria. Smear from Popliteal Gland.—No tubercle bacilli. Smear from Distended Alveolus in Left Mamma.— No tubercle bacilli seen. Smear from Disiended Alveolus in Right Mamma.— No tubercle bacilli seen. Smear from Caseo-calcareous Nodule in Right Mamma.—No tubercle bacilli seen. A few diplo- bacilli. Smear from Several Miliary Tubercles from Right Mamma.—Three tubercle bacilli seen. Milky fluid from Left Sinus.—No tubercle bacilli seen Milky fluid from Right Sinus—One (?) tuberc'e bacillus seen. Animal Inoculated. Guinea-pig 3287 was inoculated intraperitoneally with an emulsion of the right supramammary gland. It was killed after 49 days and showed slight general tuberculosis. 27676 130 GUINEA-PIGS INOCULATED WITH THE MILK OF GOAT 69. Total A No. of Mode Half Killed | Duration Date. ue of Guinea- of of Dose of Milk. or of Result, 24 Tee pig. | Injection.| Udder. Died. Life. June 3, 1908 3093 Intrap. Left 10°0 cc, | K. 83 days | Healthy. [before in- . oculation ]. 3094 5 Right 10°0 cc. K, 83 ,, ra 3095 5 Left and About 9:0 ce. K. 83, 3 Right. June 4, 1908 | Left half | 3096 Intrap Left 20°0 ce. K. 37 days | No tuberculosis. {24 hours | 70-0 ce. after inocu- 3097 5H i 10:0 ce. K. 70 4, ” ” lation }. : Righthalf| 3098 5 Right 20°0 ce. K. 37 yy ” ” 50:0 ce. 3099 ii 6 10:0 cc. K. 70 4, ” ” : Centrifuged de- 70 a No tuberculosis June 5, 1908| Left half | 3100 Intrap Left : ; K. ays o tubercu : posit of 30:0cc. plus [48 hours | 150°0 ce. 10°0 ec. of uncen- after inocu- 3101 si as [ trifuged milk K. 70 yy ” ” tion ]. Centrifuged de- |} ‘ ‘ ged de- oe 3102 és Right | positof 30-0ce. plus K. 70 5, ” ” : Ces 3103 10:0 cc. of uncen- K 70 " ” trifuged milk, : ¢ 2 a June 6, 1908 | Left half | 3105 Intrap. Left 10-0 ce. K. 72 days | No tuberculosis [3 days after} 77-0ce. inoculation ]. 3106 ‘9 9 10°0 ec. K. 72 «4, 5 ‘ Righthalf| 3107 33 Right 10:0 ce. K. 72 ~~, 5 100°0 ce. 3108 a 20 10°0 ce. K. 72 ~«,, ” ” June 8, 1908} Left half | 3109 Intrap Left 10°0 ce. K. 70 days | No tuberculosis [5 daysafter| 34:0 ce. inoculation ]. ; 3110 i 33 10:0 ce. K. 70 4, 8 i Righthalf| 3111 3 Right 10°0 ce. K. 90° 3 4 ‘ 104°0 ce. 3112 5 i 10°0 ee. K. 70 ,, ‘3 sf Junel0, | Left half | 3113 Intrap Left 10:0 ce. K. 69 days | No tuberculosis 1908 [7 days| 62°0cc. after inocu- 3114 os - 10:0 ec. K. 69 ,, ‘ “5 lation]. Righthalf| 3115 x Right 10°0 ce, K, 69 ,. 4 3 157°0 ce. 3116 i » 10-0 ce. K. 69 ., a ‘i June 17, | Left half | 3129 Intrap Left e K. 78 days | Chronic _ general 1908[14days| 60-0 ce. Centrifuged de- [ tuberculosis, not : posit of 30:0 ce. after inocu- each ( severe, lation ]. 3130 is i ‘ K. 78, No tuberculosis, Righthalf} 3131 ‘3 Right Centrifuged de- K. 1 Chronic generai 60°0 cc. { posit of 30-0 ce. tuberculosis, 3132 3 si each. K. 78 ,, Chronic general tuberculosis, net severe, June 24, | Left half | 3133 Intrap. Left 10:0 ee. K. 84 days | Chronic _ general 1908 [21days| 45:0 cc. tuberculosis. after inocu- 3134 % 5 10:0 ce. K. 84, Chronic _ general lation }. : . tuberculosis. Righthalf| 3135 75 Right 10:0 ec. K. 84, Chronic _ general 60°0 cc, tuberculosis, 3136 5 5 General tuberculosis. ‘0 cc. 3140 ” ” 10°0 ce. K. Te Chronic general tuberculosis. July 17, Left half | 3155 Intrap Left About 7:0 ce. D. 35 days | General tuberculosis, 1908 [44days) 140 ce. aga a 3156 ” ” ” i D. 19, General tuberculosis. ation]. “i ae half} 3153 i Right About 2°5 ce. Dz. 22 (, General tuberculosis. 5:0 cc. 3154 ” ” ” ” K. 71 General tuberculosis. 131 EXPERIMENT I. GOAT 61 [Adult Female]. Subcutaneous inoculation of culture derived from the original material of Virus H. 109 “MW.” (lupus), through Guinea-pig 3002. Dose—1°0 milligramme. Date of Inoculation—July 27, 1908. Killed when in good health—February 9, 1909. Clinical Notes. The"goat remained well after the inoculation. A small elongated tumour developed at the seat of inocu- lation on the left side of the neck, which afterwards disappeared, and the adjacent prescapular gland was slightly enlarged for a few weeks. The milk was inoculated into guinea-pigs at intervals ‘during the experiment (sce table). Centrifuged deposit from the milk of each side of the udder was ‘examined microscopically on two occasions, August 4 and October 7 ; no tubercle bacilli were seen. On the latter date cultures were attempted, two egg tubes being sown with the centrifuged deposit of the milk from each side of the udder; all the tubes remained sterile. _~ Nothing abnormal was felt in tle mammary gland during the experiment. The goat had given birth to a kid ten weeks before her3inoculation ; the kid, No. 71, was permitted to suck its mother from August 13 (17 days after her inoculation) until January 14, 1909. _ Temperature. The temperature was a little raised for a period of 47 days commencing eighteen days after the inocula- tion; it averaged about 39:3°C.; on. one occasion 40:0° C. and on another 40°3° C. was recorded. Subse- ‘quently the temperature was approximately normal. Tuberculin Tests November 4, 1908. [100 days after inoculation.] [197 days after inoculation. ] January 7, 1909. [164 days after inoculation.] Dose, 1:0 cc. Reacted. Rise of temperature, 1°9° C. Weights. qzs. Ibs. July 27,1908 ... wis es 3 14 February 9, 1909... wee aes 3 17 Total gain of weight.—3 lbs. POST-MORTEM EXAMINATION. The carcass was in good condition Local Lesion.—In the subcutaneous tissues at the seat of inoculation there were two nodules with fibrous walls and creamy caseo-purulent contents ; one was the size of a hemp seed, the other was slightly larger ; in the muscles near these there was a similar nodule the size of a hemp seed. Left Prescapular Gland.—The left prescapular gland was normal in size and showed no lesions on section. Lungs.—On the dorsal surface of each caudal lobe in the posterior part there were several firm grey patches and in the rest of the lungs isolated grey nodules caused by filariae. There were no definite tuberculous lesions. Mesenteric Glands.—In the cortex of one a minute greenish-yellow focus was seen. Mammary Gland.—Normal. All other organs and glands were examined and Dose, 1:0 cc. Reacted. Rise of temperature, 2°1° C. found normal. GUINEA-PIGS INOCULATED WITH THE MILK OF GOAT 61. Total : ield of | N°-°F | Mode ot | Half / Dose | Killed | poration of Date. Slat iee Guinea- Iniecti of of or Life Result. oifilk in |pig. | Mection- | vader. | Milk. | Died. . July 27, 1908 _ 3176 Intrap. | Right | 10°0 ce. K. 124 days Healthy. Before inocu- ; "tation 3177 9 Left | 10:0 cc. K, 124 ,, 53 3178 Ss Right |10°0cc.] K. 124 ,, m and left. July 28, 1908 Both 3179 Intrap. Lef 10°0 ce. K. 66 days No tuberculosis, [24 hours after halves (Milk inoculation ]. 840°0 cc. with- drawn by hand). 3180 » ” ” K. 66 ” ” » 3181 si Right ; x 66 ,, 5 7 © Baba) egy LA gs. abl cage (i SE BR. ae, 0 GuINEA-PIGS INOCULATED 132 WITH THE MILK OF GOAT 61—continued. Total ‘ i 5 yield of No. of | Mode of ner ue eee Duration of Result. sic Milk in |°™2°| Tniection. | yguer,| Milk. | Died Life, 24 hours. | P18 ne : ; July 30, 1908 _ 3187 Intrap. Left | 10°0 cc. K. 68 days | No tuberculosis. {3 days after inoculation]. 3188 ‘5 7 ” K. 68 ,, ” ” 3189 ” Right ” K, 68 ” ” ” 3190 ” ” ” K. 68 ” ” ” August 1, 1908 Both 3195 Intrap. Left | 10-0 cc K, 72 days No tuberculosis (5 days after halves inoculation]. 390-0 cc. 3196 5 ‘ 3 K. 72 5, ” ” 3193 ‘3 Right i K. 72 ” ” 3194 ‘ ai 7 K. 72 4) ” ” August 4, 1908 | Left half 3197 Intrap. Left | 10°0 ce. K, 69 days No tuberculosis [8 days after 1770 ce. inoculation ]. 3198 4 as 5 K. 69 ” ” Right half] 3199 9 Right o K, 69 5, ” ” 170°0 ce. 3200 ” ” ” K. 69 ” ” August 12, 1908 | Left half 3209 Intrap. Left | 10:0 cc. K. 69 days No tuberculosis (16 days after | 175°0 cc. inoculation ]. 3210 ” ” ” K. 69 ” ” ” Right half 3207 a Right ” K. 69 ,, ” ” 122°0 ce. 3208 ” ” ” K. 69 ” »” ” August 26, 1908 _ 3226 Intrap. Left | 10-0 cc. K, 62 days No tuberculosis [30 days after inoculation ]. 3227 ” ” ” K. 62 ” ” 3228 $3 Right 7 K. 62 ,, i ' 3229 ” ” ” K. 62 ” ” ” Sept 16, 1908 _ 3273 Intrap. Left | 10:0 ce, K, 69 days No tuberculosis [51 days after inoculation ]. 3274 54 i i K, 69 ,, 5 3 3271 ” Right ” K. 69 ” ” ” 3272 ” ” ” K. 69 ” ” ” Sept. 30, 1908 _ 3307 Intrap. Left | 10°0 ce. K. &3 days No tuberculosis. [65 days after inoculation]. 3308 7 i a K. 83 ,, Sy es 3305 ‘ Right 55 Dz 73, ‘s *f 3306 ” ” ” K. 83 ”» ” yy Oct. 26, 1908 _ 3335 Intrap Left | 10°0 ce, K. 101 days No tuberculosis [91 days after inoculation ]. 3336 is a a K, 101 ,, +4 53 3337 i Right 3 K, 101 ,, Fe a 3338 A ” ” K. 101 ,, ” ” Nov. 5, 1908 _- 3347 Intrap Left | 10°0 cc K, 93 days No tuberculosis [101 days after i inoculation]. 3348 i Right 5 K. 93 Cy, Pa ‘ Nov. 6, 1908 — 3353 Intrap. Left | 10:0 ce. K, 105 di N ic [ioe aye . ays o tuberculosis, noculation ]. 3354 ” ” ” K. 105 ,, ” ” 8351 i} Right i K, 105 ,, ‘i a 3352 ” ” ” K. 105 ” ” ” 133 GUINEA-PIGS INOCULATED WITH THE MILK or Goat 61—cuntinued. , Total ; No. of i Date. a of Guineas Mode of cae aie ae Duration of ilk in i Injection. a aS Life. Result, 24 hours. pig. Udder. | Milk. Died. Jan, 6, 1909 _ 3425 Intrap. Left | 10:0 cc, K i 16s ats ae p e: ee ; 69 days | No tuberculosis. inoculation ]. 3426 uF i is K, 69 ,, e $3 3423 ” Right ” K,. 69 ” ” ” 3424 ” ” 2 K. 69 ” a ” ) Feb. 9, 1909 Left half | 3460 Intrap. Left | 4:0 ce, K, 42 x i [is7 ee ee =o p ce days | No tuberculosis. inoculation]. 3461 Fe a 4:0 ce, K, 42 ,, e 4 Right half | 3458 si Right | 5:0 ce. K. 42 ,, y rr 9°0 ce. 8459 5 3 4°0 cc. K. 42 ,, 45 % KID 71. (Ten weeks old.) SUCKING GOAT 61. Duration of experiment—The kid was placed with Goat 61 on August 13, 1908, 17 days after her inoculation, and continued sucking until January 14, 1909, a period of 154 days; on this day the kid was killed. Clinical Notes. The kid remained well and grew normally during the experiment. Tuberculin Tests. No. 1. November 4, 1908. [83 days after com- mencement of experiment.] Dose, 1:0 cc. No reaction. Rise of temperature, 0°3° C. No.2. January 7, 1909. [147 days after inocula- tion.] Dose, 1:0cc. Rise of temperature, 0°8 O. Weights. August 13,1908 ... ees ie January 14, 1909 ... qrs. 1 wes 2 Total gain of weight.—1 qr. 3 lbs. Ibs. 6 9 POST-MORTEM EXAMINATION. The carcass was in good condition. All the organs and glands were examined and were found free from tuberculosis, EXPERIMENT J. GOAT 81 [Adult Female]. Intravenous inoculation of culture derived from the original material of Virus H. 115. “N.G.” through Guinea-pig 3087. Dose—2°0 milligrammes. Date of Inoculation—December 18, 1908. Died—January 28, 1909. [41 days after inoculation.] Clinical Notes. The goat was first noticed to be-ill during the second week in January; she lost appetite, was quiet, and looked seedy and depressed. The loss of appetite continued, the respiration became quickened, and the goat gradually became very weak and emaciated ; death ensued on the 41st day after inoculation. _ Milk was withdrawn from the udder by hand (the -ducts of the teats being too narrow to admit a cannula) at intervals from December 18 (the day of inocula- tion) until January 2 and inoculated into guinea-pigs ; after this date the udder was dry and no milk could be obtained. On January 2 milk from both halves of the udder was centrifuged and the deposit examined micro- scopically ; no tubercle bacilli were seen. On January 28, after death, 10°0 cc. of saline were injected into the milk sinuses of each half of the mamma. After manipulating the mamma some milky fluid was withdrawn. Microscopical examination of the fluid from each half of the mamma showed no tubercle bacilli; the fluid was inoculated into guinea- Pigs. _ 134 Temperature. The temperature was raised and irregular during the whole period of the experiment (maximum 40:4°C., minimum 38°3° C.). Weights. qrs. Ibs. December 18, 1908... wa we «= 2 20 January 28, 1909 ... sa waa 42 0 Total loss of weight.—20 lbs. POST-MORTEM EXAMINATION. The carcass was emaciated. Local Lesion.—In the muscles near the middle of the left jugular vein there was a small collection of caseo-pus ; the vein itself was normal. Thorax. Lungs.—The greater part of the right cephalic lobe, the whole of the right middle, the ventral portions of the left cephalic lobes, and the antero-ventral portions of the caudal lobes were firm, reddish and airless ; the rest of the lung was congested, air-containing, but much firmer and less crepitant than normal. The consolidated tissue showed on section greyish areas surrounded by reddish tissue, many of. the grey areas (which probably corresponded to lobules) having irregular ill-defined yellowish centres. The crepitant parts of the lungs were closely beset with grey almost transparent miliary tubercles. Thoracic Glands.—The bronchial and dorsal medias- tinal glands were much enlarged and congested ; they were composed of reddish tissue mottled with small ' translucent grey areas ; there was no sign of caseation. Heart.—Normal. Abdomen.. Spleen.—The spleen was soft, no tubercles were: seen on the surface or on section. Liver.—The liver was pale and firm, no tubercles were seen on the surface or on section. Kidneys.—Normal. Mesenteric Glands.—In one there was a large pea- sized calcareous nodule (not tuberculous). Mammary Gland.—The mammary gland was small, the gland substance was tough, and showed on section about half a dozen yellow bodies each about. 1 mm. in. diameter which stood up from the cut surface ; when crushed they had a fibrous wall and milky or creamy contents; no tubercle bacilli were found in smear preparations made from two; they were probably of the nature of retention cysts. There was a small quantity of milky fluid in the milk sinuses. The mucous membrane was normal. All other organs and glands were examined and found normal. Microscopical Examination. Scraping from Consolidated portion of Lung.— Tubercle bacilli numerous. : Scraping from Mediastinal scattered tubercle bacilli. Nodule from Mesenteric Gland.—No tubercle bacilli. Soft focus from the Mamma.—No tubercle bacilli. Gland. — Sparsely- GUINEA-PIGS INOCULATED WITH MILK OF GOAT 81. (The milk was withdrawn by hand.) Total , = No. of Mode Half Dose Killed 7 Date. ee bes Guinea- of of of or ne of Result. 24 hours, | Pig: | Injection.| Udder. | Milk. | Died. nes Dec. 18, 1908 3349 Intrap. | Right | 10:0 ce. K, 169 days | Healthy. [before inocu- lation]. 3400 $i Left | 10:0 ce. K. 169, 5 Dec. 19, 1908 Right 3401 Intrap. Right | 10:0 ce. K. 51 days | General tuberculosis. [24 hours after half inoculation ]. 161°0 ce. 3402 5 i 10:0 ce. K. Sly; Slight general tubercu- losis. 3403 4 ss 10°0 ec. K, 51, Slight general tubercu- losis. 3404 9% o 10°0 ce. K. 51, Very slight general tuber- culosis. Dec. 20, 1908 Right 3405 Intrap Right | 10-0 ce. K. 50 days | Slight general tubercu- '48 hours after half losis. inoculation]. 190°0 ce. 3406 5 or 10°0 cc. K, 50 yy Slight general tubercu- losis. 3407 eS 4 10:0 ec. K. 50 Early general tubercu- losis. 3408 > ; 10°0 ce. K. 50, Very slight general tuber- culosis. Dec. 21, 1908 Right 3409 Intrap Right | 10-0 ce. K. 50 days | No tuberculosis, [3 days after half inoculation]. 187°0 cc. | 3410 3) 9 10:0 ce, K. 50, 7 si Left 3411 8 Left | 10-0 cc. K, 50, : - half 185°0 ce. 3412 - i 10°0 ce. K. 50 Cy, General tuberculosis, not. severe, 135 GUINEA-PIGS INOCULATED WITH MILK oF Goat 81—continued, Total : No. of Mod i Date. yield of Guinea- of era ee : sca Duration of Milk in 7 Mates or 0 or Life Result. 94 hours: pig. | Injection.| Udder.| Milk. Died. : Dec. 26, 1908 Right 3413 Intrap. Right | 10-0 ce. K. 77 da i [3 ae after iE p g c ays | No tuberculosis. Inoculation ]. 170°0 ce, 3414 3 si 10°0 ce, K. IT Slight general tubercu- losis. Left 3415 o Left | 10:0 ce. K. TT 35 General tuberculosis, half : moderately severe. 170°0 ce. 3416 Fi a 10-0 cc. K. TE x Slight general tubercu- losis. Jan. 2, 1909 Right 3417 Intrap. | Right | 10:0 ce. K, 70 days G 1 tubere i [is re atta mer p g days eneral tuberculosis. inoculation ]. 90°0 ce. 3418 53 As 10°0 ce. K. 10° 4 Slight general tubercu- losis. ee 3419 4 Left | 10°0 ce. K, 704, No tuberculosis. a 90°0 cc. 3420 ‘5 - 10:0 cc. K. 70, Slight general tubercu- losis. Early in January the udder became dry ; January 2 was the last occasion on which milk was withdrawn. On January 28, 1909, 10-0 cc. of saline were injected into the milk sinuses of each half of the mamma. After manipulating the mamma some milky fluid was withdrawn through the catheter used for injecting the saline, 4-0 cc. from the right side and 5:0 cc, from the left. Jan. 28, 1909 | 3436 | Intrap. | Right [41 days after | inoculation]. 3437 4 3434 . Left 3435 - 4 20cc.of, K 47 days | General tuberculosis, the fluid. a K. 47 General tuberculosis. | 25 cc. off D 28 OC, General tuberculosis. the fluid. » D | 35, | General tuberculosis. I I EXPERIMENT K. GOAT 75 [Young Adult Female]. Fed once with culture derived from the original material (synovial membrane of knee-joint) of Virus H. 76. “G.M.” through Guinea-pig 2887. Dose—74:0 milligrammes. Date of Feeding—September 22, 1908. Killed when in good health—August 16, 1909. Clinical Notes. On October 14, 22 days after feeding, centrifuged deposits of milk from the right and left halves of the udder were examined microscopically ; no tubercle bacilli were seen in either. Culture tubes sown from the deposit remained sterile. The goat gave birth to two kids on May 30, 1909, eight montbs after the commencement of the experi- ment ; the kids sucked their mother for nearly two- and-a-half months, and when killed on August 10, 1909, were found to be perfectly healthy. The goat remained well during the experiment and was killed on August 16, 1909. Temperature. There was a very slight rise of temperature com- mencing 28 days after inoculation which lasted a week and reached a maximum of 40:0° C. With this exception the temperature was normal during the experiment. Tuberculin Tests. November 4, 1908. [43 days after inoculation. ] Dose, 1:0 cc. Reacted. Rise of temperature 1-6° C. January 7, 1909. [107 days after inoculation. ] Dose, 1:0 ce. Reacted Rise of temperature 2°0° C. 27676 [328 days after feeding. ] Weights. qrs. Ibs. September 22, 1908 ae aise 2 16 August 16,1909 ... nah iis 3 4 Gain of weight.—16 lbs. POST-MORTEM EXAMINATION. The carcass was in good condition. Alimentary Tract. Tongue, Pharynz, Zonsils.—Normal. Submacillary Glands.—In the cortex of the left there was a group of three or four small calcareous tubercles ; the right was normal. Retro-pharyngeal Glands.—In the right pharyngeal gland there was a calcareous tubercle and two calcareous grains ; the left was normal. Parotideal Giands.—The left contained a few minute calcareous grains ; the right was normal. Cervical Glands.—Normal. Intestines.—Normal. Mesenteric Glands.—The large terminal gland con- tained an irregular calcareous patch and a few Y 136 calcareous tubercles; one in the middle of the millet-seed sized calcareous tubercle; other portal mesentery showed a millet-seed sized caleareo-caseous glands were normal. tubercle. Other mesenteric glands were normal. Ileo-colic Glands.—Six out of seven glands were affected, but were not obviously enlarged ; three < : contained rather large calcareo-caseous patches ; two There was no sign of tuberculosis elsewhere. showed a calcareo-caseous nodule or two, and in the Mammary Gland,—Normal. other there was a single hemp-seed sized caseous Microscopical Examination. gas. Smear from the, Spleen Substance-—No tubercle bacilli. : Portal Glands.—One portal gland contained a Nodule from a Portal Gland.—A few tubercle caleareo-caseous nodule the size of a pea and a bacilli. GUINEA-PIGS INOCULATED WITH MILK OF GOAT 75. Total yield | No. of Half | Dose | Killed ; Date. of Milk in | Guinea- Me Se of of or ee of Result, 2thours. | pig. | MMO) Udder.) Milk. | Died. ; ! | E Sept. 22, 1908 | 3280 Intrap. Left | 10°0 ce. K. 256 days No tuberculosis. [before inocula- tion]. | 3281 os Right | 10-0 ce. Dz 118 ,, No tuberculosis. Death ; 3 during parturition. | 3282 <3 Mixed | 10:0 cc. K. 256 ,, No tuberculosis, | milk, ! | Sept. 24,1908 | Left half | 3288 Intrap. Left | 10:0 ce. K. 72 days No tuberculosis. [48 hours after | 70-0 ce. inoculation ]. 3284 . ” 10:0 ce. K. 72 ” ” ” Right half | 3285 ‘ Right | 100 ce. K, 12 i i “a 50°0 ec. 3286 5 7 100cc.} K. 72, ” ” Sept. 26,1908 | Left half 3291 Intrap. Left | 10-0 ce. K. 81 days No tuberculosis. [4 days after 91:0 ec. inoculation]. 3292 ” ” 100 cc,} K. 81 ,, ” » Right half | 3293 9 Right | 10-0 cc. K, 81, "y e 77-0 ce, 3294 - » |{100ce] K. 81 ,, i As Sept. 28,1908 | Left half 3297 Intrap. Left | 10-0 ce. K, 79 days No tuberculosis, [6 days after 68°0 ce. inoculation] 3298 ss + 10°0 ce. K. G9! 35 sf Sy Right half | 3295 . Right | 10-0 ce. K. 7D" 45 i 67°0 cc. , : 3296 3 10°0 ce, K. 79, 3 3 Sept. 30,1908 | Left half | 3303 Intrap. Left | 10-0 cc, K. 83 days No tub i [8 days ‘after 82:0 ec. patent inoculation ]. | 3304 3 +) 10:0 ec. K. 83 ,, 3 cs Right half| 3301 |° ,, Right | 10:0 ce. K. 83, 3 : | 70:0 ce, : 3302 3 i 10°0 ce. K, 83, - as Oct. 2, 1908 Left half | 3309 Totrap. Left | 10-0 cc. K. 96 days | Not i [10 days after 101°0 ce. fs oe inoculation]. 3310 ey $3 10°0 ce. K. 96, ‘i bid Right half| 3311 a Right | 10:0 ce. K. 96, 61:0 ec. i i 3312 3 +3 10-0 ce. K, 96, ; <5 Oct. 4, 1908 Left half | 3315 Intrap. Left | 10-0 ce. K, 93 d i [12 days after | 70°0 ce. . ae +) Me tnibenob lenis inoculation ]. 3316 a iy 10°0 ec. K. 93, i s Right half| 3313 55 Right | 10-0 cc. D. 81 , 40°0 ec, : o : 3314 is si 10°0 ec. K. 93 ,, ‘3 fe Oct. 6, 1908 Left half | 3319 Intrap. Left | 10-0 cc, K. 9 ; rid ave mien lego oa Pp 2days | No tuberculosis. inoculation }. 3320 59 $i 10'0 cc. K. 92 ,, ” ” Right half| 3317 4 Right | 10:0 ce. K. 92, 80°0 ce, ? " 3318 $5 % 10:0 ce. K, 92 , ‘ : 137 GUINEA-PIGS INOCULATED WITH THE MILK or Goat 75—continued. 2 \ Total yield | No.of | Mode Half Dose | Killed ‘ Date. of Milk in |Guinea-| of | __ of of oe | Dumtionice Result. . 24hours. | pig. |Injection.| Udder.| Milk. | Died. TatEes Oct. 14, 1998 Left half 3325 Intrap. Left | 10°0 ce. K 86d No tub losi [22 days after 80:0 ce. : ae eee inoculation]. 3326 Fe i 100cc. K, 86, 3 , Right half | 3327 os Right |100ce.| K ae ; : 70°0 ce. ; 3328 ‘i $5 10°0 ce. K. 86, ” ” Oct. 28, 1908 Left half | 3341 Intrap. Left | 10°0 ce. K. 98 days | No tuberculosis [36 days after 46:0 ce. inoculation}. 3342 zi ‘s 10°0 ec. K, 98, ¥ ey Right half] 3339 * Right | 10°0 ce. K. 98, Ps 5 About 21:0 cc. | 3340 _ » |100ce.! K, 98 # = Nov. 6, 1908 | Left half | 3350 Intrap. Left | 10:0 ce. K. 92 days | No tuberculosis. (45 days after | 11°0 ce. inoculation]. | Right half) 3349 ‘5 Right | 2°0 ce. Ke 92) is 2°0 ce. June 7, 1909 _— 3777 Intrap. Left | 10-0 cc. K. 77 days No tuberculosis. [258 days after inoculation, 3778 5 ss 10°0 ce. K. 77 3 5 8 days after giving birth to 3779 ay Right | 10°0 cc. K. qo 6 2 two kids]. 3780 7 35 10°0 ce. K, TT oss 4 ‘4 RABBITS INOCULATED WITH MILK OF GOAT 75. No. of Mode Half Dose Killed Daiation of | Date. Rabbit of of of or Life | Result. abit. | Tnjection.| Udder.| Milk. | Died. md } | Sept. 24, 1908 2016 Subcut. Left | 40-0 cc. K. 148 days | No tuberculosis. (48 hours after Intrap. 10°0 ce. inoculation]. 2017 Subcut. | Right | 30:0 ce. D. 9 4 | No tuberculosis. The cause of death was not apparent. ABSTRACTS OF THE CASES AND DETAILED RESULTS OF THE INVESTIGATIONS SUMMARISED IN PART II OF THE REPORT. Case 1—HEIFER-CALF 1479. Subcutaneous inoculation of culture derived from a guinea-pig inoculated with an emulsion of a bronchial gland from Calf 1325 (3rd passage), Virus H. 79. “JN.” Dose—100°6 milligrammes. Age at Inoculation—6 months. Killed when in good health—110 days after inoculation. ABSTRACT OF POST-MORTEM NOTES. (For fall details see the Report on Cases of Human Tuberculosis other than Lupus.) There was a cyst at the seat of inoculation and the adjacent glands contained caseous gritty masses. The suprarenal bodies contained nearly two dozen calcareous tubercles. The mesenteric. ileo-colic, and a few other abdominal glands contained discrete calcareous 27676 tubercles, the thoracic glands contained small cal- careous patches. One supramammary gland contained a calcareous grain. The mamma was normal. Some saline was injected into each of the mammary sinuses and a small quantity of turbid fluid recovered from each. No tubercle bacilli were demonstrated. in smear preparations made from the fluid. Y2 1 Q oO 8 GUINEA-PIGS INOCULATED. No. of Mode of Duration of Result = Guinea-pig. | Inoculation. Dose. Life. env Intrap. Small K. 62 days General tuberculosis. Mixed fluids from the two fore- | 3008 Durap y quarters of the mamma 3567 ‘i 5 K.62 ,, General tuberculosis, 5 Intrap. Slightl D. 58 General tuberculosis, Mixed fluids from the two hind- 3568 ALEAD laewere quarters | 3569 es ‘i K. 62 ,, General tuberculosis. ! Case 2.—HEIFER-CALF 1601. Subcutaneous inoculation of culture derived from the original material of Virus H. 143. “L.L.” through Guinea-pig 3536. Dose—68°0 milligrammes. Age at Inoculation—4 months. Killed when in good health—91 days after inoculation. ABSTRACT OF POST-MORTEM NOTES. {For full details see the Report on Cases of Human Tuberculosis other than Lupus.) There was a cyst containing thin pus and caseo- necrotic masses at the seat of inoculation; the adjacent prescapular gland contained a fibro-caseous mass, and one left prepectoral gland was fibro-caseous throughout ; and one cervical gland contained a caseo- calcareous nodule. Half a dozen minute grey tubercles with calcareous centres were seen in the lungs, and each suprarenal body contained seven or eight miliary caseous tubercles. A moderate number of minute caseous foci were seen in the ileo-colic glands. The mamma and supramammary glands were normal. Four cubic centimetres of salice solution were in- jected into each of the mammary sinuses and about 3 cc. of slightly milky fluid were recovered in each case. One or two tubercle bacilli were seen in smear preparations made from the fluids recovered from the left fore, right fore, and right hind quarters, none was seen in that from the left hind quarter. GuINEA-PIGS INOCULATED. No. of Mode of Duration of = Guinea-pig. | Inoculation. Dose. Life. Result, (The left fore quarter ... | 3931 Intrap. 3:0 ce. K. 70 days General tuberculosis. wa ae | The left hind quarter ... | 3933 #3 3 D.47 i, General tuberculosis. sai The right fore quarter ... | 3930 - v4 K.70 ,, General tuberculosis. The right hind quarter... | 3932 55 5 K.70 ., General tuberculosis. Case 3—HEIFER-CALF 1461. Subcutaneous inoculation of culture derived from the thoracic gland Virus H. 53. “ D.H.” (a). Dose—50°0 milligrammes. Age at Inoculation—6 months. of Calf 1367 (3rd passage), Killed when in good health—111 days after inoculation. ABSTRACT OF POST-MORTEM NOTES. (For full details see the Report on Lupus Viruses.) There was a cyst at the seat of inoculation con- taining turbid fluid and caseo-necrotic masses. The adjacent prescapular gland was mainly composed of caseo-calcareous tissue. One prepectoral and one cervical gland were similarly affected. In the thoracic glands there were a few scattered calcareous foci. The right supramammary gland con- tained a group of three caseous and softened tubercles. The milk sinuses contained a small quantity of opalescent fluid; in the fluid from one quarter a moderate number of tubercle bacilli were seen on microscopical examination. The mammary tissue was normal. GUINEA-PIG INOCULATED. _ No. of Mode of Duration of Guinea-pig. | Inoculation. Life. Result. With a few drops of the fluid from one quarter of the mamma. 3476 | Early general tubercu- Intrap. | Killed 33 days losis. | 139 Case 4—HEIFER-CALF 1535. Subcutaneous inoculation of culture derived from the original material of Virus H 53. “D.H.” (d), through Guinea-pig 3216.’ Dose—50:0 milligrammes. Age at Inoculation—About 3 months. Killed when in good health—103 days after inoculation. ABSTRACT OF POST-MORTEM NOTES. The supramammary glands contained altogether 10 caseous gritty nodules up to 2 mm. iu diameter. (For full details see the Report on Lupus Viruses.) : The mamma was normal; the milk sinuses of the There was a large thin-walled cyst at the seat of right fore quarter contained 2 or 3 cc. of purulent inoculation containing caseo-pus and caseous masses; fluid; the fluid contained yellow shreds which were the adjacent prescapular gland was enlarged, dense, casts of the smaller ducts; on standing these fell to and caseous throughout. The lungs showed on the the bottom, leaving a slightly turbid supernatant surface a very few minute calcareous tubercles and fluid. On microscopical examination of the pus some minute grey foci ; the spleen contained scattered tubercle bacilli were found, sparsely scattered ; one calcareous tubercles. was seen inside a leucocyte. There was no pus in the The Peyer’s patches of the small intestine con- other quarters of the udder. tained altogether several yellow foci and three small A quantity of saline solution was injected into the ulcers, slightly calcareous; several small ulcers left fore quarter and the hind quarters of the udder ; without caseation or calcification were also seen in with the fluids recovered from two of these quarters the intestines. Nearly every lymphatic gland in the guinea-pigs were inoculated. No tubercle bacilli were body contained caseous or calcareous tubercles or seen in smear preparations of the fluids recovered nodules more or less numerous. from the hind quarters. GuINEA-PIGS INOCULATED. No. of Mode of Duration of _ Guinea-pig. | Inoculation. Life. Result, With pus from the right fore quarter wee 3749 Intrap. Died 77 days Severe general tubercu- losis. With fluid recovered from the left fore 3750 Intrap. Killed 97. No tuberculosis. quarter : : With fluid recovered from the right hind (3761 Intrap. Killed 97 ,. No tuberculosis. quarter \s752 Intrap. Killed 97, No tuberculosis. Case 5.—HEIFER-CALF 1571. Subcutaneous inoculation of culture derived from the small mediastinal gland of Calf 1561 (let pas:age) Virus H. 53. “ D.H.” (6). Dose—50‘0 milligrammes. Age at Inoculation—3} months. Killed when in good health—111 days after inoculation. ABSTRACT OF POST-MORTEM NOTES. Two thoracic and five abdominal glands each contained ‘ one or more minute calcareous or caseous foci. (For full details see the Report on Lupus Viruses.) The mamma, and supramammary glands, were There was a small fibrous-walled cyst with caseous normal, and softened contents at the seat of inoculation. The adjacent prescapular gland was extensively caseo- Three cubic centimetres of saline solution were in- calcareous and softened, one cervical gland showed a jected into each of the mammary sinuses and about calcareous patch, and one prepectoral gland acalcareous _1 cc. of very slightly milky fluid was recovered in each focus. There were three small tubercles in the spleen case. No tubercle bacilli were seen in these fluids on and a caseous gritty nodule in one suprarenal body. microscopical examination. GUINEA-PIGS INOCULATED. No. of Mode of Duration of | ae | Guinea-pig. | Inoculation. Dose. Life. | Result, | | (The left fore quarter ... |. 3907 Intrap. Rather K. 62 days No tuberculosis. | less than With the 1:0 cc. fluids | The left hind quarter ... 3905 ¥ a K, 62 No tuberculosis. recovered } from— | The right fore quarter ... 3904 s is D.13~=—C*#, | No tuberculosis. | The right hind quarter... 3906 | et 4 K. 62. ,, | No tuberculosis. 140 Case 6.—HEIFER-CALF 1505. Subcutaneous inoculation of culture derived from the lung of Calf 1453, Virus H 107. “ H.H.” Dose—100 0 milligrammes. Age at Inoculation—5 months. Killed when in good health—101 days after inoculation. ABSTRACT OF POST-MORTEM NOTES. (For full details see the Report on Lupus Viruses.) There was a small fibroid local tumour containing a cavity filled with caseo-pus; the adjacent prescapular gland and one prepectoral gland were fibro-caseo- calcareous. Scattered ulcerated nodules containing yellow pus (smear, T.B.) were seen in the small intestine ; there was one in the large intestine. The mamma and the supramammary lymphatic glands were normal. Some saline was injected into each of the mammary sinuses and a small quantity of slightly turbid fluid recovered from each. No tubercle bacilli were seen in the mixed fluids from the two fore quarters or in that from the two hind. quarters on microscopical examination. GUINEA-PIGS JNOCULATED. No. of Mode of Duration of Resul ~ Guinea-pig. | Inoculation. Dose. Life. esutte \ 7 2 ‘0 ce. illed 28 d No tuberculosis. Mixed fluids from the fore-quarters } oan | Toney. | ater ieilled ay! 3 escheat a aeiclie Gb theamuarsee || 3708 ) 20cc, |Killed 44 ,. | : | 3705 Intrap. 25cc, |Killed23 ,, | F Mixed fluid from the hind-quarters} { | | 3706 , | Q5ec. |Killed 44 ,, — si { Case *,—HEIFER-CALF 1549. 6c H Dose—50°0 milligrammes. Age at Inoculation—10 weeks. Subcutaneous inoculation of culture derived from the mediastinal gland of Calf 1497 Virus H. 107. .H.” Killed when in good health—97 days after inoculation. ABSTRACT OF POST-MORTEM NOTES. (For full details see the Report on Lupus Viruses.) Slight general tuberculosis, apparently progressive in thoracic glands. There was a fibrous-walled cystic local tumour filled with caseo pus and caseous masses; the adjacent prescapular gland was enlarged and caseous. The liver, spleen and lungs contained scattered calcareous or caseo - calcareous miliary tubercles, the kidneys a few grey tubercles. The lungs also showed on the surface in certain limited areas patches of fibro-calcareous tubercles, and a few scattered caseous gritty nodules. In the right side of the heart about fifty grey nodules up to 6 mm. in diameter were counted, the majority with calcareous centres, and in the first few inches of the pulmonary artery several small fibrous tubercles were seen. The thoracic glands were much enlarged and mainly occupied by caseous gritty tracts and patches of calcareous tubercles : the portal and one coeliac gland were similar. In each suprarenal about a dozen grey nodules with caseo-calcareous centres were seen. Very numerous congested nodules each with a small central ulcer were seen in the small intestine and the Peyer’s patches were thickened, ulcerated and beset with calcareous patches and foci. The abdominal lymphatic glands not hitherto mentioned contained numerous calcareous tubercles, the mesenteric and ileo-colic glands being severely affected. An occasional tubercle was seen in the peripheral glands and in several haemo-lymph glands The marama was well-developed ; many of the milk ducts contained short plugs of purulent substance. No tubercles were seen in the organ. Saline was injected into the mammary sinuses ; the: fluid recovered was milky, and deposited on standing yellow pus composed of the plugs from the ducts. The fluids from the two fore quarters were mixed together, as were also those from the hind quarters. A few tubercle bacilli were seen in smear preparations made from the mixed fluids. No guinea-pigs were inoculated. Case 8. HEIFER-CALF 1577. Subcutaneous inoculation of culture derived from the bronchial gland of Calf 1551 (2nd passage), Virus H. 107. “ H.H.” Dose—50:0 milligrammes. Age at inoculation—3} months. Killed when in good health—103 days after inoculation. ABSTRACT OF POST-MORTEM NOTES. (For full details sce the Report on Lupus Viruses.) There was a fibrous-walled cyst with caseo-purulent contents at the seat of inoculation. The adjacent prescapular gland was extensively, and one prepectoral gland was entirely, caseous ; two other adjacent glands contained each a calcareous nodule. Each suprarenal body showed one miliary tubercle. The thoracic glands contained a few minute caseous or calcareous foci, and three abdominal glands contained one each. The mamma, and supramammary glands, were normal, ; Four cubic centimetres of saline solution were injected into each of the mammary sinuses and about 3-0 cc. of fluid (very slightly milky) were recovered in each case. No tubercle bacilli were seen in the fluids on. microscopical examination. 141 GUINEA-PIGS INOCULATED, == No. of Mode of Duration of Guinea-pig..| Inoculation. Dose. Life. Result, | ( The left fore quarter 3916 | Intrap. °} 3°0 cc. K. 60 days No tuberculosis, With the | ; finids | The left hind quarter’ ... 3914 3 K, 60 ,, No tuberculosis, recovered : from | Phe right fore quarter... 3917 | 08 a D.28 , No tuberculosis. The right hind quarter... 3915 | ‘9 “ D.16_,, No tuberculosis, Case 9. HEIFER-CALF 1583. pabenta nous saoulavion of culture derived from the mediastinal gland of Calf 1549 (1st passage). ‘Virus H. 107. “ Dose—50:0 milligrammes. Age at inoculation—11 weeks. Killed when in good health—127 days after inoculation. ABSTRACT OF POST-MORTEM NOTES. | (For full details see the Report on Lupus Viruses.) There was a small cyst containing caseous masses and turbid fluid at the’seat of inoculation, the ad- jacent prescapular gland contained caseous and caseo- calcareous nodules, and one cervical gland was beset with calcareous foci. One suprarenal body contained a minute calcareous focus. The thoracic and two or three abdominal lymphatic glands contained one or more calcareous foci, mostly minute. The udder was small and showed very little glandular tissue; the supramammary glands were normal. Four cubic centimetres of saline solution were in- jected into each of the mammary sinuses and about 3-0 cc. of fluid were recovered from each. No tubercle bacilli were seen in these four fluids on microscopical examination, GUINEA-PIGS INOCULATED. No. of Mode of Duration of = | Guinea-pig. | Inoculation. Dose. Life. Result, | | (The left fore quarter ... | 3912 | Intrap. , 3:0 ce. K. 56 days No tuberculosis, Laces | The left hind quarter ... | 3910 | 5 5 K. 56, No tuberculosis. : 5 { recovered The right fore quarter... | 3913 | ss | “ K.56 No tuberculosis. from— & f The right hind quarter... 3911 | 7" ‘ 35 K. 56 No tuberculosis. | | Case 10—HEIFER-CALF 1538. “ALR Dose—50°0 milligrammes. Age at Inoculation—About 4 months. Subcutaneous inoculation of culture derived from the mediastinal gland of Calf 1417, Virus H. 108. Killed when in good health—98 days after inoculation. ‘ABSTRACT OF POST-MORTEM NOTES. (For full details see the Report on Lupus Viruses.) Slight retrogressive generalised tuberculosis. There was a large thin-walled cyst at the seat of noculation containing caseo-pus and caseous masses ; the adjacent prescapular gland was large and composed of caseous gritty substance. The lungs contained -sixteen small nodules with caseo-calcareous centres, the spleen a moderate number of fibro-caseous gritty tubercles, the liver similar but less numerous tuber- -cles, the right kidney two caseo-calcareous tubercles, the left a minute focus, the right suprarenal ‘body two caseous gritty nodules, the left a caseous tubercle. Nearly every Peyer’s patch in the small intestine contained a few caseo-calcareous tubercles. All the lymphatic glands in the body contained caseo- calcareous tubercles more or less numerous, in some cases confluent ; several mesenteric and the ileo-colic glands were more than half caseo-calcareous. Every haemo-lymph gland examined contained a casec- calcareous tubercle. The mamma was normal. Some saline was injected into each of the mammary sinuses and small quantity of slightly turbid fluid recovered from each. No tubercle bacilli were seen in the mixed fluids from the fore quarters or in that from the hind quarters on microscopical examination. 142 GUINEA-PIGS INOCULATED. No. of Mode of Duration of Result. == Guinea-pig. | Inoculation. aeee Life. Mixed fluids from the fore quarters of 3736 Intrap. Small | Killed 72 days! Slight general tuber- the mamma . < culosis. ina Mixed fluids from the hind quarters of 3737 Intrap. Small | Killed 72 _,, No tuberculosis. the mamma Case 11.—HEIFER 80. Subcutaneous inoculation of an emulsion made Virus B. V. Dose—4,500,000 tubercle bacilli (Vol. 2°32 cc.). Age at Inoculation—About 18 months. from the original material (a mediastinal gland) of Killed when in good health—62 days after inoculation. ABSTRACT OF POST-MORTEM NOTES. (For full details see Vol. I of the Appendix to the 2nd Interim Report, page 198.) Slight generalised tuberculosis. The tumour at the seat of inoculation was mainly composed of dense caseating tissue; the adjacent glands were partly fibroid and contained caseous foci and patches. The lungs showed sparsely scattered congested nodules containing yellow foci up to a large pea in size. Caseous tubercles were scanty in the mediastinal and numerous in the bronchial glands. There were three caseating tubercles in the spleen, and five greyish nodules each containing a minute caseous focus in the liver. One pancreatic gland contained a group of small yellow tubercles. There was no sign of tuberculosis élsewhere. The uterus contained a 5 months old foetus. A guinea-pig inoculated with an emulsion of its spleen remained healthy. The mammary sinuses contained a small quantity of thick yellowish turbid fluid; a smear preparation. made from it showed no tubercle bacilli. GuInEa-PIG INOCULATED. = No. of Mode of Duration of | Result Guinea-pig, | Inoculation. Life. eeulbs With a small quantity of the fluid from the 816 Killed 88 days | General tuberculosis. mammary sinuses | Intrap. Case 12. HEIFER-CALF 1569. Subcataneous inoculation of culture derived from the original material of Virus H. 127. “R.R.” (a): through Guinea-pig 3475. Dose—50°0 milligrammes. Age at inoculation—About 12 weeks. Killed when in good health—105 days after inoculation. ABSTRACT OF POST-MORTEM NOTES. (For full details see the Report on Cases of Human Tuberculosis other than lupus.) Slight general tuberculosis, apparently retrogressive. There was an ulcerated fibro-caleareous tumour at the seat of inoculation and the adjacent glands were fibro-caseo-calcareous or contained discrete tubercles. The lungs showed a moderate number of tubercles up to 2 mm., the smaller grey the larger caseous and calcareous, and also a few consolidated lobules which were fibro-calcareous. The spleen contained a mode- rate number of miliary caseo-calcareous tubercles, the liver scattered calcareous tubercles. \ The thoracic and abdominal glands contained scat- tered calcareous tubercles and calcareous patches, the peripheral glands caseous gritty miliary tubercles. The udder was well formed but not very large ; it appeared normal. The supramammary glands were normal. Thin grey purulent fluid was expressed from the teats of three of the quarters ; the sinuses and ducts of the right hind quarter contained a large quantity but there was only a small quantity in each of the fore quarters ; microscopical examination of these fluids showed a few tubercle bacilli in each case From the left hind quarter no pus could be ex- pressed and this quarter was washed out with salt. solution ; microscopical examination of the recovered. fluid showed no tubercle bacilli. \ 143 Case 13.—GOAT 77 [Young Adult Female]. Subcutaneous inoculation of culture derived from the (sputum) through Guinea-pig 3343. Dose—25-0 milligrammes. Date of Inoculation—February 12, 1909. original material of Virus H. 118. “F.C.” Killed when in good health—June 8, 1909. [116 days after inoculation. ] Clinical Notes. A large tumour developed at the seat of inoculation on the left side of the neck. On March 8, 24 davs after inoculation it extended from the midde of the neck into the dewlap ; it was soft and fluctuating, and the skin over it was thin and red. The adjacent prescapular gland was enlarged. Twelve days later the tumour burst and its contents were evacuated. On April 5, 52 days after inoculation, the goat gave birth to two dead full-time foetuses ; on post-mortem examination their organs and glands were found to be normal ; a smear made from a portal gland in each case showed no tubercle bacilli. Milk withdrawn from the udder by means of a catheter eight hours, one day, two days, 8, 14, 28 and 59 days after the birth of the kids, was inoculated into guinea-pigs. The milk withdrawn from each half of the udder on April 5 (eight hours after parturition) was examined microscopically ; it was thick and ereamy-yellow in colour.; no tubercle bacilli were seen. Centrifuged deposit from the milk was also microscopically examined with a negative result. Culture tubes were sown with the milk ; no growth resulted. Two days after the kids were born centrifuged deposits of milk from the right and the lefi halves of the udder were again examined microscopically ; no tubercle bacilli were seen. The goat remained in good health during the experiment. Temperature. The temperature was raised for a period of 19 days after the inoculation (maximum 40°5° C. on the 7th day) ; from the 19th to the 27th day it was irregular ; subsequently it was normal, the range of variation during three months being only 1:0 C. (maximum 39-0°, minimum 38°0° C.). Weights. qrs. Ibs. February 12, 1909 aie nt 2 19 June 8, 1909 se dios ibe 2 9 Loss of weight—10 lbs. POST-MORTEM EXAMINATION. The carcass was in good condition. Local Lescon.—At the seat of inoculation there was a tumour measuring 6°/ by 4 by 2 cm. composed of caseous gritty nodules and masses set in fibrous tissue ; the skin over it showed an ulcer 4:5 by 2 cm. in area with red granular floor showing dry haemorr- hagic scabs and thickened inverted margins. Left Prescapular Gland.—The left prescapular gland measured 4°5 by 2 by 1:5 cm. and showed in the cortex scattered caseo-calcareous nodules ranging from 2 to 7 mm. in diameter. At the inferior extremity of the gland there were several discrete caseo-calcareous nodules in loose connective tissue. Right Prescapular Gland.—The right prescapular gland measured 3°3 by 1:4 by 1 cm. and showed in the cortex a small group of about half a dozen caseo- calcareous tubercles. Cervical Glands.—On the left side one of the lower cervical glands contained a pea-sized caseo-calcareous nodule and several small tubercles. The rest were normal. Lungs.—The lungs were crepitant and collapsed normally. They showed under the pleura sparsely scattered, evenly distributed, uniformly caseous gritty tubercles ranging from about 0°5 to rather more than 1: mm. in diameter. Here and there was a tubercle with a grey margin and caseo-calcareous centre. There was no sign of disease elsewhere. The mammary gland was normal. GUINEA-PIGS INOCULATED WITH THE MILK OF GOAT 77. (All the milk was drawn with a catheter.) Total 5 yield of NG: of Mode of Hale Hass. |. Eulted Duration of Date. Milk i Guinea- Tiiection of of or Life Result. Sd hous, | Be | | Widder | Milk, | Died: : April 5, 1909 Left 3677 Intrap. Left 10°0 ce. D. 35 days No Ce ) cause 52 days after 115°0 ce. of death, See 3678 7 53 10°0 ce. D. 30 No ee, Death 8 hours after probably from injuries. giving birth to Right 3679 5 Right | 10-0 ce. K. 36 C,, No tuberculosis. two dead 170°0 ce. foetuses ]. 3680 5 sj 10°0 ce. Dz 33 C,, No tuberculosis. ? cause of death. April 6, 1909 Left 3687 Intrap. Left | 10°0 ce. K, 51 days No tuberculosis, 53 days after 140°0 cc. Leonie ‘ 3688 3 5 10:0 ce. K, 51, a ‘ 1 day after ; parturition ]. Right 3685 5 Right | 10:0 ce. K. 51, A i 220°0 ce, 3686 PA = 10:0 cc. K. 51 5 3% April 7, 1909 Left 3695 Intrap Left | 10-0 ec. K. 56 days No tuberculosis. 54 days after 100°0 ce. SE aeraHan ; 3696 95 a 10°0 ce. K. 56, is ns 2 days after : parturition ]. Right 3693 ‘ Right | 10:0 ce. K. 56; ‘ os 150°0 ce. 3694 3 ai 10:0 ec. K. 56 ,, ‘ Z 27676 144 GUINEA-PIGS INOCULATED WITH THE MILK or Goat 77—continued. Total : i cee No. of Mode Half Dose | Killed : Date. eld of | Quinea- of of of or ee of Result. 24 ae pig. | Injection.} Udder. | Milk. | Died. April 13, 1909 Left 3701 Intrap. Left | 10-0 ce. K. 56 days | No tuberculosis. [60 days after 160°0 ce. inoculation ; 3702 ” ” 10°0 ce. K. 56 ” ” ” 8 days after ; parturition ]. Right 3703 ‘3 Right | 10-0 cc. K. 56, ” ” 220°0 cc. 3704 ca ” 10°0 cc. K. 56 ” ” ” April 19, 1909 Left 3722 Intrap Left | 9:0 cc. K. 53 days No tuberculosis [66 days after | 229-0 cc. inoculation ; 3723 by 100 ce. K. 53, , ” 14 days after parturition]. Right 3720 “ Right | 10-0 ce. K, 53, ” ” 280°0 ce. 3721 ‘s 10°0 ec. K. 53, ‘4 7 May 3, 1909 Left 3745 Intrap. Left | 10-0 ce. K. 74 days No tuberculosis [80 days after | 240-0 ce. inoculation ; 3746 a 10°0 ce. K. the, ” ” 28 days after parturition ]. Right 3747 3 Right | 10:0 cc. K, 74 yy ” ” 300°0 ce. 3748 “ ij 10°0 ce. K. TA: 5 4s i June 3, 1909 Left 3769 Intrap Left | 10:0 ce. K, 55 days No tuberculosis [111 days after | 190-0 cc. inoculation 3770 9 ‘ 10°0 ce. K. 55g +5 6 59 days after parturition ]. Right 3767 a Right |} 10:0 ce. K, 55, 8 3 230°0 cc. 3768 10°0 cc. K. 55, 4 45 145 INVESTIGATION OF TUBERCLE BACILLI FROM CASES OF SWINE TUBERCULOSIS BY F. GRIFFITH, M.B. AND A, STANLEY GRIFFITH, M.D, 27676 2A 146 INVESTIGATION OF TUBERCLE BACILLI FROM CASES OF SWINE TUBERCULOSIS. TABLE OF CONTENTS PAGE Introduction sa oe Ste < ee ‘igs Nee see ... 148 Consideration of Results ... — Ly ee age Bre ane .. 149 The Group from which Bovine Tubercle Bacilli were Isolated... se 149 The Group from which Human Tubercle Bacilli were Isolated ... wes LAS The Group from which Avian Tubercle Bacilli were Isolated ... ws LOO Mixed Virus... oie ane sais er ee wins dee wee LOL Viruses from which Cultures were not obtained ... ee ees ve, “151 Summary ... sa wine eve “as ie sin wee ‘eg! A Conclusions ... ae is ae vee ste Mis ae .. 152 Table of Origins ... sia ee ie ee ae dn Pha tex. 153 Cultural Characters of, and General Results of Inoculation Experiments with Tubercle Bacilli of Porcine Origin :— Cultural Characters of Tubercle Bacilli of Porcine Origin... son ss» 168 General Summary of Inoculation Experiments on Rabbits with Tubercle Bacilli of Porcine Origin ... ois ‘see an ie as we 10D General Summary of Inoculation Experiments with Tubercle Bacilli of Porcine Origin on Calves, Pigs, Guinea-pigs, and Rats... wr ise G2 Tabular Summaries of the Inoculation Experiments : On Calves with Cultures of Virulent Tubercle Bacilli of Porcine Origin ... sats ane 174 On Calves with Cultures of Slightly Virulent Tubercle Bacilli of Porcine Origin a mee 175 On Pigs with Cultures of Slightly Virulent Tubercle Bacilli of Porcine Origin aie Sis 176 On Rabbits with Cultures of Virulent Tubercle Bacilli of Porcine Origin... se oe 177 On Rabbits with Cultures of Slightly Virulent Tubercle Bacilli of Porcine Origin... a 191 On Rabbits with the Original Material of the Porcine Viruses... ae a Si 196 On Guinea-pigs with Cultures of Virulent Tubercle Bacilli of Porcine Origin nae z.. 198 On Guinea-pigs with Cultures of Slightly Virulent Tubercle Bacilli of Porcine Origin PAS 204 On Gninea-pigs with the Original Material of the Porcine Viruses ... site vee uy 205 On Rats with Virulent Tubercle Bacilli of Porcine Origin ane aoe joe dis 207 On Rats with Slightly Virulent Tubercle Bacilli of Porcine Origin sea ae ase 209 147 Charts of certain of the Viruses and the Post-mortem Notes of the Larger Animals Inoculated with them :— PAGE Of Virus P. IV. 911 » » PY. ase ass a i oy Fa VG sis ss eee 178 aa 225 yo an BE NIIL was waa ate ie oe ae es ave 227 Fie RINE de i 2 as ie me A a wee 229 ip Jy ee, ava bea 4 es ise a we 232 an ayo Re XID we o ez we sg nes ue sae we 234 wo Pe KTVisw ave wea aes “9 oes a6 de .» 240 » » DP. XXIX. aus nts aed oes dias ba sa w. 250 Win Sin “PecX Dees, ea ae aay eae oss ae evs eva 252 » » P. XLVI. oat an a ae sis a st we 261 Nore.—The remaining viruses were inoculated into Rabbits and Guinea-pigs only. These experiments are not charted ; the results will be found in the tabular summaries on pages 177-206. 27676 toa 2A2 148 INTRODUCTION. Tuis report gives the results of the inquiry into the nature of the tubercle bacilli concerned in the production of tuberculosis in swine, and is based upon the investi- gation of material from 63 cases of natural tuberculosis. The material used was taken from swine which had been found to be tuberculous at the slaughterhouse, the extent to which these animals were diseased ranging from tuberculosis seemingly limited to certain lymphatic glands to tuberculosis generalised throughout the body. Out of the total number of swine slaughtered at the Metropolitan Meat Market during the period from March, 1905, to October, 1906, those showing apparently localised tuberculosis were specially sought for to furnish material for investigation. In addition material was obtained from cases of general tuberculosis of all degrees of severity, and the cultures obtained were isolated sometimes from the submaxillary glands, sometimes from metastatic lesions. The cases which were considered to have been localised include all those in which the examination ordinarily adopted at the slaughterhouse revealed no obvious tuber- culosis elsewhere. But there must be remembered the possibility of early lesions having been overlooked, and indeed it is difficult to believe that such extensive local tuberculosis as was found in Viruses XX XTIL, XXXVI; LL, and LIL, could exist in young animals such as these without any dissemination having occurred, In 21 out of the 63 swine from which tuberculous material was obtained the disease was stated to be localised ; in nine no details were given as to the condition of the rest of the body ; the remaining 33 showed generalised tuberculosis varying in severity, This series shows clearly that natural tuberculosis in swine arises by infection from the alimentary tract, and that the disease when localised is usually limited to the glands situated near the angle of the jaw. These, the submaxillary glands, have been most often used as the source of cultures both because the disease was some- times limited to them and also because they were the glands nearest the site of entry of the tubercle bacilli to the alimentary tract. Cultures were obtained altogether from 59 of the cases ; the sources from which they were isolated are as follows :—Submaxillary glands (44), bronchial glands (4), spleen (2), mesenteric glands (2), submaxillary and bronchial gland (1), skin, bone, joint, lung, udder, and inguinal gland (each 1). From the remaining four viruses cultures were not obtained, nor did the animals inoculated with the original material develop tuberculosis. Tn certain instances, when several animalsin a single herd have been tuberculous, specimens have been taken from one or two individuals only. Metuop oF INVESTIGATION. The specimens on arrival were carefully examined and the tuberculous lesions which appeared most suitable for experimental purposes were selected. From this material emulsions for cultures and animal inoculations were made in the usual way ; the surface of each part selected was seared with a Paquelin’s cautery and pieces were removed with sterile instruments and emulsified with salt solution, the procedures being the same as those described in a previous report (Appendix to 2nd Interim Report, Vol. II, page 3). In all cases cultures were sown direct from the original material, and guinea-pigs were almost invariably and_ rabbits occasionally inoculated with emulsions of the fresh material. ’ The pathogenic effects of the various viruses have been investigated by the inoculation of culture obtained in all but four instances direct from the original material ; the inoculations of guinea-pigs and rabbits with emulsions of the original material were made with the object of obtaining a culture from them should the direct culture fail, as well as to afford some preliminary indication of the virulence of the bacillus. The virulence of the cultures of all the viruses has been tested on rabbits and guinea-pigs and cultures of fifteen viruses have also been inoculated into calves. Rabbits have been systematically employed in place of calves for testine the pathogenic properties of the cultures since it has been established in previous investigations that, when a virus shows full virulence for rabbits by the various methods i: inoculation, it will be capable also of setting up progressive tuberculosis in the calf. 149 CONSIDERATION OF RESULTS. In this investigation a study has been made of the cuitural characters and virulence ‘of tubercle bacilli obtained from 59 cases of tuberculosis occurring naturally in swine. The bacilli isolated from these cases have manifested wide variations in the ‘character of their growth on artificial media and in their virulence for animals, and -can be divided into three groups clearly distinguished from each other. The group from which bovine tubercle bacilli were isolated. In this group, by far the largest of the three, are included 40 out of the total ‘number of swine viruses investigated. The cultures isolated correspond exactly with these obtained from cases of natural tuberculosis in the ox. They vary in capacity for growth on artificial media within the same limits as the bovine tubercle bacillus and exhibit the same high degree of virulence when inoculated into calves, rabbits, and guinea-pigs. All the 50 cultures have been inoculated into rabbits (intravenously, intraperi- toneally, and subcutaneously), and guinea-pigs (intraperitoneally and subcutaneously), and have produced in these animals without exception fatal general tuberculosis. In addition seven have been inoculated subcutaneously into calves in doses of 50 milligrammes, and, with one exception, have caused severe and fatal tuberculosis within 48 days identical in all its features with that set up by bovine tubercle bacilli. The exceptional result occurred with the culture derived from Virus P. XIV. (see table, pages 240-241), which gave rise in four calves to general tuberculosis, much less severe than that ordinarily produced by 50 milligrammes of bovine tubercle bacilli. A culture isolated from one of these calves was inoculated into. two more calves, each in a dose of 50 milligrammes, and caused fatal. general tuberculosis within the usual period. Apparently the culture from the original material, which was slightly though definitely less virulent than any bovine tubercle bacillus, became possessed of the full virulence of a bovine tubercle bacillus after a single passage through a calf. The group from which human tubercle bacilli were isolated. This group contains three viruses, P. [V., P. XLIL., and P. XLVL, obtained from swine in which the disease was localised in the submaxillary glands. The cultures of these viruses grow much more luxuriantly on media containing glycerin than any of those in the preceding group and resemble closely the more easy growing cultures -of human origin. They have a correspondingly low degree of virulence for the calf and rabbit, though one, Virus P. 1V., appears to be slightly more virulent than the -other two. . ~ Virus P. XLI. and Virus P. XLVI. were inoculated subcutaneously each into two calves, the doses being 100 milligrammes and 50 milligrammes in one case, and 75 milligrammes and 50 milligrammes in the other. The calves were killed in from 98 to 105 days and each showed a local lesion and slight tuberculosis of the nearest glands only. A series of rabbits was inoculated with culture from each of the two viruses, intravenously, intraperitoneally and subcutaneously (pages 191-195) : a few of the rabbits died prematurely, the rest increased in weight and when killed after periods varying from 99-174 days were found to have slight and non- progressive tuberculous lesions. : be The culture of Virus P. IV. was inoculated subcutaneously into two calves in doses of 50 milligrammes and 34 milligrammes. The former, Calf 312, killed after ‘a period of 90-days, showed a small fibrocaxseous local lesion with caseation of - the ‘prescapular gland, tuberculous broncho-pneumonia of the dorsal parts of the caudal 150 lobes of the lungs and several tubercles in the suprarenal bodies and in a few of the lymphatic glands. In the second calf, Calf 314, killed after 135 days, there was a cystic local lesion and caseation of the nearest glands, a tubercle in a mediastinal gland, and a few gritty foci in the mesenteric glands without any disease elsewhere. A third calf, Calf 350, was inoculated with the culture after it had been a longer period in cultivation (total period 183 days) in a dose of 50 milli- grammes subcutaneously, and when killed after 90 days showed a cystic local tumour, caseation of the nearest glands and a few foci in the mediastinal and mesenteric glands without any disease elsewhere. The culture from the original material of Virus P. IV. was also inoculated into a series of rabbits intravenously, intraperitoneally and subcutaneously (pages 191-195) : one rabbit, inoculated intraperitoneally with a dose of 10 milligrammes died in 18 days of tuberculosis ; another died of chronic general tuberculosis in 333 days after an intraperitoneal inoculation of 1:0 milligramme ; the rest were killed after a period of about 90 days, and showed generalised tuberculosis rather more severe than that usually produced by a slightly virulent virus of human origin. The virulence of these three viruses was tested also on the pig. Two pigs, one, Pig 166, inoculated subcutaneously, the other, Pig 172, fed, each with 50 milligrammes of culture of Virus P. XLI. were killed after 102 and 98 days respectively : the animal that was fed showed only slight apparently retro- gressive tuberculosis ; the inoculated animal, Pig 166, had, in addition to local disease, a severe tuberculous broncho-pneumonic condition of the lungs with very little disease elsewhere. From the lung of Pig 166 a culture was isolated directly and used to inoculate rabbits and a second pig, No. 202. Pig 202 received 50 milligrammes subcu- taneously, and died after 64 days of tuberculous consolidation of the lungs, the condition being very similar to that in the first pig. From the lung of Pig 202 an emulsion and a culture were made. The emulsion was inoculated subcutaneously into Pig 228, which was killed when well after 166 days, and showed slight non-progressive tuberculosis ; the culture was inoculated into rabbits and into a third pig, No. 248. Pig 258 was inoculated subcutaneously with 50 milligrammes of the culture, and died of tuberculous broncho-pneumonia in 103 days ; from the lung a culture was isolated which was inoculated into rabbits. This final culture, as well as the intermediate cultures, had only slight virulence for rabbits, and grew in the same luxuriant manner as the original culture. This virus therefore exhibited higher virulence for the pig, though not for other animals, than the human tubercle bacillus, a virulence which was not increased by passage through the body of the pig The other two viruses (P. XLVI. and P. IV.) displayed no higher virulence for the pig than the human tubercle bacillus. Two pigs, Pig 164 inoculated subcutaneously, Pig 168 fed, each with 50 milli- grammes of Virus P. XLVI. were killed after 97 and 60 days respectively, and showed only slignt retrogressive tuberculosis. The virulence of Virus P. IV. was tested on four pigs; one was inoculated and ‘one fed with culture from the original material, and one inoculated and one fed with culture from Calf 312, the dose in each case being 1 milligramme. All were killed after about four months and showed slight retrogressive tuberculosis. Cultures from each of the three viruses in this group have also been inoculated into guinea-pigs (subcutaneously and intraperitoneally) and into fowls (Virus P. IV. by feeding, Viruses P. XLI. and P. XLVI. intravenously), All the guinea-pigs developed fatal general tuberculosis : the fowls remained well and when killed were free from tuberculosis. The group from which avian tubercle bacilli were isolated. The five viruses, P, IL, P. IIL, P. XXL, P. XXVIL, and P. XLIV., included in this group were, like those in the preceding group, obtained from swine in which the disease was seemingly limited to the submaxillary glands. Cultures, isclated in each case directly from the glands, exhibit marked differences from those in the first two groups and possess all the properties of the avian tubercle: bacillus. 151 The cultures have been compared, as regards their virulence for animals and manner of growth on artificial media, with cultures of avian tubercle bacilli isolated from birds with natural tuberculosis and found to be identical. The experiments with the different strains of avian tubercle bacilli are fully considered in a separate report. See Vol. IV. of this Appendix. Mixed Virus. In the case of Virus P. XLII. the culture isolated from a pig’s submaxillary gland was at first considered to be a pure culture of the bovine tubercle bacillus ; a fourth subculture on glycerin agar showed, however, besides dry warty colonies a few shiny translucent colonies ; by repeated subculture and incubation at 42° C. a pure culture of the avian tubercle bacillus was separated. The original culture caused fatal tuberculosis in rabbits and guinea-pigs identical with that produced by the bovine tubercle bacillus. . The avian culture was inoculated into two fowls and two guinea-pigs. Both fowls, one inoculated intraperitoneally, the other subcutaneously, died of general tuberculosis identical with that produced by cultures of avian tubercle bacilli derived from birds. One guinea-pig, No. 2489, inoculated subcutaneously with 200 milli- grammes, showed merely a scar at the seat of inoculation when kilied after 114 days ; the second, No. 2490, inoculated intraperitoneally with 1:0 milligramme, died in 80 days, and showed at the post-mortem examination softening of the sternal and portal glands and grey foci in the liver and kidneys ; the culture isolated from the sternal gland was typically avian. VIRUSES FROM WHICH CULTURES WERE NOT OBTAINED. ' Included under this head are Viruses P. XLVIII., XXVIII, XXXV., and XLV. Virus P. XLVIIL. was one of two received on the same day and obtained from a herd of swine, several of which had tuberculosis ; a rabbit inoculated intraperitoneally with an emulsion of the original materia] developed progressive general tuberculosis, but cultures were not obtained. Virus P. XXVIII. was obtained from an old sow which had enlarged calcareous submaxillary glands with no sign of tuberculosis elsewhere ; although tubercle bacilli were fairly numerous direct cultures failed, and none of the animals (two rabbits and a guinea-pig) inoculated with an emulsion of the original material developed tuberculosis. Cultures from Virus P. XX XY. also failed, nor did any of the animals (a rabbit and a guinea-pig) inoculated develop tuberculosis, although tubercle bacilli were fairly numerous in smear preparations from the original material. Cultures from Virus P. XLV. failed through ali the tubes becoming contaminated ; the rabbit inoculated from the original material died in 10 days from psorospermosis. SUMMARY. The material*used in the investigation has been obtained from 63 cases of tuber- -culosis occurring naturally in swine; in 21 of these the disease was apparently localised to the lymphatic glands (submaxillary and mesenteric) of the alimentary tract from which the material used for investigation was taken ; in 33 the disease was generalised ; in the remaining nine the condition of the rest of the body could not be ascertained with certainty. In every animal in the series with two exceptions the disease appeared to be alimentary in origin. In these two cases, P. VI. and P. XVI., the condition of the glands in connection with the alimentary tract had not been ascertained. Cultures have been isolated from 59 out of the 63 cases, either directly from the original material or through the guinea-pig ; the source of the culture was, in the majority of instances, the submaxillary glands (44), but cultures have been isolated from various metastatic lesions (15). From the remaining four cases cultures were not obtained. 152 The cultures trom the 59 cases can be divided according to their cultural characters and virulence into three groups ; in the first group there are 50 cultures: . which are identical with bovine tubercle bacilli with the exception of one, Virus. P. XIV., which is definitely less virulent for calves and rabbits than any bacillus derived from a bovine source ; the second group’ contains three, Viruses P. IV... P. XLI., and P. XLVI. which correspond to the slightly virulent tubercle bacillus of human origin (the human tubercle bacillus) ; in the third group there are five, Viruses P. IT., P. IIL, P. XXI., P. XXVIL, and P. XLIV., which exhibit all the. characters of the avian tubercle bacillus. From the remaining culture, Virus P. XLII., which was found to be a mixture, pure cultures of bovine and avian tubercle bacilli have been separated. These figures cannot be taken as representing the relative frequency with which in nature swine tuberculosis is caused by the three types of tubercle bacilli mentioned, since, as stated in the introduction, some amount of selection was exercised in obtaining material. A consideration of the extent of the disease in relation to the type of bacillus. isolated shows that in none of the swine from which either human tubercle bacilli or avian tubercle bacilli in pure culture were obtained was there evidence of generalised tuberculosis, the disease in every instance being localised in the- submaxillary glands. | ‘When the tuberculosis is slight in extent and limited to the submaxillary glands, it is impossible from the appearance of the lesions alone to decide what particular type of bacillus produced it. This will be seen from a comparison of the descriptions. of the original material of the viruses from which the avian and human tubercle bacilli were isolated with those of Viruses P. XXXVIIL, P. XXXIX., P. XLIIL, and P. XLVIL., from which bovine tubercle bacilli were obtained. Out of the total number of 50 swine from which bovine tubercle bacilli were- isolated, 32 showed generalised tuberculosis, in 9 the tuberculosis was regarded as localised, and in 9 no details as to the condition of the rest of the carcasses could be- obtained. CONCLUSIONS. 1. Natural tuberculosis in swine may be the result of infection either with the bovine. tubercle bacillus, the human tubercle bacillus, or the avian tubercle bacillus. 2: The bovine tubercle bacillus is the most common cause of tuberculosis in swine. and has been found in every case in this series where there was disseminated. tuberculosis, as well as in more than half (10 out of 18) of those in which, so. far as could be ascertained, the disease was localised. 3. The tuberculous lesions in the pig’s lymphatic glands caused by the human. tubercle bacillus or the avian tubercle bacillus cannot be distinguished from the lesions sometimes produced by the bovine tubercle bacillus. F. Grirriru. uf A. STANLEY GRIFFIru.. 27676 153 TABLE OF ORIGINS OF PORCINE VIRUSES. “purely [eIqouoig *10}OVIVYO IL[IMIS JO Vjnpou s[AuIs v puels Telqouorqg BS saynpou Asvayo 494113 pourejuoo spuels Aareypixemqns oxy, *‘pesiepue sjutol oq} puv ‘ormommneud ssun{ ‘ssid-vauiny ‘sey pue purely oYy ‘snofnoraqn, 19M SpUeTs TeUINSUT oy} 4eq) poleis sem AT “ALOS “‘VoATCW 9193") “OULAOg ‘Suqqey ‘FLVeQ Arey ixemqug GB WOIZ spuLys [VIOUCIG pue IE[[IXeUIGNS 94} 910M paatooor suatITOedy CO6L ‘8I Tady ueqpodoray ‘syey pue ‘ssid - vous) ‘OUSSI1} PUBS [VULIOU UI pappaquis sapo1eqny snosievo[eo-oasea 94010 ‘suiqqey =‘ [Mog “SIP Poj¥[OSI Poule}UOD ‘padiel[Us Jou ‘spuL[s OY, ‘eLaqMesT[I sIsOTNO I ‘Soq I ‘9%90 | “purely aq nN} JO USIS OU UMOTS OABY 0} poztodar seM TOIT PTO syyUOM UsAds “‘qoyleyy 9T99e%O ‘CIT dnory) uewunp ‘SSIg fp ‘soareg 9g Areyprxewqng qnoqe sid v Wor spuLps Are][IxeMIQns 94} 919M paalaved suautoeds eq, GOGL ‘Te Yorryl ue odorz0yE ‘Al ‘d “SqUy ‘spurs aq} JO au0 TOI] IvoTUS pue ‘s3rd - voumy @ UL pUNOF 910M T][IOVY B]OIEQN} Mey W “paartasqo atam UOLZeVdYgTOTvO ‘sqiqqvy ‘sL Mog ‘Soq Ajree Jo SUSIs YOIYM Jo o1UID oY} UL WoNvosea Jo syvorys as1v00 T ‘sqeQ 7 ‘AoHUOTT “purty TOT}O9S UO PIMOYS SPULLs soy], “a1ATMOSTS osvesIp Jo UTIs ou MMOS ‘QOYIB]L 21992O ‘URIAY I ‘stg T ‘soateg % Axey[rxeuquyg avy OF poz10ded plo syjUOMI xIs qnoqe sid v Woy spuels LIV] [IxeMQng GOBL ‘OL Youre py ueqrpodorj.eyq "TIT “ad ‘sey ‘T][Tovq epor1eqny Jo stoquanu oVIOpoOU poAMoys spurs oq Fo pure ‘ssid - voumy oud WOIJ Ivos WY ‘100J PUL SHveTys snoaseo ou WOTJOVS UO SAMOS = ‘sqIqqey ‘STMT qnq pesavjus ATQueredde you spuv[s Are[p[rxeuqns 0M4 ates pearooor oS ‘sqeg g ‘skaquoW F “purely suemloeds ey, ‘aeyMasfa WseasIp JO USTs OU UALOYS aAvy oF pa}1oder “qayIV] 9[19% “URIAY ‘SYVOD GZ ‘BSITG F FBO T Arey[ixemqug PIO syjUOM xXIS ynoqe Sid Sunod B woIy paureyqo sem [eIIe}VM sy, SO6L ‘6 Fore ueqpodoma lq ‘Il ‘d ‘SO[NPOU SNOISVd-OdIVO[VO OSIV] POUIVITI0D spuLls AIV[IXVUQNS IayIO SY, “eoURJsqns SnNoOosvd-OCaIVO[VO Jo qnoysnoryy pesodu0d puv pasiv[ws sem (OpvVU aT soINg[No 94} WOIyM wWo.1z) purrs Arey]ixvuqns sug = peqoayre A[QYSI[S Os[e o1aM ssunt 9q} yey? 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Subcutaneous. Dose : 1 mg. Killed: Jan. 12, 1906. 114 days. Small caseating tumour, with fibrous margins. One left inguinal gland con- tained a caseated mass, other inguinal glands contained small caseous and calcareous tubercles and nodules ; some were normal. There were a few tubercles in the portal and mesenteric glands. In the lungs was a moderate number of caseous and slightly gritty tubercles, up to 1 mm. 213 CALF 312. Virus P. IV. Subcutaneous inoculation of culture obtained from the original material of Virus P. IV. Dose—50 milligrammes. Date—July 19, 1905. Weight at Inoculation—36-74 kilogrammes. [Age about 4 weeks.] Killed when in fairly good health—October 17, 1905. [90 days after inoculation.] Clinical History. A week after inoculation there was a flattened slightly raised tumour 65 cm. in greatest diameter and a slightly enlarged prescapular gland. _ The tumour increased in size, and four weeks after inoculation was prominent, rounded, and perceptibly softened in the centre. Seven weeks after inoculation the tumour measured 12:5 by 7 by 2°5 cm., and was soft and fluctuant in the upper part; the prescapular gland was hard and measured 7°5 cm. in greatest length. 2 _ Subsequently both tumour and gland diminished in size. There was no change in the general health until the fifth week when the respirations were seen to be distinctly quickened ; they gradually increased in frequency, and for nearly three weeks were rapid and rather laboured, expiration ending in a short grunt ; during this time there was frequent cough and the animal lost flesh, At one time the animal looked so ill that it appeared to be going to die. The symptoms, however, abated, the respirations decreased in frequency and the general condition became much improved. At the end of the experiment the respiration had not quite resumed its normal rate, but there was every indication that the animal was making a complete recovery. Temperature. JvLy August OCTOSER, Tuberculin Test.—The calf was not tested subse- quent to inoculation. Weights. Kilogrammes. July 19, 1905 asi ais ee 36°74 August 20,1905 ... dee iee 38-09 October 17,1905... els es 43°53 Total gain of weight.—6°80 kilogrammes. The calf was weighed every week after inoculation ; during the first six weeks the weight was practically stationary ; subsequently there was a slight steady increase. POST-MORTEM EXAMINATION, Carcass.—In moderate condition. Local Lesion.—The local tumour was small, thin, and lenticular in outline, measuring 9 by 65 by 2°5 em. On section it was composed of translucent fibrous tissue, showing in the centre irregular tracts of tough yellowish white caseated tissue ; there were also, chiefly around the margins, irregular streaks and foci, slightly gritty from calcification. in the middle of the upper part of the tumour there was a cavity the size of a walnut, filled with serous fluid containing small masses of broken down caseous tissue; the walls of the cavity were formed by caseated tissue, ragged and necrotic on the internal surface. Left Prescapular Gland.—The left prescapular gland measured 6 by 3 by 2 cm. On section the greater part of the gland, about three quarters, was composed of firm homogeneous yellowish white caseated tissue, very slightly gritty from calcification. Around the margins of this mass, between it and the. 27676 capsule, was a narrow zone of more or less normal gland tissue, containing here and there irregular opaque foci, most of which were gritty. The rest of the gland was normal, except for one firm encapsuled caseous nodule containing gritty foci. Left Prepectoral Glands.—The rounded gland, the size of a pea, contained two minute opaque yellow foci ; the reniform gland was normal. Right Prepectoral Glands, Right Prescapular Gland (5 by 2 by 1 em.), Cervical Glands, and Awillary Glands.—Normal. Abdomen. Peritoneum.—There was a slight excess of fluid in the peritoneal cavity. Omentum.—Normal. Spleen.—The spleen weighed 56-6 grammes. It was normal on the surface and on section. Liver.—The liver was normal on the surface. On section two or three minute opaque yellow foci were seen (fatty ?) Portal Glands—One portal gland showed two or three minute opaque yellow foci in the cortex ; another contained one similar point. Kidneys.—The right kidney showed on the surface two minute grey points of a doubtful nature. The left was normal. Suprarenals.—The left suprarenal body showed in the cortex fairly numerous tubercles, ranging in size from about 1 to 1:5 millimetre in diameter ; the tubercles were grey, with slightly opaque centres. The right suprarenal contained similar tubercles ;. in the centre of one was a minute calcareous grain. 2L 214 Thorax. Heart—The endocardium of the right ventricle showed small patches of thickening, but no tubercles. Heart muscle and pericardium normal. ©. Pleura.—There was slight hypertrophy of the lymphatic fringes around the margin of the tendon on the right side, and small hard bodies could be felt at the base of some of them. The pleura covering the ribs was normal. Lungs.—The dorsal borders and posterior portions of the caudal lobes were firm, dark red and consoli- dated, and showed under the pleura a yellowish mottling. The yellow patches were situated in the centres of the small lobules, and showed around the margins fine arborisations, corresponding with the finer bronchioles. On section the solid lobules showed numerous small cavities (dilated bronchi) filled with tenacious muco-pus ; some of the lobules were com- posed throughout of firm reddish tissue, mottled with opaque yellow foci; in the dorsal parts of the cephalic lobes there were a few consolidated lobules. The ventral parts of the caudal lobes, aud almost the whole of the cephalic and right middle lobes were crépitant ; on the surface of the former very numerous glistening transparent subpleural tubercles were seen; they projected slightly above the surface, the majority being about one millimetre in diameter. On section through these portions similar tubercles were seen in the depth; they were not, however, so distinct and apparently not so numerous as on the surface. In the cephalic lobes on the surface a few transparent tubercles were seen here and there and several doubtful transparent grey points ; on section tubercles were not definitely visible. Left Bronchial Gland.—The left bronchial gland showed in the cortex a few minute whitish specks, some of which were just perceptibly gritty. Right Bronchial Gland, Mediastinal Glands, Laryna, and Trachea.—Normal. Alimentary Tract. Pharyna, Tonsils, and Intestines —Normal. Mesenteric Glands.—Three mesenteric glands showed each a minute white speck. All other lymphatic glands were normal. Testes.—Normal. Microscopical Examination. Smears from :— Lung (muco-pus, two smears).—Numerous tuber- cle bacilli. Left Prescapular Gland.—Moderately numerous tubercle bacilli. Right Prepectoral Gland, Mesen- teric Gland, Left Suprarenal Body, Right Suprarenal Body, and Portal Gland. Left Bronchial Gland, Liver, and b No tahesenaedlt Kidney. : . A few tubercle bacilli. CALF 314. Virus P. IV. Subcutaneous inoculation of culture obtained from the original material of Virus P. IV. Dose—34 milligrammes. Date—July 19, 1905. Weight at Inoculation—33'55 kilogrammes. [Age about 5 weeks.] Killed—December 1, 1905. [135 days after inoculation. ] [The calf was killed because it was suffering from paralysis of the hind legs (due to caries of the 12th dorsal vertebra). Clinical History. Seven days after inoculation there was an oval swelling measuring 5 by 4 cm., and a slightly enlarged prescapular gland. The swelling increased in size and on the 30th day measured 10 by 6 cm. and was distinctly softened ; the gland was moderately enlarged, measuring 8 by 5 cm. Subsequently the tumour was converted into a thin-walled prominent pendulous cyst, 15 cm. in length by 10 cm. in breadth. The prescapular gland diminished in size and became irregular and nodular in outline. An enlarged prepectoral gland was felt during life. There was no obvious change in the general condition of the animal until November 29th, when it was found lying on its side and unable to rise from the ground, the hind legs being rigid and evidently paralysed. On the next day the calf was in much the same condition and, since it was unable properly to take nourishment, was killed. Temperature.—On the 10th day after inoculation the temperature rose suddenly to 40°4°C., but fell to the normal two days later. During the remaining period of the experiment the temperature was a little raised, and at the same time irregular. The maximum range of variation within a period of 4 months was 1°6° C. (38:1° C.—39-7° C.). Tuberculin Test—November 17th, days after inoculation.] Positive reaction temperature, 1°3°C. 1905. [121 Rise of The general condition was good.] Weights. Kilogrammes, July 19, 1905 ... an sa as : August 15, 1905 eee as woe 28°57 October 18, 1905 se 8 owe «= 4444 November 21, 1905 ... we =: 89°45 December 1,1905 ... ees 46°25 Total gain of weight during experiment.—12°69 kilo- grammes. POST-MORTEM EXAMINATION. The carcass was in fair condition. Local Lesion—At the seat of inoculation there was a tense fluctuating tumour, measuring 15 by 10 by 9 cm. and weighing with skin 793 grammes. On section it was a thin-walled cyst, filled with thick creamy yellow caseo-pus; the internal surface was pale and granular, and numerous calcareous grains were felt on passing the fingers over it ; the cavity was crossed by one thick fibrous band and several slender ones. Left Prescapular Gland.—The left prescapular gland weighed a little over 28 grammes, and measured. 65 by 4 by about 2°5 cm. in greatest thickness. It was irregular and nodular externally, and showed on section the greater part of the cortex replaced by calcareo-caseous masses, containing cavities filled with light brownish or creamy caseo-pus ; the medulla was normal. 215 Left Prepectoral Glands.—The spherical gland, 14 millimetres in diameter, showed about three quarters of the substance composed of dense calcareo-caseous tissue ; the gland was very indurated and the capsule thickened. The reniform gland was normal. Cervical Glands.—Normal. Thorax. Pleura.—Normal. Lungs.—The right cephalic and right middle lobes showed some dark red patches of collapse ; otherwise the lungs were perfectly normal to the naked eye. Trachea.—Normal. Thoracic Glands.—The caudal mediastinal gland contained one yellow pinhead-sized tubercle. Other mediastinal and the bronchial glands were normal. Heart.—Normal. Abdomen. Peritoneum and Omentum.—Normal. Large Intestine and Small Intestine.—Normal. Mesenteric Glands.—The mesenteric glands were normal in size ; they showed in the cortices minute yellow calcareous tubercles ; these were most numerous in the terminal glands. Spleen.—The spleen weighed 70-8 grammes, it was normal in size and general appearance. Liver, Gall Bladder, and Portal Glands.— Normal. Kidneys and Suprarenal Bodies.—Normal. Bladder.—The bladder was greatly distended. All the lymphatic glands not hitherto mentioned were normal, Tongue, Tonsils, Pharynz, Larynz.—Normal. Eyes.—Normal. Vertebral Column.—On cutting into the spinal canal, the cord was found to be compressed at the level of the twelfth dorsal vertebra between a rounded projection of new formed bone, arising from the upper surface of the body of the vertebra, and by a recent blood clot between the laminae and the periosteum. Section through the vertebra showed most of the cancellous tissue of the body hollowed out by rare- fying osteitis, the floor of the cavity being formed by the anterior vertebral ligament,-the roof chiefly by the mass of new formed bone, and the sides by the remains of the cancellous tissue. There was no sign of caseation or suppuration in the bone surrounding the cavity. The cord showed a well marked con- striction, apparently recent ; there was much gela- tinous oedema in the posterior part of the canal between the periosteum and dura mater CALF 350. Virus P. IV. Subcutaneous inoculation of culture derived from original material of Virus P. IV. Dose—50 milligrammes. Date—September 20, 1905. Weight at Inoculation—33-10 kilogrammes. [Age about 4 weeks.] Killed when in good health—December 19, 1905. [90 days after inoculation.] Clinical Notes. Ten days after inoculation on the left side of the neck a slight local thickening could be felt; the adjacent prescapular gland was slightly enlarged. After three weeks a small slightly raised tumour had developed, which measured 6°5 by 5 cm.; the prescapular gland was still slightly enlarged, 5°5 cm. in length. No further change of any importance took place in the local conditions. The calf was killed on the 90th day. remained well during the experiment. Tt had Temperature. SEPTEMBER OCTOBER, _From November 15 to December 19 the temperature was fairly normal, though somewhat irregular. NOVEMBER The maximum range of variation was between 38:0 and 39°3° C, 27676 2L2 216 Tuberculin Test—November 19, 1905 [58 days after inoculation]. Positive reaction. Rise of tem- perature 1°8° C. Weights. Kilogrammes. September 20,1905 ... 33°10 December 19,1905 ... 54:87 Total gain of weight—21-76 kilogrammes. Rate of: gain per week.—1-67 kilogrammes. POST-MORTEM EXAMINATION. Carcass.—Carcass in moderate condition. Local Lesion.—In the subcutaneous tissues on the tight side of the neck there was a small tumour 6 by 4 by about 1°5 cm. in greatest thickness ; on section it was composed of dense fibrous tissue con- taining two small cavities filled with buff coloured cageo-pus, and numerous small irregular gritty caseous nodules, most of which were softened ; the skin and muscles were adherent but not infiltrated. Right Prescapular Gland.—The right prescapular gland measured 5 by 3 cm. by about 2 cm. On section it showed rather more than a third of the cortex dense caseous and calcareous; the rest of the gland was normal. Left Prescapular Gland.— The left prescapular gland measured 4°5 by 2°5 cm. by about 1°5 cm. and was normal on section. Prepectoral Glands—-Right and left prepectoral glands normal. Cervical Glands—Right and left cervical glands normal. Thorax. Pleura.—Normal. Lungs.—The lungs were normal in general appear- ance ; in the left caudal lobe under the pleura there was a grey almost transparent miliary tubercle; under the pleura in all the lobes there was a number of very minute dark grey points of a doubtful nature ; on section nothing abnormal was seen. Trachea,—Normal. Bronchial and Mediastinal Glands.—The bronchial and mediastinal glands were not enlarged. The caudal gland showed in the cortex a few minute yellow foci, some of which were appreciably gritty. The gland above the bifurcation of the trachea, the bronchial and infratracheal glands, showed similar yellow foci, in small number. The ventral mediastinal glands were normal. Heart.—Normal. Abdomen. Peritoneum and Omentum.—Normal. Small Intestine, Large Intestine, Gastric Glands, Mesenteric Glands, and Colic Glands.—Normal. Spleen.—The spleen weighed 141 grammes ; it was normal on the surface and on section. Liver, Gall Bladder, Portal Glands, Kidneys, and Suprarenal Bodies.—Normal. Coeliac Glands.—One coeliac gland contained a small irregular gritty tubercle; the others were normal. Lumbar Glands and Iliac Glands.—Normal. Testes.—Normal. Special Glands. Precrural, Pudic, Gluteal, Ischiatic, Popliteal, Sub- maxillary, Parotid, and Retropharyngeal.—Normal. Tongue, Tonsils, and Larynx.—Normal. Eyes.—Normal. Microscopical Examinations. Smear preparations. Prescapular Gland.— Numerous tubercle bacilli many long and beaded forms. Coeliac Gland.—T wo tubercle bacilli found. Mediastinal Giland.—No tubercle bacilli. PIG 124. Virus P. IV. Fed with culture derived from original material of Virus P. IV. Dose—1 milligramme. Date—September 20, 1905. Weight at Inoculation—10-0 kilogrammes. Killed when in good health—January 16, 1906. Clinical Notes—The pig remained in good health during the experiment. Temperature.—Normal throughout. Luberculin Test—The pig was not tested subsequent to inoculation. Weights. Kilogrammes. September 26, 1905 ... ‘ee Pre 10-0 October 18, 1905 oes eit we: 10°88 January 16,1906... sis we 33°55 Total gain of weight—23-55 kilogrammes. POST-MORTEM EXAMINATION, Carcass.—In good condition. [Age 7 weeks. ] [118 days after feeding.] Alimentary Tract. Tongue, Pharyna, Tonsils—Normal. Submasillary Glands.— The submaxillary glands, one on each side, contained a moderate number of yellow cheesy caseous nodules, slightly gritty from calcification. They were irregular in outline, and shelled out easily from the surrounding gland tissue ; in the gland on the left side they ranged in size from a pin’s head to that of a wheat grain; in that on the right side they were a little larger, up to that of a pea. Retro-pharyngeal Glands, Cervical Glands.—Normal. Stomach, Small Intestine, Large Intestine, Gastric Glands.—Normal. Mesenteric Glands.—A gland in the anterior part of the mesentery contained two irregular caseous gritty 217 ‘tubercles, the largest 1:5 millimetre in greatest ‘diameter ; other mesenteric glands were normal. lleo-Colic Glands, Colic Glands —N ormal. _ Abdomen. Omentum, Peritoneum, Spleen, Liver, Gall Bladder. -—Normal. ‘Portal Glands.—One portal gland contained two ‘pearly white spherical tubercles, each less than a pin’s head in size. Coeliac Glands, Normal. Kidneys, Suprarenal Bodies.— Thorax. Heart, Pleura.—Normal. Lungs.—In the right cephalic lobe of the lung there was a subpleural grey tubercle, the size of a pin’s head, with a yellow gritty centre. In the substance of the left caudal lobe there was a caseo-caléareous tubercle 1:5 millimetre in diameter. The rest of the lung was normal. Bronchial Glands.—On the right side, in the angle between the right anterior bronchus and the trachea, a gland contained two yellow pinhead-sized caseous tubercles, slightly gritty from calcification. The other bronchial glands were normal. Dorsal Mediastinal Glands, Ventral Mediastinal Glands.—Normal. Various Lymphatic Glands. Prescapular Glands, Prepectoral Glands, Precrural Glands, [liac Glands, Lumbar Glands.—Normal. Microscopical Examination. Lung Tubercle.—No tubercle bacilli seen. Tubercle from Bronchial Gland.—No tubercle bacilli seen. Tubercle from Portal Gland.—No tubercle bacilli seen. é PIG 126. Virus P. IV. Subcutaneous inoculation of culture derived from original material of Virus P. IV. Dose—1 milligramme. Date—September 20, 1905. Weight at Inoculation—8°5 kilogrammes. [Age 7 weeks-] Killed when in good health—January 12,1906. [114 days after inoculation. ] Clinical Notes. A very small swelling developed at the seat of ‘inoculation in the abdominal wall, and the nearest inguinal gland became slightly enlarged. Temperature.—The day after inoculation the tem- perature rose to 40-°0° C. It quickly fell to normal. On the 13th day the temperature again rose, and reached a maximum of 39°8° C., returning to normal -within a week. Subsequently the temperature re- mained normal. Tuberculin Test.—The pig was not tested sukse- . quent to inoculation. Weights. Kilogrammes. September 26, 1905 85 October 18, 1905 10:43 January 12, 1906 27°66 Total gain in weight during experiment.—19°16 kilo- - grammes. POST-MORTEM EXAMINATION. Carcass.—In good condition. Local Lesion—At the seat of inoculation near the umbilicus there was a nodule the size of 2 broad bean composed of tough yellowish white caseated tissue, gritty around the margins and softened in the centre, surrounded by a thin capsule of fibrous tissue ; the skin was not adherent. Inguinal Glands.—The glands on the right side were not enlarged ; the one nearest the local lesion contained a pea-sized caseous slightly gritty nodule, with a mul- berry like outline, and half-a-dozen caseous nodules, the largest the size of a hemp seed; the nodules -Shelled out easily from the gland substance ; the other -glands in this group were normal. On the left side the gland nearest the local lesion was a little enlarged and showed about half its substance replaced by an irregular mass of firm caseated tissue slightly gritty from calcification; a pea-sized gland near it contained a small caseous nodule and another smaller gland a caseous tubercle ; other glands in this group were normal. Abdomen. tliac Glands, Llio-Sacral Glands, Lumbar Glands, Omentum and Peritoneum.—Normal. Spleen.—The spleen showed on the convex surface a patch of cicatricial thickening ; otherwise normal. Liver.—Normal. Portal Glands.—One portal gland contained about nine yellowish tubercles, the largest the size of a pin’s head; another contained three or four similar tubercles. Kidneys, Suprarenal Bodies.—Normal. Thora. Heart, Diaphragm, Pleura.—Normal. Lungs.—The lungs were crepitant throughout and contained a moderate number of evenly distributed shotty tubercles ranging in size from about 0°5 to 1 millimetre in diameter; the tubercles were yellow caseous and slightly gritty and had narrow grey margins. Dorsal Mediastinal, Bronchial Glands, Larynx, and Trachea.—Normal. Alimentary Tract. Tongue, Pharyna, Tonsils—Normal. Stomach, Intestines.—Normal. Submaxillary, Glands.—Normal. Retro - pharyngeal and Cervical 218 Mesenteric Glands.—There were a few caseous gritty tubercles (largest 1 millimetre in diameter) in the glands in the anterior part of the mesentery and two in those in the posterior part. Lleo-Colic Glands.—Normal. Prescapular Glands, Prepectoral Glands, Coeliac Glands.—Normal. Microscopical Examinations, Portal Gland.—One tubercle bacillus seen. Mesenteric Gland.—No tubercle bacilli. Animal Inoculated. A guinea-pig inoculated with an emulsion {from the spleen died in 18 days showing no evidence of tuber- culosis. CALF 356. Virus P. IV. Subcutaneous inoculation of culture derived from the lung of Calf 312. Doge—50 milligrammes. Date—December 4, 1905. Weight at Inoculation—56°69 kilogrammes. Killed when in good health—March 19, 1906. Clinical Notes. Eleven days after inoculation on the left side of the neck there was a slightly raised swelling measuring 10 by 7°5 cm. and about 2 cm. thick. The adjacent prescapular gland was about 7 cm. in length. On the 32nd day there was a prominent fluctuating tumour at the seat of inoculation, measuring 14 by 75cm. The prescapular gland was enlarged, 6 cm. in length. On the 80th day the tumour was cystic, measuring 11 by 8 by about 5 cm. On the 103rd day the tumour burst, discharging caseo-pus. On the 105th day the calf was killed. It had shown no sign of ill-health during the experiment. Temperature.—On the tenth day the temperature rose to 40°4° CO. It remained high for ten days, and then returned to the normal. The temperature sub- sequently remained normal until the end of the experiment. Tuberculin Test—February 2nd, 1906, 60 days after inoculation. Positive reaction. Rise of tem- perature, 1°5° C. Weights. Kilogrammes. December 5,1905 ... ae we = 56°69 January 25,1906... aia we 72°56 February 21,1906... te we = 85°71 March 19, 1906 wise a 93°42 Total gain of weight during experiment. — 36°73 kilogrammes. Average rate of gain per week.—2°44 kilogrammes. POST-MORTEM EXAMINATION. Carcass.—In very good condition. Local Lesion—At the seat of inoculation on the left side of the neck there was a raised tumour measuring about 9 by 6:5 by 4 cm. At the upper extremity the skin showed a sinus, the diameter of a crow-quill, the outer opening of which was concealed by a mass of dried discharge. On section the tumour was composed of dense white fibrous tissue containing a few scattered yellow gritty tubercles ; just under the skin there was an irregular cavity 4°5 cm. in greatest diameter filled with thick yellow caseo-pus ; the cavity was lined with reddish granulation tissue and was crossed by several thick fibrous trabeculae ; this cavity communicated externally by the sinus men- tioned above ; the skin external to it was very thick except at one part where it appeared to be just on the point of breaking down, Left Prescapular Gland.—The left prescapular [Age about 12 weeks. ] [105 days after inoculation. ] gland measured 6°5 by 3 by nearly 1:5 cm., and was. only slightly larger (thicker) than the right; on section it showed in the cortex a number of yellow calcareous foci and a few soft whitish caseous tubercles, the largest a little more than 1 mm. in diameter ; there were besides two small nodules, the largest the size of a split pea, composed of brownish translucent tissue and calcareous particles ; the gland tissue was normal in appearance. Right Prescapular Gland.—Normal, Prepectoral and Cervical Glands on both sides were. normal. Thorax. Heart.—Normal. Pleura.—Normal. Lungs.—Under the pleura a few minute scattered grey foci of a doubtful nature were seen (a smear made from one showed no tubercle bacilli) ; other- wise the lungs were perfectly normal. Bronchial and Mediastinal Glands.—Normal. Abdomen. Omentum and Peritoneum.—Normal. Spleen (311 grammes).—Normal. Liver.—Normal. Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Lonsils, Pharynx, Larynx, and Trachea—Normal. Intestines.—Normal. Testes.—Normal. Eyes.—Normal. Special Lymphatic Glands. Parotid, Submazillary, Pharyngeal, Axillary, Renal, Lumbar, Mesenteric, Gastric, Colic, Iliac, Precrural, eee Gluteal, Pudic, and Ischiatic Glands.— ormal. Microscopical Examination. (Smear Preparations.) Prescapular Gland (soft tubercle).—A few tubercle - bacilli seen. Lung (grey focus).—No tubercle bacilli seen. 219 CALF 358. Virus P. IV. ‘Subcutaneous inoculation of culture derived from the lung of Calf 312. Dose—50 milligrammes. Date—December 4, 1905. Weight at Inoculation—63-50 kilogrammes. [Age about 14 weeks, ] Killed when in good health—March 7, 1906. Clinical Notes. Eleven days after inoculation on the left side of the ‘neck there was a flat swelling measuring 9 by 7°5 cm. The adjacent prescapular gland was slightly enlarged, -6°5 cm. in length. The swelling developed into a prominent softened tumour which on the 32nd day measured 9°5 by 7°5 cm.; the prescapular gland was enlarged, 7:5 cm. in length. On the 80th day the tumour was tense, prominent and cystic, measuring 9 by 7°5 cm.; the prescapular ‘gland had diminished in size and now measured about 5 cm, in length. On the 93rd day after inoculation the calf was killed. It had remained well during the experiment. Temperature.—There was a slight rise of tempera- ture on the 11th day after inoculation which lasted 11 days and reached a maximum of 39°8°C. Subse- ‘quently the temperature remained normal. Tuberculin Test.—February 2, 1906, 60 days after inoculation. Slight reaction. Rise of temperature ‘0°6° C. Weights. Kilogrammes. December 5, 1905... me : January 25,1906... eee 90-70 February 21, 1906 ... eee 102-94 March 7, 1906 at as 108-84 Total gain of weight during experiment.—45°34 kilo- -grammes. Average rate of gain per weck.—3°49 kilogrammes. POST-MORTEM EXAMINATION. General Condition —Good. Local Lesion.—At the seat of inoculation, on the left side of the neck, there was a tense egg-shaped fluctuant swelling measuring 9 by 6°5 by 5 cm. On section it was found to be a thin-walled cyst filled with thick tenacious caseo-pus and yellow watery fluid; the walls of the cyst were formed by fibrous tissue lined internally with pale granulation tissue ; the cavity was crossed by several fibrous trabeculae. Left Prescapular Gland—The left prescapular gland measured 5 by 2°5 by about 1°5 cm., and was only slightly thicker than the right. On section the cortex along the convex margin showed an irregular patch of dense tough caseo-necrotic tissue very gritty around the margins. The capsule of the gland [93 days after inoculation.] external to the caseous wass was slightly thickened, and around the margins was a narrow zone of brownish translucent tissue ; the latter and the peri- pheral parts of the caseated tissue were very gritty from calcification. The right Prescapular, the right and left Prepectoral and Axillary Glands were normal. Thorax. Pleura.—Normal. Lungs.—The lungs showed a few minute grey foci of a doubtful nature. Thoracic Glands.—A dorsal mediastinal giand anterior to the bifurcation of the trachea contained a greyish white spherical tubercle about 1 mm. in diameter. Other thoracic glands were normal. Heart.—Normal. Abdomen. Peritoneum and Omentum.—Normal. Spleen.—Normal. Liver.—Normal. Portal Glands.—One portal gland seemed a little firmer than normal, and showed on section in the cortex slightly opaque greyish-white foci in some of which were yellow points ; they were not perceptible to the touch. Kidneys.—Normal. Suprarenal Bodies.—Normal. Renal, Lumbar, and Iliac Glands.—Normal. Alimentary Tract. Tongue, Pharynx, Palate, Tonsils.—Normal. Intestines.—Normal. Mesenteric Glands.—Normal. Various Lymphatic Glands. Submasxillary, Pharyngeal, Normal. Precrural, Popliteal, Gluteal, Ischiatic, Pudie.— Normal. Parotideal, Hyoid.— Microscopic Examinations. Emulsion of left Prescapular Gland.—Tubercle bacilli in moderate numbers. Tubercles from Portal Gland.—(1) No tubercle bacilli seen. (2) One tubercle bacillus seen. PIG 132. Virus P. IV. Subcutaneous inoculation of culture derived from the lung of Calf 312. Dose—1 milligramme. Date—December 4, 1905. Weight at Inoculation—26-0 kilogrammes. Killed when in good health—April 5, 1906. Clinical Notes. A small tumour developed at the seat of inocula- -tion in the abdominal wall, and the adjacent inguinal _gland became slightly enlarged. [Age about 15 weeks. ] [122 days after inoculation. ] The pig showed no sign of illness during the experiment. Temperature.—Normal througkout. Tuberculin Test.—February 2, 1906. [61 days 220 after inoculation.] Positive reaction. Rise of tem- perature, 2°3° C. Weights. Kilogrammes. December 6, 1905... aes w. = 260 April 5, 1906 ... a8 ee wee = 68A7 Total gain of weight during experiment.—42°47 kilo- grammes, Average rate of gain per week.—2'44 kilogrammes. POST-MORTEM EXAMINATION. Carcass.—In very good condition. Local Lesion—At the seat of inoculation a little to the left of the umbilicus there was a firm nodule, slightly adherent to the skin, 2'2cm. in length and about 1‘5cm. in diameter; on section it was composed of somewhat tough yellow caseous material, slightly gritty from calcification and at one part beginning to soften, surrounded by a thin fibrous capsule. Inguinal Glands—On the left side the glands nearest the local lesion contained six caseous slightly gritty nodules ranging in size from 4 to 8 mm. in diameter ; each nodule had a thin capsule of fibrous tissue ; the caseous material in each case was friable but not softened and could be readily shelled out in a mass from its surrounding capsule ; other glands in this group were normal. On the right side the inguinal glands were normal. Precrural Glands.—Normal. Tliac and Lumbar Glands.—Normal, Abdomen, Omentum and Peritoneum.—Normal. Spleen.—Normal. Liver.—The liver showed under the capsule two small tubercles (largest about 1mm.) with grey margins and opaque yellow centres ; in the depth of the liver substance one minute yellowish white spherical body was seen. Portal Glands.—The portal glands were not enlarged, and contained discrete yellow calcareous tubercles, some very irregular in outline, the largest: about 1 mm. in diameter ; they very readily shelled out from the surrounding gland tissue which was. perfectly normal in appearance. Kidneys.—Normal. Suprarenal Bodies—Normal. Thorax. Heart and Pleura.—Normal. Lungs.—The lungs were normal in _ general appearance ; they showed just under the pleura altogether about ten minute tubercles each about G5 mm. in diameter; the tubercles were glassy and very slightly opalescent in the centre. Bronchial and Mediastinal Glands.—Each bronchial. gland showed on section one minute white point ; they were otherwise normal. The inter-bronchial,. infra-tracheal, and dorsal mediastinal glands were normal. Two ventral mediastinal glands were slightly enlarged; one contained a pea-sized gritty cheesy nodule ; the other contained a similar but larger mass which almost completely replaced the gland substance ;, each mass was surrounded by a thin fibrous capsule, Laryna and Trachea.—Normal. Alimentary Tract. Tongue, Tonsils, and Pharynx.—Normal. Intestines.—Normal. Gastric Glands.—Normal. Mesenteric and Colic Glands.—Normal. Uterus.—Normal. Thyroid Body.—Normal. Special Lymphatic Glands. Coeliac Glands.—The largest gland contained a few minute yellowish white gritty foci; another smaller gland contained one similar focus; the rest were normal. Submasillary, Pharyngeal, Cervical, Prescapular, Prepectoral, and Precrural Glands.—Normal. PIG 134. Virus P. IV. Fed with culture derived from the lung of Calf 312. Dose—1 milligramme. Date—December 4, 1905. Weight at Inoculation—20-25 kilogrammes. Killed when in good health—April 11, 1906. Clinical Notes. The pig remained in good health during the experiment. Temperature.—Normal. Tuberculin Test.—February 2, 1906. inoculation.] Positive reaction. ture, 1°3° C. [61 days after Rise of tempera- Weights. Kilogrammes. December 6, 1905 20°25 April 10, 1906 63°03 Total gain of weight during caperiment.—42°78 kilo- grammes. Average rate of gain per week.—2°35 kilogrammes. POST-MORTEM EXAMINATION. Body in good condition. [Age about 15 weeks. ] [128 days after feeding. ] Alimentary Tract. Tongue, Tonsils and Pharyne.—Normal. Submaaillary Glands.—The left submaxillary gland contained about half a dozen caseous gritty nodules, the largest nearly 3 mm. in diameter ; each nodule was surrounded by a delicate capsule of fibrous tissue from which it could be readily shelled. The right submaxillary gland contained two similar caseous nodules and a few discrete yellow gritty tubercles, irregular in outline. Pharyngeal and Cervical Glands. —Normal. Stomach.—Normal. Small Intestine.—-Normal. Large Intestine—Normal. Gastric, Mesenteric, Ileo-Colic and Colie Glands.— Normal. Abdomen. Omentum and Peritoneum.—Normal. 221 Spleen.—Normal. Liver—In the substance of the liver under the capsule on the anterior surface there was a whitish fibrous nodule, the size of a pea; the liver substance around it showed an increase in interstitial connective tissue ; with this exception the liver was normal. Portal Glands.—Normal. Kidneys, Suprarenal Bodies.—Normal. Thorac. Heart and Pleura.—Normal. Lungs.—The lungs were crepitant throughout and collapsed normally ; in the right caudal lobe under the pleura there was one grey almost transparent tubercle the size of a pin’s head ; with this exception the lungs were normal, Larynx and Trachea.—Normal. Bronchial and Dorsal and Ventral Mediastinal Glands.—Normal. Various Lymphatic Glands. Prescapular, Prepectoral, Precrural, Inguinal, Iliac and Lumbar Glands.—Normal. Special Organs. Thyroid Body, Testes and Urinary Bladder.— Normal. Microscopical Examination. (Smear Preparation.) Submaxillary Gland Nodule.—No tubercle bacilli. Animal Inoculated. A guinea-pig inoculated with an emulsion made from the nodule in the liver remained free from tuberculosis. CALF 426. Virus P. IV. Intramuscular inoculation of culture derived from prescapular gland of Calf 358 (injected with culture from Calf 312). Dose—200 milligrammes. Date—J une 1, 1906. Weight at Inoculation—29-02 kilogrammes. Died—June 30, 1906. Clinical Notes. The calf was not in very good condition when inocu- lated; it had suffered since its arrival at Walpole from severe diarrhea. Five days after inoculation on the left side of the neck a firm deep seated tumour had developed, measuring about 6°5 cm. in greatest diameter; the [29 days after inoculation. ] [Age about 8 weeks. ] skin was moveable over it. The adjacent prescapular gland was slightly enlarged. On the 20th day the tumour was about 9 cm. in diameter ; the prescapular gland was slightly enlarged ; the prepectoral gland was the size of a pea. On the 29th day the calf died. It had been in poor condition throughout the experiment; its appetite was moderately good, but there was almost constant diarrhoea. Temperature. JUNE Tuberculin Test.—The calf was not tested subse- quent to inoculation. Weights. Kiiogrammes. June 1, 1906... 29°02 June 5, 4 sg 26°30 June 25, 4, .. a 27°66 Total loss of weighi during experiment.—1°36 kilo- grammes. 27676 POST-MORTEM EXAMINATION. Carcass.—In very poor condition. Local Lesion.—In the muscles on the left side of the neck there was an ill-defined swelling, measuring roughly 10 by 7:5 by 2°5 cm.; the skin was freely moveable over it. On section the muscles were found to contain a mass of yellow caseo-necrotic tissue, the margins of which were not definitely circumscribed ; in the 2M 222 thicker part of the mass there was an egg-shaped cavity, 5 cm. in greatest diameter, filled with clear yellow serous fluid ; the internal wall was formed by soft shreddy necrotic material. Left Prescapular Gland.—The left prescapular gland was enlarged and measured 6 by 3 cm. by about 2 cm.; on section the cortex was firmer than normal, and in places indurated, and showed a fine yellow network of necrosis, more definite in some parts of the cortex than in others. Right Prescapular Gland.—The right prescapular gland measured 4 by 2 by 1 cm., and was normal on section. Prepectoral Glands—On the left side there were two enlarged glands, one the size of a broad bean, the other a little smaller ; both were indurated and showed on section a fine yellow network in the cortices. On the right side the glands were normal. Cervical Glands.—Two lower cervical glands on the left side were enlarged and on section resembled the left prepectoral glands. On the right side the glands were normal. A gland just within the entrance to the thorax was enlarged and a little firmer than normal and showed in the cortex a few very early foci of necrosis. Thoraa. Lungs.—The lungs collapsed normally and showed, chiefly in the left lung, small irregular patches of collapse ; scattered evenly throughout the substance of the lungs were moderately numerous minute dark grey tubercles which projected slightly from the cut surface ; here and there was seen a larger tubercle with a slightly opaque centre. The Bronchial and Mediastinal Glands appeared normal, Heart and Pleura —Normal. Abdomen. Omentum and Peritoneum.—Normal. Spleen.—The spleen was normal in size and appeared normal on section. Liver.—The liver was norrhal on the surface; on section the substance sbowed numerous minute greyish white tubercles. Portal Glands.—Normal. Kidneys——The capsules of the kidneys stripped badly ; the cortex was pale and diminished in thickness ; the kidney substance generally was tough and fibroid ; no tubercles were seen. Suprarenal Bodies.—Normal. Renal, Lumbar, and Iliac Glands.—Normal. Alimentary Tract. Pharynz, Tonsils.—Normal. Intestines.—Normal. Mesenteric, Gastric, and Colic Glands—Normal. Various Lymphatic Glands. Pharyngeal, Parotid, Submazillary, Axillary, Coeliac, Precrural, Popliteal, Pudic, Ischiatic, and Gluteal Glands.—Normal. 22 VIRUS 3 P. V. (April 18, 1905.) BRONCHIAL GLAND. CULTURE INOCULATIONS, AUGUST 30, 1905. The strain was derived from the original material and had (been in cultivation) a total period of 134 days. The culture used was the 7th generation, 20 days old. " CALF 336. RABBITS. Subcutaneous. 8¢ SS Dose : 50 mg. Number. | Method. | Dose. | Duration of Life. | Result. Died : Sept. 28, 1905. 29 days. Intrav. O-lmg. | D. 21d G. T. P.M.—General Tuberculosis, severe. eee fee a | a 394 | Intrap. 1:0 mg. | D2 ag G.T. 399 ! Intrap. 1:0 mg. D.13,,, G. T. | 398 | Intrap. | O-Lmg. | D. 29, G.T. | 400 | Intrap. | 001mg. | D. 30 ==", G. T. 395 | Subeut. | 10mg. | D. 88, Gf, 396 Subcut. Olmg. | D.58 ,, | G.T. | I CALF 336. Virus P. V. Subcutaneous inoculation of culture derived from the original material of Virus P. V. Dose—50 milligrammes. Date—August 30, 1905. Weight at Inoculation—29:48 kilogrammes. Died—Serptember 28, 1905. Clinical Notes. Seven days aiter inoculation on the left side of the aeck there was a flattened local thickening, infiltrating the muscles, measuring 7°5 by 6:5 cm.; there was slight oedema in the loose tissues around the lower extremity. The prescapular gland was very slightly enlarged. On the 23rd day a tumour had developed at the seat of inoculation, which measured 11-5 by 9 em. and was [Age about 6 weeks. ] {29 days after inoculation.] The prescapular gland The general con- not more than 2 cm. thick. was enlarged, 7°5 cm. in length. dition of the calf was unchanged. On the 29th day the calf died. It was not a very strong animal at the beginning of the experiment ; it was thin and did not thrive ; after the 23rd day it began to get thinner and weaker, and on the morning of the 29th day was found on the ground in a dying condition and unable to rise; the respirations on the previous day were not obviously increased. Temperature. August SEPTEMBER 27676 2M2 224 Tuberculin Test.—The calf was not tested subse- quent to inoculation. Weights. Kilogrammes. August 30,1905 ... ves 29°48 September 28, 1905 23°12 Total loss of weight.—6°36 kilogrammes. Average rate of loss per week.—1°58 kilogramme. POST-MORTEM EXAMINATION. Carvass.—Emaciated. Seat of Inoculation—In the subcutaneous tissues on the left side of the neck there was a flattened mass of dense yellow caseo-necrotic tissue, 10 by 5 by 15¢m.; the mass was adherent to the skin and muscles, both of which were infiltrated to a slight extent. Left Prescapular Gland.—The left prescapular gland measured 6°5 by 4 by 25 cm. On sevtion the greater part of the substance was composed of dense yellow caseated tissue ; whilst the rest, chiefly around the hilum, was closely beset with small caseous tubercles. Prepectoral Glands, Right Prescapular Gland.—The prepectoral glands on both sides and the right prescapular gland showed discrete irregular caseous tubercles in the cortices. Cervical Glands.—On the left side a small gland near the inferior extremity of the tumour was caseous throughout ; two others in the middle of the neck showed part of their cortex caseating. The superior cervical gland, near the mastoid, showed a reddish grey caseating patch and numerous discrete caseous tubercles. The glands on the right side showed small caseous tubercles or caseous foci. Abdomen. Omentum.—On the inferior surface of the omentum were numerous grey tubercles, many loosely attached and deeply hemorrhagic. Peritoneum.—Normal. Spleen—The spleen was slightly enlarged, and weighed 226°5 grammes. On section the pulp was packed with opaque miliary tubercles, which gave the cut surface a coarsely granular appearance. Liver—The liver was mottled on the surface with purplish patches. The substance contained moderately numerous tubercles, ranging in size from a mere point up to about 1 millimetre in diameter ; the larger ones were opaque in the centre. Gall Bladder—The gall bladder showed three opaque yellow foci under the mucous membrane. Portal Glands.—The portal glands were slightly enlarged, and showed the cortex firm, grey, and in a state of early caseation ; the glands were soft and oedematous in the centre. Kidneys.—The kidneys contained sparsely scattered tubercles, the largest a little over a millimetre in diameter ; they were slightly opaque in the centre. Suprarenals.—The right suprarenal body contained an opaque tubercle the size of a pin’s head. Thorax. Heart, Parietal Plewra.—Normal. Diaphragm.—On the pleural surface of the diaph- ragm there was one pinhead-sized caseous tubercle. Lungs.—The lungs showed only a moderate number of small tubercles, the largest about a millimetre in diameter. The tubercles had congested grey margins and opaque caseous centres. There were no large areas of consolidation, but in the left lobes a few lobules were dark and collapsed (not recent). In the latter lobes many of the tubercles or small groups of tubercles were surrounded by little angular patches of red hepatisation. Bronchial and Mediastinal Glands.——The bronchial and mediastinal glands were moderately enlarged ; they showed in the cortices grey or reddish grey trans- lucent tissue, containing a fine yellow caseous network. The ventral mediastinal glands were closely beset with yellow caseous tubercles, irregular in outline. Larynx, Trachea.—Normal. Alimentary Tract. Tonsils.—The tonsils contained each a few caseous tubercles. Tongue, Pharynx.—Normal. Small Intestine—All the Peyer’s patches contained minute yellow caseous tubercles; they were very numerous in the long Peyer’s patch, and not so numerous in the others. In the mucous membrane a few sparsely scattered tubercles were seen ; in the middle part the mucous membrane showed three or four haemorrhagic erosions. Large Intestine —Normal. Mesenteric Glands.—The mesenteric glands were slightly enlarged ; the cortex of each was composed throughout of grey translucent tissue containing a fine caseous network. In the medullary parts of the glands were discrete pinhead-sized caseous tubercles. Ileo-Colic Glands.—The ileo-colic glands were simi- larly, but rather less severely, affected. Colic Glands, Gastrie Glands.—These glands con- tained numerous caseous foci. Various Lymphatic Glands. Precrural, Popliteal, Pudic, Gluteal, Asillary.— All contained numerous small irregular caseous tubercles. Ischiatic.—These glands showed a few minute caseous foci. Submaaillary, Retropharyngeal.—T hese glands were very closely beset with minute caseous tubercles. Coeliac.—The coeliac glands were slightly enlarged, and showed early caseous foci in the cortex. A few haemolymph glands contained small caseous tubercles. Testes, Eyes.—Normal. Microscopical Examination. Tubercle from Suprarenal Body.—No tubercle bacilli seen, 225 VIRUS P. VI. (April 28, 1905.) INGUINAL GLAND. ‘CuLtuRE InocuLaTIoNns, SEPTEMBER 1, 1905. The strain was derived from the original material through G.P. 1515, and had been in cultivation a total period of 126 days. The culture used was the 6th generation, 20 days old. CALF 326. RABBITS. Subcutaneous. Dose :* 50 mg. Number. | Method. Dose. Duration of Life. Result. Killed when very ill : Sept. 30, 1905. 29 days. 403 | Intray. | 04 me. D. 20 days ar P.M.—General Tuberculosis, severe. 401 Intrap. 1:0 mg DAS ay G.T. 404 Intrap. 1:0 mg. | D.20 ==, G. T. 405 Inotrap. O-L mg. D..33> 4; G.T. 407 Intrap. | 0-01 mg. D. 36 C=. G. T. 402 Subcut. 1:0 mg. D.71_—s,, G.T. 406 Subcut. 0-1 mg. D. 88 Ca, G.T. CALF 326. Virus P. VI. Subcutaneous inoculation of culture derived from the original material of Virus P. VI through G.P. 1515. Dose—50 milligrammes. Date—September i, 1905. Weight at Inoculation—41-27 kilogrammes, Killed when very ill—September 30, 1905. Clinical Notes. Five days after inoculation on the left side of the neck there was a local tumour measuring 7 by 5 em. by about 2 cm. in thickness ; the prescapular gland was enlarged, 7 cm. in length. On the 12th day the tumour measured 14 by 10 by about 4 cm. ; it was prominent and firm except in the centre where a sense of deep fluctuation was obtained. The prescapular gland was considerably enlarged, 10 cm. in length. [Age about 64 weeks. ] [29 days after inoculation. ] On the 21st day the tumour measured 16°5 by 12:7 cm., and the tissues around the inferior extremity were oedematous. The calf was unwell; there was some loss of flesh, its coat was staring, and there was slight increase in respiration. On the 29th day the calf was killed. During the previous week it had continued to lose flesh and to become weaker ; the respirations were also obviously increased in frequency but were not very rapid ; the animal would probably have lived two or three days longer. Temperature. SEPTEMBER 2 Tuberculin Test.—The calf was not tested subse- quent to inoculation. Weights. Kilogrammes. September 5, 1905... ae vee 41°27 September 19, 1905 ... 43°53 September 30, 1905 ... 39°45 Total loss of weight during experiment.—1'82_ kilo- gramme. POST-MORTEM EXAMINATION. ¢ Carcass.—In poor condition. Local lesion—At the seat of inoculation on the left side of the neck there was a tumour, weighing with the skin and muscles to which it was attached 793 grammes, and measuring 14 by 11'5 by 6cm. On section it showed between the skin and muscles a mass of firm pinkish yellow caseo-necrotic tissue, contain- ing in the upper part a cavity, 6°5 by 4 by 2°5 cm., filled with clear reddish serous fluid. The muscles internal to the mass were fibroid, and infiltrated for a considerable depth with yellow caseous nodules ; the nodules were for the most part arranged in lines which ran parallel to the muscle fibres. The skin was firmly adherent, and closely beset with yellow tubercles; opposite the cavity it was deeply congested. There was a moderate amount of oedema fluid in the tissues around the local tumour, particularly near the inferior part. Left Prescapular Gland.—The left prescapular gland weighed 170 grammes, and measured 10 by 5 by 4 cm. On section it was composed throughout of dense tissue, chiefly uniform and pinkish yellow in colour, but in parts greyish and translucent, and mottled with a close yellow network. At the inferior extremity of the prescapular gland there was a. gland the size of a small marble, composed of translucent tissue, moderately well advanced in caseation. Prepectoral Glands.—The rounded prepectoral gland on the left side, the size of a Barcelona nut, was tense and closely beset with minute whitish foci. The kidney-shaped prepectoral gland was much enlarged and firmer than normal, but showed only one or two casecus foci; a smaller gland more posterior was similarly affected. Cervical Glands.—Along the left side of the trachea there was a chain of half a dozen glands, the largest the size of a pigeon’s egg ; several resembled the left prescapular, the others were slightly less advanced in caseation. The left upper cervical gland, the size of a walnut, was composed throughout of dense yellow caseous tissue ; a large gland near the latter showed a caseating mass in one part of the cortex, and in the rest scattered greyish white tubercles. Just within the entrance to the thorax, on the left side, there was a large gland, composed of dense translucent tissue, containing a close yellow caseous network. In the middle of the neck, on the right side, there was a gland the size of a small bean, composed of grey translucent tissue with a yellow network. All other glands on this side were slightly enlarged, and showed irregular caseous foci. Thorax. Heart.—On the endocardium of the right ventricle one grey point was seen; otherwise the heart was normal, Pleura.—On the pleural surface of the diaphragm there were about eight nodules, the largest the size of a wheat grain; one was loosely attached and haemorrhagic. There were a few similar nodules on the fringes along the margins of the ribs ; also here and there clusters of minute grey granules. Lungs.—The ventral part of the left cephalic lobe and the ventral portions of the right cephalic and middle lobes were, with the exception of a few lobules along the margins, dark red and quite airless ; in the anterior parts of the caudal lobes there were a few isolated solid lobules. The lung tissue was moderately closely beset with grey tubercles, slightly opaque in the centre, the largest about one millimetre in diameter. The tubercles appeared to be rather more 6 numerous in the solid than in the crépitant portions.. The bronchi in the collapsed areas were filled with muco-pus. Bronchial and Mediastinal Glands.—The bronchial and mediastinal glands were moderately enlarged. The cortices were composed of firm grey tissue, con- taining an opaque yellow caseous network. One infratracheal gland resembled the bronchial ; another was enlarged and oedematous, but showed no- caseous foci. * Trachea.—The mucous membrane of the anterior end of the trachea showed three or four slightly raised elongated greyish tubercles containing caseous points. Abdomen. Omentum.—On the inferior surface there were fairly numerous lenticular nodules, the largest the size of a wheat grain. All were yellow in the centre, and the majority were loosely attached, some being deeply hemorrhagic. Parietal Peritoneum.—Normal. Spleen.—The spleen weighed 283 grammes. The pulp was closely beset with yellow caseous tubercles, somewhat irregular in outline, and ranging up to about 2 millimetres in diameter. Liver.—The liver contained very numerous tubercles, evenly distributed throughout the substance. The tubercles were grey, and ranged in size from a mere point up to about 1 millimetre in diameter, the larger ones showing opaque yellow centres; some of the tubercles on the surface, under the capsule, were flattened out tc a greater diameter. Gall Bladder—Under the mucous membrane of the gall bladder there were three yellow pinhead-sized tubercles. E Portal Glands.—The portal glands were enlarged and very oedematous ; they showed in the cortices grey translucent tissue, chiefly in the form of nodules, speckled with white caseous foci. Kidneys.—The left kidney showed on the surface, as well as in the depth of the cortex, moderately numerous grey tubercles, the largest about 1 millimetre in diameter, the smallest just visible ; the larger ones were definitely opaque in the centre. The right kidney showed similar tubercles. Suprarenal Bodies.—In the cortex of the left suprarenal three minute grey tubercles were seen. In the right suprarenal two or three very minute grey points were seen. Alimentary Tract. Tongue.—On the right side of the tongue there was one minute grey tubercle. Pharynx.—Normal. Tonsils.— Each tonsil contained a few caseous tubercles. Small Intestine—The Peyer’s patches contained sparsely scattered opaque whitish tubercles, smaller than a pin’s head. Large Intestine.—Normal. Mesenteric Glands, Gastric Glands, Colic Glands.— These glands contained scattered discrete caseating miliary tubercles. Testes—Normal. Eyes.—Normal. Various Lymphatic Glunds. Precrural, Popliteal, Gluteal, Ischiatic, Pudic, Lumbar, liac.— These glands contained fairly numerous discrete tubercles, somewhat irregular in outline, ranging in size up to 15 and 2 millimetres ; the tubercles were all caseous in the centre. Right Prescapular, Right Prepectoral, Asillary (Right and Left)—They contained similar discrete caseating tubercles. Parotid, Submazxillary, Pharyngeal, Renal.—In the parotid, submaxillary, and pharyngeal glands, and in the renal gland, the tubercles were rather more numerous. Coeliac.—The coeliac glands were a little enlarged, and showed in the cortices early caseous foci in a grey translucent matrix. Several haemolymph glands contained small caseous tubercles. 227 VIRUS P. VIII. (May 11, 1905.) SUBMAXILLARY GLAND. CuLTuRE InocuLations, SepremBeR 5, 1905. The strain was derived from the original material, and had been in cultivation a total period of 117 days. . The culture used was the 5th generation, 20 days old. CALF 328. RABBITS. Subcutaneous. ne Visetioh ae | F Ro caer eee 4 oe : Number. | Method. Dose. | Duration of Life. | Result. Dose : 50 mg. | Died October 3, 1905. eet a =il a= ck eee x oe Ne shes ee 28 days. 427 Intrav. | Olmg. ! D. 18 days. G. T. P.M.—General Tuberculosis, severe. 424 Intrap. 1:0 mg. | D.18 =, G. T. | 426 Intrap. 0-1 mg. D.27~—, G. T. 429 Intrap. | 001mg.) D.32 ,, G. T. 405 | Baten, | Mia, | Dee ,, G. T. 428 Subcut. 0-1 mg. D. 87 _~,, | G. T. | CALF 328. Virus P. VIII. Subcutaneous inoculation of culture derived from the original material of Virus P. VIII. Dose—50 milligrammes. Date—September 5, 1905. Weight at Inoculation—34-0 kilogrammes. [Age about 6 weeks. ] Died—October 3, 1905. [28 days after inoculation. ] Clinical Notes. it was pear shaped in outline, and about 2°5 em. thick. The prescapular gland was very large and measured 12'5 cm.in length. Up to this time the general con- dition of the animal had been good ; it now began to Hight days after inoculation on the left side of the neck, there was a local tumour measuring 9 by 5°5 cm. ; the ee ea gland was slightly enlarged, Joc¢ weight, and became thin and ill. The progress of ‘65 em. in length. the disease was very rapid and the animal died on the On the 17th day the tumour measured 14 by 9 cm., 28th day after inoculation. Temperature. SEPTEMBER O2TSBER 228 Tuberculin Test. The calf was not tested subsequent to inoculation. Weights. Kilogrammes. September 5, 1905 ... ie ow. 34:0 September 19, 1905 ... ioe .. 36°28 September 26, 1905 ... wee 35°37 October 3, 1905 aig oe gee, BLED. Total loss of weight.—2°71 kilogrammes. POST-MORTEM EXAMINATION. Local Lesion.—At the seat of inoculation in the subcutaneous tissues on the left side of the neck there was a flattened tumour, weighing 340 grammes, aud measuring 14 by 9 by 4cm. It was composed of a dense yellowish network of caseous tissue, softening in the centre, where there was a small cavity contain- ing turbid fluid. Left Prescapular Gland.—The left prescapular gland weighed 170 grammes, and measured 10 by 6°5 by 4°5 cm., and was adherent to the subjacent muscle. It was firm, and the cortex was infiltrated with a yellow caseous network, the hilum containing dis- crete tubercles. Left Prepectoral Gilands.—The left prepectoral (reniform) gland measured 2°5 by 2 cm. It was oedematous, and contained in the cortex four yellowish foci about a pinhead in size. The round prepectoral gland was firm, and irfiltrated with irregular caseous foci. Right Prescapular Gland.—The right prescapular gland contained fairly numerous opaque yellow tubercles, about 1 millimetre in diameter. Cervical Glands.—The cervical glands on the left side in the mid cervical region contained numerous very minute opaque foci, A lower cervical gland, three centimetres long, contained at one extremity a few isolated tubercles, and at the other numerous minute caseous foci. On the right sile the cervical glands contained fairly numerous minute caseous foci in the cortices. Thorax. Heart.—In the muscle of the heart there was one grey tubercle; the valves and endocardium were normal. Lungs.—The left lung was closely filled with grey tubercles, opaque in the centre, varyiug in size up to 1:5 millimetre, and becoming confluent in patches. The anterior lobe was solid and sank in water. The substance of the right lung was similarly filled with tubercles, and the greater part of its anterior and middle lobes, and the apex of the posterior lobe were solid and airless. Bronchial Glands—The right bronchial gland was firm and infiltrated with yellow caseous foci. The left bronchial gland contained numerous minute foci in the cortex. Mediastinal Glands.—The posterior mediastinal glands were infiltrated throughout the cortex with yellow caseous foci. Four other glands contained foci numerous but still discrete. Thyroid.—On the posterior part of the thyroid cartilage internally were nine flattened translucent tubercles, caseous in the centre, and up to 2°5 milli- metres in diameter. Trachea.—There were four similar tubercles down the trachea. Abdomen, Omentum.—On the omentum were sparsely scattered flattened grey tubercles, opaque in the centre, about 15 millimetre in diameter. Spleen.—The spleen weighed 283 grammes, and was closely packed with greyish opaque miliary tubercles. Liver.—In the liver were numerous grey tubercles, caseous in the centre, varying in size from a pinpoint. up to 1°5 millimetre. Portal Glands.— The portal glands contained numerous minute opaque foci in the cortices, giving them a yellowish granular appearance. Gall Bladder.—In the mucous membrane of the gall bladder were ten opaque tubercles, the largest the size of a pinhead. Kidneys.—The kidneys contained sparsely scattered grey tubercles with caseous centres, froma point up to 1 millimetre in diameter. Alimentary Tract. Pharynx, Tongue.—On each side of the pharynx, on the folds at the root of the tongue, were four small. opaque tubercles. Tonsils.—The tonsils were normal. Intestines—Sparsely scattered over the mucous membrane of the small intestine were minute yellowish tubercles. There were a few in the Peyer’s patches. Large intestine normal. Mesenteric Glands.—In all the mesenteric glands there was a moderate number of opaque slightly yellow tubercles, varying in size up to a small shot. Lieo-Colic Glands.—In the ileo-colic glands a few pinhead-sized opaque tubercles were visible, and the substance felt firm. Lymphatic Glands. Gastric, Coeliac, Precrural, Popliteal, and Gluteal.— They contained numerous minute opaque yellow tubercles, Awillary, Iliac, Sacro-Iliac, Lumbar, and Retro- pharyngeal. — All contained rather larger caseous tubercles. ; Parotid.—In the parotid lymphatic glands there were several yellow tubercles, the largest almost equal. to a wheat grain in size. The haemolymph glands contained caseous foci. 229 VIRUS P. IX. (May 18, 1905.) SUBMAXILLARY GLAND, CuLturE Inocunations, SEPTEMBER 20, 1905. The strain was derived from the original material, and had been in cultivation a total period of 125 days. The culture used was the 7th generation, 20 days old. CALF 334, salle Subcutaneous. Dose : 50 mg. Number. | Method. Dose. Duration of Life. | Result. Died : November 4, 1905. 45 days. 469 Intrav. | 0-1 mg. D. 16 days G.T. P.M.—General Tuberculosis, severe. 466 Intrap. 1-0 mg. D.20 , G.T. 468 Intrap. O'1'mg. *| D.30 =, G. T. 465 Subeut. | 10-0 mg. ‘|’ D. 28 ,, G. T. 467 Subcut. 1:0 mg. D.51 =, G. T. 470 Subcut. 0-1 mg. D. 80 =, G. T. CALF 334. Virus P. IX, Subcutaneous inoculation of culture derived from the original material of Virus P. TX, Dose—50 milligrammes. Date—September 20, 1905. Weight at Inoculation—41'72 kilogrammes. [Age about 9 weeks. ] Died—November 4, 1905. [45 days after inoculation.] Clinical Notes. Ten days after inoculation on the left side of the neck an elongated local tumour had developed, extending into the dewlap ; it measured superficially 15 by 75 cm. The adjacent prescapular gland was enlarged, 7°5 cm. in length; the prepectoral gland was the size of a large marble. On the 20th day the tumour was very large, extending from the middle of the neck to below the level of the trachea ; it measured 18 by 12:5 cm. and projected considerably. There was a large amount of 27676 oedema in the dewlap and around tho inforior part of the tumour. The prescapular gland was 12'5 cm. long, and the prepectoral gland was very large; the animal was beginning to lose flesh and its coat was rough, its respiration was not much increased. On the fortysecond day the tumour measured about 23 by 12°5 cm. and was very promixent and thick, extending into the dewlap and surrounded at the lower part by much oedema. The animal was much emaciated, the appetite poor, and the respiration increased though not toa marked degree. Weakness and emaciation increased, and the calf died three days later. aN 9 ol 30 Temperature. SEPTEMBER, OCTOBER 38 Tuberculin Test.—The calf was not tested subse- quent to inoculation. Weights. Kilogrammes. September 20, 1905... aa 41-72 September 26, 1905 43°08 October 5, 1905 aoe sis 41-72 October 10,1905 ... 43-08 November 4, 1905 40°36 Total loss of weight during experiment.—1°36 kilo- gramme. POST-MORTEM EXAMINATION. Carcass.—Emaciated. Local Lesion—At the seat of inoculation on the left side of the neck there was a very large tumour measuring 20°5 cm. vertically, 12-5 cm. from side to side and 5°5 cm. in greatest thickness ; it extended into the loose tissues of the neck and under the trachea slightly to the opposite side ; on removal it weighed with the infiltrated skin and muscles 907 grammes ; on section it showed between the skin and muscles a mass of dense yellow caseo-necrotic tissue, spongy and juicy in the centre, 3 cm. in greatest thickness ; the skin was thickened thoroughly adherent to the necrotic mass and extensively infiltrated with caseous tubercles ; the epidermis was cracked and could be peeled off from the greater part of the tumour exposing a red granular surface ; the muscles for a depth of 2:5 cm. were closely studded with yellow caseous nodules and showed a great increase in interstitial connective tissue. Between the local tumour and the nearest glands there were several thickened caseous lymphatic vessels. The tissues around the lower part of the tumour were oedematous. Left Prescapular Gland.—The left prescapular gland weighed 170 grammes, and measured 9°5 by 4°5 by 5°5 cm. ; on section it was composed throughout of dense pinkish yellow caseated tissue very slightly gritty from calcification (the muscles around the gland were tightly adherent to the capsule but not infiltrated). Left Prepectoral Glands.—The rounded prepectoral gland was a little over 2°5 cm. in diameter and resembled the prescapular. The reniform gland, 3°5 cm. in length, was oedematous and showed part of the cortex composed of reddish grey tissue beset with early caseous foci. Cervical Gilands—On the left side the superior cervical gland showed part of the cortex composed of greyish red tissue beset with caseous foci; a mid- cervical gland, the size of a thrush’s egg, was indu- rated and caseating throughout (not so advanced as the prescapular) ; two smaller glands nearer the thorax were indurated and showed streaks and foci of caseation; a gland, the size of a pigeon’s egg, close to the entrance to the thorax was dense yellow NOVEMBER and caseous throughout ; the corresponding gland on the opposite side was enlarged and showed a small patch of the cortex yellow and caseous, the rest translucent and closely beset with yellow caseous foci : other cervical glands on the right side contained each a few caseous nodules. Abdomen. Omentum and Peritoneum.—The omentum showed numerous discrete flattened nodules ranging in size from a millet seed to a split pea; they were firm, yellowish in the centre and greyish red around the margins. On the parietal peritoneum there was one loosely attached lenticular nodule about the size of a split pea. Spleen.—The spleen was very firm and much enlarged (weight 538 grammes). On the surface there were two flattened caseating nodules, the largest 5 mm. in diameter : on section the pulp was packed with irregular yellow caseous nodules with reddish margins ranging up to about 5 mm. in diameter. Liver.—The liver was enlarged and pale; on the surface under the capsule numerous nodules ranging in size from 1 to about 5 mm. in diameter were seen; the larger ones were slightly raised above the surface and on section were thin and compressed; in the depth of the’ liver there was numerous evenly dis- tributed yellow caseous tubercles, the largest about 2 mm. in diameter. Gall Bladder.—The gall bladder showed one sub- mucous caseous tubercle. Portal Glands—The portal glands were enlarged and showed in the cortices irregular coalescing nodules with caseous centres and grey margins. Kidneys.—The left kidney showed in the cortex moderately numerous tubercles ranging in size from a pin’s head to a wheat grain; they were yellow and caseous in the centre and had grey translucent margins ; scattered about was a number of minute points just visible to the eye; most of these were grey, a few were yellowish. : The right kidney contained similar tubercles ; they were however not so numerous and generally smaller. Suprarenal Bodies—The suprarenal bodies were pale ; no tubercles seen, Thorax. Heart.—Under the pericardium covering the right auricle there was a caseous tubercle a little smaller than a wheat grain ; otherwise the heart was normal. Pleura.—Along the margins of the ribs chiefly in the dorsal regions were lines of vascular vegetations containing flattened caseating tubercles up to 1:5 mm. in diameter. On the pleural surface of the diaphragm there were numerous patches of slightly raised reddish vegetations containing small yellowish white tubercles ; there were similar patches on the caval fold of pleura and on the pleura covering the pericardium, 231. _ Lungs.—The lungs were heavy; the cephalic and right middle lobes were dark-red and almost com- pletely solid; on the surfaces of the right cepbalic and middle lobes were some small patches of yellow slightly adherent lymph ; the antero-ventral parts of the caudal lobes were dark red and consolidated, and in the rest of these lobes there were irregular patches of consolidation. The lung substance was moderately closely beset with yellow caseating tubercles ranging in size from a pin’s head to a wheat grain; they appeared to be most numerous in the solid parts of the lung; the subpleural tubercles in the anterior lobes were considerably flattened out. The cut puns of the caudal lobes exuded frothy oedema uid. Thoracic Lymphatic Glands.—The caudal media- stinal gland was much enlarged and showed the cortex composed of firm translucent grey tissue mottled with irregular yellow patches of caseation in places form- ing a network ; the medulla was deeply congested and oedematous. The other dorsal mediastinal glands and the bronchial glands were similarly affected but a little more advanced in caseation. The ventral mediastinal and the vertebral glands and a gland in the caval fold of pleura contained discrete yellow caseous nodules. Trachea.—The trachea was filled with froth and Showed on the mucous surface three slightly raised congested tubercles. Alimentary Tract. Tongue, pharynx, and larynx.—Normal, Tonsils.—The tonsils showed each one or two small caseous nodules. Small Intestines—In the long Peyer’s patch there was a moderate number of softened caseous tubercles, the largest about 1°5 mm. in diameter ; in the rest of the intestine they were sparsely scattered. Large Intestine.—Normal. The Gastric, Mesenteric and Colic Glands resembled the precrural. : Lleo-colic Glands.—The ileo-colic glands were en- larged, and showed the cortices composed of firm greyish red tissue beset with irregular caseous foci. Various Lymphatic Glands.—The precrural, popli- teal, gluteal, pudic, parotid, submaxillary, axillary, right prescapular and right prepectoral glands con- tained a moderate number of irregular yellow caseous nodules ranging up to 3°5 or 4 mm. in diameter. The left axillary gland was deeply congested, oedematous, and much enlarged; the right pres- capular was slightly enlarged and congested. The lumbar and right renal glands were enlarged and closely beset with caseous nodules becoming confluent. The coeliac glands were enlarged and composed throughout of very dense translucent tissue mottled with irregular yellow patches of caseation. All the haemolymph glands contained caseous nodules. Testes.—Normal. 27676 2N 2 232 VIRUS P. XI. (June 1, 1905.) SUBMAXILLARY GLAND. CuLTURE INocULATIONS, SEPTEMBER 6, 1905. The strain was derived from tke original material, and had been in cultivation a total period of 97 days. The culture used was the 4th generation, 20 and 21 days old. an 338. RABBITS. Subcutaneous. Durati ¢ Lif Resul Dose: 46°5 mg. Number. | Method. Dose. uration of Life. esult, Died : October 13, 1905. 37 days. 438 Intrav. 0-1 mg. D. 19 days G. T. P.M.—General Tuberculosis, severe. 436 Intrap. 1:0 mg. D.17_,, G.T. 439 Intrap. 0-1 mg. D.23 G. T. 440 Intrap. 0-1 mg. D. 27 =, G. T. 442 Intrap. | 0°01 mg. D. 29 =, G.T. 437 Subcut. 1:0 mg. D. 120 ,, G. T. 441 Subcut. 0-1 mg. D. 62 «4, G.T. CALF 8338. Virus P. XI. Subcutaneous inoculation of culture derived from the original material of Virus P. XI. Dose—46'5 milligrammes. Date—September 6, 1905. Weight at Inoculation—31:75 kilogrammes. [Age about 6 weeks. ] Died—October 13, 1905. [37 days after inoculation. ] Clinical Notes. Seven days after inoculation on the left side of the neck there was a firm local tumour adherent to skin and slightly adherent to muscle, measuring 7°5 by 5 cm., and about 1:25 em. thick. The adjacent prescapular gland was slightly enlarged, 65 cm. in leugth. On the 24th day the tumour was elongated, measuring 15 by 7 cm. by about 2°5 cm. in thick- ness ; the prescapular gland was moderately enlarged, 9 cm. in length ; the prepectoral gland was slightly enlarged. The general condition of the animal was not very good. On the 34th day the animal’s condition was poor, its coat was rough, and there was slight emaciation ; respiration 48, somewhat jerky. Three days later the calf died. Temperature. SEPTEMBER. OG TOBER 233 Tuberculin Test—The calf was not tested sub- sequent to inoculation. ' Weights. Kilogrammes. September 6 ise si vi B75 September 12 sir sels ee 33°55 S»ptember 19 ose ais es 34°91 September 26 ix ees vee = 82°65 October 13 eee wea 26°30 Total loss of weight.—5-43 kilogrammes. Rate of loss per week.—1°04 kilogramme. POST-MORTEM EXAMINATION. Carcass.—In poor condition. _ Local Lesion—The tumour at the seat of inocula- tion on the left side of the neck was lenticular in outline, measuring 12-5 by 7 om. by a little over 25 cm.in greatest thickness, and weighing with skin and muscles 226 grammes, It was composed of the asual pinkish yellow caseo-necrotic tissue, showing in the centre a series of small spaces filled with yellow serous fluid. ' The muscles were infiltrated for a short distance ; the skin was thickened, and studded with yellow tubercles, particularly over the centre of the tumour. Left Prescapular Gland.—The left prescapular gland weighed 141 grammes, and measured 9 by 6 by 4 cm. On section it consisted throughout of dense pinkish yellow caseated tissue. : Left Prepectoral Glands.—One, the size of a pigeon’s egg, showed the cortex firm and in a state of early caseation ; the medulla was distended with oedema fluid. Another gland was enlarged, oedematous, and contained discrete yellow tubercles. Cervical Gilands.—A small gland in the middle of the neck on the left side showed the greater part of the cortex in a state of early caseation ; other cervical glands contained discrete caseous tubercles. Thorax. Heart.—Normal. Parietal Pleura.—The parietal pleura showed a few flattened caseating nodules, and here and there a slight hypertrophy of the lymphatic fringes, at the base of some of which minute grey granules were seen. Diaphragm.—On the pleural surface of the dia- phragm there was one small flattened caseating nodule, and slight hypertrophy of the lymphatic fringes. Lungs.—The lungs were moderately closely beset with yellow caseous tubercles, ranging in size from a small point to a little over 1 millimetre in diameter ; they were somewhat irregular in outline, and showed a tendency to occur in small groups. The right cephalic lobe, with the exception of the thin dorsal border, and the ventral portion of the right middle lobe, were extensively hepatised, being firm, heavy, and dark red. In the ventral portion of the left cephalic lobe and the antero-ventral portions of the caudal lobes there were patches of consolidation, irregular in outline, and not sharply defined, pieces from which sank in water. On the dorsal surface of the right caudal lobe there was a large patch of collapse, which appeared to be of old standing. In other parts of the lung single tubercles and groups of tubercles were surrounded by angular red patches. One of these patches showed early tuberculous infil- tration, é Mediastinal Glands.—The caudal mediastinal gland was moderately enlarged, and showed the cortex dense, yellowish, and far advanced in caseation, very little grey translucent tissue remaining ; the medulla was congested and oedematous. Other mediastinal and the bronchial glands were similarly affected. They weighed together 85 grammes, _ The infratracheal, oesophageal, ventral and medias- tinal glands, and the glands in the caval fold, contained ‘discrete yellow caseous tubercles. Trachea.—On the mucous membrane in the anterior part of the trachea there was one caseous tubercle. Abdomen. Omentum.—On the inferior surface of the omentum were scattered lenticular loosely attached nodules, ranging up to 2 millimetres in diameter; all had yellow centres, and some were congested. Spleen.— The spleen was enlarged, weight 282 grammes. The pulp was closely packed with yellow caseous nodules with greyish red margins, the largest about 2 millimetres in diameter, Liver—The liver contained moderately numerous opaque caseous tubercles, ranging in size from 0°5 or less up to 1 millimetre in diameter ; they were evenly distributed throughout the substance. Gall Bladder—The gall bladder showed one pin- head-sized submucous tubercle, and three or four small ulcers with caseous floor and congested margins. Portal Glands—The ‘portal glands were enlarged. They showed in the cortices nodular masses of aggre- gated caseating tubercles. Kidneys.—The left kidney showed on the surface fairly numerous tubercles with grey margins and yellow centres, the majority the size of a millet seed, some barely visible to the eye. On section many of the larger surface tubercles extended for some distance down into the cortex. A few tubercles were seen in the depth of the cortex. In the right kidney were similar, but less numerous tubercles. These did not show the same tendency to extend downwards into the cortex. left suprarenal showed one Suprarenals. — The The right was normal. minute grey tubercle. Alimentary Tract. Tongue.—The mucous membrane at the base of the tongue showed two small ulcers with yellow necrotic floors. Tonsils—Each tonsil contained a few caseous tubercles. Small Intestine—Almost all the Peyer’s patches contained pinhead-sized caseous tubercles in small number ; they were most numerous in the long Peyer’s patch. Caseous tubercles were seen also here and there under the mucous membrane. Large Intestine.—Normal. , Mesenteric Glands.--The mesenteric glands were not enlarged. They contained discrete caseous tubercles and caseating patches in the cortices. Gastric’ Glands and Colic Glands.—These glands contained discrete tubercles. Various Lymphatic Glands. Coeliac Glands.-—T wo coeliac glands were enlarged, firm, and on section showed yellow caseous patches ; other coeliac glands contained discrete nodules. Renal Gland.—The renal gland was enlarged, and closely beset with yellow caséous tubercles, up to 2 millimetres in diameter. Precrural, Pudic, Popliteal, Gluteal, Ischiatic, Axillary, Right Prescapular, Right Prepectoral, and Parotid. — These glands contained yellow caseous nodules, somewhat irregular in outline, ranging in size up to about 2 millimetres. Iliac, Itio-Sacral, Submasillary and Pharyngeal.— There were similar, but rather more numerous nodules in these glands. : Testes and Eyes,—Normal. Sec ARR sen stamens a 234 VIRUS P. XII. (June 26, 1905.) SUBMAXILLARY GLAND. CuLture Inocuxations, DecEMBER 14, 1905. The strain was derived from the original material, through G.P. 1636, and had been in cultivation a total period of 101 days. The culture used was the 5th generation, 20 days old. BITS. CALF 376. Han Subcuta i iia Number. | Method. Dose. Duration of Life. Result. Dose: 50 mg. Killed when dying: Jan. 2, 1906. 19 days. 597 Intrav. | 0-1 mg. D. 19 days G. T. P.M.—General Tuberculosis, severe. 598 Intrap. 1:0 mg. D. 16 =, G. T. 600 Intrap. | 0-1 mg. D. 18 «yy G. T. 602 Intrap. | 0:01 mg. D. 75 ,, G. T. 599 Subeut. | 1:0 mg. D. 97 ~OCy, G.T. 603 Subcut. | 0-1 mg. D.105-— G. T. 601 Subcut. | 0:01 mg. D. 89 G. T. CULTURE INOCULATIONS, JUNE 29, 1906. The strain was derived from the original material, through G.P. 1636, and had been in cultivation a total period of 298 days. The culture used was the 9th generation, 21 days old. * CALF 444, CALF 450. Subcutaneous. Subcutaneous. Dose : 50 mg. Dose : 50 mg. Killed when dying : Died: Aug. 9, 1906. Aug. 16,1906, 48 days. 41 days. P.M.—General Tuberculosis, severe. P.M.—General Tuberculosis, severe. CALF 376. Virus P. XII. Subcutaneous inoculation of culture derived from the original material of Virus P. XII through G.P. 1636, Dose—50 milligrammes. Date—December 14, 1905. Weight at Inoculation—39-46 kilogrammes. [Age about 6 weeks.] Killed when dying—January 2,1906. [19 days after inoculation.] length; the prepectoral gland was the size of a Clinical Notes. walnut. On the 18th day the calf became suddenly ill Seven days after inoculation on the left side of the refusing food. On the following morning it ae neck there was a local tumour measuring 10 by 5cm.; found in a moribund condition, and was therefore the adjacent prescapular gland was enlarged, 6 cm. in killed. 235 Temperature. ECEMBER fa Tuberculin Test.—The calf was not tested subse- quent to inoculation. Weights. Kilogrammes. December 14, 1905 aes 39°46 January 2,1906 ... : 33°55 Total loss of weight.—5'91 kilogrammes. POST-MORTEM EXAMINATION. General Condition.—Moderate. Local Lesion.—At the seat of inoculation there was a tumour, which measured 9 by 5°5 cm. in superficial area. On section it consisted of congested yellow necrotic tissue, broken down in the centre to form thin watery fluid, turbid with caseous flakes. At this part the necrotic tissue infiltrated the muscles, and was about an inch in thickness. The greater part of the tumour, however, was thin, and situated wholly in the subcutaneous tissue. Two thickened caseous lymphatic vessels extended from the Jocal tumour towards the prescapular gland. Left Prescapular G'land.—The left prescapular gland weighed 56 grammes, and measured 8 by 4°5 by 25cm. Onsectionthe cortex was composed practically throughout of dense pinkish yellow caseo - necrotic tissue. Right Prescapular Gland.—The right prescapular gland appeared normal. Prepectoral Glands. — The rounded _prepectoral gland measured nearly 2°5 cm. in diameter, and was composed throughout of dense pinkish yellow caseo- necrotic tissue ; the reniform gland was enlarged, and showed part of the cortex firm, and in a state of early caseation. The prepectoral glands on the right side appeared normal. Agillary Glands—The axillary glands appeared normal. Cervical Glands—Normal. Thorax. Pleura.—Normal. Lungs.—The lungs showed in various parts firm red patches of collapse with a lobular outline ; they were largest in the right cephalic and middle lobes. The lung substance contained numerous grey translucent tubercles, mostly about 0°5 millimetre in diameter ; the tubercles appeared to be most numerous in the anterior lobes. Trachea.—Normal. Thoracic Glands.—The caudal mediastinal gland was slightly enlarged and firmer than normal. On section the cortex showed in some places a fine yellow network, in others irregular caseous foci, singly or in small groups. Other mediastinal and the bron- chial glands were in a similar condition, but a little more extensively caseated. Heart.—Muscle and valves normal. Abdomen. Omentum.—The omentum showed on the under surface a few minute congested tubercles. In the omental sac there was some yellowish turbid fluid together with flakesof lymph. Between the omentum, along its attachment, and the stomach, there was a band of yellow fibrinopurulent exudation, which contained clear serous fluid in the centre. Stomach.—Near the pylorus, on the ventral surface of the stomach, there was an oval aperture, covered by omentum, which adhered to the margins, On the mucous surface the opening was seen to be situated in the centre of an ulcer, one inch in greatest diameter The ulcer appeared to be of old standing, the margins being raised, thickened, and somewhat puckered ; the base was thin, and showed in the centre the opening described above. Near the large ulcer there were two smaller ulcers with thickened margins. Intestines.—Normal. Mesenteric Glands.—There were a few doubtful tubercles in the mesenteric glands. Spleen.—T he spleen weighed 170 grammes, On the convex surface there was a thin layer of fibrinous lymph. On section the pulp was dark red, moderately firm, and showed no tubercles. Liver.—The liver was firm and dark red, and showed throughout the substance moderately numerous opaque minute tubercles. Gall Bladder—Normal. Portal Glands—The portal glands were slightly enlarged, and beset in the cortices with irregular caseous foci. Kidneys. — Each kidney showed in the cortex numerous grey ill-defined tubercles, the largest about 1 millimetre or less in diameter. Suprarenals.—The right suprarenal body contained several minute pearly grey tubercles. The left appeared normal. Lumbar Glands.—The lumbar glands contained a moderate number of irregular caseous foci. Coeliac Glands.—One coeliac gland showed part of the cortex firm and grey, with early caseous foci. Iliac Glands.—Kach iliac gland showed one or two greyish white tubercles, 236 Genito-Urinary System. Testes.—Normal. Bladder.—The bladder was greatly distended ; its mucous membrane was normal. Special Glands. Right Precrural.—Contained two caseous foci. Left Precrural.—Normal. Pudic.—One pudic gland contained a caseous focus ; the other was normal. Gluteal and Left Popliteal—Each contained a few caseous foci. Ischiatic, Submazillary, and Parotid.—Normal. Retropharyngeal.—Each contained one or two small grey tubercles. Tongue, Tonsils, Pharynx, and Laryne.—Normal. Eyes.—Normal. Microscopical Examinations. Pharyngeal Gland.—Numerous tubercle bacilli. Gluteal Gland.—Numerous tubercle bacilli. Mesenteric Gland.—No tubercle bacilli. CALF 444. Virus P. XII, Subcutaneous inoculation, on left side of neck, of culture derived from original material of Virus P, XII through G.P. 1636. Dose—50 milligrammes. Date—June 29, 1906. Weight at Inoculation—37'19 kilogrammes. Killed when dying—August 16, 1906. Clinical Notes. Seven days after inoculation on the left side of the neck a flat subcutaneous thickening could be felt, -10 by 6°5 cm. in area ; the adjacent prescapular gland was enlarged, measuring 7°5 cm. in length. On the fourteenth day there was a firm prominent [Age about 7 weeks.] [48 days after inoculation. ] infiltrating local tumour measuring 14 by 9 cm. ;: the prescapular gland measured 10 cm. in length. The tumour continued to increase in size ; on the fortieth day it measured 20 by 10 cm., and the gland was 12:7 cm. in length. The calf was unwell, the respiration was distinctly increased, and it was apparently losing flesh. The progress of the disease was subsequently very rapid, and the calf was killed when in a dying condition 8 days later. Temperature. ~ GUNE * ’ Jv ne yy ul ip we 4 by Tuberculin Test—The calf was not tested subse- ‘quent to inoculation. Weights. Kilogrammes, June 29, 1906 1 aa bus aia 37°19 August 16,1906... ies oes 36-73 Total loss of weight during experiment.—460 grammes, POST-MORTEM EXAMINATION. General Condition —Thin. Local Lesion —At the seat of incculation, on the - PUGUST ) = left side of the neck, there was a large firm tumour weighing, with skin and muscles to which it was attached, 538 grammes, and measuring 19 by 9 cm., by nearly 5 cm. in preatest thickness. On section it was lenticular in outline, and composed of pinkish yellow caseo-necrotic tissue a little spongy and juicy in the centre but containing no large cavity. The skin was thickened, universally adherent and beset with yellow caseous tubercles principally over the central part of the tumour. The muscles were infiltrated and caseous, to a greater extent and depth under the upper part of the tumour than under the lower. Left Prescapuiar Gland.—The left prescapular gland weighed 170 grammes, and measured 9 by 5 by 5:5 cm. 237 and was composed throughout of dense pinkish yellow caseo-necrotic substance. Right Prescapular Gland.—The right prescapular gland, not enlarged, contained several yellow caseous nodules, the largest 4 mm. in diameter. Prepectoral Glands.—The rounded prepectoral gland on the left side, the size of a pea, was indurated and caseous around the cortex. The kidney shaped gland on the same side was much enlarged and firmer than normal, and showed besides a number of small caseous foci several irregular caseous nodules up to a hemp seed. The prepectoral glands on the right side contained several small caseous nodules. Left Cervical Glands.—The glands in the middle of the neck were slightly enlarged, one was indurated and showed a few caseous foci in the cortex, the others contained discrete caseous nodules. The superior cervical gland contained a number of nodules. The mastoid gland was slightly enlarged and con- tained discrete caseous nodules up to a hemp seed, and one firm caseous nodule the size of a French bean. Right Cervical Glands—The cervical glands on the right side contained nodules similar to those in the peripheral lymphatic glands. Axillary Glands.—The axillary glands contained nodules similar to those in the right prescapular. Thorax, Pleura.—The costal pleura was normal. On the pleura covering the diaphragm there was a loosely attached caseating nodule the size of a hemp seed. Lungs.—The lungs were moderately closely beset with firm greyish yellow caseating nodules ranging in size from a pin’s head to a small pea. The nodules were most numerous in the ventral portions of the lobes while in several places they were aggregated together and confluent. The nodules under the pleura were slightly raised above the surface and flattened. On section similar nodules were seen throughout the parenchyma; here and there in the depth were caseating masses occupying some part of a lobule; the ventral portions of the cephalic lobe and the antero- ventral portions of the caudal were diffusely consoli- dated but still air-containing ; elsewhere in the lung single nodules and small groups of nodules were surrounded by angular red zones of consolidation. Thoracic Glands.—J ust within the entrance to the thorax there was a firm gland the size of a pheasant’s egg which was dense yellow and caseous throughout. The bronchial and mediastinal glands showed a moderate degree of enlargement. On section the cortices were firm and far advanced in caseation, only a small amount of grey translucent tissue remaining between the caseous areas. Heart and Pericardium.—Normal. Larynx and Trachea.—Normal. Abdomen. Omentum and Peritoneum.—On the ventral surface of the omentum there were three firm flattened lenti- cular nodules with yellow centres, the largest about 2 mm. in diameter. There was no hypertrophy of the fringes. The peritoneum was normal. Spleen.—The spleen was small, weighing 113 grammes. the pulp a moderate number only of yellow firm tubercles, the largest a little larger than a millet seed. Liver.—The liver at first sight appeared normal ; on close inspection, however, a few very sparsely scattered opaque greyish white tubercles, the largest not more than 5 mm. in diameter were seen. On It showed evenly distributed throughout - section similar tubercles were sparsely scattered throughout the substance. Gall Bladder.—Normal. Portal Glands.—The portal glands were moder- ately enlarged and closely beset with coalescing caseous nodules arranged principally in groups. Kidneys—The left kidney showed on the sur- face four greyish-white translucent pinhead sized tubercles ; none was seen in the depth. On the surface of the right kidney there were two. or three tubercles similar to those on the left, and also a few minute grey foci. In the depth of the cortex one pinhead-sized tubercle was seen. Suprarenal Bodies—The left suprarenal body contained one pinhead-sized tubercle. The right appeared normal. Coeliac G'lands.—T wo coeliac glands were slightly enlarged, they were indurated and showed their cortices extensively caseated ; other coeliac glands contained discrete caseous nodules up to a hemp seed in size. The Iliac, Ilio-sacral, and Lumbar Glands were slightly enlarged and contained discrete caseating nodules. Renal Gland—The renal gland was distinctly enlarged, and closely beset with caseous nodules. Alimentary Tract. Pharynx.—Normal. Tonsils—One tonsil contained a pea sized caseous nodule, the other three or four, the largest the size of a hemp seed. The Submasillary, Pharyngeal, and Parotideal Glands contained numerous firm yellow caseous nodules, the largest 5 mm. in diameter. Gastric Glands—The gastric glands contained discrete caseous nodules. Intestines—All the Peyer’s patches in the small intestine contained yellow caseous nodules up to about 2 mm. in diameter ; they were most numerous in the long patch where several showed the mucous membrane over them ulcerated. There were a few nodules under the mucous membrane as well. Large Intestine : Beneath the mucous membrane of the caecum and the early part of the colon there were numerous raised congested softened caseous nodules, the largest a little larger than a millet seed. Meserteric Glands.—The mesenteric glands were slightly enlarged and showed their cortices composed practically throughout of firm tissue in an advanced stage of caseation, but not so advanced as in the thoracic glands. Colic Glands.—The colic glands contained discrete caseous nodules similar to those in the gastric glands. Testes.—Normal. Eyes.—Normal. Various Peripheral Lymphatic Glands. The Precrural and Pudic Glands contained numerous discrete firm yellow caseating nodules somewhat irregular in outline, the largest about the size of a hemp seed. The Popliteal and Gluteal Glands resembled the precrural. One Ischiatic gland contained a caseating nodule. Haemo-lymph Glands all over the body contained single caseous nodules up to a hemp seed in size. 27676 20 9 a 38 CALF 450. Virus P. XII. Subcutaneous inoculation of culture derived from the original material of Virus P. XII, through G.P 6. Dose—50 milligrammes. Date—June 29, 1906. Weight at Inoculation—36°74 kilogrammes. Died—August 9, 1906. Clinical Notes. Seven days after inoculation on the left side of the neck there was a slightly raised firm local tumour measuring 7°5 by 55cm. The prescapular gland was slightly enlarged, about 5 cm. in length. On the fourteenth day the tumour measured 10 by 6 cm. ; the adjacent muscles were slightly infiltrated, [Age about 7 weeks. ] [41 days after inoculation.] prescapular gland was considerably enlarged, 10 cm. in length. The tumour continued to increase in size; on the twenty-eighth day it measured 125 by 7-5 cm. by about 1 cm. in thickness. On the thirty-first day the respiration of the calf was increased in frequency ; on the thirty-eighth day the animal was acutely ill; it was wasting and lay on the floor most of the day, the respiration was laboured but not very frequent. Three days later the calf was and there was some oedema of the dewlap. The dead. Temperature. JUNE = Tuberculin Test—The calf was not tested subse- ‘quent to inoculation. Weights. Kilogrammes. June 29, 1906 36°74 August 9, 1906 34:46 Total loss of weight during experiment—?2°28 kilo- ‘grammes. PGST-MORTEM EXAMINATION. General Condition.—Thin. Local Lesion —At the seat of inoculation, on the left side of the neck, there was an oval tumour measur- ing 12°5 by 7:5 em., by 3:2 cm. in greatest thickness, composed of yellow congested caseo-necrotic tissue infiltrating to a slight extent the muscles. The interior part of the tumour was spongy and juicy and softer than the peripheral part. The skin was every- where adherent, and over the central part was infil- trated up to the surface and completely replaced by softening caseous substance; elsewhere the skin showed discrete yellow tubercles. Left Prescapular Gland.—The left prescapular gland weighed 141 grammes,and measured 9 by 65 by 4cm.; on section the cortex, except for a small patch at one extremity which contained discrete caseous tubercles, was composed of dense yellow somewhat congested caseous tissue. Right Prescapular Gland.—The right prescapular gland contained discrete yellow caseous nodules, Left Prepectoral Glands.—Two left prepectoral glands, the size of walnuts, were composed throughout of very dense yellow caseated tissue slightly congested and not perceptibly gritty. Another gland, kidney shaped, 4 cm. in length, was oedematous and showed the cortex firmer than normal and beset with very minute early caseous feci. Right Prepectoral Glands.—Normal. Cervical Glands.— A left lower cervical gland, slightly enlarged, and firmer than normal, showed in the cortex numerous minute caseous points. Other cervical glands on this side, and all those on the right, contained discrete yellow caseous nodules. Asillary Glands.—The left axillary gland was slightly enlarged, and on section similar to the kidney- shaped left prepectoral gland. The right axillary gland contained discrete yellow caseous nodules. Thorax. Pleura.—Along the margins of the ribs were small clusters of minute tubercles surrounded by reddish tissue forming incomplete lines. On the pleural surface of the diaphragm there were a few small congested villous tufts with no tubercles. Lungs.— The lungs were heavy ; the left lobes were extensively consolidated, very little air containing tissue remaining. The right lobes were more diffusely 239 consolidated and were of a terra-cotta colour mottled with irregular dark red patches of consolidation. The lung parenchyma was closely and evenly beset with yellow caseous tubercles varying in size from a mere point up to that of a millet seed. Thoracie Glands—The bronchial and mediastinal glands were moderately enlarged, weighing together about 112 grammes. On section the cortices were firm and composed of greyish red translucent tissue well advanced in caseation. The caseous areas formed irregular patches and in some places a coarse network. Heart (muscle and valves).—Normal. Larynx.—There was a miliary caseous tubercle in the mucous membrane of the larynx just above the left vocal cord. Trachea.—The mucous membrane in the upper part of the trachea showed some scattered minute tubercles. Abdomen. Omentum and Peritoneum.—On the ventral surface of the omentum there were about half a dozen flattened’ congested nodules slightly opaque in the centre, the largest slightly larger than a millet seed. The parietal peritoneum was normal. Spleen.—The spleen was soft and slightly atrophied, it weighed a little over 141 grammes. On section the pulp contained a moderate number of discrete firm yellowish tubercles ranging in size from a pin’s head to a millet seed. Liver.—The liver was normal in size, and contained, evenly distributed, a moderate number only of greyish white tubercles ranging in size from a mere point to that of a pin’s head. Portal Glands—The portal glands were slightly enlarged and closely beset in the cortices with caseating tubercles showing a tendency to form groups. Kidneys.—The kidneys contained rather numerous grey tubercles, the largest 1 mm. in diameter, with minute opaque yellow centres. Some of these tubercles were elongated in a radial direction. Suprarenal Bodies——The left showed two greyish white tubercles in the cortex; the right contained one. The Lumbar and Renal Glands were slightly en- larged and closely beset with caseous nodules tending to coalesce. Coeliac Glands.—One coeliac gland, slightly enlarged, was composed of firm grey tissue mottled with irregular yellow caseous foci. Other coeliac glands contained more or less discrete yellow caseous nodules. Alimentary Tract. Tongue.—Normal. Pharyne.—The pharynx was deeply congested, but contained no tubercles. Tonsils—Each tonsil contained a few caseous nodules, the largest the size of a hemp seed. The Pharyngeal, Submaxillary and Parotid Glands were slightly enlarged, and contained numerous yellow caseous nodules varying in size up to that of a hemp seed. Intestines.—The small intestine showed throughout its whole length numerous caseous tubercles, the largest the size of millet seed. These occurred chiefly in Peyer’s patches and were most numerous in the long patch. There were a few caseous nodules in the caecum and the first part of the colon. Gastric Glands.—The gastric glands were slightly enlarged and beset with caseous tubercles. Mesenteric Glands.—The mesenteric glands con- tained numerous caseating nodules, which in some had coalesced. Lleo-Colic Glands.—T he ileo-colic glands were closely beset with tubercles. Colic Glands—The colic glands were slightly enlarged and beset with caseous tubercles. Testes.—Normal. Eyes.—Normal. Various Lymphatic Glands. The Precrural Glands contained moderately nume- rous discrete yellow caseous nodules somewhat irregular in outline with narrow grey margins, the largest about the size of a hemp seed. The Pudic, Iliac, [lio-sacral, Popliteal, and Gluteal Glands contained similar nodules. Several haemo-lymph glands contained discrete caseous nodules. 27676 20 w 240 VIRUS P. XIV. (July 13, 1905.) SUBMAXILLARY GLAND. CULTURE INOCULATIONS. (a) June 29, 1906. The strain was derived from the original material, through G.P. 1623, and had been in cultivation a total period of 290 days. The culture used was the 12th generation, 21 days old. CALF 446. CALF 448. RABBITS. Subcutaneous. Subcutaneous. | | a Dose : 50 mg. Dose : 50 mg. Number. | Method. | Dose. | Beat a Result. Died : Sept. 17, 1906. Killed when moderately well : | : 80 days. October 11, 1906. \ P.M.—Slight general tuber- 104 days. 694 | Intrav. | 0-1 mg.| D. 19 days | G. T. cuiosis. Death from cystitis P.M.—General progressive ‘é and nephritis, caused by urinary fhbseoutons P ae 695 Intrap.| 10 mg.| D.10 ,, Karly G. T. uleols pee - 699 | Intrap. | O-1mg.| D.19 ,, | G.T. 698 Subcut. | 0-Olmg.| D. 58 ,, G. T. CULTURE INOCULATIONS. April 16, 1907. The strain was derived from the lung of Calf 448 and had been in cultivation a total period of 187 days. The culture used was the 8th generation, 21 days old. CALF 536. CALF 538. RABBITS. Subcutaneous. Subcutaneous. Dose: 50 mg Dose : 50 mg. Number. |Method.| Dose. _ Pan at Result. | Died: June 1, 1907. Died : May 22, 1907. oe 46 days. 32 days. | General tuberculosis, severe. General tuberculosis, severe. 1304 Intrav. | 0-1 mg. | D. 19 days. | G. T. 1306 Subcut. |10 Img.} D. 78 G.T. ” 1305 Subcut.| 10mg. D. 42. ,, | G.T. CULTURE INOCULATIONS. July 23, 1907. The strain wus derived from the prescapular gland of Calf 538 and had been in cultivation a total period of 62 days. The culture used was the 4th generation, 19 days old. RABBITS. Method. Dose. Duration of Result. Number. Life 1426 Intrav. | 1:0 mg.| D. 16 days. | G. T. 1427 Intrav. | O-l mg.} D.17_ ,, G. T. | 1428 | Subcut./10-0mg.| D.61 ,, | GT. i 24] VIRUS P. XIV.—continued. CoLTURE INOCULATIONS—continued., (b) ApRix 16, 1907. The strain was derived from the original material, through G.P. 1623, and had been in cultivation a total period ‘of 581 days. The culture used was the 21st generation, 21 days old. RABBITS. CALF 532. CALF 534. Subcutaneous. Subcutaneous. Number. | Method.| Dose. | meas of Result. Dash OU aie: Doser 20 mg. ife ; Died: July 1, Killed: July 12, 1907. 1907. 1307 | Intrav. | 0-1 mg.| D. 67 days. | GT. 70 dave. Gf cya, P.M. — General P.M. — General 1308 Subcut.} 1-0 mg.| D. 72, G.T. tuberculosis; severe progressive suber- in the lungs. culosis of moderate 1292 Subcut. | 10-0 mg.| D.99_,, | GT. severity. CULTURE INOCULATIONS. OctTosBrER 14, 1907. The strain was derived from the spleen of Calf 534, and had been in cultivation a total period of 94 days. The culture used was the 4th generation, 21 days old. RABBITS. | | 4 Somber, Method.| Dose. | es of Result. | ife fo | 1518 | Intrav. | 10 mg. | D.19 days | G.T. 1517 | Intrav. | 01mg. | D.17_,, ae : 1519 | Subcut. | 10-0 mg., D.58 ,, | G.T 1516 | Subeut. | 10mg. | D.80 ,, | GT. { I (ce) Juuy 16, 1907. The strain was derived from the original material, through G.P. 1623, and had been in cultivation a total period of 672 days. The culture used was the 23rd generation, 21 days old. RABBITS. Number. Method. Dose. ee of Result. | 1407 Intrav. | 1:0 mg.| D. 21 days; G.T. 1406 Intrav. | O-lmg.| D. 45 ,, | G.T. 1409 Subcut. | 10-0 mg.! D. 201 ,, G. T. 1408 Subcut.| 10 mg.| D.177 ,, | GT. 242 CALF 446. Virus P. XIV. Subcutaneous inoculation of culture derived from the original material of Virus P. XIV, through G.P. 1623. Dose—50 milligrammes. Date—June 29, 1906. Weight at Inoculation—36-74 kilogrammes. [Age about 5 weeks.] Died—September 17, 1906. [80 days after inoculation.] Clinical Notes. Seven days after inoculation on the left side of the neck there was a flat local thickening measuring 7°5 by 5 em.; the adjacent prescapular gland was very slightly enlarged. On the fourteenth day a firm slightly raised tumour had developed, which measured 10 by 6cm. The gland was slightly enlarged, 5 cm. in length. On the fortieth day the local conditions were unchanged. The general condition of the calf was fair. On the fifty-fifth day the tumour was firm and measured 11°5 by 6:2 cm., by about 7:5 cm. in thick- ness; the gland was 6cm.in length. The calf was not in very good condition, and seemed a little weak, but there was no increase in the respiration, and the appetite was good. Three weeks later the local condi- tions were unchanged; the calf was thin but apparently not ill, Three days later it died. Temperature.—The temperature was irregular during the experiment but was at no time very high, the maximum range of variation being from 38-2° C. to 39°8° C. Tuberculin Test—The calf was not tested sub- sequent to inoculation. Weights. Kilogrammes. June 29, 1906 eye ose aa 36°74 September 17, 1906 ce apes 34-01 Total loss of weight during experiment.—2°73 kilo- grammes. POST-MORTEM EXAMINATION. General Condition —Fair. Local Lesion.—At the seat of inoculation on the left side of the neck there was a firm flattened tumour measuring 11 by 5:7 cm., by 17 cm. in greatest thickness, composed of dense yellowish white caseo-necrotic tissue adherent both to skin and muscle but no} infiltrating either to any extent. Left Prescapular Gland.—The left prescapular viand measured 5-7 by 4 cm. by about 2 cm.; on section the greater part of the gland was composed of dense pinkish yellow caseated tissue showing no sign of calcification. A little normal gland tissue remained chiefly around the periphery and this con- tained more or less discrete yellow caseous tubercles. Right Prescapular Gland.—The right prescapular gland contained one yellow caseous nodule 1-5 mm. in diameter. Prepectoral Glands—One on the left side, the size of a sparrow’s egg, was dense yellow and caseous throughout ; its capsule was thickened. Other prepectoral glands were normal. Left Cervical Glands.—One of the left lower cer- vical glands, a little over 1 cm. in diameter, was indurated and mottled with caseous foci which formed a more or less continuous line around the periphery of the gland ; the capsule was much thickened. A mid-cervical gland contained a yellow caseous focus. The superior cervical gland contained a yellow caseous nodule a little larger than a millet seed. Right Cervical Glands.—The right upper cervical gland contained one yellow pinhead tubercle. Other cervica! glands on this side were normal. Mastoid Glands.—The left showed at one extremity a group of yellow caseous tubercles. The right was normal, Avillary Glands.—Normal. Thorax. Pleura.—The parietal pleura on thetlast few ribs on the right side showed a number of small connective tissue outgrowths, in the apices of which small hard tubercles, some caseous some hemorrhagic, could be felt. Lungs.—The lungs collapsed normally and showed scattered throughout, not very numerous, firm nodules. ranging in size from a pin’s head to a hemp seed ; on. section they were yellow and caseous, and in the majority of cases surrounded by a small angular dark red zone; they appeared to be most numerous in the caudal lobes. In the anterior lobes there were some large areas of collapse. In the posterior parts of the caudal lobes there were some small consolidated lobules which were either homogeneously red through- out or showed early caseous infiltration. Thoracic Glands.—The bronchial and mediastinal glands were not apparently enlarged ; they contained fairly numerous more or less discrete yellowish white- caseous tubercles ranging up to about 2 mm. in diameter. Heart.—Normal. Abdomen. The peritoneal cavity contained a small quantity of yellow turbid fluid suspended in which were flakes of yellow pus. The bladder was greatly distended, the serous sur- face was markedly injected and covered with a layer of yellowish fibrino-purulent material. The serous membranes in contact with it, i.e. of the rumen and left kidney, were congested and covered with yellow fibrinous lymph. On incision the bladder was found to be filled with dark red bloody fluid, and to contain rather a large quantity of gravel. The mucous mem- brane was congested and deeply hemorrhagic; th: wall of the bladder was thickened. Kidneys.—The left kidney was enlarged, the pelvis and calyces were dilated, their walls being thickened, and contained numerous concretions ; the cortex was tough, most of the lobules showing on the surface a greyish mottled appearance; on section these grey areas extended into the cortex as radial streaks. In several places the cortex showed yellow purulent areas. The capsule stripped badly and the perirenal tissue was much increased in amount. The right kidney was slightly larger than normal, and showed an oedematous condition of the pelvis and calyces, with a few scattered concretions. The surface was speckled with petechial hemorrhages, and the cortex showed a marked increase of interstitial connective tissue, with here and there some greyish radial streaks. No tubercles were seen in either kidney. Suprarenal Bodies.—Normal. Spleen.—The spleen was normal in size, and showed on section sparsely scattered firm yellowish tubercles, the largest the size of*a millet seed. Liver.—The liver appeared normal on the surface and on section. Portal Glands.—The portal glands, little if at all enlarged, contained discrete firm yellow caseous. nodpine ranging in size from a pin’s head to a hemp. seed, 243 Coeliac Glands——One coeliac gland was slightly enlarged, and closely beset with firm irregular yellow caseous nodules up to a hemp seed in size. Lumbar Glands.—The lumbar glands were enlarged. The largest contained three or four yellow millet seed sized tubercles and a few smaller greyish yellow foci. Alimentary Tract. Tongue.—Normal. Tonsils—In one tonsil a soft yellow tubercle was found. Submasillary G'lands.—Hach contained three or four yellow caseous nodules, the largest 1°5 mm. in diameter. Pharyngeal Glands.—EHach contained numerous yellow caseous tubercles, the largest a little larger than a millet seed. Intestines.—The terminal Peyer’s patch of the small intestine contained about a dozen softened caseous nodules up to a hemp seed in size, over which the mucous membrane was congested and ulcerated. Another patch just in front of this contained one similar nodule. ‘he large intestine was normal. Mesenteric Glands—The terminal mesenteric glands were slightly enlarged and showed on section trm yellow caseous masses which extensively replaced the gland substance; the other glands contained scattered caseous nodules up to about lcm. in diameter. Lleo-colic Glands.—One ileo-colic gland showed several large caseous areas, another showed a small caseous patch. Colic caseous, Glands.—Two colic glands were partly Pharyns, Larynz.—Normal. Testes.—Normal. Eyes.—Normal. Various Peripheral Lymphatic Glands. Popliteal Glands.—The left showed in the cortex a pin-head sized yellow caseous tubercle. The right was normal. Precrural Glands.—Normal. Gluteal and Ischiatic Glande.—Normal. Pudie Glands.—Normal. Microscopic Examinations. Tubercle from Tonsil. — Tubercle bacilli numerous, Tubercle from bacilli seen. Tubercle from numerous. very Lumbar Gland.—Three tubercle bacilli Intestine.—Tubercle very CALF 448. Virus P. XIV. Subcutaneous inoculation of culture derived from the original material of Virus P. XIV, through 23. G.P. 1623 Dose—50 milligrammes. Date—June 29, 1906. Weight at Inoculation—31-75 kilogrammes. [Age about 5 weeks. ] Killed when moderately well—October 11, 1906. [104 days after inoculation.] Clinical Notes. Seven days after inoculation on the left side of the neck there was a flat local thickening 7°5 by 5 cm. in area ; the adjacent prescapular gland was very slightly enlarged. On the fourteenth day there was a slightly raised firm flattened local infiltration mea- suring 9 by 6cm.; the prescapular gland was slightly enlarged, 5cm. in length. On the fortieth day the tumour at the seat of inoculation was small and slightly raised, measuring 95 by 55cm. ; the gland was 75cm. in length. The left prepectoral gland was enlarged. On the seventy-seventh day the tumour was smaller (7-5 by 45cm.) and softening in the centre. The calf was thin, but not ill, Twenty-four days later it was killed ; it was then in poor condition : there was some inspiratory dyspnoea and an occasional cough. Temperatwre.—The temperature was irregular during the experiment, but at no time was it very high, the maximum range of variation being 1:8° C. (38:0° C.— 39°8° C.). Tuberculin Test—The calf was not tested subse- quent to inoculation. Weights. Kilogrammes. June 29, 1906 ven oes ees 31°75 October 11,1906 ... ae we 45°35 Total gain of weight during experiment—13°60 kilo- grammes. Average rate of gain per week.—907 grammes. POST-MORTEM EXAMINATION. Carcass.—Condition fair. Local Lesion—At the seat of inoculation in the subcutaneous tissues of the left side of the neck there was a flattened pear-shaped tumour 8°3 by 5 by 2 cm. On section it consisted of yellow caseo-necrotic sub- stance with scattered calcareous streaks ; the centre was beginning to break down forming a small cavity. It was adherent to skin and muscle, in both of which there were scattered caseous tubercles. Left Prescapular Gland.—The left prescapular gland measured 5°8 by 3 by 4 em. On section about one third of the gland consisted of semi-translucent caseated tissue with yellowish white calcareous streaks. In the rest of the gland the cortex was in part caseous and in part contained discrete caseous tubercles. The medulla showed scattered caseous nodules. Right Prescapular Gland.—Normal. Prepectoral Glands.—A left prepectoral gland was replaced by caseated tissue, with yellow calcareous streaks and foci mainly around the periphery. A second gland was enlarged and had the cortex filled with confluent caseous gritty nodules. Right prepec- toral glands normal. Aciliary Glands.—Normal. Cervical Glands.— A left lower cervical gland was enlarged and caseous throughout with calcareous streaks. Of the left middle cervical glands one was almost entirely caseous and two others contained caseous nodules. The left superior cervical gland was normal. The right superior cervical gland contained a single irregular caseous nodule with calcareous foci. One right middle cervical contained two caseous nodules. The right inferior cervical gland was normal, ad Thora. Pleura and Diaphragm.—Normal. Lungs.—The anterior and middle lobes of the right lung contained numerovs nodules which projected from the surface and measured up to 1 cm. in diameter. In places they had become confluent forming larger masses. The majority showed a tendency to soften and from some the whole soft caseous centre could be picked out, leaving a trans- lucent capsule. In the lung substance between the nodules were dark congested ureas. The posterior lobe presented much the same appearance, but in many parts the nodules had run together, forming large caseous masses. ‘The softening was more marked, some being caseo-purulent throughout and showing the early stages of cavity formation. The left lung was similar to the right except that in the centre of the posterior lobe there was a caseo- purulent mass replacing about one-third of the substance. Thoracic Glands.—The long mediastinal gland was large and the greater part of the cortex was filled with irregular confluent caseo-calcareous noduies ; the rest of the gland showed discrete tubercles and nodules. Two other mediastinal glands were more than half replaced by firm caseated tissue with calcareous streaks and foci. The right bronchial glands were enlarged and almost entirely caseous. The left were filled with caseated tissue similar to that in the mediastinal glands. Heart and Pericardium.—Normal. Abdomen. Omentum and Peritonewm.—Normal. Spleen.—The spleen weighed 141 grammes. Beneath the capsule could be seen scattered tubercles project- ing slightly from the surface. On section the pulp contained moderately numerous discrete yellow caseous tubercles up to a hemp seed in size. The majority of these contained fine calcareous streaks in the centre. Liver.—On the anterior convex surface there were three small slightly projecting tubercles with filmy marginal expansions. Just beneath the surface and sparsely scattered throughout the substance were tubercles varying from a grey point up to a millet seed. The largest were caseous with grey margins and gritty centres. Portal Glands.—The portal glands (four in number) were enlarged and the cortices were closely filled with 44 firm caseous nodules with grey margins and yellow calcareous centres. Kidneys.—In the cortices there were a few scattered minute grey foci and an occasional grey tubercle with an opaque calcareous centre. Suprarenal Bodies——The right showed two caseous tubercles the size of rape seed with calcareous centres. The left contained three similar tubercles. Intestines—In the first part of the small intestine there were scattered nodules up to 5 mm. in diameter occurring both in the Peyer’s patches and in the mucous membrane. They projected internally and in the centres of some there were small openings. On section the majority consisted of soft caseous substance. The rest were of translucent tissue with soft yellow centres. In the lower part of the intestine only occasional nodules were met with in the Peyer’s patches. One had a small ulcer in the centre, the base of which was formed by soft caseous tissue. Mesenteric Glands—The mesenteric glands con- tained numerous nodules varying in size from a pea to a pigeon’s egg. They were separated from each other by normal gland tissue or tissue with a few caseous foci. The nodules consisted of a caseous network embedded in firm translucent tissue similar to that in the bronchial glands. ’ Coiie G'lands—The colic glands were enlarged and their cortices were almost entirely replaced by a fibro-caseous network with calcareous foci. Gastric Gland.—One gastric gland contained a caseous nodule. The Tongue, Larynx, Pharynx, Tonsils, and Trachea were normal. Special Glands. Coeliac Glands——One gland was replaced by confluent caseo-calcareous nodules. Two _ others showed irregular caseo calcareous tubercles. Lumbar Gland.—One lumbar gland was filled with caseous gritty nodules. Renal, Left Iliac, Right Precrural, Gluteal, Left Popliteal, Left Ischiatic, Posterior Pharyngeal Glands.—These glands contained a varying number of discrete caseous nodules. Right Iliac, Left Precrural, Right Popliteal, and Left Parotid Glands were normal. CALF 532. Virus P. XIV. Subcutaneous inoculation of culture derived from the original material of Virus P, XIV. through. Guinea-pig 1623. Dose—50 milligrammes. Date of Inoculation—April 16, 1907. [Age about 11 weeks.] Died—July 1, 1907. [76 days after inoculation.] Temperature. The temperature began to rise about the 7th or 8th day after inoculation and reached 39:9 on the 17th day; subsequently there was irregular pyrexia (max. 39-9). Weights. Kilogrammes. April 16, 1907 sis ass ie 39°90 duly 1, 1907 ao aes is 39°90 POST-MORTEM EXAMINATION. Condition.—Poor. Local Lesion —At the seat of inoculation on the left side of the neck there was a tumour measuring 127 by 75 by 5cm. On section it was found to be a cyst filled with a clear serous fluid with a few flakes and small masses of 'caseo-necrotic substance ; the inner walls were shaggy and lined by a thin layer of caseo-necrotic substance slightly gritty. There was a little infiltration of both skin and muscle. 24 Left Prescapular Gland.—The left prescapular gland measured 8 by 3:8 by 3 cm., the periglandular tissue was oedematous. On section it showed about two-thirds of its cortex completely cascous with a tendency to central softening, and in the other third a close network of yellow caseous tissue. Right Prescapular Gland.—The right prescapular gland measured 5 by 2:2 by 1:2 cm. On section it was congested and contained a few scattered caseous nodules and a smali collection at one extremity. Left Prepectoral G'lands.—The left spherical prepec- toral gland was enlarged and caseous throughout. The left reniform gland was enlarged and oedematous and showed the cortex beset with caseous tubercles. Left Cervical Glands.—The left inferior cervical gland showed discrete caseous tubercles. The left superior cervical gland was oedematous and contained one small collection of caseous tubercles. The Right Prepectoral and Cervical Glands were oedematous but contained no tubercles. The Axillary Glands on both sides were oede- matous but otherwise normal. Thorax. Pleura.—Normal. The Tissues of the Posterior markedly oedematous. Lungs.—The lungs were voluminous and did not collapse ; they were congested and the anterior lobes on both sides, the right lobe, and the ventral margins of the caudal lobes were solid and airless; they were reddish grey in colour and were fairly closely beset with yellow caseous nodules and patches. The posterior portions of the caudal lobes were also severely affected, containing large firm pneumonic patches beset with caseous nodules ; in some of these whole lobules were caseous throughout. Trachea.—Normal. Thoracic Glands.—The long mediastinal gland was enlarged, measuring 10°5 by 3:°2 by 2cm. On section the cortex was firm and was closely beset, but not entirely replaced, by irragular waxy caseous nodules The other mediastinal and bronchial glands were similar to the long mediastinal gland. Heart and Great Vessels appeared normal. Mediastinum were Abdomen. Omentum.—In the omentum there was one grey submiliary tubercle. Peritoneum.—Normal. Spleen. — The spleen weighed 70 grammes. On section it contained moderately numerous discrete yellow caseous nodules varying from a rape-seed up to a hemp seed in size ; the majority were of the larger size and were softened in the centre, Liver.—On the convex surface beneath the capsule there was one minute grey tubercle. On the inferior surface there were two caseating miliary tubercles. py o On section through the substance there was an occasional minute grey tubercle. The Gall Bladder was normal. Portal Glands.—The cortices of the portal glands were closely filled with waxy caseous nodules up toa hemp seed in size. Kidneys.--On the surface of both kidneys there were scattered foci of congestion. In one there was a grey miliary tubercle caseous in the centre and on section slightly elongated, and also a minute grey focus. On the surface of the other kidney there were four grey tubercles with caseous centres varying up to a millet seed ; section revealed one more tubercle. Suprarenal Bodies.—The suprarenal bodies were normal, Lumbar Glands—The lumbar glands contained disurete waxy caseous nodules. Iliac Glands.—Normal. Alimentary Tract. Tongue, Tonsils, Larynx, and Pharync.—Normal. Large Intestine. —Normal. Small Intestine—In two of the Peyer’s patches there were small ulcers with caseous floors. Gastric Glands.—Several of the gastric glands showed the margins of the cortex caseous. Mesenteric Glands.—T wo or three of the mesenteric glands at each extremity of the mesentery were firm and almost replaced by a yellow caseous network. In the rest of the chain of glands there occurred at intervals caseous nodules varying in size from a wheat grain up to a nodule measuring 2 by 1:5 cm, Various Lymphatic Glands. Coeliac Glands.—The coeliac glands were to a con- siderable extent replaced by a network of yellow caseated tissue. Submaxiliary Glands.—The left submaxillary gland showed one hemp seed sized caseous nodule. The right was normal. The Parotid Glands showed caseous nodules. Precrural, Popliteal, Gluteal, Ischiatic, and Posterior Pharyngeal Glands.—Normal. The caseous lesions showed a greater tendency to soften than to calcify. Microscopical Examination. (Smear Preparations.) Tubercle from Liver.—A few tubercle bacilli. Bronchial Gland.—Moderately numerous tubercle bacilli. CALF 534. Virus P. XIV. Subcutaneous inoculation of culture derived from the original material of Virus P. XIV. through Guinea-pig 1623. Dose—50 milligrammes. Date of Inoculation—April 16, 1907. Temperature. _ The temperature began to rise on the 10th day after. inoculation, reaching 40:2 on the 14th, and remaining 27676 [Age about 11 weeks. ] Killed—July 12, 1907. [87 days after inoculation.] above forty for four days; it then fell and sub- sequently fluctuated slightly above normal. Tuberculin Test. The calf was not tested during the experiment. 2P 246 Weights. Kilogrammes. 54:87 April 16, 1907 nea vee bee 72°10 July 12, 1907 aa vee sae Total gain in weight.—17°23 kilogrammes. POST-MORTEM EXAMINATION. General Condition. Good. Local Lesion.—In tbe subcutaneous tissues on the left side of the neck there was an elongated tumour weighing with skin and muscle 396 grammes, and measuring 20 by 7:5 by 4cm. On section it was found to be composed of caseo-necrotic gritty substance with an irregular central cavity containing serous fluid ; it infiltrated both skin and muscle. Left Prescapular Gland.—The left prescapular gland measured 9°5 by 5 by 45cm. On section it was composed almost throughout of caseo-necrotic tissue with gritty foci ; at one part there was some softening with the formation of a small cavity. Right Prescapular Gland.—The right prescapular gland was normal. Left Prepectoral Glands.—The left reniform pre- pectoral gland was enlarged and closely filled with caseous streaks and foci. Tbe cortex of the left rounded prepectoral gland showed numerous caseous foci. Cervical Glands.—The left inferior cervical gland was enlarged measuring 3°8 by 25cm. On section about one-third of its cortex was replaced by a caseating gritty nodule; the remainder was firm grey and teset with caseous foci. The left middle and superior cervical glands were normal. The Right Cervical Glands were normal. The Right Prescapular and Right Prepectoral ‘Glands were normal. Thoran. Pleura.—The lymphatic fringes in the intercostal spaces were enlarged and congested, and several of them contained flat grey translucent nodules, Lungs.—The lungs did not collapse normally ; they were crepitant, but a little congested and oede- matous. Scattered throughout the substance were moderately numerous irregular grey nodules with a caseous network, varying considerably in size, the largest being about 2 by 3cm.in area. The majority were surrounded by a zone of congestion. Trachea.—Normal. Thoracic Glands.—The long mediastinal gland measured 15cm. in length by 4°3cm. in greatest thickness ; on section the cortex consisted of firm grey tissue with a yellowish caseous network slightly gritty in parts; the small amount of cortical tissue left was deeply congested. The other mediastinal glands were similar. The Bronchial Glands resembled the mediastinal glands, but there was more firm grey tissue and less caseation. _A Gland at the Entrance to the Thorax on the left side contained a fine caseous yellowish network. One on the right showed a few opaque foci. The Heart and Great Vessels appeared normal. Diaphragm.—On the pleural surface of the dia- phragm the lymphatic fringes were prominent, form- ing shaggy grey growths in which could be felt minute hard grey tabereles ; on the peritoneal surface there were a few nodules similar to those in the omentum, Abdomen. Omentum.—In the omentum there were scattered translucent nodules varying in size up to about a wheat grain; the largest were flattened and loosely attached, and a few were congested. Peritonewm.—Normal. Intestenes.—The small intestine ; in the first part in the mucous membrane there was a circular nodule lem. in diameter with a crater-like ulcer and a caseous base. Along the intestine there were four similar but smaller nodules, two of them being in Peyer’s patches. In the large intestine there were two congested circular ulcers resembling those in the small gut. The Mesenteric Glands were not severely affected ; many of them showed large single firm grey nodules with a caseous gritty network. Spleen.—The spleen weighed 170grammes. Onsection it contained scattered nodules (30 counted) up to 5 mm. in diameter. On section these consisted of soft cagseous substance enclosed within grey translucent capsules. Liver.—On the ventral surface there were four slightly projecting nodules with fibrinous fringes : four flattened nodules were visible on the convex surface. Attached to the sharp margin but not affecting the substance, there was a pea sized grey nodule caseous in the centre. On section there were sparsely scattered throughout the substance nodules similar to those in the spleen and varying up toa hemp seed in size. The Gall Bladder was normal. Portal Glands.—The portal glands contained in the cortices discrete grey nodules caseous in the centre. Kidneys.—The left kidney showed in the cortex just beneath the surface five minute opaque tubercles, The right kidney contained one tubercle, On section no more were revealed. Suprarenal Bodies—The left suprarenal body showed three gubmiliary caseous tubercles. The right suprarenal body was normal. The Renal, Lumbar, and Iliac Glands were normal. Tongue, Tonsils, Larynx, and Pharynz were normal. Various Lymphatic Glands. Coeliac Glands.—Three were normal; in a fourth. the cortex was beset with caseous miliary tubercles. The Precural, Right Gluteal, Popliteal, Submazillary, Parotid, and Ischiatic Lymphatic Glands were normal, The Left Gluteal Gland contained a few irregular caseous nodules. Microscopical Examination. (Smear Preparation.) Vicer from Large Intestine —Moderately numerous tubercle bacilli. CALF 586. Virus P. XIV. Subcutaneous inoculation of culture derived from the lung of Calf 448. Dose—50 milligrammes. Date of Inoculation—April 16, 1907. [Age about 13 weeks.] Died—June 1, 1907. [46 days after inoculation. ] Clinical Notes. The course of the disease was similar to that usually seen in animals acutely infected. 247 Temperature. MAY Tuberculin Test.—The calf was not tested subse- quent to inoculation. Weights. Kilogrammes. April 16, 1907 ies oe 58°50 June 1, 1907 ... eee ws 53°05 Total loss of weight.—5'45 kilogrammes. POST-MORTEM EXAMINATION. Carcass.—Emaciated. Local Lesion.—The tumour at the seat of inocula- tion on the left side of the neck weighed with the attached skin and muscle 368 grammes, measuring 13°5 by 10 by 6°5 cm. It was in close proximity to and adherent to the gland, and the surrounding tissues were oedematous. On section the superficial balf consisted of pinkish caseo-necrotic tissue, while the deeper part was compose of caseous tissue infiltrating the muscle and the oedematous tissues. Left Prescapular Gland.—The Jeft prescapular gland measured 8 by 4°6 by 4 cm. The tissues were adherent to the thickened capsule and on section the gland was composed throughout of firm caseo-necrotic tissue with scattered yellowish white gritty streaks. Right Prescapular Gland.—The right prescapular gland measured 4:5 by 2 by 1:2 cm. On section it showed moderately numerous irregular caseous tubercles up to a hemp-seed in size, and also a few iarger collections. Left Prepectoral Glands.—The left round _prepec- toral gland, the size of a large pea, was diffusely caseous throughout. ‘The left reniform gland con- tained discrete caseous nodules in the cortex. Left Cervical Glands. — The left inferior cervical gland had nearly half the cortex diffusely caseous, while the rest was firm and contained numerous caseous foci. A second deeper gland was caseous throughout. A left middle cervical gland was diffusely caseous almost throughout. The left superior cervical glands contained discrete irregular caseous nodules. Right Prepectoral Glands.—A few hempseed sized nodules were found in the reniform gland, and smaller nodules in the rounded gland. The Right Axillary, and the Superior, Middle, and Inferior Cervical Glands were similar to the right pre- scapular gland. The left axillary gland contained numerous irregular caseous nodules up to 4 mm. Thorax. Pleuva.—On the pleural surface of the pericardium there was a single flattened congested nodule. On the parietal pleura there were ten flattened nodules varying in size up to 10 mm. 27676 Lungs.—The lungs were large and did not collapse normally. The substance was closely filled with caseous tubercles with grey margins, for the most part discrete and measuring up to 2mm. ‘They were most numerous in the anterior and middle lobes where there was considerable lobular congestion, although there still remained a fair amount of normal lung tissue. The ventral part of the right posterior lobe bd much more congested, quite airless, and sank in water. : Trachea.—Down the trachea there were a dozen flattened congested nodules with caseous centres. Thoracic Glands.— The long mediastinal gland measured 15 by 4°5 by 2°3cm. On section almost the whole cortex was composed of firm grey tissue with a close caseous network. The other mediastinal glands were similar. The remaining part of the cortices of these glands was deeply congested and contained caseous foci. The bronchial glands were enlarged and ressmbled the long mediastinal gland. The glands within the entrance to the thorax on the left side showed the cortices firm and in a con- dition of early caseation. Those on the right side, and the ventral mediastinal glands, showed numerous irregular caseating nodules with grey margins. Heart and Great Vessels——Normal. Abdomen. Omentum.—On the omentum there were moderately numerous flattened nodules up to 3 mm. in diameter. The majoritv were loosely attached, many were deeply congested ; they were firm and translucent and almost all showed some caseation in the centre. Peritoneum.—On the parietal peritoneum there were four flat grey nodules. Spleen.—The spleen weighed 255 grammes. The vessels on the capsule were injected and there was one flattened grey nodule. On section the substance was closely filled with yellowish caseous nodules with grey margins, varying up to a hempseed in size. Liver.—The surface of the liver was studded with numerous grey tubercles with irregular caseous centres varying up to 15 mm. Some projected from the surface and were flattened and mushroomed. On section the substance showed similar tubercles dis- tributed throughout. Gall Bladder.—The vessels of the mucous mem- brane of the gall bladder were injected ; on it were two caseous tubercles, and a number of congested spots the centres of which were a little firm ; these were not definite tubercles. Portal Glands.—The portal glands were large and the greater part of the cortices was replaced by a network of caseated tissue. Kidneys.—The cortices of the kidneys, on the 2Pp2 248 surface and on section, showed scattered grey tubercles with opaque centres up to 1°5 mm. in diameter. Suprarenal Bodies——The right was normal; the left showed one opaque submiliary tubercle. Lumbar, Iliac, and Renal Glands.—The lumbar and iliac glands contained numerous caseous nodules. In the renal glands these had become confluent, almost entirely replacing the gland tissue. Alimentary Tract. Tongue.—Normal. Tonsils—In the tonsils there was a moderate number of caseous tubercles. On the laryngeal surface of the epiglottis there was a congested circular ulcer 3 mm. in diameter. Small Intestines—Beneath the mucous membrane along the whole length of the small intestine, mainly in Peyer’s patches, there were fairly numerous irregular caseous tubercles varying up to a rapeseed in size but here and there attaining the size of a hemp- seed. Over many there was a small aperture in the mucous membrane. Large Intestine.—In the large intestine many of the solitary follicles were opaque in the centre. Gastric Glands.—These glands contained irregular caseous nodules. Mesenteric Glandsi—The mesenteric glands were somewhat enlarged and contained numerous irregular caseous nodules surrounded by firm grey tissue. Genito-Urinary Systems. Testes. — Each testis showed a caseous miliary tubercle. Various Lymphatic Glands. Coeltac.—The coeliac glands were enlarged. In the largest the cortex was replaced by a caseous network, and the three small ones were similarly though rather less affected. Precrural Glands.—Showed a moderate number of irregular caseous nodules up to 3 mm. Pudie Glands——Enlarged and similar to the pre- crural, Popliteal, Gluteal, Ischiatic, and Parotideal Glands all contained irregular caseous nodules. Retro - pharyngeal Glands. — Closely filled with caseous nodules. Submazillary Glands.—The left contained numerous irregular caseous nodules up to 4 mm. in diameter, The right was similar. Microscopical Examinations. Foci from Trachea.—Tuhercle bacilli numerous. Nodule from Epiglottis—Tubercle bacilli scanty. Focus from Testis.—Tubercle bacilli numerous. Nodule from Small Intestine. — Tubercle bacilli numerous. Follicle numerous. Congested Mucous Membrane of Gall Bladder.—Two tubercle bacilli seen. of Large Intestine. — Tubercle bacilli CALF 5388. Virus P. XIV. Subcutaneous inoculation of culture derived from the lung of Calf 448. Dose—50 milligrammes. Date of Inoculation—April 16, 1907. Died—May 22, 1907. [Age about 7 weeks.] [32 days after inoculation. ] Clinical Notes. The course of the disease was similar to that usually seen in animals suffering from acute tuberculosis. Temperature. MAY Tuberculin Test.—The calf was not tested subsequent to inoculation. Weights Kilogrammes. April 16, 1907 ed a cas 43-99 May 22, 1907 se pee ets 39-00 Totul loss of weight.—4-99 kilogrammes. POST-MORTEM EXAMINATION, Carcass emaciated. _ Local Lesion.—At the seat of inoculation on the left side of the neck there was a slightly prominent tumour measuring 16 by 8 by 5 em. and weighing 396 grammes. It was adherent to and infiltrated the skin and muscle 249 around. On section it was composed of firm pinkish grey caseo-necrotic tissue. At one extremity of the ear there was a smnall cavity containing clear serous uid. Left Prescapular Gland.—The left prescapular gland measured 11 by 5°5 by 5°5 cm. and weighed 170 grammes. On section it was composed throughout of dense pinkish yellow necrotic tissue. The capsule was thickened and adherent to the muscles. The tissues and muscles around the local tumour and prescapular gland were greatly thickened and oedema- tous, and contained numerous caseous foci and nodules. Right Prescapular Gland.—The right prescapular gland measured 5 by 2 by 1 cm. On section it was found to contain a moderate number of yellow caseous nodules 5 mm. in diameter. Left Prepectoral Glands.—These giands were en- larged and contained a few caseating nodules in the cortices. Cervical G'lands.—The left inferior cervical gland was composed practically throughout of dense yellow necrotic tissue similar to that of the left prescapular gland. The left middle and left upper cervical glands were firm and congested, and showed in the cortices a close network of yellow caseating tissue. _ The right cervical glands contained a few irregular ‘caseous tubercles. Agillary Glands—The left showed two yellow -caseous nodules 5 mm. in diameter. The right con- tained a few small nodules with caseous foci. Right Prepectoral Glands——These were similar to the right axillary gland. Thorax. Pleura.—The lymphatic fringes along the margins of the ribs were congested, and showed numerous minute grey tubercles. On the caval fold and on the right side of the pleural surface of the diaphragm there were some congested fringes containing minute grey tubercles. Lungs.—The lungs were very heavy. The left cephalic and right middle lobes and the ventral portions of the right cephalic lobe were dark red and hepatised. There were a few large patches of conges- tion in the caudal lobes. The lung parenchyma was closely beset with grey and translucent tubercles with opaque caseous centres, ranging in size from a mere point up to 4 mm. in diameter, the larger ones being formed by the aggregation of a few small tubercles. The trachea showed a few congested caseous foci. ‘Some of the smaller bronchi were blocked by yellow purulent material. Thoracic Glands.—The mediastinal glands were en- larged, firm, and hemorrhagic, and showed in their cortices about half of the substance replaced by a network of grey nodules with caseous foci. The bronchial glands were similarly affected. A gland at the entrance to the thorax was composed practically throughout of pinkish yellow necrotic tissue, the capsule of the gland being thickened. Heart.—There were two grey tubercles in the heart ‘muscle. Abdomen. Omentum.—On the inferior surface of the omentum there were moderately numerous nodules up to a hemp seed in size, some of whicn were deeply con- gested, with caseous centres; others were grey and translucent. Peritoneum.—On the peritoneal surface of the diaphragm there was one flattened grey nodule with -a caseous centre. The rest of the peritoneum was normal. Spleen.—The spleen weighed 198 grammes. It was normal on the surface; on section it was closely beset with caseating tubercles 2 to 3 mm. in diameter. Liver.—The surface of the liver was closely beset with grey tubercles with caseous centres, ranging in size from about 0°5 up to 3 and 4 mm. On section similar tubercles were seen throughout the substance. Gall Bladder.—In the gall-bladder there were five submucous caseous tubercles the size of pin heads. Portal Glands.—The cortices of the portal glands were closely beset with small irregular greyish yellow caseating tubercles in places forming a close network. Kidneys.—Each kidney showed in the cortex mode- rately numerous grey translucent tubercles, the majority having opaque centres ranging in size from a mere point to a millet seed. Suprarenals.—The left showed one, the right five, small grey tubercles with minute caseous centres. Liac, Lumbar, and Renal Glands.—These showed a moderate number of irregular caseous tubercles up to 4 mm. in diameter. : Alimentary Tract. Tongue.—Normal. Tonsils—Each tonsil contained one or two small caseous foci. There was one flattened nodule on the epiglottis. Intestines.—The small intestine showed just beneath the mucous membrane a moderate number of small caseous tubercles ; these were most numerous towards * the ileum. A few of the lymphoid follicles of the large intestine were caseous. Gastric Glands.—These glands showed a moderate number of yellow caseating tubercles. Mesenteric Glands—All the mesenteric glands showed a moderate number of yellow caseating nodules. Colic and Ileo-colic Glands.—These showed a few small caseous tubercles. Genito-Urinary System. Testes —Normal. Urinary Biadder.—There were four small tubercles with caseous centres in the bladder. Various Lymphatic Glands. Submaxillary, Parotid, and Retro-pharyngeal.—These glands showed a moderate number of discrete caseating tubercles ranging in size from a mere point to that of a millet seed. : Coeliac.—The coeliac glands showed in the cortices a greyish yellow caseous network and a few discrete nodules. Pudic.—The pudic glands were deeply congested, and contained a moderate number of irregular caseating tubercles. Precrural, Popliteal, Gluteal, Ischiatic.—Hach showed a few discrete caseating tubercles. Microscopical Examinations. Suprarenal body, tubercle from.—A few tubercle bacilli seen. Heart, tubercle from.—No tubercle bacilli seen. Emulsion of Prescapular Glund.—Numerous tubercle bacilli. 2 0 VIRUS P. XXIX. (December 20, 1905.) SUBMAXILLARY GLANDS. Cu.ture InocuLaTions, January 7, 1907. The strain was derived from the original material, and had been in cultivation a total period of 383 days. The culture used was the 12th generation, 24 days old. CALF 454. RABBITS. Intravenous. Dose : 50 mg. Number. | Method. Dose. Duration of Life. | Result. Died : Jan, 23, 1997. 16 days. 320 | Intrav. | 0-1 mg. D. 17 days. G. T. P.M.—General Tuberculosis. 394 Titian, Ol wag. D. 18 , GT. 825 Tntrap. 0-1 mg. D. 29 =, G.T. 821 Subcut. 1:0 mg. D. 7 a, G. T. 822 Subcut. 0-1 mg. D.126_—s=o«, G. T. 823 Subcut. | 0°01 mg. D. 80 =, G. T. CALF 454. Virus P. XXIX. Intravenous inoculation (also partly subcutaneous) of culture derived from original material of Virus P. XXIX. Dose—50 milligrammes. Date—January 7, 1907. Weight at Inoculation—131-98 kilogrammes. Died—-January 23, 1907. Clinical Notes. The injection was begun in the vein (median) of the left ear, but owing to the restiveness of the animal the needle came out several times and finally the right ear was used. About 60 milligrammes of culture were used, part of which was lost and part went beneath the skin. Two days later the calf was noticed to be always [Age about 8} months. | [16 days after inoculation. ] lying down ; appetite good. On the third day it was. ‘very quiet, was breathing quickly (54 per minute), and panting a little ; its urine was not so abundant. On the tenth day the respirations were laboured (60 per minute) ; the animal’s nose was hot, and it lay down a great deal. On the 14th day it was breathing hard, and looked ill and thin ; two days later the difficulty in breathing had become more evident (70 per minute), and the anima] died during the day. Temperature. JANDARY a i = <£ 9 ad Tuberculin Test.—The calf was u not tested subse- ‘quent to inoculation. Weights. Kilogrammes. January 7,1907 ... aie 131-98 January 23,1907... is 97°04 Total loss of weight during eaperiment.--34:94 kilo- ‘grammes. POST-MORTEM EXAMINATION, General Condition.—Very thir. Local Lesions.—In the subcutaneous tissues of each ‘ear at the seat of inoculation there was a patch of yellowish necrotic substance. Parotid Lymphatic Glands—About half of each parotid gland was replaced by greyish early’ caseous substance which in parts was becoming distinctly yellow. Thorax. Lungs.—The lungs weighed 5:78 kilogrammes. They, were large and did not collapse normally. There was considerable subpleural emphysema on the dorsal ‘surface of the lungs, but none on the ventral surface. The tongue-shaped lobe of the right lung was reddish grey and almost completely solid ; the free half of it sank in water ; on section it was found to be closely ' packed with minute irregular grey foci. In the attached half, which floated in water, there were small parts of the lung in which the foci were not’ quite so numerous. Part of the middle lobe was solid and presented a similar appearance. Similarly the anterior and ventral portions of the left lung were dark, congested, and in parts airless, and on section they presented the same appearance. The rest of the substance of the lungs, that is the greater part, was oedematous but not airless, and was closely filled with minute grey foci varying slightly in number in different lobules. Bronchial and Mediastinal Glands—The caudal mediastinal gland was enlarged, measuring: 11°5 by 5 by 25cm. The cortex was firm and the outer part closely filled with small irregular grey patches becoming discrete as it extends inwards, so that there was almost a complete thin margin of early caseation. , The next mediastinal gland was firm and congested -and showed numerous similar irregular grey patches 1 51 forming a network. A third gland showed a similar early caseous margin and network. The smaller mediastinal glands were congested and showed grey foci. The bronchial glands resembled the mediastinal. Pleura.—Normal. Heart.—Normal. The tissues around the heart were very emphy- sematous, there was also considerable emphysema of the posterior mediastinum. Abdomen. Omentum and Peritoneum.—Normal. Spleen.—The spleen weighed 541 grammes. There was a tendency to become diffluent, otherwise nothing abnormal could be seen. Liver.—On section of the liver the cut surface showed the interlobular grey network well marked. No tubercles were seen in the organ. Kidneys.—The kidneys showed on the surfaces alternate pale and congested patches. In the pale areas there were sparsely scattered minute irregular heemorrhagic foci. Suprarenal bodies.—Normal. Renal, Lumbar, and Lliac Glands.—Normal. Portal Glands.—Normal. Alimentary Tract. Tongue, Pharynx, Palate, Tonsils—Normal. Intestines —The small intestine was deeply con- gested in the greater part of its extent. The large intestine was normal. Mesenteric Glands.—Normal. Various. Lymphatic Glands. Pharyngeal, Submawillary, Precrural, Popliteal, Gluteal, Ischiatic, Prescapular, Awillary—No tubercles seen. {The glands on the left side of the neck and shoulder were all dark and congested. The tissues between the muscles of the neck showed dilated small vessels—congestion due to hanging of the animal’s head during life.] 255 CALF 478. Virus P. XLI. Subcutaneous inoculation of culture derived from the original material of Virus P. XLI. Dose—50 milligrammes. Date of Inoculation—November 15, 1906. Weight at Inoculation—73'48 kilogrammes. [Age about 18 weeks. ] Killed when in good health—February 22, 1907. Clinical Notes. 14 days after inoculation on the left side of the neck there was a prominent tumour firm and adherent to the skin, measuring 12:5 by 9 cm. The left prescapular gland was considerably enlarged, measuring 10 by 5 cm. Both the tumour and gland slowly diminished in size, and the former becime soft and fluctuating ; on the 54th day it measured 10 by 6:3 cm. The calf remained well during the experiment. Temperature.—The temperature rose to 39:9° C, on the 13th day. At the close of the third week it had returned to the normal, and it remained normal subsequently. Tuberculin Test—February 19th, 96 days after imoculation. Positive reaction. Rise of temperature, 2°0° C. Weights. : Kilogrammes. November 15, 1906 ... 73°48 February 22, 1907 96°14 Total gain of weight.—22'66 kilogrammes. Rate of gain per week.—1-62 kilogrammes. POST-MORTEM EXAMINATION. General Condition.—Good. Local Lesion.—At the seat of inoculation on the left side of the neck there was a prominent tumour measuring 14 by 83 by 7 em. and weighing 453 grammes. On section it consisted of a thick fibrous- walled cyst distended with a mucoid caseo-purulent substance. Left Prescapular Gland.—The left prescapular gland measured 7 by 3 by 15cm. On section it contained a caseo-calcareous nodule 1°5 by 0°75 cm. ; two slightly smaller nodules consisting of soft slightly gritty caseous substance; and also, just beneath the capsule, scattered tubercles, some soft and caseous, others caseo-calcareous. Near the prescapular gland was a small pear-shaped nodule 1:75 cm. in length. Right Prescapular Gland (6 by 3 cm.).—Normal. Prepectoral and Axillary Glands.—Normal. {99 days after inoculation. ] Thorax. Pleura.—Normal. Lungs.—Normal. Thoracic Glands.—Normal, Heart.—Normal. Abdomen. Peritoneum.—Normal. Spleen (368 grammes).—Normal. Liver.—The liver contained beneath the capsule and in the substance numerous closely placed irregular grey nodules, the majority with a yellowish tint and many surrounded by congested areas and foci. They varied in size up to roughly 5 mm. in greatest diameter. On section they were soft. Portal and Coeliac Glands.—Normal Kidneys.—Normal. Suprarenal Bodies.—Normal. Renal, Lumbar, and Iliac Glands.—Normal. Alimentary Tract. Tongue, Pharynz, Palate, Tonsils——Normal. Submazillary, Pharyngeal, Hyoid, and Parvtid Glands.—Normal. Intestines.—Normal. Mesenteric Glands.—Normal. Testes.—Normal. Eyes.—Normal. Various Lymphatic Glands. Precrural, Popliteal, Gluteal, Ischiatic, Pudic.— Normal. Microscopic Examinations. Smear from Liver—No tubercle bacilli. Emulsion of Liver Tissue-—No tubercle bacilli. Emulsion of nodule near Prescapular Gland.— Tubercle bacilli in moderate numbers. CALF 480. Virus P. XLI. Subcutaneous inoculation of culture derived from the original material of Virus P. XLI. Dose—100 milligrammes. . Date of Inoculation—November 15, 1906. Weight at Inoculation—68°90 kilogrammes. [Age about 17 weeks. } Killed when in good health—February 21, 1907. ‘Clinical Notes. 14 days after inoculation on the left side of the neck there was a prominent rounded tumour, ad- herent to the skin and infiltrating the subjacent muscles, and measuring 15:3 by 12:5 cm. The dependent tissues were oedematous. The left pres- capular gland was not palpable, being overlapped by the tumour. 27676 (98 days after inoculation. ] On the 35th day the tumour measured 10 by 7 cm.; it was soft in the centre and firm round the margins. The prescapular gland was now quite evident, and measured 9 cm. in length. The tumour afterwards became soft and fluctuating all over, but did not ulcerate. The gland diminished in size until it was no larger than its fellow on the right side. Temperature.—There was a period of slight pyrexia 2R 256 ‘commencing on the 12th day after inoculation, and lasting a week. The highest temperature recorded during this period was 40:1°C. Subsequently the temperature remained normal. ' Tuberculin Test.—February 19, 96 days after inocula- tion. Slight reaction. Rise of temperature 0'8° C. Weights. Kilogrammes. November 15, 1906 ... wee sis 68°90 February 21,1907 ... aes wee 98°40 Total gain of weight.—29°50 kilogrammes. Rate of gain per week.—2'10 kilogrammes. POST-MORTEM EXAMINATION. General Condition—Good. Local Lesion.—In the subcutaneous tissues on the left side of the neck there was a tumour measuring 10 by 7-5 by 5em. On section it consisted of a cyst filled with’ turbid fluid in which masses of curdy caseous substance were floating ; it was lined by grey granulation tissue to which flocculi were adherent. It did not infiltrate either skin or muscle. Left Prescapular Gland.—The left prescapular gland measured 7°5 by 3°5 by 1:3 cm. On section it con- tained a caseo-calcareous nodule 2°5 by 1:3 cm. anda second the size of a pea; otherwise the gland was normal. Prepectoral Glands.—The rounded prepectoral gland on the left side was slightly enlarged and firm, con- taining a network of translucent tissue with caseo- calcareous streaks. Other prepectoral glands were normal. Right Prescapular Gland.—Normal. Aaillary Glands.—Normal. Thorax. Pleura.—Normal. Lungs.—Normal. Thoracic Glands.—Normal. Feart.—Normal. Abdomen. Peritoneum.—Normal. Spleen.—Normal. Liver.—On the surface of the liver just beneath the capsule about seven scattered grey slightly irregular tubercles were seen, not larger than a rape seed. On section they were soft and could be picked out. An occasional focus was seen on section in the substance of the liver. Gall Bladder.—Normal. Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Renal, Lumbar, Iliac, and Coeliac Glands.—Normal Alimentary Tract. Tongue, Pharynx, Palate, Tonsils.—Normal. Pharyngeal, Submazillary, Hyoid, and Parotid Glands.—Normal. Intestines.—Normal. Mesenteric Glands.—Two showed foci of congestion, otherwise the glands were normal. Testes.—Normal. Eyes.—Normal. Various Lymphatic Glands. Precrural, Popliteal, Gluteal, Ischiatic, Pudic.— Normal. Microscopic Examinations. Liver, Tubercle from.—No tubercle bacilli seen. Prepectoral Gland, Emulsion of—No tubercle bacilli, seen. PIG 166. Virus P. XLI. Subcutaneous inoculation of culture derived from the original material of P. XLI. Dose—50 milligrammes. Date of Inoculation—January 25, 1907. Killed when in good health—May 7, 1907. Clinical Notes. In the subcutaneous tissues 18 days after inoculation there was a rounded prominent soft tumour 6°5 cm. in diameter; this ulcerated after 84 days and dis- charged soft caseous substance. Temperature.—The temperature during the whole of the experiment varied very little, at no time rising above 39°7° C. Tuberculin Test.—74 days after inoculation. tive ; Rise of temperature 2:7° C. Weight at Death.—31-29 kilogrammes. Posi- POST-MORTEM EXAMINATION. General Condition.—G ood. Local Lesion.—At the seat of inoculation to the left of the umbilicus the skin over an area 3°8 cm. square was pigmented and brownish red ; in it there were two small ulcers with thin overhanging margins, from which a little pus could be expressed, and also two pigmented papille, which, on section, were fibrous and [Age 8 weeks. ] [102 days after moculation.] obviously a consequence of healed ulcers. The skin on the under surface was thickened, fibrous, and pigmented ; there were in the subcutaneous tissues two small sinuses in connection with the ulcers, and seattered soft yellow gritty miliary nodules within grey fibrous capsules. Left Inguinal Glands.—One left inguinal gland measured 3 by 2 cm, and contained a nodule, re- placing nearly half its substance, and consisting of soft yellow caseo-pus, which could be evacuated, leaving a smooth pinkish grey wall ; in the other half of the gland there was a pea-sized nodule similarly constituted, and in the substance between were a few opaque yellowish tubercles. Three other glands were a little firm but otherwise normal. Right Inguinal Glands—The glands on the right side were normal. . Left Ventral ediastinal Gland.—The left ventral mediastinal gland was enlarged, and about half re- placed by yellow caseo-purulent substance similar to that in the left inguinal gland. _Precrural Glands.—The precrural glands on both sides were normal. 257 Thorax. Pleura.—The lymphatic fringes on the parieta pleura were prominent ; along these were moderately numerous flat fibroid nodules, some in the fringes, others loosely attached to the pleura. At the apex of the right pleural cavity there was an area of parietal pleura fairly thickly sown with small hard grey tubercles ; the largest were up to a wheat grain in — and several were opaque and yellowish in the centre. Lungs.—The lungs did not collapse normally. The right posterior lobe showed on the surface, more markedly in the posterior half, numerous raised irregular translucent nodules, varying when discrete up to about 3 mm. in diameter; they extended a little way into the substance. In addition there were similar nodules beneath the pleura not raised from the surface. The two sets were so close to- gether as to form a coarse network of grey nodules. Around the margins of the lobes the nodules had run together, so that the margins were hard and nodular in*places nearly 2 cm. in width. On section through the lung substance there were scattered discrete grey miliary tubercles. In the right anterior lobe there was the same distribution of grey nodules on the surface, but also the whole of the right anterior lobe was firm, grey, and solid and sank in water. The middle tongue-shaped piece was about half grey and solid. The left posterior lobe resembled the right, but the nodules were even more closely placed, leaving only small islands of pink lung tissue. The left anterior lobe and the middle tongue-shaped piece were almost entirely grey and solid and sank in water. Nowhere was there any definite caseation, but occasionally there was some slight central opacity in the nodules, and more especially in the discrete tubercles. The Large Bronchi and the Great Vessels about the Heart were normal. The Small Bronchi and Bronchioles in the solid grey areas were filled with muco-purulent fluid. The Bronchial and Dorsal Mediastinal Glands were normal in appearance. Trachea.—Normal. Heart.—Normal. Diaphragm.—The lymphatic fringes on the pleural surface of the diaphragm were prominent, and several of them contained hard flat translucent grey nodules, slightly irregular but roughly the area of a millet seed. On the right side of the peritoneal surface of the diaphragm there were about thirty somewhat flat- tened translucent nodules, similar to those on the pleural surface ; one was a little opaque in the centre. On the left side there were three similar nodules, Abdomen. Omentum.—Normal. Peritonewm.—Normal. Large and Small Intestines.—Normal. Mesenteric Glands.—Normal. Spleen.—In the spleen there was a single yellowish. tubercle a little larger than a millet seed, which on section was caseous in the centre. Liver.—Normal. Portal Glands.—Normal. Kidneys.—In the cortices of the kidneys there were moderately numerous discrete grey nodules, yellowish towards the centre. On section they extended a short way through the cortex as a streak ; the largest was about the size of a millet seed. A few were met with in the medulla. Suprarenal Bodies.—In the right suprarenal body there was a firm grey tubercle, opaque in the centre, the size of a rape seed. The left suprarenal body was normal. The Coeliac, Lumbar, and Iliac Glands were normal. Alimentary Tract. Tongue, Tonsils, Laryna, and Pharyna.—Normal. Various Lymphatic Glands. Prescupular, Prepectoral, Cervical, Submasillary,. and Parotid Lymphatic Glands were normal. Microscopical Examinations. (Smear Preparations.) Nodule on Diaphragm.—No tubercle bacilli found. Pneumonic Lung Substance.—Four tubercle bacilli seen. Muco-pus from Bronchiole—Three tubercle bacilli seen. Marginal Nodule from Lung.—A moderate number of tubercle bacilli. Right Suprarenal Body Nodule.—No tubercle bacilli found. Right Kidney Nodule.—No tubercle bacilli found. Bronchial Gland Substance—Two tubercle bacilli seen. Animals Inoculated. Three guinea-pigs inoculated with an emulsion of the muscle remained healthy. Guinea-pigs inoculated with emulsions of the lung, liver, spleen, and kidney became tuberculous. PIG 172. Virus P. XLI. Fed once with culture derived from the original material of Virus P. XLI. Dose—50 milligrammes. Date of Feeding—January 25th, 1907. Killed’ in good health—May 3, 1907. [98 days after feeding. ] Clinical History. There was no sign of illness during the experiment. Temperature—The temperature remained normal throughout. Tuberculin Test.—74 days after feeding. Positive. Rise of temperature 2°3° CO. Weight at death.—32°19 kilogrammes. 27676 POST-MORTEM EXAMINATION. General Condition.—Good. Tongue.—Normal. Tonsils.—Normal. Pharynz.—Normal. Submaxillary Glands.—Left submaxillary glands. Two small glands were normal. A third was enlarged and contained about a dozen discrete nodules up to 3 mm., which easily shelled out of the surrounding gland tissue ; they consisted of grey tissue becoming opaque towards the centre where there were whitish slightly gritty streaks and foci. Right submaxillary glands. One was normal. A 2R2 258 second was enlarged and contained about fourteen nodules, similar to those on the other side, up to 5 mm. in diameter. Pharyngeal Glands.—Normal. Cervical Glands.—Normal. Abdomen. Peritoneum.—Normal. Omentum.—Normal. Stomach.—Normal. Intestines.—Small intestine normal. Large intestine showed the follicles prominent, otherwise normal. Gastric Glands.—Normal. Mesenteric Glands.— Almost all the mesenteric glands were beset with irregular waxy caseous nodules with fine opaque white streaks in the centre ; they varied in size up to a small pea, the majority being about the size of a hemp seed; they shelled out leaving smooth walled cavities in apparently normal gland tissue. Spleen.—Normal. Liver.—In the liver just beneath the capsule there was a yellow caseous tubercle within a grey capsule rather larger than a millet seed. Portal Glands.—In one portal gland there was one opaque spherical tubercle the size of a rape seed ; ina second gland there were five similar tubercles which were soft and not appreciably gritty ; the rest were normal. Kidneys.—In the cortex of the left kidney there was a pale area with the surface slightly depressed below the normal level and the substance a little firmer than the rest of the kidney. The right kidney appeared normal. Suprarenal Bodies.—Normal. Renal, Lumbar, and Iliac Glands.—Normal. Thora. Pleura.—Normal. Lungs.—On the surface of the lungs just beneath the pleura there were a few scattered minute trans- lucent tubercles (eight counted), one slightly opaque in the centre ; there were also two rather larger, the size of millet seeds, and perfectly translucent. Section of lungs revealed no more. Thoracic Glands appeared normal. Pericardiwm.—Normal. Heart.—Normal. Diaphragm.—On the left side of the diaphragm on the abdominal surface there were about thirty fibroid translucent hard nodules somewhat flattened; the majority were of the size of millet seeds, but a single large one measured 4 mm. in diameter. Testes.—Normal. Prescapular, Prepectoral, Parotid, Cervical, Gluteal, Precrural, Inguinal, and Popliteal Lymphatic Glands appeared normal. Vicroscopical Examinations. (Smear Preparations.) Lung, Small Tubercle.—No tubercle bacilli seen. Lung, Large Tubercle——No tubercle bacilli seen. Diaphragm (Nodule).—No tubercle bacilli seen. Follicle of Large Intestine—No tubercle bacilli seen, Liver Tubercle.—No tubercle bacilli seen. Portal Gland Tubercle—One tubercle bacillus seen, Animals Inoculated. Four guinea-pigs from the muscle, and one from the kidney, remained healthy. Two guinea-pigs from the became tuberculous. mesenteric glands PIG 202. Virus P. XLI. ‘Subcutaneous inoculation of culture derived from the lung of Pig 166. Dose—50 milligrammes. Date of Inoculation—September 20, 1907. Died—November 23, 1907. Temperature. The temperature remained normal until the fifty- ninth day, five days before death, when it rose slightly, reaching 40°3° C. on the sixty-second day. Tuberculin Test. October 29, 1907. 39 days after the experiment began. Dose of tuberculin (Avian) 4:0 cc. Rise of temperature 0-4° C. Weights. Kilogrammes. September 20, 1907 29:02 November 23, 1907 29°02 POST-MORTEM EXAMINATION. Local Lesion.—In the subcutaneous tissues on the right side of the abdomen there was a circular tumour 6 cm. in diameter. On section it was found to bea eyst filled with caseo-purulent substance. Tt was [Age 19°5 weeks. ] [64 days after inoculation. ] adherent to the muscle in which was a number of discrete soft caseous miliary tubercles. Inguinal Glands—One right inguinal gland was enlarged and almost entirely replaced by softening caseous slightly gritty substance. A second gland contained two caseous tubercles, The left inguinal glands were normal. Thoraa.. Lungs.—The lungs were heavy and did not collapse ; ‘the substance was firm. The anterior lobes were, except for emphysematous margins, reddish grey and airless. On section the solid lung substance was keset with discrete caseous gritty granules. The posterior lobes were extensively consolidated and presented a similar appearance ; the pneumonic tissue was how- ever not continuous, leaving small islands of crepitant lung tissue. Thoracic Lymphatic G'lands.—The bronchial and mediastinal glands contained sparsely-distributed minute opaque caseous tubercles. The right ventral mediastinal gland was enlarged and entirely replaced by softening caseous slightly gritty substance. © 259 Abdomen. Spleen.—Normal. Liver.—Normal. Kidneys.—In the cortices of the kidneys there were some areas slightly paler than the rest of the substance. . There was no tuberculosis of any other organ or gland in the body. Microscopical Examinations. (Smear Preparations.) Lung (grey patch).— A moderate number of tubercle bacilli. Spleen (substance).—No tubercle bacilli. Liver (substance).—No tubercle bacilli. Kidney (substance).—A few tubercle bacilli. PIG 228. Virus P. XLI. Subcutaneous inoculation of an emulsion of lung of Pig 202. Dose—9°0 cubic centimetres. Date of Inoculation—November 23, 1907. Temperature. The temperature remained normal during the experiment. Tuberculin Test. January 31,1908. [69 days after the experiment began.] Dose of tuberculin (Bovine), 2°0 cc. Rise of temperature, 0:7° C. Weights. Kilogrammes. November 25, 1907 wee 19°95 May 7,1908 ... cid ee ve» 63°49 Total gain in weight.—4404 kilos. POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion—In the subcutaneous tissues on the right side of the abdomen there was a tumour about the size of a hen’s egg, which on section was a fibrous walled cyst filled with soft caseous substance con- taining gritty foci. Inguinal Glands—One right inguinal gland con- tained a softening caseo-necrotic nodule 1 cm. in diameter. In the other right inguinal glands there [Age 14:5 weeks. ] Killed when in good general health—May 7, 1908. [166 days after inoculation.] were a few sofi caseous tubercles. A left inguinal gland contained a softening caseous nodule 6 mm. in diameter ; the rest were normal. Thoraa. Lungs.—In the lungs, which were pink and crepitant, there were beneath the pleura scattered minute trans- lucent foci and a few translucent nodules up to a wheat grain in size (parasitic). There was also an occasional grey tubercle with an opaque centre. Abdomen. Spleen.—Normal. Liver.—Normal. Portal Glands.—In one portal gland there was a minute opaque calcareous tubercle. There was no tuberculosis elsewhere in the bedy. Microscopical Examinations. (Smear preparations.) Lung (translucent nodule).—No tubercle bacilli. Lung (opaque nodule).—No tubercle bacilli. PIG 258. Virus P. XLI, Subcutaneous inoculation of culture derived from the lung of Pig 202. Dose—50 milligrammes. Date of Inoculation—March 12, 1908. Died—-June 23, 1908. Clinical History. The animal did not grow, remaining the same size as when it was inoculated. It was listless and was usually lying down; its appetite was poor. The breathing was rapid during the last weeks of life. Temperature. The temperature reached 40°1° Centigrade on the 23rd day and then quickly fell; it remained normal until shortly before death. Its highest point (40°6°) was reached 3 days before death. [Age 15°5 weeks. ] [103 days after inoculation.] Weights. Kohlogrammes. 18-14 15:42 March 12th, 1908 June 23rd, 1908 Total loss in weight.—2°62 kilogrammes. POST-MORTEM EXAMINATION. General condition poor. Local Lesion.—In the subcutaneous tissues of the abdomen to the left of the umbilicus, there was a baggy cyst about the size of a goose’s ego; the skin 260 over it was intact and the tumour could be readily dissected from the under surface. On section it was filled with shreds of necrotic substance and yellow caseo-pus, Inguinal Glands.—One left inguinal gland the size of a pigeon’s egg, and a second half that size, both consisted of thin walled cysts filled with caseo-pus. In two or three smaller glands the centres of the lymphoid follicles were irregular, opaque, and greyish white ; some were a little firmer than the surrounding tissue, others were softer. The right inguinal glands resembled the small glands onithe left side. Precrural Glands.—The precrural glands were beset with ill-defined opaque, yellowish-white patches. Thorax. Lungs.—The convex dorsal surfaces of the posterior lobes were alone air-containing, and just sustained the lungs in water, the anterior and inferior portions of the posterior being quite solid. On section the solid portions of the lungs presented a mottled appearance, the lobules consisting of greyish translucent tissue with occasional soft yellowish foci surrounded by reddish pneumonic lung tissue. In the anterior lobes the greater part was fibroid tissue ; in the posterior the fibroid and pneumonic tissues were about equal. The crepitant portions of the lungs, small in amount, contained discrete nodules similar to those described above. OAUoA The lymphatic fringes on the surfaces of the lungs were prominent. Thoracic Lymphatic Glands.—The bronchial glands were a little firmer than normal, and showed greyish- white streaks and foci. aAv08 The left ventral mediastinal gland was enlarged _to about the size of a partridge’s egg and consisted of a cyst filled with caseo-pus. Abdomen. Mesenteric Glands.—The majority of the mesenteric. glands were normal in appearance, several showed opaque yellowish foci just beneath the capsules. Intestines.—Normal. Spleen.—Normal, Liver.—The liver was congested, but otherwise normal. Kidneys.—In the cortices of the kidneys there were scattered minute perfectly translucent foci. Suprarenal Bodies.—Normal. Various Lymphatic Glands.—The lumbar, iliac, pre- scapular, and the majority of the submaxillary glands contained opaque patches consisting of a definite caseous network. Tonsils—The tonsils showed a few foci of soft yellowish substance. Tongue, Larynx, Pharynx, and Trachea.—Normal. Microscopical Examinations. (Smear preparations.) Lung.—-Very numerous tubercle bacilli. Spleen.—Moderately numerous tubercle bacilli. Inguinal Gland.—Numerous tubercle bacilli. Bronchial Gland.—Numerous tubercle bacilli. Lumbar Gland.—A few tubercle bacilli. Precrural Gland.—A few tubercle bacilli. Tonsil.— A few tubercle bacilli. 61 VIRUS P. XLVI. (July 2, 1906.) SUBMAXILLARY GLAND. CULTURE INOCULATIONS. I.—November 15, 1906. The strain was derived from the original material, and had been in cultivation for a total period of 136 days. The culture used was the 7th generation, 21 days old. CALF 484, Subcutaneous. Dose : 50 mg. Killed : Feb. 25, 1907. 102 days. Fibrous - walled cyst filled with caseo-pus at seat of inoculation. In the left prescapular gland was a single caseous and softened nodule. The liver showed some irregu- Jar grey foci. Result. { Slight T. of lungs, kidneys, ; spleen, and mamma. Slight T. of lungs, kidneys, Slight T. of peritoneum, kidneys, and lungs. One Slight T. of peritoneum, kidneys, and lungs. One CALF 486. RABBITS. Subcutaneous. — eee Dose : 75 mg. Number. | Method.} Dose. ee of Killed : Feb. 28, 1907. nhes 105 days. Fibrous-walled cyst at 1045 Intrav. | 1:0 mg.| K. 100 days seat of inoculation. The left prescapular gland 1046 Intrav. | 0-1 mg.| K. 100 ,, een ee contained a single fibro- : , : caseous nodule. One 1048 Intrap. |10-°0mg.| K. 100 ,, Trowel . - oN ; tubercle in liver. careous vodule. Two 1047 Intvap. | 1:0 mg.| K. 100 ,, oe ee ee tubercle in spleen fated inte ts 200 1050 | Subeut. | 600mg. K.100 ,, | Local lesion. in lung. One tubercle II.—January 24, 1907. The strain was derived from the original material, and had been in cultivation for a total period of 206 days. The culture used was the 9th generation, 21 days old. PIG 164. Subcutaneous. Dose : 50 mg. Killed : May 2, 1907. 97 days. P.M.— Two minute ulcers and caseo-purulent cyst at seat of inocula- tion. Caseation of left inguinal and _— siernai glands. Four _ small tubercles in the lungs. No tuberculosis _ else- where. PIG 168. Fed. Dose : 50 mg. Killed : March 26, 1907, 60 days. P.M.— Caseation of submaxillary, mesenteric and colic glands ase oe D u is ati ti Peete Cultivation. | Rabbit. | Grammes, mB: Died. Ree (a) BEFORE LOSS OF VIRULENCE. i is { 595 1,190 10:0 Died 23 days General tuberculosis. months ... 596 1,220 10 3 49 days General tuberculosis. (0) AFTER LOSS OF VIRULENCE, | 804 1,270 1:0 Killed 52 days , Slight chronic tuberculosis of peri- #3 ionthe nz toneum and lungs. Uterus filled | with tuberculous deposit. 805 1,380 10 5 92 days Slight tuberculosis of peritoneum, lungs, and kidneys; very retro- gressive in the former. ( 828 900 10 3% 37 days Slight chronic tuberculosis of peri- 16} months ad toneum, lungs and kidneys. (Injec- tion partly intracecal.) 829 920 O1 fA 145 days = of tuberculosis in peritoneum only. t 924 1,400 10-0 : 82 days Slight tuberculosis of peritoneum and | ungs. ‘ | 9295 1,420 10:0 ‘ 82 days Local lesion ; slight tuberculosis of 194 mcnths ... } ; lungs. (Injection subcutaneous.) | 926 1,310 10 5 82 days Slight tuberculosis of peritoneum, ne A ; lungs, kidneys, and uterus. u) §Z7 1,250 10 +5 82 days Slight tuberculosis of peritoneum, | lungs, kidneys, and uterus, 313 VIRUS H. 17, “Sp.B."—continued. r INJECTIONS OF RABBITS; MADE TO DETERMINE WHETHER THE VIRUS CONTAINS A STRAIN OF BACILLI, BELONGING TO GROUP IL, AND CAPABLE OF BEING ISOLATED BY RAISING STRAINS FROM SINGLE COLONIES; BUT SHOWING INSTEAD A LOSS OF VIRULENCE OF THE STRAIN EVEN WHEN GROWN CONTINUOUSLY ON PURE SERUM. CULTURE DERIVED FROM CALF 539 [THIRD PASSAGE], THROUGH RABBIT 181. Separation of Colonies on a Glycerin-Serum Medium. Cultures 3 weeks old. Rabbits inoculatedtintraperitoneally. Length of time during | yoainm on etsucion ad which the which final Dose Number | Killed | Duration strain has in Milli- of or of Result, Source of Culture, been in culture was grammes, | Rabbit. Died. Life. ° artificial Brown. cultivation, Culture derived from | 15} months | Pure serum 10-0 95 Died 95 days | Chronic progressive large wrinkled tuberculosis. colony on a gly- 10°0 196 Killed } 166 days | Slight tuberculous peri- cerin-serum tube ' tonitis. Chronic tuber- of 28-11-05. ' culosis of lungs and kidneys, the former rather severe. 10 497 5 166 days | Slight tuberculosis of peritoneum and kid- neys. 10 198 ‘ij 166 days | Slight tuberculosis of peritoneum and kid- neys. (Injection partly subcutaneous.) sa ” 16 months | Pure serum 10°0 800 Killed 92 days | Chronic fibrous tubercu- losis of peritoneum, lungs, and kidneys. 10-0 801 Died | 36 days | Slight tuberculous peri- tonitis, Tuberculosis of lung. Trace in kidneys. 10 802 Killed 92 days | Abscess at seat of inocu- lation. A few fibrous tubercles in lungs. In- jection intramuscular. 10 803 33 92 days | Slight tuberculosis of lungs and peritoneum. Trace in kidneys. Culture derived from | 164 months | Pure serum 1:0 828 i 37 days | Slight chronic tubercu- small colony on a losis of peritoneum, glycerin - serum lungs, — and kidneys, tube of 28-11-05. (Injection partly in- tracecal.) O1 829 sy 145 days | Trace of tuberculosis io peritoneum only. Serum Series. [Controls.] Culture 3 weeks old, Rabbits inoculated intraperitoneally. 10th generation of | 16 months | Pure serum 1-0 804 Killed | 52days | Slight chronic tubereu- culture on pure losis of peritoneum serum, and lungs. Uterus filled with tuberculous deposit. 10 805 6 92 days | Slight tuberculosis in peritoneum, lungs, and kidneys; very retro- gressive in the former. 27676 BAZ 314 MODIFICATION OF THE VIRULENCE OF THE VIRUS H. 13. “A.D.” The third instance of apparent modification oc- curred in the case of Virus H. 13. “A.D.” This came from a male child, aged four years, who died on June 6, 1908, of meningitis and miliary tuberculosis, affecting the lungs, liver, and spleen. The mesenteric glands were normal, but the bronchial glands were caseous, and were almost certainly the seat of the oldest lesion which was found after death. The largest, which measured 1 by 3 in., contained caseating nodules, one of which had already broken down into grey purulent matter. Two other bronchial glands, which were whitish, opaque, and rather dense, con- tained caseating foci. There were numerous tubercle bacilli seen in a preparation from one of these glands. An emulsion was made from the gland most affected, and from the most affected portions of the two other bronchial glands, and since this did not seem sufficient for the purpose there was added two or three grammes of spleen. It was not very rich in tubercle bacilli, only 1,600 tubercle bacilli per cubic centimetre being estimated. Two calves, Nos. 119 aud 129, were injected subcu- taneously with 10 cc. of this emulsion, on June 7, 1903. The estimated dose, 16,000 bacilli, has been shown by other experiments to be too small to pro- duce general tuberculosis with certainty in the calf, even if the virus be virulent for this species of animal (see Appendix to 2nd Interim Report, Vol. II, page 1035). Therefore, the fact that in the cases now under consideration, minima! lesions, which did not spread beyond the nearest lymphatic glands, were found when the animals were killed some 9 or 19 weeks later, cannot be accepted as conclusive evidence that the virus was not virulent for the calf, and belonged to our Group II. Unfortunately, no rabbits were inoculated at this stage of the experiment Another calf, however, was subcutaneously injected with a larger dose of bacilli contained in an emulsion made from tbe calcareous prescapular gland of Calf 119, the estimated number of bacilli injected in this case being about 3,500,000. The result waz even less than in the former calves, for when the animal was killed 89 days later there was only a pea- sized calcareous nodule at the seat of inoculation, and no tuberculosis elsewhere. Even in this case the dose was not large enough to be quite satisfactory. Nevertheless there has occurred no instance in our experience where a virus of Group I. has produced a local lesion only, however small the dose; and in mapy cases general tubercu- losis has been caused by injection of far smaller quantities of tubercle bacilli than were used in this case. The nearest approach to this result which has been produced here by a virus of Group I. was afforded by Calf 223 (H. 20. “F.L.”), in which a dose of 2,300,000 bacilli produced minimal calcareous lesions which did not spread beyond the nearest lymphatic glands. This calf, however, was nearly twice as large as No. 225." It cannot be maintained that the virulence may have deteriorated while the virus was subsisting in a calcareous lesion in a resistant animal, for our experience here affords striking evidence to the contrary.t It must therefore be held that the virus “ A.D.” at this stage had a low degree of virulence for the calf, and consequently that it belonged to our Group II. At a later stage it developed high virulence for this animal. The steps by which this modification oc- curred were as follows: When Calf 129, one of the two animals injected with the emulsion of haman tissue, was killed on August 12, 1903, the lesions found in it were so small that there was very little tuberculous material available for the injection of other animals. The virus was accordingly passed through guinea-pigs with the objects of obtaining larger quantities of tubercle bacilli. Three sets of guinea-pigs were injected in succession, and from the tuberculous organs of the last of these an emulsion was made containing over 27,009,000 tubercle bacilli per cubic centimetre. This was injected in doses of 20 cc. (541,300,000 bacilli), into Calves 299 and 301. In the case of No. 299 the injection was intra- peritoneal. In spite of the size of the dose the animal remained well, and when killed 87 days after injection, the disease was found to be limited to the peritoneum and to a few lymphatic glands. It was nowhere severe, and none of the great organs were affected. Even allowing that this may have been a highly resistant individual, it is certain that the virus had not yet attained to the virulence of those be- longing to our Group L.} : : The virus, however, at this stage had attained a virulence for the calf far in excess of that of a member of Group II, for the calf, No. 301, which was injected subcutaneously, developed severe miliary tuberculosis, and was very ill when killed 33 days after injection. A rabbit injected with the same emulsion died of general tuberculosis in 24 days. In contrasting Calf 301 with No. 299, it seems probable that it had a very low degree of individual resisting power. From the posterior thoracic gland of Calf 301, an emulsion rich in tubercle bacilli was made and injected subcutaneously into three calves. No. 315 received a dose estimated to contain 15,000 bacilli, which was intended to be approximately tne same, or a little smaller than that given to the original pair of calves. It produced only small lesions at the seat of inoculation and in the nearest lymphatic glands, and although there were one or two minute tubercles in the thoracic glands, and one or two also in the lungs, liver, and spleen, such lesions have been produced by many of our Group II. viruses, and it cannot be maintained that this animal atfords any certain proof of increase of virulence. This, however, is not the case with the two other calves injected at the same time. No. 321 received a dose estimated to contain 3,500,000 tubercle bacilli, the same number as was contaired in the dose given to No. 225. The calf rapidly developed severe general tuberculosis, and was very ill when killed 63 days after injection. Calf 325, which received a dose estimated to contain 500,000,000 tubercle bacilli, which was slightly smaller than the number injected into No. 301, developed very acute miliary tuberculosis, and was in a dying state when killed 24 days after the injection. Rabbits injected with the same emulsion also died of general tuberculosis. ; We may conclude from these results that at this stage the virus possessed all the virulence for the bovine and rabbit species of the viruses belonging to Group I. The fact that No. 315 did not develop general tuberculosis is not in conflict with this con- clusion, because, as already stated, even typical viruses of Group I. cannot be relied upon to produce general tuberculosis with this dose. To sum up these results, we may say that the virus derived from a case of meningitis and general tuber- culosis in a young child, which probably arose in a bronchial gland, possessed, in all probability, when first tested on bovine animals, a low degree of viru- lence for them characteristic of our Group II. It is not possible to make a more definite statement, because the doses used at this stage of the experiment were too small. It is more certain that after the virus had passed through a series of three guinea-pigs and a calf its virulence for the ox species had become greatly augmented, and that it then possessed the high bovine virulence characteristic of our viruses of Group I., and that at an intermediate stage, when it was injected into Calf 301, its virulence was notably higher than that of viruses of Group II., but had not yet reached the standard of a virus of Group I. CuLTURE INJECTIONS. Unfortunately, no cultures were obtained either directly or indirectly from the human material, nor from the calves which were injected in the earlier stages of the experiment. ‘he first culture obtained was sown from the posterior thoracic gland of Calf 301. It was eugonic. Its virulence was not tested * It is quite possible that the “ F.L.” virus, when it was injected into Calf 223, that is when it first came from man, had not the full degree of bovine virulence which it afterwards attained ; modification. + See Calf 223 (H. 20.“ F.L."). Calf 797 (H. 49, “1.C.”). Heifer 11 (H. 2. “ Sp.A.”). in other words, that “F.L.” affords another instance of Bull Calf 35 (H. 10. “ B.S.”). + For the effect of injecting a virus of Group II. into the peritoneal cavity, see Calf 309 (H. 11. “ E.D.”). dl. ‘until after it had been fifteen months in artificial culture on bovine serum. At this period it was injected in 50 mg. doses into two calves, and also into rabbits. Calf 761 developed a tumour at the seat of inocula- tion, consisting of a fibrous walled cyst with softened ‘caseous contents. The nearest prescapular gland con- tained a large caseous nodule, and this also was softened. These lesions are characteristic of viruses of Group II. ‘There were, however, in the lungs some small lesions slightly more severe than those commonly produced by viruses of Group II. These consisted of about a ‘dozen pea-sized fibrous nodales with large irregular caseating nuclei. One of these nodules was } in. in diameter, and there were also smaller tubercles, each with a caseous centre. It is in the existence of ‘this central caseation as well as in their size, that these lesions differ from those sometimes produced by viruses of Group II. A corresponding calf, No. 765, developed minimal lesions which did not extend beyond the nearest lymphatic glands. The rabbits also developed mini- mal lesions. Thus if the virulence at this stage differed at all from that of a virus of Group IL. the difference was ‘slight only. The virulence must undoubtedly have Y ov been higher fifteen months earlier when the virus existed in the body of Calf 301, which suffered from acute general tuberculosis. This conclusion is sup- ported by the fact that an emulsion of the thoracic gland of this animal produced general tuberculosis when injected into a rabbit. We are therefore bound to conclude that this strain was once virulent for rabbits and calves, and that its virulence was unstable, and disappeared during the course of fifteen months’ cultivation on artificial media. A parallel instability of virulence has already been described in one strain obtained in each of the two previously described modification experiments. To return to the virus “A.D.” A second culture obtained from Calf 321, injected in a dose of 50 mg. into Calf 825, provoked acute miliary tuberculosis, which proved fatal in 25 days, thus confirming the opinion already expressed, that in the final stage of the passage experiment the virus had attained the high bovine virulence characteristic of cur Group I. Injected in 10 milligramme doses into Calves 821 and 823, it produced severe tuberculosis fatal in 50 days in one, and slight tuberculosis only in the other. This culture was distinctly dysgonic, and showed as much reluctance to grow on glycerin media as any of our viruses of Group I. CALVES INJECTED WITH THE VIRUS H. 13. “A.D.” I—tTissue Emulsions. | | Mode Estimated dose — | of of of Tubercle ! Animal. Injection. Bacilli. ' | Fate of Animal and | Condition at Death, | Amount and Range of Tuberculous Lesions at Death. CALVES INJECTED WITH EMULSION OF HUMAN BRONCHIAL GLANDS. ed Calf 19 | S. 16.000 os | Calf 199 S. 16,000 | Killed when well | Local tuberculosis, | after 67 days. | Killed when well | after 66 days. Local tuberculosis. CALVES INJECTED WITH THE VIRUS DESCENDED FROM CALF 129. ; Calf 299 DB 541,333,000 Ist Passage ... Calf 301 8. 541,333,000 (Calf 315 Ss. 15,000 und Passage... | 4 Calf 321 s. 3,500,000 Calf 325 De 500,000,000 Killed when well after 87 days. Killed when very ill after 33 days. Killed when well after 70 days. Kiiled when very ill after 63 days. Killed when dying after 24 days. Slight tuberculosis. General tuberculosix, severe. Slight tuberculosis. General tuberculosis. severe, — General tuberculosis, severe. CALF INJECTED WITH THE VIRUS DESCENDED FROM CALF 119. 1st Passage ... | Calf 225 | Se i 3,400,000 Killed when well Local tuberculosis. ‘ after 89 days. { \ | S = Subcutaneous. P = Intraperitoneal. RABBITS INJECTED WITH THE VIRUS H. 18. “A.D.”} I.—Tissue Emulsions. I Source of Number | Estimateddose} Kilied es ies aad Material used for of of Tubercle or a tate Result, Injection. Rabbit. | Bacilli. Died. ys. Guinea-pigs 905-907. { 98 135,330,000 | Died 24 General tuberculosis. (Third series from | - : Calf 129.) l.a9 r a 12 No tuberculosis. Se 2 ; | (Subcutaneous). ; ‘Calf 301 ges si { 37 99,332,000_ 4 a7 General tuberculosis. (Lhoracic gland.) ar | ; . t 38 37,000,000: 33 18 Lungs solid, Tubercle bacilli swarm- | (Intravenous). ing in all organs. . Calf 801. . (Through 107 fr 16 Tuberculosis of peritoneum. Rat 15.) " ! Tubercle bacilli in organs. + All injections were intraperitoneal, except where otherwise mentioned. 316 CALVES INJECTED WITH THE VIRUS H. 13. “A.D.” II.—Cultures. Immediate Total Number | Weight Fate of Animal Amount and Range —= Source Duration of of in an of Tuberculous Lesions of Culture. Cultivation. Calf. Kilos. | Condition at Death. at Death. A.—DoseE: 50 MILLIGRAMMES, Subcutaneous. Ist Passage ... Calf 301 15 months 61 62°58 Killed when well | General tuberculosis, after 90 days. slight. . ” 4s $5 165 65°75 Killed when well Local tuberculosis. after 91 days. 2nd Passage... Calf 321 153 months 825 44-44 Died General tuberculosis, after 25 days. severe, B.—DosE: 10 MILLIGRAMMES. Subcutaneous. Ist Passage ... _ _- _ _ _ — 2nd Passage... Calf 321 154 months 821 41°27 Dying General tuberculosis,. , after 50 days. severe. ” ey o 823 43°54 Killed when well Slight tuberculosis. after 90 days. RABBITS INJECTED WITH THE VIRUS H. 13. “A.D.” I1.—Cultures. Immediate Total Number | Weight Killed | Duration of — Source Duration of of in or Life Result. of Culture. Cultivation. Rabbit. |Grammes. Died. in Days, A.—DosE: 10 MILLIGRAMMES. Intraperitoneal. Ist Passage ... Calf 301 15 months 372 1,070 Killed 90 Slight tuberculosis of lung, peritoneum, and kidneys, ? liver.. i Cnjection ? intra- cecal.) 2nd Passage... Calf 321 1534 months 518 1,240 Died 132 Chronic tuberculosis. (Injection intracecal.) B.—DosE: 1 MILLIGRAMME, Intraperitoneal, Ist_Passage ... Calf 301 15 months 373 800 Killed 90 Chronic general tuber- culosis. : 2nd Passage... Calf 321 153 months 519 1,220 Died 33 General tuberculosis. ” ” fe 520 1,200 55 36 General tuberculosis. FA 5 24 months 856 660 5 17 General tuberculosis. ” ” ” 857 630 i 19 General tuberculosis. The hypothesis that one was dealing with a mixture of bacilli belonging to Groups I, and IT. was tested in this case as was done in the case of the first modifica- tion experiment. The first virulent culture, namely that from Calf 321, was grown for several generations on glycerin serum, a method which, as already stated, has sometimes been successful in eliminating the virulent (dysgonic) element from an artificial mixture. In this case the strain was grown for six generations during ten months and finally on pure serum. A parallel series of cultures was kept up on pure serum. ‘When the final test was made the strain was two years old. Both series of culture proved to be fully virulent for rabbits. The result of these experiments lent no support to the hypothesis that we had to deal with a mixture of. bacilli of Groups I. and IT. 317 INJECTION OF RABBITS; MADE TO DETERMINE WHETHER THE VIRUS CONTAINED A STRAIN OF BACILLI, BELONGING TO GROUP II., AND CAPABLE OF BEING ISOLATED BY MEANS WHICH PROVED SUCCESSFUL IN SEPARATING;AN ARTIFICIAL MIXTURE OF GROUPS I. AND IL, NAMELY, GROWTH ON GLYCERIN MEDIA. CULTURES DERIVED FROM CALF 321, DIRECT [SECOND PASSAGE]. Glycerin-Serum Series. Cultures 3 weeks old. Rabbits inoculated intraperitoneally. Length of . time during | Medium on ‘ Generation /sentcheteain | which Hual | , Woe | Number | seateg | Daehn oO has been in @ulbnre in Milli- of or Died of Life Result. Culture. artificial was grown grammes, | Rabbit. fi in Days. cultivation. after 6 2 fiom in fe Pure serum 10°0 852 Died 15 days General tuberculosis. er 6 on months glycerin on glycerin 10:0 853 5 12 days Early general tubercu- serum ]. serum ]. losis. 1:0 854 i 21 days General tuberculosis. 1:0 855 3 21 days General tuberculosis, Serum Series. [Controls.] Cultures 3 weeks old. Rabbits inoculated intraperitoneally. 14th 2 years Pure serum 10 856 Died 17 days General tuberculosis. on pure serum. 10 857 4 19 days General tuberculssis, MODIFICATION OF VIRULENCE OF THE VIRUS H. 49. “T.C.” In the hitherto described instances the modification has been complete, that is to say, the viruses had at the start the low degree of bovine virulence cha- racteristic of Group IL., and they ended with the high degree of bovine virulence characteristic of Group I. In the case now about to be described the original virulence at first fell only a little short of the virulence of Group I., and it gained the full virulence cha- racteristic of this class after a single passage through a calf. The case presents several points of interest. It is the only one* in which the original virulence (for ox -and rabbit) of a virus obtained from man was in any sense intermediate between our Groups I. and II. As already stated it was much nearer Group I. than Group II. And of all the viruses which were virulent for the ox and rabbit it was the only one which came from an adult. The fact that the least virulent of the virulent viruses came from the oldest patient, in whom, moreover, the disease was of long standing, may be taken as evidence that the bacillus of Group I. becomes modified in the direction of Group I. by residence in man. Tf this kind of modification can be established, it supports strongly the evidence of the contrary kind of modification, namely, that of Group IT. into Group I. in the ox. T.C. was a youth who died at the age of eighteen of tuberculosis of thelungs. There were small cavities in those organs, but there was evidently much older disease in the mesenteric glands, one of which was nearly as hard as stone. Unfortunately no strain of tubercle bacilli was obtained from the lung. All the animals injected with an emulsion made from it died of some acute infection; but a strain of culture was obtained directly from the mesenteric glands. This strain was injected when only two months old in doses of 50 mg. of bacilli into two calves. One of these, No. 787, died 81 days after injection of severe tuberculosis of the lungs, with a few small tubercles in other organs. The other, No. 797, killed after 83 days, had only small lesions near the seat of inoculation, and a few little tubercles in the lungs and thoracic glands. Even allowing that this animal was a highly resistant individual it cannot be doubted that this result indicates a degree of virulence which is distinctly below the standard of Gronp I. The only instance in which 50 milligrammes of a true Group I. virus failed to produce fatal tubercuiosis within three months occurred in the case of Calf 667, H. 10. “ B.S.,” and in this animal the lesions were far more severe and very widespread. But in order to leave no doubt as to the degree of virulence of the virus at this stage four aclditional calves were subcutaneously injected with a culture of the same strain, on November 3, 1905, two receiving 50 milligrammes and two 10 milligrammes. The former pair, Nos. 957 and 959, died in 61 and 53 days respectively of general tuberculosis. This is distinctly longer than the usual duration of illness after 50 milligrammes of a virus of Group I., and therefore afford additional evidence of the failure of the virus at this stage to reach the standard of that group. Of the two animals which were injected with 10 milligrammes, No. 953 died of general tuberculosis after 108 days, and No. 945 was killed when in good health after 96 days, and showed, in addition to a * Except the lupus virus H. 53. “D.H.” and possibly H. 20.“ F.L” 2 o local lesion, and tuberculosis of the nearest lymphatic glands, only a few small lesions scattered in the lungs, liver, spleen, and in the thoracic and portal glands. That these results indicate that the virulence for the calf was not quite up to the standard of Group I, can be best seen by referring to the diagram* which sets out the duration of life of calves which received subcutaneous injections of 50 and 10 milligrammes of culture of Group I. viruses. The four calves above mentioned are placed in this table for comparison. Stronger evidence that the virulence of the virus was not up to the standard of Group I. was afforded by the injection of rabbits. These animals died of tuberculosis, but after a disease which was protracted decidedly beyond the usual limits. Thus even 50 milligrammes injected intraperitoneally proved fatal only in 39 days, 10 milligrammes in 69 and 35 days respectively, 1 milligramme in 87, 59, and 88 days respectively, and 0:1 milligrammes in 79 and 60 days respectively. The unusual slowness with which the disease in these cases approached a fatal termination can be best appreciated on reference to the table of culture injections into rabbits. These injections of calves and rabbits indicate, I think, sufficiently clearly that the virus H. 49. “ T.C.” when first obtained from man had a degree of viru- lence for these animals which approximated to, but fell somewhat short of, the standard of viruses of Group I. Full bovine virulence was, however, obtained by passage through a single calf. From the prescapular gland of No. 797, the calf which was the least affected of those which were in- jected with culture from the original material, a culture was made, and this was injected subcutaneously into four calves, two receiving 50 milligrammes and two 10 milligrammes. : All four suffered from very acute and fatal general tuberculosis. Nos. 931 and 949 (50 milligrammes) died in 21 and 39 days respectively, and Nos. 929 and 933, which received 10 milligrammes, died one in 39 days and the other actually earlier than either of the 50 milligramme calves, namely, in 19 days. That this was a real increase of virulence I think cannot be denied. The rabbits also, injected at the same time as the calves just mentioned, gave similar results. All died very rapidly of general tuberculosis, in a much shorter time than those which received corresponding doses of the culture obtained direct from the human mesenteric glands. The experiment with this virus therefore falls to some extent into line with the modification experi- ments previously described. In them the original virulence for calves and rabbits was that of an 18 ordinary member of Group II. In this it was high. for these animals, but not up to the standard of Group I. In all, the virulence eventually rose to the. full standard of Group I. The patient from which the virus was obtained in this case was a young adult, the only one from whom a virus has been. obtained which was originally virulent for calves and rabbits. In the mesenteric glands of this youth there was tuberculous disease of very old standing. He died of more recent tuberculosis of lung. Here then we seem to have gota case of mesenteric infection with a bovine tubercle bacillus, probably in childhood, and when the disease proved fatal the bacilli had already to some slight but definite extent lost their virulence for calves and rabbits ; and full virulence was rapidly restored by passage through a calf. It is much to be regretted that we failed to get a strain of tubercle bacilli from the lungs of this patient. The virulence for calves and rabbits of the original strain disappeared under artificial culture, as the following experiments will show. The strain cultivated from the human mesenteric glands had, as already stated, been tested on calves and rabbits and found virulent. These tests had been made when it had been two months under cultivation on pure serum. They were repeated when it had been growing for seven months; the strain was still virulent. It was then grown for three genera- tions on glycerin serum, a control strain being kept up on pure serum. These cultures were tested when the strain had been nine months under artificial cultivation. The result was unexpected. Not only had the cultures of the glycerin series lost their virulence for the rabbit, but those of the control series on pure serum also. About two months later the strain was again tested. From a culture grown for the first time on glycerin serum two strains were raised, one from a small colony and the other from a large colony ; the former was grown again on pure serum, and the latter. on glycerin serum [for the second time]. Finally pure serum cultures were sown from each and injected into rabbits. Neither culéure proved virulent. In order to leave no doubt about this unexpected loss of virulence the culture was again tested on Calf No. 1097 and three rabbits. The strain had then been thirteen and a half months under artificial cultivation, always on pure serum, and had reached its thirteenth subculture. These animals were killed three months after in- jection, and only the minimal lesions, characteristic of an injection with a virus belonging to our Group IL., were found. SUMMARY. These results may be summarised as follows. A strain of culture was raised from the mesenteric glands of a youth, the subject of mesenteric and pulmonary tuberculosis, the former being evidently of very long standing. When first tested two months after isolation from man this strain of tubercle bacilli was virulent for calves and rabbits, and produced fatal tuberculosis in them. But the virulence for these animals was de- finitely below the standard of our Group I. Passed through a single calf the virus at once reached this standard ; and proved as rapidly fatal to calves and rabbits as any we have dealt with. The same virus, however, lost its virulence for these animals during artificial cultivation ; and when tested nine and again thirteen months after isolation, it was. found to possess then no more virulence for them than avy virus belonging to our Group II. Of these facts there is ample evidence, a sufficient number of animals having been injected on each. occasion to eliminate any uncertainty which might arise in consequence of the known difference of sus- ceptibility of different individual animals. These results are of great interest, for they reveal a virus unstable in both directions ; capable of being increased in virulence for the bovine species when passed through a calf, and of losing whatever virulence it possessed for these species during continued growth on artificial media. * Appendix to!2nd Interim Report, Vol. LI. page 1029. + Appendix to 2nd Interim Report, Vol. II, page 1091. O19 CALVES INJECTED WITH THE VIRUS H. 49. “T.C.” Cultures. [No INJECTIONS WERE MADE WITH ‘TISSUE EMULSIONS. ] i t i I Immediate Total : ( . Amount and Range of — Source Duration of | Number | Weight | Fate of Animal and | “qypercnlous Testes of Culture. Cultivation of Calf. | in Kilos. | Condition at Death. at Death .* A.—Dost: 50 MILLIGRAMMES. Subeutancous. _ Human mesen- 2 months 187 44°90 Died General tuberculosis, teric glands, after 81 days. moderate. a= direct. = af % "97 44:90 Killed when we Slight tuberculosis. after 83 days. _ 53 7 months 957 44 90 Died General tuberculosis, after 61 days. severe. _ 5 r 959 42°18 Died General tuberculosis, . after 53 days. severe. 1st Passage Calf 499 2 months....|__.931 56°24 Died General tuberculosis, | after 21 days. severe. | is OnoTsInG: AND NECESSARINS (including Materials). Vo” Priced Vocabulary of. 1911. 13, © ENGINEERING. PiELD, Manual of. 1911, od, Equipment REGULATIONS :-— Part 2. Sec. xq. Signal Company (Cable). (Regular Army). 2d. 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Investigations into the Tuberculosis occurring naturally in Certain Animals other than Man: and Modification Experiments. F. GRIFFITH, M.B. Presented to both Houses of Parliament by Command of His Majesty. LONDON: PUBLISHED BY HIS MAJESTY’S STATIONERY OFFICE. To be purchased, either directly or through any Bookseller, from WYMAN anp SONS, L@D., FeTTER LANE, E.C., and 32, ABINGDON STREET, S.W.; or OLIVER and BOYD, TWEEDDALE COURT, EDINBURGH ; or E. PONSONBY, Lrp., 116, GRAFTON STREET, DUBLIN. PRINTED BY DARLING AnD SON, LTpD., BACON STREET, E. : 1911. E [Cd. 5894.] Price 4s. 5d. No 3436 PREFACE. Tuts volume records the results of five investigations carried out at Walpole Farm the titles of which are given on the opposite page. Much of the preliminary work on two of the investigations, namely those on Equine and Avian Tubercle Bacilli, was carried out conjointly with Dr. A. Srantey Grirrira. Iam much indebted to Mr. C. F. Fox for his assistance in the prepara- tion of the reports. EG: CORNELL UNIVERSITY. THE Roswell P. Flower Library THE GIFT OF ROSWELL P. FLOWER FOR THE USE OF THE N. Y. STATE VETERINARY COLLEGE. 1897 LIST OF INVESTIGATIONS CONTAINED IN THIS VOLUME. PAGE Investigation of Tubercle Bacilli derived from ‘Tuberculosis occurring naturally in the Horse na sae wine we ‘i ie — 4 Investigation of Viruses obtained from Casual Tuberculosis of Various Mammals 149 Investigation of Avian Tubercle Bacilli obtained from Birds and Swine wie! LOT Modification Experiments with Tubercle Bacilli derived from Animals other than Man _... wea oss ses ai re ine ‘si i 98D Investigation of Artificially Mixed Cultures... Bis 2 be we 487 2000, Wes, T.L. 277/03 & 13670/04-5. 12/08. D&S. 32. 27676E Az INVESTIGATION OF TUBERCLE BACILLI DERIVED FROM TUBERCULOSIS OCCURRING NATURALLY IN THE HORSE. BY F. GRIFFITH, M.B. INVESTIGATION OF TUBERCLE BACILLI DERIVED FROM TUBERCULOSIS OCCURRING NATURALLY IN THE HORSE. TABLE OF CONTENTS. PAGE Introduction hie sia or ee en i e sate wee ‘General Results :— (a) Experiments with cultures from the original material... ray OE (b) Passage experiments se ak fue bebe ha we os Cultural characters ee se ke sia ete feb va ~ O Table of Origins of the Equine Viruses ... ae ae sek si van 10 Summaries of Results of Inoculation Experiments with each Virus :— Virus E. I. ans axe ai ste ae we se Seg. Virog HeNIT. ans wie’ si Kit wie ccd ne Fa, “Te Virus E. V. sa ses sin me ee ah ie va 12 Virus E. II. (a) Cultures from the original material oe as day. |e (b) Cultures after passage wae sini si me wae LS Virus E. IV. (a) Cultures from the original material ee wus sve Ld (b) Cultures after passage oe aie sa i we 14 Details of the Investigation. PART I. EXPERIMENTS WITH CULTURES FROM THE ORIGINAL MATERIAL. ‘Tabular Summary of Inoculation Experiments on Calves ... ite sale ai = we 17 . i a Monkeys, Baboons and Lemurs... ats --. «18 - 3 B Horses. ... sas es ‘i or we = 19 Pigs. te i Aisa vii w= =18 Tabular Summaries of Inoculation Experiments on Rabbits :— With Virus E.I. ... ue ae ee a vee sie ee w» =. 20 With Virus E. II.... ase oe si sae “es a se se 21 With Virus E. III. ae es a ia seh hs ss we 26 With Virus E. IV. aes ee tee sgt ae a site we 29 With Virus E. V.... aes “ex aes ene ae et sie a §=—38 ‘Tabular Summaries of Inoculation Experiments on Guinea-pigs :— With Virus E.I. ... ie ai bis ace ee ei ae dae Oe With Virus E. I.... de ae aie ies a a sing we 26 With Virus E, III. sie ie ate ote weg ts se ., 28 With Virus E. IV. seg uae i ss ae ds si cag: TBE With Virus E. V.... ae ae ne i ae sia = wa 38 Descriptive Charts of the Experiments with each Virus, and full Post-mortem Notes on the larger Experimental Animals :— Virus E. I. E. If. E. III. E. IV. E. V. PART II. PassaAGE EXPERIMENTS. Tabular Summaries of Inoculation Experiments on Calves (Viruses B. II. & E.IV.) ... Monkeys (Viruses E. II. & E. IV.) Rabbits :— Virus E. II. ... Virus E. IV. ... ” ” y ” ” ” Guinea-pigs :— Virus E. IT. ... Virus E. IV. ... Descriptive Charts of the Experiments, and full Post-mortem Notes on the larger Experimental Animals :— Virus E. IT. Virus E. TV. Control Experiments with Bovine Tubercle Bacilli on Horses and Monkeys :— Summary of Results of Inoculation Experiments on Horses with Bovine Tubercle Bacilli... Control Experiments on Monkeys with Bovine Tubercle Bacilli Descriptive Charts and Post-mortem Notes :— Viruses B. XI. and B. XII... Plate.—Photograph of Cultures of Bovine and Equine Tubercle Bacilli Index to Animals PaGE 39: 47 67 73 92 94 95 96 100 99 104 105. 123 137 137 138 147 446. “I INVESTIGATION OF TUBERCLE BACILLI DERIVED FROM TUBERCULOSIS OCCURRING NATURALLY IN THE HORSE. . INTRODUCTION. Tuberculous material has been obtained from five cases of tuberculosis occurring naturally in the horse, and the characters and virulence of the cultures isolated from this material have been made the subjects of investigation. The original material of virus E. III. came from a horse which had enormously enlarged tuberculous mesenteric glands ; that of viruses E. I. and E. V. was obtained from horses affected with general tuberculosis. Virus E. II. was derived from the spleen of a horse which had tuberculosis of the mesenteric glands, and virus E. LV. from the spleen of a horse in which no lesions were visible elsewhere. The results of the investigation are presented in sequence thus. This introduc- tion is followed by the general results of the experiments under two headings and by a short statement as to the characters of the cultures. On page 10 are the Tables of Origin of the Equine Viruses, which contain, together with other details, a description of the material from which the cultures were obtained. On page 12 are summarised the results of the inoculation experiments on animals with the cultures from the original material of each virus, and in addition the results of the animal passage experiments with virus H. IL. and virus E. IV. The rest of this Report, which contains the tabular summaries of inoculation experiments on different species of animals, the descriptive charts showing the inoculation experiments made with each virus, and the detailed accounts of the post- mortem examinations of the larger animals, is divided into two parts. In the first part are the details of the experiments with the cultures from the original material ; in the second part are the details of the passage experiments. At the end of the Report there are included control experiments with bovine tubercle bacilli on horses and monkeys. GENERAL RESULTS. (a) EXPERIMENTS WITH CULTURES FROM THE OriciINAL MATERIAL, Cultures from three viruses, HE. I, E. III. and E. V., correspond exactly in their characters on artificial media and in their high virulence for rabbits and guinea-pigs with cultures isolated from bovine animals. Viruses E. I. and E. III. have in addition been tested by the subcutaneous inoculation of culture on calves and pigs and have caused in all these animals fatal general tuberculosis such as follows the inoculation of virulent tubercle bacilli of bovine origin. The cultures from the remaining two viruses, E. II. and E. IV., grow on artificial media like bovine tubercle bacilli, alike when freshly isolated and after three years of residence on artificial media ; with this individual difference that the first has the characters of the most difficult growing bovine tubercle bacillus, and the second resembles an easy growing bovine bacillus. The culture of virus E. IL. was inoculated subcutaneously into seven calves all of which remained in good ,health and developed merely retrogressive tuberculosis, slight in extent and similar to that produced in calves by a tubercle bacillus of low virulence derived from man. For rabbits virus E. II. is moderately virulent, causing fatal general tuberculosis by the intravenous and intraperitoneal methods of inoculation, and setting up after subcutaneous inoculation tuberculosis varying in degree but never so severe as that produced by a bovine tubercle bacillus. The tuberculosis produced in monkeys by virus E. II. is definitely less severe than that following the inoculation of bovine tubercle bacilli. After subcutaneous inoculation this animal develops slowly progressive disease although fatal tuberculosis is invariably produced. After feeding, the disease in 8 monkeys is still more chronic and in one case was only slightly generalised even: after 101 days. In pigs and guinea-pigs fatal general tuberculosis has followed subcutaneous inoculations in every case but the fatal result in these animals has. been usually somewhat retarded as compared with that resulting from the inoculation of bovine tubercle bacilli. The culture from virus E. IV. was inoculated subcutaneously into six calves. Five of these remained in good health and showed when killed after about three months slight non-progressive tuberculosis. The sixth (No, 498) died of general tuberculosis of an irregular type, 89 days after inoculation ; it showed a severe tuber- culous pneumonic condition of the lungs with very little disease elsewhere. This exceptional result may be due either tc greater susceptibility of the particular animal or to accidental introduction of a portion of the culture into a vein at the time of inoculation ; of the two suppositions the latter appears the more probable. For rabbits the culture of virus E. IV. is moderately virulent and the different methods of inoculation have caused tuberculosis of varying degree the results being similar to- those obtained with virus E. JI. Two young baboons were tested, one by subcutaneous inoculation the other by feeding. The former, Baboon 72, succumbed to moderately severe general tuberculosis after a somewhat prolonged period, the latter, Baboon 74, remained well and when killed after 448 days showed only a slight non-pro- gressive tuberculosis. For monkeys the virulence was lower than that of a bovine: tubercle bacillus. In pigs and guinea-pigs fatal general tuberculosis was produced. after periods usually longer than after the inoculation of bovine tubercle bacilli.* The virulence of three of the equine viruses has been tested on horses. The culture of virus E. I. inoculated intravenously in a dose of 10 milligrammes caused fatal tuberculosis in 17 days. The horses inoculated intravenously with equiva- lent doses of viruses E. If. and E. IV. died of general tuberculosis in 40 and 98 days respectively. The culture of virus E. II. did not cause progressive tuberculosis after subcutaneous inoculation: a horse inoculated subcutaneously witn virus KE. [V. was killed when very ill after 125 days and had general tuberculosis. The experiments on horses with bovine tubercle bacilli have not resulted in the production of progressive tuberculosis after either subcutaneous inocula- tion or feeding with moderately large doses, but by the intravenous inoculation of 10 milligrammes acute tuberculosis was set up fatal in 20 days (see page 137). (b) Passage EXPrrRIMENTs. Since the cultures of viruses E. II. and E. LV. resembled bovine tubercle. bacilli so closely in their cultural characters and in some respects in their patho- genic properties, it was considered possible that they might be cultures of bovine tubercle bacilli which had become attenuated in their virulence by residence in the horse or before gaining entry into the horse. An attempt was made to. throw light on this latter question by inoculating horses with fully virulent bovine tubercle bacilli, subsequently recovering the bacilli by culture and testing the virulence of these cultures on rabbits and a calf. There was however no: alteration in virulence or characters after in one case 124 days and in another 132 days residence in the body of the horse (see page 187). A sufficient number of experiments has been performed for the determination of the virulence for calves of the cultures from the original material of viruses E. Il. and E. IV. and out of 13 calves inoculated 12 have shown no more disease: than may follow the subcutaneous inoculation of slightly virulent tubercle bacilli . of human origin (see Table, page 17). With the culture of virus E. II. four passage experiments on calves were performed (see pages 105-109). In the first experiment, beginning with Calf 404, one of the two final calves showed rather more generalised tuberculosis than had occurred with the original culture. In the second, beginning with Calf 406, there- were two branches, in one of which the result was negative and in the other the full virulence of a bovine tubercle bacillus was attained. In each of the other two passage experiments the virulence was increased for calves and rabbits. Two monkeys were tested with passage cultures from Calves 506 and 404 of which one had increased in virulence for the calf and the other was unchanged ; the * A series of Charts illustrating diagrammatically these experiments will be found in Volume VII. of the Appendix. 9 former culture showed increased virulence for the monkey and the latter produced n10 more disease in the monkey than that caused by the original culture. A culture was recovered from a rabbit, No. 1330, which had died 265 days after subcutaneous inoculation with 1 milligramme of culture from the original material of Virus H.IL., and was inoculated into a calf, in which it produced severe general tuberculosis. . _ ‘With the culture of virus E. IV. three passage experiments were performed (see pages 123-126). The first, beginning with Calf 408, was entirely negative. The second, beginning with Calf 498 (the animal which developed severe lung tuber- culosis), was also negative. In the third passage experiment there was an increase in virulence up to that of a fully virulent bovine tubercle bacillus. The culture derived from the lung of the first calf in this third passage, No. 502, was fully virulent, but the culture from the suprarenal body was not so virulent and produced in a calf only a chronic tuberculosis after 90 days. With the culture isolated from the mediastinal gland of this latter calf fatal general tuberculosis was set up in another calf in 39 days. In these experiments with virus E. IV. as with virus E. II. there was increased virulence for a monkey in a culture which had increased in virulence for the calf, while a culture that was unchanged in virulence for the calf showed no increase in virulence for a monkey. A culture, recovered from a baboon, No. 74, which had been fed with culture of E. IV. and had been killed after 448 days, was tested in calves and rabbits ; there was no increase in virulence or change in characters (see page 75). From Baboon 72, which died of general tuberculosis 144 days after subcutaneous inoculation of the same virus, cultures, derived both directly and through a guinea- pig, were definitely increased in virulence for rabbits. CULTURAL CHARACTERS. The cultures of viruses E. I., IL, TIL, 1IV., and V., do not differ in their cultural characters from cultures of bovine origin; they show merely slight variations such as have been demonstrated among different strains of bovine tubercle bacilli, which fall according to their cultural characters into three classes. Viruses E. II. and E. Ill. grow with the most difficulty and correspond with bovine cultures of Class I. Virus E. I. belongs to Class II. and viruses E. IV. and E. V. grow the most readily and fall into Class III. Cultures have been isolated from a number of experimental animals in- cluding those in the various passage experiments, but no definite alteration in their manner of growth could be detected. Cultures were recovered and were found to be unchanged from the following animals :—Calves 404, 406, 408, 502, 504, 506, 582, 584, 588, 596, 598, 616, 620, and 638, Baboons 72 and 74, Monkeys 182, 186, and 202, Pony 22, and Rabbits 1320 and 1330. FE. GRirFira. 34684 B 10 ‘deeds oy} wozz oporeqng Lord oy UI PUNO; IEA T{TIONG e[OTEqny ‘ [BULIOU eLeM SpULTS pUv sUBSIO JOyYO [Te § puss Iequimny euo ur qutod enbedo eynuim @ pure ‘qstmorjes r0q}0 oy3 ‘Lar13 ou0 ‘aeeyds oy} Ul saporoqny ArerIu omy $ uIN|MEMIA eYy UI saporeqny queonysuely F AO ¢ SIOM OIOY} | MOTWeTNOOU! 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VIRUS E. I. Calves, Pig, and Pony. Three calves have been inoculated subcutaneously each with 50 milligrammes of culture of virus H. I. One, No. 436, inoculated when the culture had been in cultivation a total period of 188 days, died prematurely in 16 days of diarrhoea ; at the post-mortem examina- tion there was local tuberculosis with early generalised tuberculosis. The other two calves, 522 and 528, in- oculated with culture of a total age of 482 days, died of general tuberculosis in 30 and 36 days respectively. Pig 192 was inoculated subcutaneously with 10 milli- grammes of culture and died in 48 days of general tuberculosis. Pony 16 inoculated intravenously with 10 milli- grammes of culture died in 17 days of acute tubercu- losis, the lungs alone showing macroscopic evidence of disease ; there was no great multiplication of tubercle bacilli in the organs and death was evidently due to a toxaemia rather than to a true tuberculosis. Rabbits and Guinea-pigs. Cultures varying in total age since isolation from 188 days to 782 days were inoculated intravenously, intra- peritoneally or subcutaneously into 15 rabbits, all of which succumbed to general tuberculosis of the type and severity of that following the inoculation of bovine tubercle bacilli. Guinea-pigs developed general tuber- culosis both after intraperitoneal and subcutaneous inoculations. VIRUS E, III. Calves, Pig, and Lemur. Two calves, Nos. 440 and 508, were inoculated subcutaneously each with 50 milligrammes of culture and both died of general tuberculosis in 72 and 73 days respectively. Pig 194 died of general tubercu- losis in 63 days after a subcutaneous inoculation of 10 milligrammes and Lemur 14 in 28 days after a sub- cutaneous inoculation of 1:0 milligramme. Rabbits and Guinea-Pigs. Cultures of virus E. IET., of a total age of 218 and 410 days respectively, were inoculated into 10 rabbits, the intravenous, intraperitoneal and subcutaneous methods being used. All died of general tuberculosis, but the results in two or three cases after subcutane- ous inoculations were not quite so severe as after the similar inoculation of bovine tubercle bacilli. Guinea- pigs died of general tuberculosis after both intra- peritoneal and subcutaneous inoculations. VIRUS E. V. The original material obtained from the horse consisted of lung, spleen and a mesenteric gland, from each of which emulsions were made and inoculated into rabbits and guinea-pigs. All the animals developed severe general tuberculosis of the type following the inoculation of bovine tubercle bacilli, Cultures were isolated direct from the original material (lung) and were inoculated into five rabbits and two guinea-pigs, all of which died of general tuberculosis. VIRUS E. IU. A.—CULTURES FROM THE ORIGINAL MATERIAL. Calves. Two calves have been inoculated each with 10 milligrammes of culture, one with 20 milli rammes, three each with 50 milligrammesand one with 100 milli” grammes. They were all killed when well after periods varying from 80 to 100 days and all showed slight retrogressive tuberculosis. At the seat of inoculation there was formed a fibrous walled cyst filled with soft caseous substance. The prescapular gland on the side of the local lesion was slightly enlarged and either was partly replaced by caseating substance or contained soft caseous nodules. The dis- seminated tuberculosis consisted of a few caseous tubercles and gritty foci in various lymphatic glands, a few foci or tubercles in the lungs and occasionally a tubercle in the liver and intestinal mucous membrane. Rhesus Monkeys and Lemurs. Five rhesus monkeys were inoculated subcutaneously, one, No. 168, with 10 milligrammes, three, Nos. 166, 180 and 216, each with 1:0 milligramme, and one, No. 184, with 0-1 milligramme. Monkey 168 died pre- maturely in 51 days and showed early general tuber- culosis: monkey 166 dead after 19 days showed local tuberculosis only. The remaining three died of general tuberculosis in 66, 98 and 101 days. Three monkeys were fed with culture. Monkey 186 received a dose of 1-0 milligramme from a pipette and died in 139 days of tuberculosis, most severe in the lungs (probably part of the dose was inhaled). Monkey 202 was fed naturally with 60 milligrammes in sterile milk, receiving 10 milligrammes per day for six days, and when killed after 101 days was well and in good condition ; the post-mortem examination showed very slight generalised tuberculosis. Monkey 208 received 50 milligrammes of culture in sterile milk and died in 115 days of general tuberculosis of mode- rate severity, most severe in the intestines and mesenteric glands. Lemurs 16 and 18, inoculated and fed respectively, each with 1:0 milligramme of culture, died of general tuberculosis after 35 and 76 days respectively. Pigs. Two pigs were inoculated subcutaneously, No. 184 with 10 milligrammes and No. 186 with 50 milli- grammes. Pig 186 was killed when ill after 174 days and Pig 184 when well after 268 days: both showed general tuberculosis severe in the lungs. Ponies. Pony 18 was inoculated intravenously with 10 milligrammes of culture and died of general tubercu- losis in 40 days, Pony 20 was inoculated subcutaneously with 50 milligrammes and was killed when well 130 days later. Tt showed a small local lesion and caseation of the adjacent prescapular glands, a nodule in the spleen, a few in the lungs, and no disease elsewhere. , Rabbits. Thirteen rabbits were inoculated intravenously, six with 1:0 milligramme, six with 0-1 milligramme, and one with 0-01 milligramme. All died of general tuber- culosis, those inoculated with 1-0 milligramme in from 20 to 57 days, those with 0-1 milligramme in from 22 to 127 days, and that with 0-01 milligramme in 36 days. The rabbit, No. 1331, which died after 127 days, had only moderately severe tuberculosis of a chronic type ; in the rest the disease was severe and extensive. Three rabbits were inoculated intraperitoneally. Two, each receiving 1-0 milligramme, died in 29 and 88 days respectively, and one, after 0-1 milligramme, in 43 days, all with general tuberculosis, a Fourteen rabbits were inoculated subcutaneously with doses ranging from 0-1 nilligramme up to 50 milligrammes, of which eight were Killed in from 102 to 178 days and five died in from 37 to 110 days. In the remaining one, rabbit 1330, the inoculation was partly 13 intraperitoneal and death occurred after 265 days of chronic general tuberculosis, severe in the lungs. In these rabbits the tuberculosis varied very considerably, but in none was the disease of the severity of that produced by bovine tubercle bacilli and in a few cases it was as slight as after a similar inoculation of slightly virulent human tubercle bacilli (Group II). The following is a summary of the lesions found. At the seat of inoculation, usually in the subcutaneous tissue of the back, there was formed a caseous tumour which almost invariably softened and became cystic. The adjacent lymphatic glands contained curdy caseous nodules easily shelling out and leaving cavities in the apparently normal gland tissue that remained. In the lungs there was in several casesa single nodule ora small number of minute translucent tubercles, in others moderately numerous caseating tubercles and caseating nodules in the margins of the lobes. The kidneys, after large doses, were pitted on the surfaces and showed a few translucent tubercles in the cortices and caseous streaks in the pyramids. The kidneys of one animal, Rabbit 1468, the most severely infected, were studded with proje:ting caseating nodules up to a pea in size, such as are often found after the inoculation of bovine tubercle bacilli. Guinea-pigs. One guinea-pig, inoculated intraperitoneally with 0-1 milligramme, died in 40 days of general tuberculosis. Five guinea-pigs inoculated subcutaneously, three with ‘0'1 milligramme and two with 1:0 milligramme, died in from 44 to 212 days of general tuberculosis. The tuberculosis in several instances was of a very chronic ‘type. B.—CuLTuRES oF Virus E. II. AFTER PASSAGE. PassaGE u. This passage experiment (see Table, page 105) began with the culture from the prescapular gland of Calf 404 (which had been inoculated with the culture from the original material). It was inoculated into the calves Nos. 590 and 598, of which the former died in seven days of broncho - pneumonia and diarrhoea, and the latter was killed after 91 days when it showed only slight disease. A culture was obtained from the spleen of Calf 590 and inoculated into Calf 618, which, when killed after 84 days, showed slight disease only. A culture was isolated from the prescapular gland of Calf 598 and was inoculated into Calf 640, which developed slight general tuberculosis. ‘There was perhaps a slight increase of virulence shown for Calf 640, though it cannot be considered as definite. The rabbits inoculated with the culture from Calf 618, the final calf of the other branch of the passage, were more severely affected than any of the other series of rabbits and judging from those results it might certainly be expected that the culture would have been more virulent for calves (the virulence of the culiure for calves was however not tested). The tests on rabbits -of the cultures from Calves 404, 590, and 598 showed no great increase over what had occurred with the original material culture. A rhesus monkey, No. 212, was fed with the culture from Calf 404, and showed after 82 days very slight tuberculosis. PassaGe pf. This passage (sce Table, page 106) began with the culture from the prescapular gland of Calf 406 (which was inoculated at the same timeand from the same cultureas Calf 404). It was inoculated into two calves, 596 and 600, and into a series of rabbits ; the rabbits showed that the culture had the virulence of that of the original material. Calf 596 developed chronic general tuber- culosis not at all severe, and Calf 600 had very slight tuberculosis. The culture from the lung of Calf 596 was inoculated into Calf 638 and rabbits ; the calf died of general tuberculosis in 40 days and the rabbits all developed the severe tuberculosis characteristic of the action of the bovine tubercle bacillus. The culture from the prescapular gland of Calf 600 was inoculated _ into Calf 648 and intoa series of rabbits ; the results showed that the culture was like that of the original matetial. Here then had occurred a very marked in- crease of virulence for calves and rabbits in one branch of the passage and not in the other. PassaGE y. This passage (see Table, page 107) began with Calf 504, which had been inoculated with the culture from the original material after it had been in cultivation a total period of 405 days. The animal developed only slight disease after 100 days. A culture isolated from the prescapular gland caused general tuberculosis in a calf, fatal in 54 days, and was fully virulent for rabbits. A culture from the lung was fully virulent for rabbits, and caused fatal general tuberculosis in one calf in 29 days, while the fellow calf had generalised tuberculosis not severe and ap- parently retrogressive. In this experiment there was a definite increase in virulence almost up to the highest bovine virulence after residence in a single calf for 100 days. PassaGe 0. Calf 506, inoculated at the same time as Calf 504, was at the beginning of‘this passage experiment (see Table, page 108). This animal developed slight tuber- culosis after 99 days. A culture derived from the lun, was found to be increased in virulence for rabbits, an ata later period it was inoculated into another series of rabbits and into a calf (No. 594), which however died prematurely and showed early tuberculosis. The culture was again tested in a calf and also in rabbits and amonkey. ‘Che rabbits and the monkey (No. 206) died of general tuberculosis but the calf (No. 628) lived for 90 days and when killed had only slight genera] tuber- culosis. The culture from the lung of this latter calf was inoculated into another calf (No. 646), which died of general tuberculosis in 33 days, and into a series of rabbits all of which developed general tuberculosis. In this case there was a very definite increase in virulence after residence in the first calf for 99 days as shown by the effects on the rabbits and the monkey ; there was required however another calf passage before the full bovine virulence was acquired. PASSAGE «. A rabbit, No. 1330, was inoculated partly sub- cutaneously, partly intraperitoneally and died of chronic generalised tuberculosis after 265 days. A culture was derived from a caseating growth on the diaphragm and was found to be increased in virulence for the calf (Calf 632) and for rabbits (see Table, page 109). VIRUS E. IV. A.—CULTURES FROM THE ORIGINAL MATSRIAL, Calves. Six calves were inoculated subcutaneously, two with 10 milligrammes of culture and four with 50 milli- grammes respectively. Five were killed when well after periods ranging from 88 to 100 days. One calf, No. 498, died 89 days after inoculation. The five which were killed showed only slight non- progressive tuberculosis. At the seat of the inocula- tion there was found a non-infiltrating caseous tumour which softened in most cases becoming cystic; the ajacent lymphatic glands were either partly caseous and softened or contained soft caseous nodules. In one case there was no disseminated disease and in another there were tubercles in the mesenteric glands ; in two out of the other three there were tubercles in the suprarenal bodies and tubercles in the lungs and various mesenteric glands. The result in the case of Calf 498 was very irregular ; the animal died from a severe tuberculous pneumonic condition of the lungs ; there was also local tuberculosis but elsewhere the disease was extremely slight. Baboons and Monkeys. Baboon 72 was inoculated subcutaneously and Baboon74 was fed, each with 1-0 milligramme of culture. The inoculated animal, No. 72, died of general tubercu- losis in 144 days. Baboon 74 was killed after 448 days ; it was in excellent condition and appeared perfectly well. _The post-mortem examination revealed very slight, apparently non-progressive tuberculosis ; there were a few caseo-purulent nodules in the mesenteric glands and in one deep cervical gland; the lungs. 14 liver, and kidneys showed a very few small tubercles. A culture was isolated from the mesenteric glands in which there was no change in cultural characters and no increase in virulence for calves. Four rhesus monkeys were inoculated subcu- taneously. Two, which received 1:0 and 10-0 milli- grammes, died in 24and 7 days respectively and showed Iocal tuberculosis only. A third (No. 218, receiving 1-0 milligramme) was killed after 95 days and showed general tuberculosis, severe in the spleen, the fourth (No. 178, receiving 1:0 milligramme) died in 89 days of general tuberculosis. Two monkeys, Nos. 182 aud 210, werefed. Monkey 182 received 1:0 milligramme of culture administered by a pipette and was killed in poor health 392 days later. It showed only slight general tuberculosis ; there wasa purulent nodule in the subcutaneous tissue of the neck, where during life an abscess had developed. Monkey 210 was killed when dying after 77 days ; the only disseminated disease was a single tubercle in the lung. Death was due to severe tuberculosis of the intestineand mesenteric glands. Pigs. Two pigs, 188 and 190, were inoculated subcu- taneously, the former with 10 the latter with 50 milli- grammes of culture. Pig 188 was killed after 268 days when apparently well and had general tuber- culosis. Pig 190 was killed after 175 days and had general tuberculosis, not severe. Rabbits. Twelve rabbits were inoculated intravenously, six with 1:0 milligramme and five with 0°1 milligramme and one with 0:01 milligramme. Those inoculated with 1:0 milligramme died in from 14’ to 32 days and those with 0-1 milligramme in from 18 to 79 days and the one with 0-01 milligramme in 53 days, all of general tuberculosis. Three rabbits were inoculated intraperitoneally. One received 0°1 milligramme and died in 47 days ; the other two received 1:0 milligramme each, one dying in 28 days, and the other in 96 days of general tuberculosis (in the latter the inoculation was parily intra-caecal). Sixteen rabbits were inoculated subcutaneously with doses of culture varying from 0:1 to 50 milligrammes. Eleven of them died, the majority from causes other than tuberculosis, and five were killed after periods varying from 100 to 265 days. The extent, to which the animals inoculated subcu- taneously were diseased, depended apparent!y upon the individual susceptibility rather than upon the dose. In none was the disease as extensive as that caused by the bovine tubercle bacillus. In general, there was formed at the seat of inoculation in the subcutaneous tissue, usually of the back, a caseous tumour, which became softened, cystic and occasionally ulcerated. The adjacent lymphatic glands contained softened daseous nodules easily shelling out of the gland sub- stance. The lungs contained caseous tubercles and nodules varying from a few up to a moderate number. The kidneys showed a varying amount of disease ; at the most there was a moderate number of soft caseous tubercles and depressed patches in the cortices, with caseous streaks in the pyramids. The mediastinal and portal glands occasionally contained caseous foci or tubercles ; the spleen and liver were rarely affected. The rabbit inoculated with the largest dose—50 milligrammes—gained weight and appeared well when killed ; the post-mortem examination showed only slight retrogressive tuberculosis. In order to test upon what depended the great variation in amount of disease produced in rabbits by this virus, a series of 16 was inoculated subcutaneously with culture derived from single colonies. These single colonies were obtained from the sternal gland of a guinea-pig inoculated with culture from the original material. as it was considered more probable that the bacilli would be single in emulsions than in cultures and that a single colony might be derived from a single bacillus. Each rabbit received the culture from a serum-tube (about 10 milligrammes), and two rabbits were used for each colony. The results showed the same variation in extent of disease as has been de- ascribed above (see Table, page 34). Guinea-pigs. One guinea-pig was inoculated intraperitoneally with 0-1 milligramme of culture, and died of general tuber- culosis in 30 days. Six guinea-pigs were inoculated each with 1:0 milligramme, and died of acute tuber- culosis in from 6 to 14 days. Five guinea-pigs were inoculated subcutaneously ; two received 1:0 milli- gramme each, and died of general tuberculosis in 97 to 144 days respectively ; three received 0:1 milli- gramme each, and died of general tuberculosis in 62, 167, aud 60 days respectively. One guinea-pig, inocu- lated cutaneously, died in 88 days of general tubercu- losis. B.—Cutrures FRomM Virus H. IV. AFTER PASSAGE, PASSAGE a. Calf 408, inoculated with culture from the original material, developed local tuberculosis only after 109 days, and a culture was isolated from the prescapular gland (see Table, page 123). This was inoculated into two calves, 582 and 584, three series of rabbits, and a monkey, No. 220. One calf, 582, had a little more generalised tuberculosis than had occurred after inoculation with culture from the original material ; but the resulis in the other animals showed that the virulence had not materially altered. From each of the latter two calves, another calf was inoeu- lated ; in one case the culture was derived from the spleen, in the other from the suprarenal body. Both calves 636 and 642 had only slight disease after over 90 days, but the rabbits inoculated with the culture from Calf 582 bad more extensive disease than that usually caused by the original culture. The result, however, as regards increase in virulence for the calf, was negative. PassaGE 6. Calf 498, inoculated with the culture from the original material, developed severe tuberculosis of the lungs (see Table, page 125). From the lungs a culture was isolated with which were inoculated two calves, Nos. 586 and 588, and a series of rabbits. The results showed no increase of virulence. From each calf a culture was re- covered and inoculated into a calf and a series of rabbits. These two calves, Nos. 634 and 644, developed only sligbt disease. In this case also no increase in virulence could be recorded. PassaGE y. Calf 502 was incculated at the same time as Calf 498, but the tuberculosis was only slight in degree ; and whereas no increase in virulence occurred with the culture derived from Calf 498, the culture from Calf 502 became as fully virulent as a bovine tubercle bacillus. The following are the details (see Table, page 126) : From the lung of Calf 502 a culture was gained which caused fatal general tuberculosis in two calves, Nos. 614 and 630, ina monkey, and in three series of rabbits. A culture was also obtained from the suprarenal body,and it was much less virulent than the culture from the lung. It caused in Calf 602 tuberculosis generalised, but not severe, and was not fully virulent for rabbits. From the mediastinal gland of Calf 602 a series of rabbits was inoculated, and a culture was isolated. The rabbits inoculated with emulsion developed general tuberculosis, and the culture produced in Calf 650 and a series of rabbits fatal general tuberculosis of great severity. 15 PART I. EXPERIMENTS WITH CULTURES FROM THE ORIGINAL MATERIAL. (i). Tabular summaries of Inoculation Experiments with the cultures from the original material of Viruses E.I1., E. IL, E. II., E. IV., and EB. V. (ii). 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I.—continued. CULTURE INOCULATIONS—continued. Iil—Junz 20, 1907. The strain was derived from the original material through a guinea-pig, and had been 545 days in artificial cultivation. The culture used was the 20th generation, 21 days old. PIG 192. Subcutaneous. Dose: 10:0 mg. Died: August 7, 1907. 48 days. P.M.—Genera! tuberculosis. IV.—FEeEsRuaRyY 11, 1908. The strain was derived from the original material through a guinea-pig, and had been 782 days in artificial cultivation. The culture used was the 26th generation, 22 days old. cont 16. RABBITS. Intravenous. Durati f Dose : 10°0 mg. Number.) Method.} Dose. Life, ” Result. Died : February 28, 1908. 17 days. 1639 | Intrav.] 0-1 mg. | D. 24 days] Acute T. P.M.—The lungs were congested and oedematous, and showed a diffuse grey in- 1638 | Intrav. | 0°01 mg.} D. 32 ,, | G.T. filtration which in places could be resolved j ; into collections of just visible foci. There 1640 | Subcut. | 10:0 mg.} D.120 ,, | Chronic severe was no macroscopic tuberculosis elsewhere ; G. T. moderately numerous T.B. were seen in a 1641 | Subcut.|10°0 mg.| D. 64 ,, | T., moderately smear from the spleen. pee in lungs only. 34684 42 CALF 436. Virus E. I. Subcutaneous inoculation of culture derived from the original material through a guinea-pig. Dose—50 milligrammes. Date of Inoculation—June 27, 1906. [Age about 7 weeks.] Died ~July 13, 1906. [16 days after inoculation.] [Death occurred prematurely from diarrhcea.] Temperature. Temperature rose to 39°7° on the 8th day and re- mained about 39°4° until death Weights Kilos. June 27, 1906 29:02 July 13, 1906 29°47 Gain in weight.—0'45 kilos. POST-MORTEM EXAMINATION. General condition fair Local Lesion.—On the left side of the neck there was a lenticular-shaped tumour measuring 75 by 2cm. On section it consisted of caseous tissue surrounding a central cavity containing fluid and flakes of caseous substance. The skin and muscles were infiltrated. Left Prescapular Gland—The left prescapular gland measured 6:3 by 3-7 cm. On section about half the cortex consisted of firm grey tissue with a caseous network ; in the rest of the gland there were small early caseous tubercles. Right Prescapular Gland.—Normal. Prepectoral Glands.—The left round prepectoral gland was slightly enlarged and caseous throughout The left reniform prepectoral gland appeared normal, except for slight oedema. The right prepectoral glands appeared norma!. Cervical Glands.—Normal. Thorax. Pleura.—Normal. Lungs.—The lungs were mottled by patches of collapse. Scattered throughout the substance there were moderately numerous foci, the smallest of which, that is the majority, were grey and translu- cent ; some about 5mm. had minute opacities in the centre. Mediastinal and Bronchial Glands.—Normal. Heart and Pericardium.—Normal.' Abdomen. Omentum and Peritoneum.—Normal. Intestines.—Normal. Mesenteric Glands—Each of two contained a minute opaque focus. Spleen.—The spleen appeared normal ; the Mal- pighian bodies were not prominent. Liver.—Scattered throughout the liver there were moderately numerous minute grey foci. Portal Glands.—Normal. Kidneys.—In the cortices of the kidneys there were sparsely scattered minute grey foci. Suprarenal Bodies.—Normal. Renal, Coeliac, Lumbar, and Iliac Glands.—Normal. Tongue, Tonsils, Larynx, Pharynx, and Trachea.— Normal. Various Lymphatic Glands. Parotid, Submawillary, Retro-pharyngeal, Acillary, Precrural, Popliteal, Gluteal, and Ischiatic Glands. —All appeared normal. CALF 528. Virus E. I. Subcutaneous inoculation with culture derived from the original material through a guinea-pig Dose—50 milligrammes. Date of Inoculation—April 17, 1907. Died—May 17, 1907. Temperature. The temperature rose from 38:2° to 39°7° on the 3rd day after inoculation. It remained above 39° until 14 days after inoculation, when it rose to 40°3°. It subsequently fluctuated and fell to 39:7° at which it remained for a week before death. Weights. Kilos. April 17, 1907 3 ie 39°45 May 17, 1907 ite ‘ie 36°73 Lota! loss of weight.—2:72 kilos. POST-MORTEM EXAMINATION. General condition poor. Local Lesion.—At the seat of inoculation in the subcutaneous tissues on the left side of the neck there was a firm tumour measuring 11 by 9 by 4 cm. and [Age 11 weeks.] {80 days after inoculation. ] weighing 226 grammes. It was rounded externally and on section was composed of pinkish yellow caseous necrotic substance infiltrating both the skin and muscles. In the centre of the tumour were some small spaces filled with serous fluid. Prescapular Glands.—The left prescapular gland measured 8 by 5 by 4cm. and weighed 56 grammes. On section it was composed practically throughout of dense yellow caseated tissue. The right prescapular glind was deeply congested and oedematous, and showed a number of early caseous tubercles irregular in outline. Prepectoral Glands.—The left prepectoral glands were only slightly enlarged, but deeply congested. The rounded gland showed early tuberculous infiltra- tion in the cortex ; the reniform prepectoral gland contained a few discrete greyish yellow tubercles. The right prepectoral glands showed a few irregular discrete caseous tubercles. The Cervical, Submawillary, Parotid, Retro- pharyngeal, and Azillary Glands each showed a moderate number of discrete irregular tubercles, with slight caseation, ranging in size from 0°5 mm. up to 2mm. in diameter, resembling those in the right prescapular gland. Thorax. Pleura.—Along the margins of the ribs were small clusters of minute grey tubercles surrounded by vascular connective tissue; there were also two flattened greyish red nodules. There were several flattened caseating nodules on the pleura covering the pericardium. Similar nodules (about 12 in number) were seen on the pleura cover- ing the diaphragm around the central tendon. Lungs.—The right cephalic lobe, right middle and ventral portions of the right caudal lobe, were dark red and quite airless. The left cephalic lobe was partly consolidated. The remainder of the long showed small patches of hepatisation ; the parenchyma was closely and evenly beset with grey miliary tubercles with opaque centres. Thoracic Lymphatic Glands.—The mediastinal and the left bronchial glands were moderately enlarged and showed their cortices composed of reddish grey tissue, infiltrated with yellow caseous foci and patches (in places forming a network). The right bronchial aud the infra-tracheal glands were deeply congested and closely beset with caseous miliary tubercles. A gland just within the entrance to the thorax on the left side was enlarged and resembled the mediastinal glands. The corresponding gland on the right side contained discrete tubercles. Heart.—In the heart muscle half-a-dozen tubercles were seen ; five of these were yellow and caseous, the sixth was grey and not obviously caseous. Abdomen. Omentum.—The omentum showed on the ventral surface scattered tubercles ranging up to 2 mm. in diameter ; some were deeply congested, others had a pinkish grey colour ; none showed definite caseation. Peritoneum.—Normal. Intestines.—In the long Peyer's patch of the small intestine there were fairly numerous submucous caseating tubercles. There were similar, but much more sparsely scatterei tubercles in the rest of the intestine. . : : The lymphoid follicles in the large intestine were enlarged but not caseous. Mesenteric Glands.—These glands were notenlarged; they were beset with tubercles (the majority being 43 minute) in a state of early caseation ; they were most numerous in the middle mesenteric glands which, under the capsules, had a speckled appearance. In the terminal glands they were more sparsely distributed and were also generally larger. Lleo-Colic and Colic Glands.—These resembled the terminal mesenteric glands. Spleen.—The spleen weighed 198 grammes. On sectiva the pulp had a coarsely granular appearance, and was very closely beset with greyish red tubercles with yellow caseous ceatres. Liver —The liver was evenly and closely beset with grey tubercles varying in size from a mere point up to rather more than one millimetre in diameter ; the larger were distinctly yellow and caseous in the centre. Gall Bladder.—In the mucous membrane of the gall bladder there were fairly numerous miliary caszous tubercles. Portal Glands. — The portal glands were slightly en- larged and showed in the cortices grey patches and separate nodules infiltrated with either yellow cas:ous foci or an irregular network. Kidneys—The kidneys showed io their cortices moderately numerous grey tubercles varying up to 1-5 mm.in diameter, some of which were slightly opaque and yellow in the centre. Suprarenal Bodies.—In the cortex of each were several grey miliary tubercles. The urinary bladder showed three or four early caseous tubercles the size of rape seeds. The Coeliac Glands resembled the portal glands. The Pudic, Popliteal, Giluteal, Ischiatic and Pre- crural Glands were deeply congested, and showed a moderate number ef discrete, irregular, caseating tubercles. The upper part of larynx, pharynx and tongue appeared normal. In the lower part of larynx there were a few slightly raised, flat congested tubercles ; they were moderately numerous along the whole length of trachea ; they were congested and the majority had minute pits in the centre. Microscopical Examinations. (Smear preparations. ) Prescapular Gland.—Very numerous tubercle bacilli. Heart (tubercle), Tonsil (focus), Small Intestine (caseous nodule) anu Large Intestine (follicle).—A few tubercle bacilli in each case. PIG 192. Virus E. I. Subcutaneous inoculation of culture derived from the original material through a guinea-pig. Dose—10°0 milligrammes. Date of Inoculation—June 20th, 1907. [Age 14 weeks.] Died—August 7, 1907. Temperature. The temperature rose gradually from 38°5° until it reached 40°6° on the 30th day after inoculation. For a fortnight it remained above 40°, the maximum being 40°6°, until two days before death, when it fell suddenly. Weights. : Kilos. June 21, 1907 ... 24:03 August 7, 1907... 20°40 Loss of weight.—3°63 kilos. POST-MORTEM EXAMINATION. Condition fair. 34684 [48 days after inoculation.] Local Lesion.—To the left of the mid-line of the abdomen there was a cyst measuring 5 by 4 cm., the skin over which was reddened but not thinned or ulcerated ; it was composed of a thick fibrous capsule filled with serous fluid containing curdy caseous masses. The internal wall of the cyst was roughened by adherent caseous substance. The muscles beneath the tumour were adherent and infiltrated with caseous substance. Inguinal Glands.—The left inguinal glands formed a mass measuring 6 by 5 by 4:3 cm., which, on section was caseous throughout with central softening. The right inguinal glands measured 4°5 by 2°5 by 10cm. On section they showed a moderate number of yellow caseous nodules the size of millet seeds. Precrural Glands —The precrural glands showed, F 2 44 nodules similar to those in the right inguinal glands, but rather larger. The Iliac, Lumbar, and Renal Glands showed caseous nodules with grey margins. Thoraz. Pleura.—The lymphatic fringes along the margins of the ribs were deeply congested and filled with clusters of minute translucent tubercles. The diaphragm showed a moderate number of flattened translucent nodules about 2 mm. in diameter. Lungs.—The lungs did not collapse on opening the chest-wall and filled the pleural cavities. The sub- stance was closely beset with confluent translucent nodules with central caseation forming a close network leaving small islands of congested lung tissue. Many of these nodules projected slightly on the pleural surfaces and had attached to them loose fibrinous fringes. Bronchial Glands——The bronchial glands were enlarged and showed numerous irregular caseating nodules with fine opaque yellow lines in the centre. Mediastinal Glands.—The left ventral mediastinal gland, the size of a walnut, was almost entirely com- posed of soft yellowish caseo-pus. Tongue, Tonsils, Larynx, Pharyna, and Trachea.— Normal, The thyroid body showed a few small caseous tubercles. Abdomen. Omentum and Peritoneum.—Norwal. Spleen.—The spleen was nodular on the surface and was closely filled with grey nodules with caseous centres ranging in size from a mere point up to 5 mm. Liver—The liver showed numerous grey tubercles with caseous centres from a point up to a rape seed in size. Portal Glands.—The portal glands were practically replaced by a coarse network of caseated tissue. Kidneys.—Each kidney showed one or two grey tubercles, the size of rape seed, with caseous centres. Suprarenal Bodies —The suprarenal bodies showed in their cortices one or two small grey tubercles. Intestines.—Normal. Mesenteric Glands.—All the mesenteric glands showed small yellow caseous nodules chiefly about 1 mm. in diameter; in some glands these were discrete, in others they formed a coarse network. The colic glands showed similar nodules. Submazillary, Pharyngeal, Cervical and Prescapular Lymphatic Glands.—Each showed a moderate number of yellowish caseous nodules up to a hemp seed in size, but the majority about the size of rape seed. Each ovary showed several opaque whitish foci. Microscopical Examinations. (Smear preparations. ) Emulsion of Left Inguinal Gland showed numerous tubercle bacilli. Diaphragm (nodule) Ovary (focus) ps } No tubercle bacilli seen. PONY 16. Virus E. I. Intravenous inoculation with culture derived from the original material through a guinea-pig. Dose—10 milligrammes. Date of Inoculation—February 11, 1908. Died—February 28, 1908. [Age —Yearling. ] [17 days after inoculation. ] Clinical History. The animal quickly became very ill with great respiratory distress. Temperature. During the first eleven days after the inoculation the temperature was very irregular, ranging from 37°8 to 39°7 ; on the thirteenth day 40°1 was recorded ; the subsequent daily temperatures were 40:2° C., 39°5° C., and 40°4° C.; on the 17th day the pony was dead. Weights. Kilos. February 11, 1908 ... ave sae 87°52 February 28, 1908 ... 61:22 Loss in weight.—26'30 kilos. ‘POST-MORTEM EXAMINATION. Carcass thin and emaciated. Thorax. Lungs.-—The lungs were voluminous and felt firm. On section they were congested and oedematous and a close inspection revealed a diffuse infiltration of grey tissue which in places could be resolved into collec- tions of translucent foci just visible. Abdomen. Intestines.—Normal. Mesenteric Glands.—Normal. Spleen.—The spleen weighed 254 grammes; on section the pulp appeared normal. Liver.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Many of the lymphatic glands were congested and almost all were oedematous, but none showed any sign of caseation. Microscopical Examinations. (Smear preparations.) Lung.—Numerous tubercle bacilli. Spleen.—Moderately numerous tubercle bacilli. Liver.—No tubercle bacilli. Kidney.—Two tubercle bacilli. Mesenteric Gland.—No tubercle bacilli. Intestine-—No tubercle bacilli. 45 CALF 522. Virus E. I. Subcutaneous inoculation with culture derived from the original material (through a guinea-pig). Dose—50 milligrammes. Date of Inoculation—April 17, 1907. [Age 8 weeks.] Died—May 23, 1907. [36 days after inoculation.] Temperature. '- The temperature rose to 40° C. on the sixth day after inoculation. Subsequently it fluctuated, but remained high, maximum 40-4°, for a fortnight, after which it fell to 39°2°. It quickly rose to 40° C. and a a few days above and then slowly fell until eath. Weights. kilos. April 17, 1907 ats ts 42°62 May 23, 1907 mie Be 38°54 Total loss in weight.—4‘08 kilos. POST-MORTEM EXAMINATION. General condition poor. Local Lesion.—In the subcutaneous tissues on the left side of the neck there was a slightly prominent tumour, measuring 14 by 9 by 5 cm. and weighing 481 grammes. The skin over it was adherent and just beginning to break down, showing a small crack from which a little blood-stained fluid was escaping. On section the tumour was composed of dense pinkisb- yellow necrotic tissue. which infiltrated both skin and muscle. It had a small irregular cavity in the centre. Prescapular Glands.—The left prescapular gland measured 11 by 6 by 4°5 cm. and weighed 169 grammes. On section it was composed practically throughout of dense pinkish-yellow caseous necrotic tissue. The capsule of the gland was thickened. The tissues around the local tumour and the left prescapular gland were greatly thickened and oedematous. The right prescapular gland measured 6 by 2°5 by 1 cm. and showed moderately numerous caseous nodules 2 to 3 mm. in diameter. Prepectoral Glands.—The left rounded prepectoral gland measured 2 by 1:5 cm. and was composed of translucent tissue with a fine caseous network. The left reniform prepectoral gland showed a few discrete yellow caseous tubercles. The prepectoral glands on the right side showed a few discrete caseous tubercles irregular in outline. Cervical Glands.—The left inferior cervical gland measured 4°5 by 3 by 15cm. On section it showed the greater part of the gland substance replaced by a fine yellowish caseous network. At one extremity of the gland there were two or three discrete caseous nodules 2 mm. in diameter. The other cervical glands on the left side showed a moderate number of discrete irregular yellowish caseous nodules. The right inferior cervical gland showed a few discrete caseous tubercles irregular in outline. The other cervical glands on the right side contained a few discrete caseous tubercles, the largest 2 mm. in diameter. Thorax. The lymphatic fringes along the margins of the ribs were deeply congested and showed thick clusters of minute pearly grey tubercles with an occasional flattened nodule up to 4 mm. in diameter ; these latter contained caseous centres. Diaphragm.—The peritoneal surface of the dia- phragm showed a few small tubercles with opaque cenires. On the pleural surface there were some es of small grey tubercles similar to those seen on e ribs. Lungs—The lungs were very heavy. The right middle, right cephalic, left cephalic and ventral portions of the caudal lobes were congested ; there was a small patch of congestion at the base of the left caudal lobe. The lung parenchyma was closely and evenly beset with grey tubercles with caseous centres, varying from a mere point up to 2 mm. in diameter ; in places these tubercles had run together to form larger nodules. The lung tissue of the anterior lobes and ventral portions of the posterior lobes between the nodules was reddish grey and solid, leaving only small patches of crepitant lung. The dorsal parts of the posterior lobes contained much more crepitant Jung tissue and the nodules were less closely placed. Thoracic Lymphatic Glands.—The mediastinal glands were enlarged and showed in their cortices masses of greyish yellow caseous network, practically replacing the cortices. The bronchial glands closely resembled the media- stinal glands. A gland at the entrance to the thorax and the ventral mediastinal glands showed irregular yellowish caseous nodules. Heart.—In the muscle of the heart there was an opaque greyish nodule 2 mm. in diameter. The heart was otherwise normal. Abdomen. Peritoneum.—Normal. Omentum.—On the ventral surface of the omentum there were numerous congested nodules, ranging in size from a speck up to 3mm. in diameter. The majority of these nodules had caseous centres. Intestines.—Scattered throughout the whole length of the small intestine, beneath the mucous membrane, were fairly numerous small yellow caseous tubercles. The large intestine appeared normal. Gastric Glands.—The gastric glands showed in their cortices small caseous tubercles, some discrete, others forming a network. Mesenteric Glands.—All the mesenteric glands were beset with numerous irregular caseous nodules sur- rounded by a margin of grey tissue, some discrete, some forming a caseous network. Colic Glands.—The colic glands showed small dis- crete caseous tubercles. Spleen.—The spleen weighed 481 grammes. It was swollen. The surface was normal. On section it was closely packed with grey nodules with caseous centres up to 3mm. in diameter. Liver —The liver contains numerous grey nodules with caseous centres varying from ‘5 up to 25 mm. in diameter: there was also a number of grey foci. Some of the tubercles projected above the surface. They were evenly distributed throughout the substance but in slightly fewer numbers than just beneath the capsule. Portal Glands.—The portal glands were enlarged and showed their cortices closely beset with a yellowish grey caseous network. The gall bladder showed about 18 small yellow caseous tubercles, over two of which there was ulcera- tion of the mucous membrane. Kidneys.—Each kidney contained in the cortex a moderate number of grey tubercles, ranging in size from a mere point up to 3 mm. in diameter, the larger ones being caseous in the centre. On section similar tubercles were seen in the depth of the cortex. Suprarenal Bodies.—Hach suprarenal body showed a few yellowish grey caseous tubercles. Urinary Bladder.—Normal. Coeliac Glands.—Of the coeliac glands, one snowed about one half of its cortex replaced by yellow caseous tissue. The others each showed a moderate number of yellow cascous nodules forming a coarse network. The Iliac, Lumbar, and Renal Glands contained a moderate number of discrete yellowish caseous nodules up to 5mm. in diameter. The Popliteal, Gluteal, Pudic, Precrurul and Ischiatic Glands showed discrete caseous nodules from 2 to 4mm. in diameter. Awillary Glands —Each showed in the cortex yellow caseous nodules 3 mm. in diameter. Tonsils—In each tonsil there was a moderate number of opaque foci. Along the trachea was a moderate number of flat grey congested tubercles with opaque centres. The laryox showed two similar tubercles and the epiglottis one. 46 In the oesophagus were a few small yellow caseous. tubercles. Tongue.—Normal. The submaxillary, retro-pharyngeal, and parotid glands contained a moderate number of discrete yellow caseous nodules, 3 to 4 mm. in diameter. Microscopical Examinations. (Smear preparations.) Suprarenal Body (tubercle). — Showed numerous tubercle bacilli. Heart (nodule).—Showed fairly numerous tubercle bacilli. 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"Bal 0.01 ‘anoqng “yuoqns "ynoqug 1801 G80T e801 “yynsey “SDId-VONIND ‘OUT jo uoneinqg “asoq “pony “1aquan yy “SLICAVA “SOUTySO} UL 04} Ul Sieo[N poeteyyvos [[ems o10M olay} pue ‘payvesvo pue pesiepuo A[snowious eam spur[s o11oyueseut eg, ‘siso[noreqny [eieusH— Gg ‘skep 92 “LOGT ‘g Axenaqey : porq ‘BU Q.T : 980q “pd ‘ST UNNTT “sIsO[NoIEqny, oynoy— J qT ‘skep gg “906T ‘9% tequieveq : parq ‘BUI (.[ : o80q *snosueqnoqng ‘OT UANAT "pio skep 1% ‘morjeieued YIOT 94} SVAL psn OIN4TNI ONY, ‘uoeaTy|Nd yelogtya ur skep GEE “906E ‘IZ AdaNZAON—IT *panuyuog—SNOILVIOOONT FANLTAD ‘panuyuoo— TT “HT SNUIA ueeq pty pue ‘sid-voumad v qsno1g) [else}eU [LUISIIO 944 WOIF poAop SeA UIeIys aq, 49 VIRUS E. Il.—continued. CouLtuRE INOCULATIONS—continued. 1II.—Janvary 30, 1907. The strain was derived from the original material through a guinea-pig, and had been 405 days in artificial cultivation. The culture used was the 19th generation, 21 days old. CALF 504 Subcutaneous. Dose: 50:0 mg. Killed : May 10, 1907. 100 days. P.M. — Tibrous-walled cyst at seat of inoculation. The left prescapular gland was three - parts occupied by caseo-necrotic masses beginning to break down in the centre. There were four or five tubercles in the lungs, one in one kidney, and a number of soft yellow nodules in the small intestine (T.B. seen). The thoracic and portal glands showed an occasional minute focus, the renal, mesenteric and ileo-colic glands scattered gritty tubercles. 34684 CALF 506. Subcutaneous. Dose : 50:0 mg. Killed: May 9, 1907. 99 days. P.M. — Fibrous-walled cyst at seat of inoculation. The left prescapular gland contained two caseo- calcareous nodules and scattered tubercles; one cervical gland was caseo- calcareous. The lungs contained two hard grey nodules, a collection of minute grey tubercles, and an occasional trans- lucent focus. One bron- chial gland and one portal gland contained one or more small foci. RABBITS. Number.| Method.| Dose Duration of Result er ere Life. ; 1221 | Intrav. | 1:0 mg.| D. 34 days | G. T. 1222 | Intrap.| 1:0mg.] D.88 ,, | Slight G.T., and severe pseudo-T. of spleen. 1223 | Subcut. }10°0 mg.} D. 37 ,, Loca] lesion only; death from pleu- . Tisy. G 841 days RHE Fed D 08. P.M.—Gen. in | orous walls of inocula- gland con- r tubercles. sewhere. 945 days in ITS. Life. D. 20 days D. 33 Cy, RABBITS. Number. | Method. | Dose: Bomee of Result. 1724 Intrav. O1lmg. | D. 58days | G. T. 1722 Intrav. | 0-Olmg.' D. 36, |G.T. 1723 Subcut. | 100mg. , D. 59 ,, Local T. Very slight T. of lungs and kidneys. (?) cause of death. 1725 Subcut. | 100mg. D. 45 ,, Slight T. Cause of death not apparent. 1726 Subcut. | 100mg. | K.178 ,, Local T., and very slight T. of lungs and kidneys. Duration of G. T. G. T. Result. 53 CALF 404, Virus E. Subcutaneous inoculation of culture derived from the original material through a guinea-pig _ Dose—10 milligrammes, Date of Inoculation—April 30, 1906. [Age about 10 weeks.] Killed—July 28, 1906. [89 days after inoculation. ] Clinical History. |The animal showed no sign of ill-health during the course of the experiment, and after the first month there was a considerable diminution in the size of the tumour. Temperature. The temperature was normal throughout. Tuberculin Test. July 26, 1906. [87 days after inoculation.] Rise of temperature, 0°7° C. Weights. kilos. April 30, 1906 ... one vee vow 37°64 July 28, 1906 ... see eee vee 45°80 Total gain in weight.—8'16 kilos. POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion.—At the seat of inoculation on the left side of the neck there was an oval tumour, 3 cm. in greatest diameter, which on section was composed of curdy caseo-pus surrounded by a fibrous wall, 5 mm. in greatest thickness; on the internal surface of the wall there was some adherent caseous gritty material. Prescapular Glands.—The left prescapular gland was firmer than normal and measured a little over 5 cm. in length, rather less than 2:5 cm. in breadth, and 1:3 cm. in thickness ; under the capsule numerous discrete yellow tubercles, the largest 1 mm. in diameter, were seen ; on section, similar tubercles were seen in the depth of the cortex ; they were soft and slightly gritty from calcification ; the cortex also contained a firm yellow caseous patch, 2 by 2 by 0°7 cm., gritty around the margins; the capsule over this was thickened. The right prescapular gland was slightly enlarged, 5 by 2°5 by nearly 1°3 cm., and oedematous (recent tuberculin test). Prepectoral Glands.—Normal. Cervical Glands.—Normal. Thorax. LLungs.—The lungs showed throughout the paren- chyma scattered petechial points of haemorrhage ; no tubercles were seen. Thoracic Lymphatic Glands——The bronchial and mediastinal glands were not enlarged. The long caudal gland showed in the cortex scattered yellowish- white foci, some distinctly gritty ; there were similar yellow points in the bronchial glands; the gland above the bifurcation of the trachea showed mode- rately numerous discrete yellow foci. Heart and Pleura.—Normal. Abdomen. The peritoneal cavity contained a slight excess of fluid. Spleen.—Normal. Liver.—In the depth of the liver there was a nodule the size of a hemp-seed with grey fibrous margins and an irregular opaque yellow centre, not perceptibly cal- careous. Portal Glands.—Normal. Kidneys and Suprarenal Bodies.—Normal. Tongue, Tonsils, Pharyna and Intestines.—Normal. Testes.—Normal. Various Lymphatic Glands. Submazillary, Parotid, Axillary, Gastric, Mesenteric, Colic, Lumbar, Coeliac, Iliac, Precrural, Popliteal, Gluteal and Ischiatic Glands.—Normal. The right renal gland contained about half a dozen opaque yellowish-white foci, just perceptibly gritty. Microscopical Examination. (Smear preparations.) Left Prescapular Gland.—Moderately numerous tubercle bacilli. Liver (nodule).—No tubercle bacilli. Renal Gland.—T wo tubercle bacilli. Caudal Mediastinal Gland.—One clump containing about 20 tubercle bacilli. CALF 406. Virus E. II. Subcutaneous inoculation of culture derived from the original material through a guinea-pig. Dose—10' milligrammes. Date of Inoculation—April 30, 1906. [Age about 10 weeks.] Killed—July 30, 1906. [91 days after inoculation.] Clinical History. The animal showed no sign of ill-health during the experiment. The local tumour diminished in size after the first month. Temperature. The temperature was normal throughout. Tuberculin Test. July 26, 1906. [87 days after inoculation.] Rise of temperature, 1:0" C. 34654 Weights. kilos, April 30, 1906 sine eon sas 33-55 July 30, 1906 vin cate. cae eee Total gain in weight.—10°89 kilos. POST-MORTEM EXAMINATION. Carcass in good condition Local Lesion.—In the subcutaneous tissues jon the left side of the neck there was a small oval tumour, measuring 4°5 by 3 by 2cm. On section it showed in G2 54 the centre an irregular cavity filled with an opalescent watery fluid, in which caseous flakes were suspended, and surrounded by dense yellowish-white caseous slightly gritty substance; the whole was enclosed within a fibrous capsule. Prescapular Glands.—The left prescapular gland was enlarged for about three quarters of its length and measured 6 cm. in length, and 4 cm. by a little over 1°2 cm. at its broadest part. The enlarged part was indurated, and on section irregularly caseated ; the caseous areas were dense, yellow, and slightly gritty from calcification. At the narrow extremity of the gland there was a small indurated patch in the cortex, closely beset withjyellow caseous slightly gritty tubercles. -The right prescapular gland measured 51 by 1°6 by 1:3 cm., and was slightly oedematous, . Prepectoral and Cervical Glands.—Normal. Thorax. Lungs.—In the left cephalic lobe there was a grey translucent miliary tubercle, and one or two doubtful grey points. In the left caudal lobe under the pleura there was a tubercle, a little larger than a millet seed with a yellow centre and a reddish grey margin ; there were ‘also one or two minute translucent grey tubercles. In the right lobes under the pleura there were six nodules, ranging in size from a millet toa hemp seed, with yellow centres and grey margins , there were besides a few small semi-transparent polenl No tubercles were seen on section of the ung. Thoracic Lymphatic Glands.—The left bronchial gland ‘contained a millet-seed sized tubercle with a yellow centre ; the tubercle readily shelled out from the gland substance. The right bronchial and the dorsal mediastinal glands appeared normal. ; Pleura and Heart.—Normal. Abdomen. Omentum and Peritoneum.—Normal. . Spleen.—Normal. , Liver.—The liver showed on sectiona few minute yellow foci (no T.B.) ; otherwise normal. Portal Glands.—Normal. Kidneys.—Patches of the cortex of the left kidney were pale and fibroid ; no tubercles were seen in either kidney. Suprarenal Bodies.—Normal. Tongue, Tonsils, and Pharyna.—Normal. One retro-pharyngeal gland contained a millet-seed sized grey tubercle, in the centre of which were yellow points; the other pharyngeal gland was normal. Small Intestine—The mucous membrane in the lower part of the small intestine, mainly that covering the long Peyer’s patch, showed numerous shallow pits or erosions with well-defined margins and somewhat congested floors ; there was no thickening of the submucosa. Large Intestine—All the lymphoid follicles in the large intestine were slightly enlarged and congested ; the mouths of the follicles were irregularly expanded but not definitely ulcerated. Gastric Glands. —Normal. Mesenteric Glands.—The terminal mesenteric glands (not énlarged) showed in the cortices fairly numerous firm irregular yellow tubercles, many slightly gritty from calcification, ranging in size from a mere point up to that of a millet seed; the glands as far as the middle of the mesentery were also affected but con- tained here and there only a minute yellow focus. In the anterior half of the mesentery the glands were normal. The Ileo-Colic Glands contained numerous similar tubercles ; those in the largest gland of the group were at one part aggregated together, forming a kind of network. Colic Glands.—Several of the colic glands contained tubercles similar to those in the mesenteric glands. Submasillary, Avillary, Renal, Lumbar, Coeliac, Iliac, Precural, Popliteal, Gluteal, and AIschiatic Glands.—Normal. Testes.—Normal, CALF 504. Virus E. II. Subcutaneous inoculation of culture derived from the original material through a guinea-pig. Dose—50°0 milligrammes.' Date of Inoculation—January 30, 1907. [Age about 7 weeks.] Killed when well—May 10, 1907. Clinical Notes. - - At the seat of inoculation there was after 13 days a firm circular tumour about 10 cm. in diameter, which subsequently became soft and fluctuant. The right prescapular gland slowly increased up to 10 cm. in length. The condition of the animal was fair throughout. Temperature. The temperature remained normal throughout. Tuberculin Test. February 19, 1907. [20 days after inoculation.] Dose of tuberculin, 4-0 ec. (avian). Rise of tempera- dure, 06°C. - ‘ ’ Weiglits. - kilos, January 30, 1907 ae “ vee 60°76 May 10, 1907 ... ai toe vee 92°06 Gain in weight.—31°30 kilos, POST-MORTEM EXAMINATION. The carcass was in good condition. [100 days after inoculation.] Local Lesion—At the site of inoculation on the right side of the neck there was a prominent tumour 75 cm. in diameter and 5:7 cm. in thickness. On sec- tion it was a fibrous-walled cyst, filled with curdy caseo-pus and serous fluid. The caseous masses were attached to the inner wall, but were easily scraped away leaving a smooth wall crossed by fibrous strands. Right Prescapular Gland.—The right prescapular gland measured 9 by 5:1 by 3°8 cm. It was replaced to about three-quarters of its extent by masses of caseo-necrotic substance slightly pinkish in the centre ; two had broken down forming small abscess cavities. The periphery of the nodules was made up of a close network of caseo-calcareous tissue and what remained of the gland substance contained discrete tubercles, Left Prescapular Gland.—Measured 5°8 by 2°6 by 14cm. Normal on section. - _Prepectoral, Cervical, and Axillary Glands on both sides.—Normal. Thoras. Pleura and Diaphragm.—Normal. Lungs.—The left lung showed beneath the pleura three or four minute transparent tubercles. In the - 55 margin of the posterior lobe there was i nodule the size of a wheat-grain, aoe » The right lung was normal.. Thoracic Glands.—The bronchial and mediastinal glands were normal except for an occasional minute yellowish focus. Heart.—Normal, Abdomen. Omentum and Peritoneum.—Normal. Spleen (weighed 268 grammes).—Normal. Liver and Gall-bladder.—Normal. Portal Glands.—In one portal gland there was a minute gritty focus. The rest were normal. Kidneys:—In the left kidney, at the junction of the cortex with the medulla, there was a minute trans- Iucent tubercle with a gritty centre. The right kidney was normal, ; Suprarenal Bodies.—Normal. Coeliac, Lumbar, and Iliac Glands.—Normal. Renal Gilands.—The right renal lymphatic gland showed eight slightly irregular yellowish tubercles up to a rape seed in size, some soft, some slightly gritty. ‘The left renal gland was normal. Alimentary Tract. Tongue, Pharynx, Tonsils—Normal. Submazillary, Parotid, Retro-pharyngeal Glands.—Normal. Small Intestine.—In about the first two feet of the -smal] intestine there was a moderate number of soft and yellow nodules, raised beneath the surface of the mucous membrane. They soon became sparsely dis- ees and finally were absent towards the lower part. Large Intestine.—Normal. Gastric Glands—One gastric gland contained a gritty focus. Mesenteric Glands.--The majority of the mesenteric glands contained in the cortices a few yellowish tubercles ranging up to a rape seed in size ; nearly all were slightly gritty, a few were soft. _ Tleo-Colic Glands.—The ileo-colic glands showed in the cortices moderately numerous yellow gritty tubercles. Testes.--Normal. Various Lymphatic Glands. Precrural, Popliteal, Gluteal, Ischiatic, and Pudic.— Normal, Larynz and Trachea.—Normal. Microscopical Examinations, (Smear preparations.) Lung Emulsion.—A few tubercle bacilli seen. Foci from the Left Kidney.—One long and two short tubercle bacilli seen. Foci from the Mesenteric Gland.—One long tubercle bacillus seen. Nodule from Small Intestine.—A few tubercle bacilli seen. CALF 506. Virus E. ITI. Subcutaneous inoculation of culture derived from the original material through a guinea-pig. Dose—50 milligrammes. Date of Inoculation—January 30, 1907. Clinical History. The health of the animal remained good. The ‘tumour soon softened and became fluctuant, but there was no ulceration of the skin. Temperature. The teraperature remained normal throughout. Tuderculin Test. February 19, 1907. [20 days after inoculation. ] Rise of temperature, 1:2° C. Weights. : kilos. January 30, 1907 ote wee ses 82:08 May 9,1907 ws. ssn ewe tee 10294 Gain in weight.—20°86 kilos. POST-MORTEM EXAMINATION. General condition good. Local Lesion—On the right side of the neck there _was a somewhat prominent rounded tumour which “inéasured 6°3 by 5:1 cm.; it did not infiltrate either skin or muscle. On section it was a fibrous walled cyst, traversed by cords of translucent tissue and filled with masses of soft caseous substance in a clear ‘serous fluid. .. te ‘ Prescapulax Glanils—The right prescapular gland measured 6°3 by 25 by 1:2cm. Onsection it contained [Age 7 weeks.] Killed—May 9, 1907. [99 days after inoculation.] two nodules, one at each extremity, replacing about a third of the gland ; they were composed of collections of caseo-calcareous granules. In the rest of the gland there were a few scattered tubercles, The left prescapular gland measured 5°5 by 2bylcm., and on section was normal. Prepectoral Glands.—Normal. Cervical Glands.—The right inferior cervical gland measured about 1 cm. in diameter, and was almost replaced by a collection of caseo-calcareous granules. Other cervical glands appeared normal. Agillary Glands.—Normal. Thorax. Pleuru.—Normal. Lungs.—The lungs were crepitant. On the surface of the left posterior lobe, just beneath the pleura, there was a hard grey nodule 3:5 mm.in diameter. In ‘the margin of the right anterior lobe there was a similar hard grey nodule 45mm. in diameter. On the surface of left lung, close to the spinal column, just dorsal to the entrance of the bronchi, there was a collection of minute grey tubercles twenty to thirty in number, a few of which were opaque in the centre, but not gritty. In the rest of the lungs an occasional translucent focus could be seen. On section, the parencbyma appeared normal. Mediastinal Glands.—Normal. Bronchial Glands—There was a translucent cal- careous focus in one bronchial gland ; other bronchial glands appeared normal. Heart and Pericardium—Normal. Abdomen. ~ Omentum, Peritoneum, and Diaphragm.—Normal. Intestines.—Normal. Mesenteric and Colic Glands.—Normal. Spleen—The spleen weighed 198 grammes. The surface and substance were normal. - Liver.—Normal. Portal Glands.—In the largest of the portal glands there were some scattered yellowish foci just visible, which, when crushed, cracked like egg-shells. ‘Kidneys.—Normal. Suprarenal Bodies—Normal. Renal, Coeliac, Lumbar, and Iliac Glands—Normal. Tongue, Tonsils, Larynx, and Pharyne.—Normal. 56 Various Lymphatic Glands, Parotid, Submazillary, Retro-pharyngeal, Precrurat, Popliteal, Gluteal and Ischiatic Glands.—Normal. Microscopical Examinations. (Smear preparations.) Portal Gland (foci).—One group of tubercle bacilli (7 or 8 in number). Lung Nodule (Emulsion I.).—Moderately numerous tuberele bacilli. Lung Nodule (Emulsion II.).—Numerous tubercle bacilli. Prescapular Gland.—Numerous _ tubercle (with a moderate number of small clumps). bacilli CALF 606. Virus E. II. Subcutaneous inoculation with culture derived from the original material through a guinea-pig. Dose—50-0 milligrammes. Date of Inoculation—April 10, 1908. [Age 14 weeks.] Killed when in good general health—July 8, 1908. [89 days after inoculation.] Clinical History. There was no of ill-health during the hei sign experiment. Temperature. The temperature was normal throughout. Weights. kilos. April 10, 1908 ... ine ‘ss w. 45°80 July 8, 1908 on see 71:19 Gain in weight.—25°39 kilos. POST-MORTEM EXAMINATION. General condition good. Local Lesion—At the seat of inoculation on the left side of the neck there was a small irregular ulcer in the skin, leading into a sinus lined by granulation tissue; it was surrounded by thick fibrous tissue. In the underlying muscle there were five fibrous nodules up to a pea in size with purulent centres. Prescapular Glands.—The left prescapular gland measured 4°3 by 2°4 by I-1cm. On section it contained two nodules, the largest 1-2 cm. in diameter, composed of yellowish purulent substance enclosed in sharply limited fibrous capsules. The right prescapular gland measured 4:2 by 2 by lcm. On section it was normal. Prepectoral Glands.—Normal. Cervical Glands.—Normal. Thorax. Lungs.—In the left lung, just beneath the pleura, there was a single grey submiliary tubercle with a calcareous centre. Mediastinal Glands.—The caudal mediastinal gland was a little thickened at one part by a caseo-calcareous nodule 2 cm. in length and 1 cm. broad; the nodule easily shelled out from the gland substance and was surrounded by discrete caseous gritty tubercles which also easily shelled out. In a second gland there were several caseous gritty tubercles up toa hemp seed. Ina third gland there was a single caseo-calcareous miliary tubercle. Bronchial Glands.—Normal. Abdomen. Stomach.—Loosely attached to the stomach were two fibrous nodules containing: firm opaque streaks, Liver.—In the depth of the liver there was a nodule the size of a hemp seed with a yellowish centre. Other organs and lymphatic glands showed no tuberculosis. Microscopical Examinations. (Smear preparations.) Prescapular Giland.—Numerous tubercle bacilli. Lung (tubercle).—A few tubercle bacilli. Mediastinal Gland—Numerous tubercle bacilli, Liver (nodule).—No tubercle bacilli. Stomach (nodule).—No tubercle bacilli. CALF 608. Virus E. II. Subcutaneous inoculation with culture derived from the originai material through a guinea-pig. Dose—100 milligrammes. Date of Inoculation—April 10, 1908. [Age 14 weeks.] Killed—June 29, 1908. [80 days after inoculation,] Clinical History. 39-9° C. on the 19th day ; the temperature was other- wise normal. There was no sign of ill-health during the experi- ment. Temperature. There were a few slight rises, the highest being Weights. . kilos. April 10, 1908 tee ass vee 39-90 June 29, 1908 ow eae er 45°35. Gain in weight.—5-45, kilos. POST-MORTEM EXAMINATION. Carcass in fair condition. Local Lesion.—In the subcutaneous tissue on the left side of the neck there was a prominent tumour 13 by 7 by 6 cm. On section it was a thick fibrous walled cyst containing masses of softening yellow caseo-necrotic tissue partly adherent to the walls and partly free in a straw-coloured serous fluid. There was a caseo-calcareous cord connecting the local tumour with the prescapular gland. Prescapular Glands.—The left prescapular gland measured 6 by 3:5 by 25cm. On section, about two- thirds was replaced by firm yellow necrotic substance, softening a little at one part and beset with minute calcareous granules. The right prescapular gland measured 5:3 by 2°7 by 13cm. On section it was normal. Prepectoral Glands.—Normal. Cervical Glands.—Normal. Thorax. Pleura.—Normal. Lungs and Bronchial Glands.—Normal. Mediastinal Glands. —In the caudal mediastinal gland—which was not enlarged—just beneath the ‘capsule there was a caseo-calcareous nodule 5mm. in diameter, contained within a thin fibrous capsule from which it easily shelled out. Other mediastinal glands were normal. Abdomen. Intestines.—Along the whole length of the small intestine there were numerous slightly yellowish 57 nodules, palpable and prominent on. the mucous surface varying up to3 and 4 mm. in diameter and occasionally aggregated into collections. At the place where they were most numerous about fifty were counted in 10 cm. length of gut. Mesenteric Glands.—Normal. Spleen.—Normal. Liver.—Normal. _ Portal Glands.—In the cortices there were a few irregular calcareous granules. Suprarenal Bodies.—The right suprarenal body contained a single yellow caseous tubercle. The left was normal. All other organs and lymphatic glands showed no tubereulosis. The animal had been paralysed for about a fort- night in the hind legs. On examination of the lumbar region of the spinal canal there was found a longi- tudinal fracture of a vertebra, with displacement ventrally, resulting in constriction of the cord at that spot. Microscopical Examinations. (Smear preparations.) Emulsion of Prescapular Gland.—Numerous tubercle bacilli. Emulsion of Suprarenal Body Tubercle.—Moderately numerous tubercle bacilli. Intestinal Nodule I.—A few tubercle bacilli. Intestinal Nodule II—A few tubercle bacilli. CALF 610. Virus E. II. Subcutaneous inoculation with culture derived from the original material through a guinea-pig. Dose—20°0 milligrammes. Date of Inoculation—April 10, 1908. [Age 14 weeks.] Killed when in good general health—July 10, 1908. [91 days after inoculation.] Clinical History. The animal showed no evidence of ill-health during the experiment. POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion.—In the muscle at the site of inocula- tion there was a pear-shaped tumour measuring 7 by 3cm. On section it was a cyst with fibrous walls filled with caseo-pus. Prescapular Glands.—The left prescapular gland contained a few soft caseous gritty tubercles. The right prescapular gland was normal. All organs and other lymphatic glands showed no tuberculosis. Weights. kilos. April 10, 1908 ... abe a vee 42:17 July 10,1908 ... ee te vee 78°80 Gain in weight.—36°63 kilos. PONY 18. Virus E. II. Intravenous inoculation with culture derived from the original material through a guinea-pig. Dose—10°0 milligrammes. Date of Inoculation—February 17, 1908. [Age— Yearling.] Died—March 28, 1908. [40 days after inoculation.] Clinical History. The animal speedily began to show considerable respiratory distress and emaciated rapidly, Zemperaiure, There was a slight rise between the 17th and 26th days, the highest point reached being 39°6° Centigrade. The temperature was otherwise normal. Weights. ‘ kilos. February 17, 1908 ... vee ss 76°18 March 28, 1908 ... eae ade 63°49 Total loss of weight.—12°69 kilos. 58 POST-MORTEM EXAMINATION. -Carcass emaciated. Thorax. Lungs.—The lungs were heavy and did not collapse normally ; they floated as a whole, but small pieces sank in water. They were irregularly congested on the surface. On section they cut firm and showed the substance closely filled with a network of grey translucent tissue, leaving only small patches of emphysematous and oedematous lung tissue in between. The bronchi were full of fluid. In the right anterior lobe there were two yellowish necrotic firm nodules and small patches of similar substance, though less firm, were scattered throughout the lungs. These nodules when squeezed exuded purulent fluid. Thoracie Lymphatic Glands.x—The bronchial and mediastinal glands were firm and in the cortices there were bright yellow necrotic patches. Abdomen. Mesenteric Glands.—Many of the mesenteric glands were dark red with blood ; they showed no areas of necrosis. Intestines, Stomach, and Colic Glands.—Normal. Spleen—The spleen weighed 169 grammes; on section it was normal. The glands in the hilum of the spleen were a little firm, and on section the cortices showed in parts indefinite yellowish patches, while the rest was grey and translucent. Liver—The liver had on the surface a mottled appearance from alternate light and dark red areas. On section the light areas were homogeneous in appearance, and the dark were congested and showed the interstitial tissue well marked, forming a translu- cent network. Beneath the surface there were sparsely scattered opaque yellowish nodules up to a millet seed. In the substance there were two small purulent nodules, Portal Glands.—The ‘portal glands resembled the splenic glands except that they were congested and showed more numerous yellow patches in the cortices, some of which were almost entirely yellowish. > Kidneys.—They showed scattered minute grey translucent foci, a few apparently a little opaque, they were not well defined from. the rest of the substance ; they were irregularly distributed, and in parts were numerous. On section they could be seer in the deeper parts of the cortices. Coeliac Glands—The coeliac glands resembled the splenic glands. : Iliac Glands.—In the iliac glands there were dis- crete translucent nodules, resembling lymphoid tissue. The Posterior Pharyngeal Glands were a little congested. Other organs and glands appeared normal. Microscopical Examinations, (Smear preparations.) Lung (necrotic patch).—Numerous long slender beaded tubercle bacilli. Lung (dark patch).—Numerous long slender beaded tubercle bacilli. Spleen (substance).—Moderately numerous tubercle bacilli. Liver (substance).—Moderately numerous tubercle bacilli. Kidney (substance).—A few tubercle bacilli. Bronchial Gland (cortex).—Numerous tubercle: bacilli, PONY 20. Virus E. II. Subcutaneous inoculation with culture derived from the original material through a guinea-pig. Dose—50°0 milligrammes, Date of Inoculation—-February 17, 1908. Killed when in good general health--June 26, 1903. Clinical History. A week after the inoculation there was a prominent swelling about 5 cm. -in diameter in the shape of a cone, the apex of which was on the point of ulcerat- ing. Within a few days the tumour ulcerated. There was no surrounding infiltration. On June 26 the ulcer had healed. The animal at no time showed any sign of ill-health. Temperature. The temperature remained normal throughout the experiment. Tuberculin Test. May 13, 1908. [86 days after the experiment began.] Dose of tuberculin (bovine), 3:0 cc. Rise of temperature, 20° Centigrade. Weights. : 4 kilos. February 17,1905 ... oa eos 65°30 March 16, 1908 eae sn sian 73°46 June 26, 1508 as vs es 89°33 Total gain in weight.—24-03 kilos. [Age—Yearling. ] [180 days after inoculation. ] POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion -At the seat of inoculation in the- skin on the right side of the neck there was a whitish- scar, pigmented around its margins. In the sub- cutaneous tissues beneath, there were a few discrete yellow caseous softening nodules with fibrous margins, the largest about the size of peas. Prescapular Glands.—In the right prescapular group of glands there were two glands enlarged up to the size of a thrush’s egg, which on section were almost replaced by firm yellow caseous substance ; two smaller glands were partly caseous. Two others contained caseous tubercles and patches; the rest were- normal, oo of Result. nA 1743 O-l mg. {| Intrav. | D.18 days |{G. T. Intes- 1741 |0-000:0001] Intrav. | D. 74 ,, G. T. mg. JULY 23, 1908. The strain was derived from the original material. 1,473 days. MONKEY 224. Subcutaneous. Dose: 1:0 mg. Died: August 21, 1908. 29 days. P.M.—General tuberculosis. The culture used was the 41st generation, 13 days old, and had been in cultivation a total period of RABBITS. Number. Dose. Method, pees of Result. 1847 1:0 mg. | Intrav. | D. 15 days | G. T. 1848 | 10mg. | Intra. |D14 , | GT. 141 _PONY 12. [Yearling.] Virus B. XI. Subcutaneous inoculation of culture derived from the original material. Dose—50 milligrammes. Date of Inoculation—January 24, 1908. Killed when in good general health—May 27, 1908. [124 days after inoculation.] Clintcal History. An ulcer formed, from which there was a purulent ‘discharge, 7 days after inoculation. On February 11, 18 days after inoculation, there was an indurated area 125 cm. in greatest diameter, ill-defined and highest near the ulcer. Later the breathing was quiet but a little jerky. Temperature. The temperature was slightly raised during the five weeks following the inoculation, reaching 40°4° C. on the 36th day; it then quickly fell and remained normal until death. Weights. kilos. January 24,1908 ... 268 87-07 March 16, 1908 a8 ees 87-98 May 27, 1908 ie ; 99°31 Total gain in weight.—12°24 kilos. Tuberculin Test. May 18, 1908 [109 days after the experiment began]. Dose of tuberculin, 3:0 cc. (bovine). Rise of temperature, 2°5° Centigrade. POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion.—The skin at the seat of inoculation ‘on the left side of neck was thickened by cicatricial tissue and showed three small holes about 2 mm. across. In the subcutaneous tissues there was an area about 6 cm. in diameter and about 1 cm. in thickness, composed of firm white scar tissue beset with small caseo-calcareous nodules. The scar tissue was adherent to the muscle, which contained moderately numerous ‘grey tubercles with opaque gritty centres up to a millet seed in size. Prescapular Glands.—Along the anterior margin of the left scapula there was a chain composed of numerous glands, the largest about the size of a thrush’s egg. Several were completely replaced by softening caseous gritty substance, others were uniformly enlarged and contained a few caseous tubercles and nodules. . In a line with these there was a mass just anterior to the first rib measuring 6 by 4 by 2°5 cm. On section this was composed of discrete caseo- calcareous nodules set in a matrix of fibrous tissue. Posterior to the latter, just within the thorax beneath the first rib, there was a group of glands, of which some were entirely replaced by yellowish caseo- necrotic tissue with more opaque gritty streaks and others contained caseous gritty nodules. In one right prescapular gland there was a small collection of caseous tubercles; the others were normal, : A gland on the left side of the lower third of the neck, the size of a robin’s egg, was caseous throughout. Thorax. Lungs.—The lungs were crepitant. Numerous hard tubercles could be felt beneath the surface especially in the margins, and on close examination could also be ‘seen beneath the pleura; a few projected slightly. They varied up to about 2 mm. in diameter, but the majority were submiliary. On section through them they were composed wholly of firm translucent tissue, some few had minute opaque centres. On section through the parenchyma numerous similar tubercles were distributed throughout, but the majority, being quite transparent, were difficult to see. Mediastinal Glands.—In the dorsal mediastinal glands, which were not obviously enlarged, there was an occasional grey tubercle with an opaque centre. Bronchial Glands.—In the bronchial glands, which were not obviously enlarged, there were a few similar tubercles and an occasional softening yellow caseous nodule. Abdomen Spleen.—The spleen weighed 283 grammes. It showed, both on and in the capsule, numerous firm fibroid tubercles varying up to nearly 2 mm. in diameter ; the majority were small and grey, the largest were greyish-white. On section through the substance there were sparsely scattered grey trans- lucent tubercles with slight central opacities up to a hemp seed in size, the largest of which were calcareous in the centre. Liver.—On the surface of the liver, just beneath the capsule, there were numerous discrete tubercles varying from a point up to 2 mm. in diameter, a few of which projected; the smallest were quite trans- lucent, the largest were a little opalescent and opaque yellow in the centre. On section through the sub- stance of the liver the tubercles were less numerous and smaller than on the surface. Portal Glands.—The portal glands contained mode- rately numerous firm grey tubercles with opaque centres ranging up to a millet seed in size. Kidneys.—In the cortex of the right kidney there were three grey rather ill-defined translucent tubercles, with slightly gritty opaque centres, up to a millet seed in size. In the left kidney there were a few scattered similar tubercles. Lumbar Glands—The lumbar glands contained discrete nodules up to a hemp seed in size, some of which were translucent and not sharply defined, others were grey with yellow caseous or gritty centres. Iliae Glands.—In the iliac glands there were a few minute caseating tubercles. Testes.—Normal. The right epididymis was swollen and contained two cystic nodules filled with yellow pus, the largest 5 mm. in diameter. There was no tuberculosis of any other organ or gland in the body. Microscopical Examinations. (Smear preparations.) Emulsion of Prescapular Gland.—Numerous tubercle bacilli. Epididymis (nodule).—A few tubercle bacilli. Kidney.—No tubercle bacilli. A few blue staining bacilli seen. Lung (tubercle) Liver 4 t No tubercle bacilli. Spleen ,, Nots.—Extensive dissemination but apparently retrogressive. The liver and lung tubercles were fibrous ; those in the spleen were gritty. 142 CALF 652. Virus B, XI. Subcutaneous inoculation with culture derived from the prescapular gland of Pony 12. Dose—50 milligrammes. ‘Date of Inoculation—August 5, 1908. {Age 12 weeks. ] Died—September 18, 1908. [44 days after inoculation.] Temperature. The temperature rose gradually, attaining 39-9°C. 17 days after inoculation; it remained with slight fluctuations about this level, but never rising higher, until death. Weights. kilos. August 5, 1908 we vite we =59°85 September 18,1908 ... eae wee 49°42 Total loss in weight.—10°43 kilos. POST-MORTEM EXAMINATION. Carcass emaciated. Local Lesion.—At the seat of inoculation in the subcutaneous tissue on the left side of the neck there was a prominent tumour measuring 18 by 12 by 6 cm., and weighing with skin and muscle 963 grammes. On section it consisted of congested caseo-necrotic tissue. It included at one extremity the left prescapular gland, which measured 9 by 6 by 6 cm., and consisted of caseous tissue throughout. The right prescapular gland contained discrete caseous nodules up to a pea in size, confluent in parts and forming caseous patches. Prevectoral Glands.—A left prepectoral gland showed caseous patches in the cortex. A second gland showed discrete grey foci. Cervical Glands.—The left inferior cervical glands were enlarged and showed discrete grey foci set in firm translucent tissue. A middle cervical gland was half caseous. The right inferior, middle, and superior cervical glands contained a varying number of caseous nodules. Agillary Glands.—The left axillary gland contained two or three caseating nodules and numerous grey foci. The right axillary showed a few caseous nodules up to 5 mm. in diameter. Thorax. Pleura.—The lymphatic fringes along the ribs con- tained numerous small flattened translucent tubercles. Lungs.—The lungs were heavy and did not collapse PONY 26. Fed once with culture derived from the original material. Dose—100-0 milligrammes. Date of Feeding—March 21, 1908. normally, they sank completely in water. On section the lung parenchyma was dark red, especially on one side, and was closely filled with irregular caseous nodules. Mediastinal Glands.—The caudal mediastinal gland measured 15 by 6 by 3°7 cm. On section the cortex was composed of firm reddish-grey tissue with a close yellow caseous network. The other mediastinal and the bronchial glands were enlarged and resembled the caudal gland. Abdomen. Intestines— The small intestines were congested and the Peyer’s patches showed an occasional caseous nodule. Mesenteric Glands.—The mesenteric glands were a little enlarged and were firm ; a considerable part of the cortices was replaced by caseous nodules and areas. Spleen.—The spleen weighed 283 grammes. On section it was fairly closely filled with caseating nodules up to a pea in size. Liver.—In the liver there were numerous grey nodules with opaque centres, varying much in size, from less than a rape seed up to a pea. Portal Gilands.—These glands were enlarged ; the cortices were firm and closely filled with confluent grey caseating nodules. Kidneys.—In the cortices of the kidneys there were sparsely scattered grey nodules, caseating in the centre, up to about a hemp seed in size. Suprarenal Bodies.—Hach suprarenal body contained two caseating tubercles. The lumbar and iliac glands contained discrete cageous nodules up to a hemp seed in size. Tongue, Laryna and Pharynz.—Normal. Tonsils.—The tonsils showed several caseous nodules. Trachea.—Along the trachea there were a few- flattened congested nodules. The posterior pharyngeal, submaxillary, precrural, popliteal and gluteal glands contained varying numbers of discrete caseous nodules up to a pea in size. [Yearling.] Virus B. XI. [Given in its drinking water.] Killed when in good general health—September 3, 1908. [166 days after the experiment began. ] Clinical History. There was no sign of ill-health during the experi- ment. Temperature. Normal. Weights. kilos. March 20, 1908 ais 66°66 September 3, 1908 ... 15551 Total gain in weight.—88'85 kilos. Tuberculin Test. May 13,1908. [53 days after the experiment began. ] Dose of tuberculin, 3-0cc. Rise of temperature, 1°6° C. POST-MORTEM EXAMINATION. General condition good. Submawillary Glands.—In one submaxillary gland there was a yellow caseo-calcareous granule. Cervical Glands.—Two or three superior cervical glands on each side contained an occasional bright yellow small ciseo-calcareous patch or streak. Abdomen. Mesenteric Glands.—Almost all the mesenteric 143 glands contained from one to six bright yellow, irregular, caseo-calcareous tubercles up to about a millet seed in size. Colic Glands.—A few of the very numerous colic glands contained from one to four similar though slightly smaller tubercles. Liver.—Just beneath the capsule of the liver, there were two nodules composed of soft grey translucent tissue, the size of a millet seed and a hemp seed respectively. Portal Glands.—There was an opaque focus in a portal gland. Suprarenal Bodies,—There were two small yellow bodies projecting from the left suprarenal body. Thorax. Lungs.—In the right lung just beneath the pleura there was a firm grey translucent nodule the size of a small pea. There was no tuberculosis of any other organ or gland. Microscopical Examinations. (Smear preparations.) Submazillary Gland.—A few tubercle bacilli. Superior Cervical Gland.—A few tubercle bacilli. Mesenteric Gland.—Numerous tubercle bacilli. Colic Gland.—Moderately numerous tubercle bacilli. Liver (nodule).—No tubercle bacilli. Suprarenal Body.—No tubercle bacilli. PONY 30. {[Aged.] Virus B, XI. Intravenous inoculation of culture derived from the original material. Dose—10-0 milligrammes. Date of Inoculation—March 20, 1908. Died—April 9, 1908. Clinical History. The animal showed respiratory distress and loss of appetite for more than a week before death. Temperature. The temperature rose to 39°5° on the fourth day and 39°6° on the sixth day. It fell for a few days and again rose, the highest point reached being 40-0° C. two days before death. Weights. kilos. March 20, 1908 309°33 April 9, 1908 ... 287-10 Total loss in weight,—22'23 kilos. POST-MORTEM EXAMINATION. Carcass in fair condition. Thorax. Lungs.—The lungs did not collapse normally ; they felt firm and on section were oedematous, the left lung being also congested. The lung parenchyma was closely beset with translucent tubercles slightly opaque in the centre, the majority about the size of rape seed. Plewra.—On the visceral and parietal pleura and on the pleural surface of the diaphragm there were numerous loosely attached whitish fibrous tissue [20 days after inoculation.] | growths, quite homogenous, the largest about 5°8 by 5 cm. in area. Abdomen. Spleen.—The spleen weighed 907 grammes. substance was firm but no foci could be seen. Liver.—The liver was congested but showed no tubercles. The Kidneys.—In the kidneys, just beneath the surface, could be seen an occasional minute grey tubercle. In the wall of the mesenteric artery there was a necrosed patch, in which lime salts were deposited, and in which several small worms were demonstrated. There was a similar condition in an iliac artery and in the lumen of the vessel there was an ante-mortem clot. The majority of the lymphatic glands were oede- matous, and in some the lymphoid follicles were well marked. Microscopical Examinations. (Smear preparations.) Lung (substance).—Moderately numerous tubercle bacilli. Bronchial Gland (substance).— A few tubercle bacilli. Spleen (substance).—A few tubercle bacilli. Liver (substance).—A few tubercle bacilli. Small Intestine (scraping).—No tubercle bacilli. MONKEY 214. Virus B. XI. ‘Fed with culture derived from Rabbit 1587. Dose--50 milligrammes. [Administered in milk, 10 mg. a day for five days.] Date of Feeding—April 17, 1908. Died—-May 19, 1908. POST-MORTEM EXAMINATION. Tongue, Tonsils, Larynx, Pharynx and Trachea.— Normal. Submazillary Glands.—One on the left side con- tained a softening yellow caseous nodule the size of a 34684 (32 days after the first feeding. ] hemp seed. On the right side there was a gland containing two caseous tubercles the size of rape seed. Retro-pharyngeal Glands.—The gland on each side was a little enlarged and almost replaced by softening caseous substance. x ‘144 Cervical Glands.—Normal. Stomach.—In the mucous membrane there was a small caseous ulcer. Small Intestine—All the Peyer's patches were thickened and caseous, and the internal surfaces were bright red from submucous haemorrhages A few only showed slight superficial ulceration. The mucous membrane in between the patches was beset with numerous caseous tubercles, so closely in parts as to form almost a continuous chain. In parts also there was a tendency for the nodules to encircle the gut. Large Intestine-——Beneath the mucous membrane there were three small caseous nodules: Mesenteric. Glands——-The central glands in the mesentery had ccalesced into a triangular mass, whose sides measured each about 4 cm., and which was 16cm. in greatest thickness. On section the mass consisted of soft yellow caseous substance. Mesentery—The mesentery was contracted and contained numerous caseating tubercles and nodules up to 5mm. in diameter. There were also caseous streaks (thickened lacteals) passing from the intes- tines to the mesenteric glands, many of which were continuously caseous, while others were in parts broken up into discrete tubercles. Colic Glands——The majority of the colic glands were normal, but an occasional one contained a caseous patch. There were caseous patches in the ileo-colic glands. Abdomen. Slight excess of fluid in the peritoneal cavity. Spleen.—In the pulp there was a single minute grey tubercle. Liver--On the surface, just beneath the capsule, there were sparsely scattered opaque slightly yellowish tubercles (15 counted) varying up to a rape seed in size. On section through the substance the tubercles were rather more sparsely scattered. Kidneys.--Normal. Thorax. Lungs.—The lungs did not collapse normally ; the parenchyma was oedematous and emphysematous, and the right lobes were congested. In the right lung there were visible from the surface eight caseous nodules up to 3mm, in diameter. On the left side there were seven, on the whole smaller except for one which was slightly irregular and measured about 5 mm. in greatest diameter. On section there were similar nodules scattered throughout the substance, Bronchial Glands.—They were a little enlarged, and all coutained caseous patches replacing a considerable part of the substance. Parotid Lymphatic Glands—The glands on the right side contained an opaque miliary tubercle. There was no tuberculosis elsewhere. RHESUS MONKEY 224. Virus B. XI. Subcutaneous inoculation with culture derived from the original material. Dose—1-0 milligramme. Date of Inoculation—July 23, 1908 Died—August 21, 1908. POST-MORTEM EXAMINATION. Local Lesion.—At the seat of inoculation in the subcutaneous tissues of the back there was a soft baggy cyst with five small holes in the skin, from which soft caseo - pus of a reddish - brown colour exuded. On section it was filled with brownish pus containing caseous flakes. The-.walls were rough with caseous material adherent to them. Axillary Glands.—The axillary glands on both sides were enlarged and about the size of large beans, and on section were almost entirely replaced by soft yellow caseous substance. The scapular and lower cervical glands were enlarged, caseous, and softened. Thoraa. Lungs.—The lungs were pink and crepitant; they showed a few small transparent tubercles, also a few grey tubercles with caseous centres, the largest the size of a millet seed. Bronchial Gilands.—The bronchial glands were slightly enlarged and showed soft yellow caseous nodules. Atthe junction of the ribs with the spine on the right side there were five soft yellow caseous nodules up to a pea in size and on the left one. [29 days after inoculation. ] Abdomen. Omentum.—The omentum contained one or two minute transparent glistening tubercles. Mesenteric Glands.—There were one or two soft yellow caseous nodules in the mesenteric glands. Intestines.—Normal. Spleex.—The spleen was perhaps slightly enlarged, measuring 5 by 3 by 1:3 cm.; the substance was purple in colour and closely packed with greyish opaque tubercles, from a mere point up toa millet seed in size. Liver.—The liver was closely beset with yellowish- grey tubercles ranging from a point up to a rape seed in size. Portal Glands.—The portal glands were enlarged, yellow, caseous and softening in places, Kidneys.—Each kidney showed one greyish opaque tubercle the size of a rape seed. Submasxillary, Iliac, and Lumbar Glands.—Normal. Longue, Tonsils, Laryne, and Pharynx.—Normal. 145 PONY 6. [Aged.] Virus B. XII. Subcutaneous inoculation of culture derived from the original material. Dose—50°0 milligrammes. Date of Inoculation—September 6, 1907. Killed when in good general health—January 16, 1908. [132 days after inoculation.] Clinical History. The day after inoculation a small tumour was -present, the inoculated fluid not having been absorbed. “This increased in size to about a partridge’s egg ; it was indefinite around the margins. Four days after inoculation the skin burst, and turbid yellowish serum flowed out, the tumour as a result almost entirely disappearing. Temperature. Normal throughout. Weights. kilos. September 9, 1907 ... 355°60 January 16,1908 ... age we 362°40 Total gain of weight.—6'80 kilos. Tuberculin Tests. September 26, 1907. [20 days after inoculation. ] Dose of tuberculin (bovine), 4:0 cc. Rise of tem- perature, 14° Centigrade. October 11, 1907. [35 - days after inoculation.] Dose of tuberculin (bovine), 4:0 cc. Rise of temperature, 1°5° Centigrade. POST-MORTEM EXAMINATION. The animal was fat and in good condition. Local Lesion.—In the muscle at the seat of inocula- tion on the left side of the neck there was an elongated tumour, measuring 3°5 by ‘8cm. On section this was composed of firm yellowish caseated tissue surrounded by translucent fibroid tissue. Prescapular Glands——One of the group of left prescapular glands was enlarged and firm, measuring 12cm. in diameter. On section it was composed almost throughout of firm caseated tissue ne appreciably gritty. Two of the smaller glands contained a few firm caseous nodules in the cortices. The right prescapular glands were normal. Bronchial Glands.—In a bronchial gland there were just visible two or three minute whitish foci of a doubtful nature. Liver.—In the liver, just beneath the surface, there were two grey nodules with opaque centres, the size of hemp seed. There was no tuberculosis of any other organ or gland in the body. Microscopical Examinations. (Smear preparations.) Local Lesion.—Three tubercle bacilli seen. Bronchial Gland (focus).—No tubercle bacilli. ‘Liver (nodule).—No tubercle bacilli. PONY 8. [Aged.] Virus B. XII. Fed once with culture derived from the original material. Dose—100°0 milligrammes. Date of Feeding—September 6, 1907. [Given in its drinking water. ] Killed when in good general health—January 16, 1908. [182 days after the experiment began.] Clinical History. There was no sign of ill-health during the experi- ment. Temperature. Normal. Weights. kilos. September 9, 1907 ... es we = 37782 sanuary 16,1908 ... sug ww. 37556 Total loss of weight.—2°26 kilos. Tuberculin Tests. __ September 26, 1907. [20 days after the experiment began.] Dose of tuberculin (bovine), 4°0cc. Rise of temperature, 1°1° Centigrade. October 11, 1907. [35 days after the experiment began.] Dose of tuberculin (bovine), 4°0cc. Rise of ‘ temperature, 1°3° Centigrade. POST-MORTEM EXAMINATION. The carcass was very fat and in good condition. Cervical Glands.—A right superior cervical gland showed about half its substance firm and semi-opaque. Other cervical glands were normal. Abdomen. Jntestines.—Beneath the mucous membrane of the small intestine there were four hard yellowish nodules, varying up to a pea in size. Caecum.—At the extremity of the caecum, just beneath the peritoneum, there were two projecting fibroid nodules each of which contained on section a frm dirty-yellow necrotic centre. There were several similar nodules scattered about the peritoneal fat. Mesenteric Glands.—In one mesenteric gland there was a calcareous nodule about the size of a millet seed. A second showed a smaller opaque yellow gritty nodule. Omentum.—In the omentum there were three cal- careous nodules up to a hemp seed in size. Spleen.—In the spleen just beneath the surface there was an opaque yellow nodule 3 mm. in diameter. Liver.—Projecting from just beneath the surface of 146 the liver were sparsely scattered irregular stony nodules, resembling those in the lung and also those described in the other horses ; they were also seen on section through the substance. Thorax. Lungs.—Sparsely scattered throughout the lungs, both just beneath the surface and in the depth were hard stony nodules varying up to a large pea in size. On section they consisted of a thin grey capsule from which the nodule easily shelled out; the next coat consisted of a cartilaginous layer becoming gritty and finally calcareous in the centre. Bronchial Glands.—Several of the bronchial glands, which were not enlarged, contained similar nodules pro- jecting from the surface ; thellargest nodule measured 5 mm. in diameter. There was no evidence of tuberculosis in any other organ or gland. Microscopical Examinations. (Smear preparations.) Lung (nodule).—No tubercle bacilli. Liver (nodule).—No tubercle bacilli. Omentum (nodule).—No tubercle bacilli. Intestine (nodule).—No tubercle bacilli. Caecum (nodule).—One tubercle bacillus seen. Mesenteric Gland (nodule).—Two tubercle bacilli seen. Cervical Gland (opaque substance).—Six tubercle bacilli seen. “nbn wwaah)h uo wpovg aposaqn} aurnba Jo ynouy ‘6 ‘ ‘oypjod ursaoh6 uo rwprang ajasaqny auenba pun awaog “TT ‘Or ‘DIpIwW ULLIdk)B uo uoyvanInIQns pajynadas fo poaffa Humoys :auvbv u2rsavhj6 Uo 2]I0Nq ajoLaqQn} aumog *¢ ‘ ‘wnias aind uo sasnjjnd wosf umos :4nbo urtaoh76 uo 2/200q 9]9LagN} aURLOG "FE "apiong 9jausqne (AT “a sni24) auenba pun aurmog fo bba uo saunyyno funung “4£ ‘ 147 ™ MH © CO TT ‘Or 6 S 4b “9 G v ‘S % T STO SHNOWED Wig SainOW-O "G70 SHINO Wg | ee ; i i pagveanss SS: woryra9u95 38 pyniin) ABA Baatund : 3 OA } 3 gilt 4: nae | SLY oq ony EAN lagen qn “270 SHLNOy'9 a0 SHINO WY’ 9 a0 SHLWO WO | : uaa dy 6 _waursieg 8 moywaaua $01 | Pralvn WHIIIZS TPA WN ADD rH YG) Eon | | a0 SHLNOW "9, t seamen by “EH tecany ALA SA, ‘DIOSHLNOW"D “O10 smi NOW" AINLINS AUWWIEY bee a5 OU "Ty SMU 9S gatavdwny 4b nosy) ee ITIIOVE DIOUAANL ANINOA GNV WNIAOM JO- SaunLTAd 34684 149 INVESTIGATION OF VIRUSES OBTAINED FROM CASUAL TUBERCULOSIS OF VARIOUS MAMMALS. BY F. GRIFFITH, M.B. 34684 i id 150 INVESTIGATION OF VIRUSES OBTAINED FROM CASUAL TUBERCULOSIS OF VARIOUS MAMMALS. TABLE OF CONTENTS. PaGE Introduction ... eae he this sas bak ais ase wm Lol Summary of Results... clas wits si wa sae ase vx Lol Virus Z.I. (Gnu.) Chart of the Inoculations 152 Description of the Virus and Summary of Experimental Results 153 Post-mortem Notes of Calves 386 and 388 153-155 Virus Z. III. (Antelope.) Chart of the Inoculations “ aie 157 Description of the Virus and Summary of Experimental Results . 156, 159 Post-mortem Notes of Calves 510 and 516 and Pigs 170 and 174 as 159-161 Virus Z.IV. (Rhesus Monkey.) Chari of the Inoculations as 162 Description of the Virus and Summary of Experimental Results a oes oes 162 Virus Z.V. (Chimpanzee.) Chart of the Inoculations 163 Description of the Virus and Summary of Experimental Results 164 Virus Z. VI. (Cat.) Chart of the Inoculations ... ei 164-165 Description of the Virus and Summary of Experimental Results eae 165 151 INVESTIGATION OF VIRUSES OBTAINED FROM CASUAL TUBERCULOSIS OF VARIOUS MAMMALS. A few cases of tuberculosis casually cccurring in various mammals other than the ox, the horse, and the pig, have been investigated. The animals were a gnu, antelope,-rhesus monkey, chimpanzee, and cat. The tuberculous material from the gnu and the antelope was sent by Dr. Seligmann from the Zoological Society’s Gardens, and that from the cat by Sir John McFadyean ; the rhesus monkey and the chimpanzee had been obtained for purposes of experiment ; one was killed and the other died at the isolation station. SUMMARY OF RESULTS. From a Gnu which died of general tuberculosis in the Gardens of the Zoological Society a culture was obtained which resembled in growth on artificial media a human tubercle bacillus of Group II. Cultures were obtained from tuberculous lesions in three parts of the body of this animal—the lung, the mediastinal and the portal glands—and all were identical in cultural characters. The portal gland culture alone was tested on animals. This was tested on two calves subcutaneously in doses of 50 milligrammes and produced in 90 days local tuberculosis only. For rabbits it had relatively slight virulence compared with the bovine tubercle bacillus. It caused fatal general tuberculosis in two guinea-pigs. An Antelope, which was killed in the same Gardens when ill, was found to have very extensive chronic tuberculosis of the lungs; the lung substance was riddled with communicating cavities which contained breaking down caseous substance. Cultures were isolated from this lung direct and through Guinea-pig 2164 and Rabbit 1041. All these three strains were identical and resembled a culture of a human tubercle bacillus of Group II. The direct culture was tested in doses of 50 mg. subcutaneously in two calves and caused local tuberculosis only, after 96 and 119 days respectively. Two pigs, one fed with 100 milligrammes, the other inoculated subcutaneously with 50 milligrammes, developed very slight non-progressive tuberculosis after 100 days. For rabbits it had relatively slight virulence ; in four guinea-pigs it caused fatal general tuberculosis ; it was not virulent for fowls. The possibility of the tuberculosis in the Antelope being due to a mixed infection of tubercle bacilli was considered and two passage experiments on rabbits and guinea- pigs were carried out, beginning with the rabbit and guinea-pig inoculated with the original material. The longest experiment, in which 12 rabbits and 6 guinea-pigs were used, lasted nearly two years from October 2, 1906, to September 1, 1908. But no alteration in cultural characters or increase in virulence for the rabbit was observed in any of the cultures recovered. See Report on Modification Experiments. The two results recorded above are of interest in showing that two animals, the gnu and the antelope, belonging to the order of ruminants, have acquired casually severe tuberculosis due to infection with tubercle bacilli, which experimentally had only slight virulence for calves, and could not be distinguished culturally from human tubercle bacilli of Group II. Cultures of slight virulence for rabbits, with the cultural characters of human tubercle bacilli of Group II., have been obtained from a Chimpanzee and from a Rhesus Monkey, also the subjects of casual tuberculosis. — The Chimpanzee had acquired the disease by ingestion, the post-mortem examination showing several ulcers in the small intestine and enlargement and caseation of the mesenteric glands. The immediate cause of death was acute miliary tuberculosis of the lungs. The Rhesus Monkey had chronic pulmonary tuberculosis and slight disseminated disease. From the mesenteric gland of a Cat affected with casual tuberculosis, a culture was isolated which grew with difficulty on the various: differential media, resembling a bovine tubercle bacillus of Class 2. 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Original material—The specimens received were portions of the lung, liver, and dorsal mediastinal glands, a portal and a gastric gland, of a gnu which had died in the Gardens of the Zoological Society. DESCRIPTION OF MATERIAL RECEIVED AT THE FARM. The *haterial was taken from a female gnu which died of general tuberculosis on November 19, 1905. The piece of lung was quite solid, very dense and was composed of closely packed opaque white nodules embedded in a matrix of fibrous tissue ; the nodules varied in size up to a small pea and were dense, caseous, and slightly gritty in the centre; a few were softened. The dorsal mediastinal gland was very dense and similarly beset with fibro-caseous gritty nodules. The liver contained scattered hard caseo-calcareous nodules, ranging in size from a millet seed to a wheat -grain. The portal gland was enlarged and showed numerous hard discrete spherical whitish nodules, the largest the size of a pea; they projected above the surface and were surrounded by thin fibrous capsules from which the tough caseated tissue could readily be shelled ; the nodules were appreciably gritty from calcification. The gastric gland was enlarged and contained coalescing caseating tubercles. CULTURES. Cultures were obtained direct from the lung, mediastinal gland, and portal gland. Cultures were also isolated from Guinea-pig 1779 and Rabbit 520, inoculated with the lung and mediastinal gland respectively. Growth occurred readily in primary culture, both on glycerin serum and glycerinated potato.* : The subcultures obtained directly from the lung, mediastinal gland and portal gland, as well as those from the rabbit and from the guinea-pig, were tested in the usual way on the various differential media and all grew in an identical manner. The growth on glycerin serum was thick, wrinkled, and more luxuriant than that on pure serum ; on glycerin agar and potato it was thick, wrinkled, and slightly pigmented; on glycerin gelatin and broth opaque wrinkled and warty pellicles were formed. INOCULATION EXPERIMENTS. (a2) With the original material from a Gnu. Rabbits —Rabbit 519 was inoculated intraperi- toneally with an emulsion made from the lung. It was killed after 41 days, in good condition. The omentum was slightly thickened by reddish-grey tissue, beset with caseous foci; there were scattered whitish miliary tubercles on the mesentery, mesocolon and meso-caecum. There were yellowish tubercles up to 1 mm. in the spleen and a few grey tubercles in the liver and kidneys, those in the latter caseating. The lungs showed moderately numerous grey miliary tubercles with little central caseation. Caseous foci or patches in the inguinal, lumbar, coeliac, portal, and bronchial glands. Rabbit 520 was inoculated intraperitoneally with an emulsion of the mediastinal gland and died after 181 days. The omentum contained numerous caseo- calcareous and glassy tubercles; a few also on the mesentery. Spleen normal. Hach kidney showed about a dozen tubercles with caseous centres up to 3 mm. in diameter ; there were a few caseous streaks in the pyramids. The lungs did not quite collapse; they contained numerous irregular caseous gritty nodules and the margins of all the lobes were caseated. A few tubercles in the liver and one in each suprarenal body. Caseous foci in the mesenteric, bronchial, and axillary glands. Guinea-pigs. —Guinea-pig 1779 was_ inoculated intraperitoneally with an emulsion of the lung of the Gnu and died in 43 days of general tuberculosis. Guinea-pig 1780 was inoculated intraperitoneally with an emulsion of the mediastinal gland and died of general tuberculosis in 37 days. (b) With culture of Gnu material. Calves.—Calves 386 and 388 were inoculated sub- cutaneously each with 50 mg. of culture derived from the portal glind of the Gnu. They were killed in good health after 90 days. Each showed at the seat of inoculation a non-infiltrating cystic tumour filled with caseo-pus, and slight tuberculosis of the nearest lymphatic glands. The animals were otherwise healthy with the exception of a few transparent tubercles in the lungs and two tubercles in the mesenteric glands of Calf 388. Rabbits. — Two rabbits were inoculated éintra- venously, No. 759 with 1:0 mg., No. 758 with 0-1 mg. of culture derived from the portal gland. Rabbit 759 died in 16 days of acute tuberculosis (numerous tubercle bacilli in smears from the liver and spleen substance). Rabbit 758 was killed 91 days after inoculation, and had fairly severe chronic tuberculosis. The lungs contained numerous discrete tubercles aggregated in the margin and on the dorsal surfaces of the caudal lobes beneath the pleura. Spleen showed moderately numerous yellowish tubercles. The surfaces of the kidneys were pitted and beset with caseous foci, and the medullae contained caseous streaks; some of the calyces of the right were filled with caseo-pus. Moderately numerous grey tubercles in the liver. The majority of the lymphatic glands contained discrete caseous tubercles, and there were scattered tubercles in the muscles. Rabbit 761 died of acute tuberculosis 23 days after the intraperitoneal inoculation of 1:0 mg. of culture from the portal gland of the Guu. Rabbits 760 and 762 were inoculated subcutane- ously, each with 1°0 mg. of this culture, and were killed after 91 days. Both showed caseous cystic local lesions and soft caseous tubercles in the nearest glands. Their lungs contained scattered firm caseat- ing nodules up to a pea in size. There was no tuber- culosis elsewhere. Guinea-pigs.—Two guinea-pigs, Nos. 1905 and 1906, were each inoculated with 0:1 mg. of culture from the portal gland ; the former subcutaneously, the latter intraperitoneally. Guinea-pig 1905 died in 37 days, and Guinea-pig 1906 in 13 days, both of general tuberculosis. CALF 386 Virus ZI. Subcutaneous inoculation of culture derived from the portal gland of a gnu. Dose—50 milligrammes. Date—February 15, 1906. Weight at inoculation—67'57 kilogrammes. Killed when in good health—May 16, 1906. Clinical Notes. A week after the inoculation on the left side of the [Age about 14 weeks. ] [90 days after inoculation. ] neck, there was a slightly-raised local thickening measuring 7°5 by6cm. The adjacent prescapular and prepectoral glands were slightly enlarged. * Cultures with which the animal inoculations were made were grown primarily on egg and subcultivated on pure serum. 154 On the 23rd day the local swelling measured 10-9cm.; it was slightly raised, being about 2°5 cm. in thickness. The prescapular gland was enlarged, 6°5 cm. in length ; the prepectoral was also enlarged to about the size of a walnut. The swelling subsequently became soft and fluctuating. The calf showed no sign of ill-health during the experiment ; it was killed 90 days after inoculation. Temperature. There was a slight rise of temperature, commencing on the fourteenth day, reaching a maximum of 40-0°C. on the twentieth day, and lasting under three weeks. Subsequently the temperature was quite normal. Tuberculin Test. The calf was not tested subsequent to inoculation. Weights. Kilos. February 15,1906... ive oe 67°57 March 20, 1906... é sc .» 78:00 April 18, 1906 ... 85°25 May 15,1906 |. wees aes OBI Total gain of weight during eaperiment.—24 94 kilos. Average rate of gain per week.—1°91 kilos. POST-MORTEM EXAMINATION OF CALF 386. Carcass in very good condition. Local Lesion.—In the subcutaneous tissues on the left side of the neck there was an oval fluctuating swelling, measuring 12:5 by 10 by 7°5 cm., and weighing with the skin attached to it about 453 grammes. On section the swelling was found to be an abscess with fibrous walls, varying in thickness up to 6mm., and with purulent and caseo-purulent contents. The cavity was crossed by numerous fibrous trabeculae, the largest the diameter of a cedar-wood pencil; these and the internal surface of the cyst wall were covered by a thin layer of pale granulation tissue, over which were distributed numerous calcareous grains. The outer wall of the abscess was adherent to the muscles and to the skin, the latter being slightly thickened. Left Prescapular Gland.— The left prescapular gland was a little enlarged, and measured 6 by 3cm. by a little over 1cm.; on section towards one ex- tremity of the gland a small part of the cortex was firm, brownish, and translucent, and closely beset with calcareous foci ; the capsule about the middle of this patch showed a dense white fibrous thickening, which penetrated into the cortex in a wedge-shaped manner ; the rest of the gland, except for numerous petechial haemorrhages, appeared normal. 2 Right Prescapular Gland.—The right prescapular gland measured 5 by 2°5 by 1:°2cm., and was normal on section. Prepectoral Glands.—On the left side the spherical gland was enlarged to the size of a Barcelona nut ; on section the peripheral part of the gland was composed of tough caseated tissue, gritty around the: margins, the central part of brownish translucent and whitish fibroid tissue beset with calcareous foci. Two other glands on this side were normal. The glands on the right side were normal. Cervical Glands.—Normal. Thorax. Heart and Pericardium.—Normal. Pleura.—Normal. Lungs.—Normal. Bronchial and Mediastinal Glands.—Normal. Larynsz and Trachea.—Normal. Abdomen. Omentum and Peritoneum.—Normal. Spleen.—The spleen weighed 339 grammes, and was normal on the surface and on section. Liver—In the depth of the liver substance there was one minute yellow focus ; otherwise the liver was normal. Gall-bladder.—N ormal. Portal Glands.—Normal. Kidneys.—Both kidneys showed on the surface a number of minute grey foci, not apparently of a tuberculous nature. Suprarenal Bodies.—Normal. Special Organs. Tongue, Pharynx, and Tonsils.—Normal. Small and Large Intestines.—Normal. Testes.—Normal. Hyes.—Normal. Various Lymphatic Glands. Submaxillary, Parotid, Pharyngeal, Axillary, Coeliac, Renal, Lumbar, Iliac, Precrural, Pudic, Popliteal, ee Ischiatic, Gastric, Mesenteric, and Colic Glands. —Normal. CALF 388. Virus ZI. Subcutaneous inoculation of culture derived from the portal gland of a gnu. Dose—50 milligrammes. Date—February 15, 1906. Weight at Inoculation—62°58 kilogrammes. Killed when in good health—May 17, 1906. Clinical Notes. A week after the inoculation on the left side of the neck there was a flattened swelling measuring 75 by 6cm. The adjacent prescapular and prepec- toral glands were slightly enlarged. On the 15th day the swelling was elongated 16°5 by 8cm. The prescapular gland was definitely enlarged, 7°5 cm. in length. On the 54th day the swelling was cystic and measured 15 by 7:5 cm.; the prescapular gland measured 75 cm. in length, and the prepectoral gland was the size of a large pea. [Age about 13 weeks.] [91 days after inoculation.] No further change of any importance took place in the local conditions. The calf showed no symptoms of illness during the experiment. Temperature. There was a slight rise of temperature commencing on the 12th day, and lasting three weeks. The highest temperature recorded was 39:8°C. Subsequently the temperature was quite normal Tuberculin Test. The calf was not tested subsequent to inoculation. 155 Weights. Kilos. February 15,1906... tee see 62°58 March 20, 1906 a are wee 70°74 April 18, 1906 ... a6 ae wee 79°36 May 15,1906 ... ws ies we 88°43 Total gain of weight during experiment.—25°85 kilos. Average rate of gain per week.—1°98 kilos. POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion—At the seat of inoculation on the left side of the neck there was a prominent fluctuating swelling, measuring 14 cm. in length and 9 ecm. in breadth and thickness, and weighing with the attached skin 537 grammes. On section, the swelling was found to be a cyst, with fibrous walls up to 7 mm. in thickness, filled with greenish-yellow pus in which were suspended a few small, partially softened, caseous masses ; internally, the cavity was lined by a thin layer of pale smooth granulation tissue, the surface of which was covered with small gritty particles ; cross- ing the cavity, chiefly at the extremities, were a few bands of fibrous tissue. Left Prescapular Gland.—The left prescapular gland measured 6 by 4 cm. by about 2 cm.; on sec- tion, about half the substance of the gland was com- posed of calcareo-caseous material with a scanty matrix of brownish translucent fibroid tissue ; several of the more caseous parts showed foci of softening ; the rest of the gland was normal. Right Prescapular Gland.—The right prescapular gland measured 5 by 2°5 by 1:2 cm. and was normal on section. Prepectoral Glands.—On the left side one gland, the size of a swallow’s egg, was indurated, and on section composed of fibroid tissue beset with calcareo- caseous foci closely aggregated around the periphery of the gland. Other glands on this side as well as those on the right were normal. Cervical Glands.—Normal. Thorax. Heart and Pericardium.—Normal. Pleura.—Normal. Lungs.—The lungs were normal in general appear- ance. In the right anterior lobe under the pleura two minute (each about 0:5 mm. in diameter) glassy tubercles were seen. In each of the caudal lobes there were half-a-dozen transparent subpleural tubercles, the largest about 0°75 mm. in diameter ; the majority of the latter tubercles had a minute yellow calcareous centre. In the other lobes, and on section of the lung, no tubercles were seen. Bronchial and Mediastinal Glands.—Normal. Laryns: and Trachea.—Normal. Abdomen. Omentum and Peritoneum.—Normal. Spleen.—The spleen weighed 226 grammes and was normal on the surface and on section. Liver.—Normal. Gall-bladder.—Normal. Portal Glands.—Normal. Kidneys.—Normal. Suprarenal. Bodies.—Normal. Special Organs. Tongue, Pharynz, and Tonsils.—Normal. Small and Large Intestines.—Normal. Testes.—Normal. Eyes.—Normal. Various Lymphatic Glands. Submasillary, Parotid, Retro-pharyngeal, Aaillary, Coeliac, Renal, Lumbar, Iliac, Precrural, Popliteal, Gluteal, Ischiatic, Gastric and Colic Glands.—Normal. Mesenteric Glands.—A gland in the anterior part of the mesentery showed in the cortex a minute yellow point ; one in the posterior part contained a hempseed-sized grey tubercle, slightly opaque in the centre. ’ 156 VIRUS ZIII (Antelope). Original material—The specimen received was the lung of an antelope, (Oryx Beisa), which was killed ae the Zoological Society Gardens on October 2, 1906. The anima] had been at the Gardens since July 1, 1898. DESCRIPTION OF MATERIAL RECEIVED AT THE FARM. _ The specimen sent was the lung. It was exten- sively diseased, the greater part of it being riddled with communicating cavities, separated by trabeculae of congested tissue ; the cavities contained muco-pus in the centre, and a more solid caseo-pus on the walls ; the tissue around was grey and solid. Other parts of the lung were crepitant, but deeply congested and oedematous, showing also scattered grey foci. In many of the grey areas mentioned above, a caseous yellow network was seen. A smear from the muco-pus showed numerous tubercle bacilli, also other organisms. CULTURES. Cultures were obtained direct from the lung, and from Guinea-pig 2164 and Rabbit 1041 inoculated with the original material. All the strains were identical, and grew luxuriantly on the differential media; they produced thick pig- mented, wrinkled, and warty growths on glycerin agar and glycerinated potato, and thick opaque wrinkled pellicles on glycerin broth and glycerin gelatin. INOCULATION EXPERIMENTS. (a) With the original material from the Antelope. Rabbit.—Rabbit 1041 was inoculated intraperitone- ally with an emulsion made from the lung, and was killed 97 days later. There were a few tubercles in the omentum and on the peritoneum, and a purulent nodule loosely attached to the caecum. In the kidneys there were a few caseatizg tubercles up to a millet seed. Spleen and liver normal. The lungs were congested and contained numerous grey nodules with caseous streaks up to 3mm. in diameter. Caseous foci in lumbar, iliac, mediastinal and mesenteric glands. Guinea-pigs—Guinea-pig 2164 was inoculated sub- cutaneously, and Guinea-pig 2165 intraperitoneally, with an emulsion of the lung. Guinea-pig 2164 died in 43 days of general tuberculosis. Guinea-pig 2165 was killed after 76 days, and had chronic general tuberculosis. (6) With culture from Antelope material. Calves.—T wo calves, Nos. 510 and 516, were inocu- lated subcutaneously, each with 50mg. of culture. Calf 510 was killed when well after 96 days, and showed at the post-mortem examination a cystic caseo-purulent local lesion and slight tuberculosis of the nearest glands without any disease elsewhere. Calf 516, killed after 122 days, was similarly slightly affected with the addition of a focus in a mesenteric and in a submaxillary gland. Pigs.—Pig 174 was inoculated subcutaneously with 50 mg. of culture ; it died without obvious cause 111 days after inoculation. There was a cayeo-purulent cyst at the seat of inoculation, and caseation of the inguinal, precrura], and ventral mediastinal glands, without any disease elsewhere. Pig 170 was fed with 100 mg. ; when killed in good health after 119 days it showed discrete caseous gritty nodules in the submaxillary and mesenteric glands, and tubercles in the coeliac and portal glands. There were three minute tubercles in the lungs, but no tuberculosis elsewhere. Rabbits—The culture was inoculated into rabbits after it had been subcultured a total period of 90 days. Two rabbits were inoculated intravenously. Rabbit 1166 received 0-1 mg., and died in 74 days of general tuberculosis ; at the post-mortem examination the lungs did not collapse normally, and contained numercus irregular caseous nodules which at the caudal extremities became confluent and replaced the lung tissues by diffusely caseous masses. The spleen con- tained a moderate number of minute opaque tubercles, and the liver contained greyish white foci. Moderately numerous caseous tubercles up to a rape seed in size in the kidneys. Caseous tubercles in the majority of the lymphatic glands, in the muscles of thighs and back, and in the tips of the vertebral spinous. processes. Rabbit 1167 received 10mg. and died after 201 days. The lungs were congested and oedematous, and at the caudal extremities there were caseo-purulent cystic nodules replacing a considerable part of the suostance, and smaller whitish gritty nodules scattered about the rest of the lungs. The kidneys were pitied and scarred, and on section there were caseous streaks and tubercles in the pyramids; both pelves contained soft caseous substance. In the tissue beneath each nipple there were collections of caseo-purulent cysts, and the spinous processes and the crests of the iliac bones contained caseous areas. Two rabbits were inoculated intraperitaneally with culture. Rabbit 1169 received 10 mg. and died of acute tuberculosis in18 days. Rabbit 1168 wasinoculated with 1:0 mg., and was killed 391 days later ; in the omentum there was a single small caseous nodule, and a few on the peritoneum. The lungs did not collapse normally ; the caudal half of the right posterior lobe was almost replaced by cystic nodules filled with muco-pus, while the rest of the lung contained fairly numerous discrete grey nodules with caseous centres. There were a few submiliary tubercles in the kidneys. The spleen was normal. A few doubtful tubercles were seen in the liver. The testis and vas deferens on each side were greatly enlarged and replaced by soft caseous substance. Three rabbits were inoculated subcutaneously. Rabbit 1172 received 50 mg. and was killed 111 days later ; the local lesion was large and cystic, the skin over it being intact ; adjacent lymphatic glands were normal. The kidneys contained a few minute grey tubercles with opaque centres. The lungs showed on the surfaces moderately numerous grey nodules with caseous foci. There was no tuberculosis elsewhere. Rabbit 1171 received 10 mg. and was killed after 184 days ; it showed a cystic local lesion and a single tubercle in the lungs without any disease elsewhere. Rabbit 1170 died 93 days after the inoculation of 1-0 mg. and had slight tuberculosis. There was a caseo-purulent local lesion but no tuberculosis of the adjacent lymphatic glands. In the lungs, which were pink and crepitant, there were about two dozen caseating nodules ranging up to a wheat grain in size. Three rabbits were tested with the culture after it had been in cultivation for a total period of 126 days. Rabbits 1239 and 1240 each received 10 mg. intraperitoneally. Rabbit 1240 died in 21 days of acute tuberculosis. Rabbit 1239 was killed after 95 days having gained 1,000 grammes in weight. At the post-mortem examination the omentum contained caseo-purulent nodules up to a haricot bean in size, and there were similar nodules scattered about the peritoneal cavity. The surfaces of the kidneys were irregular and pitted and the cortices were closely filled with grey nodules with caseous centres ; there were soft caseous streaks in the pyramids, and the pelvis of one kidney was filled with caseo-pus. The lungs did not collapse normally and were severely diseased ; the dorsal surfaces of both- lungs were replaced by a network of caseous tissue ; the anterior lobes were reddish and solid. Only in the centre of the ventral portions of the caudal lobes was there any crepitant lung tissue remaining. There was no tuberculosis of spleen or liver. Caseation of lumbar and mediastinal glands and tubercles in the areolar tissues—axillary, inguinal, &c. Rabbit 1241 was fed with 50 mg. and killed after 245 days. At the post-mortem examination the intestines and mesenteric glands were normal. There was a minute tubercle in a kidney. The lungs showed at both anterior extremities grey wedge-shaped nodules beset with caseous foci; there were caseous, gritty, marginal nodules and scattered nodules in the rest of the substance. There was no tuberculosis elsewhere. (Cultures obtained from the lung proved identical with that used for feeding.) RABBITS. Method of | Duration of mber.| Dose. Inoculation. Life. Heat, 239 =| 10-0 mg. Intrap. D. 95 days | Chronic G. T. Severe in| lungs. 240 10-0 mg. Intrap. D. 21 ~«,. + | Acute T. | 241 | 50-0 mg. Fed K.245 ,, | Caseousnodules | in the lungs | : only. | i? | | GUINEA-PIG 2733. Intraperitoneal. Lung nodule from Rabbit 1241. | Died : 40 days. | P.M.—General tubercu- | losis. III. The culture was the 4th generation, 5 days old, and had in cultivation a total period of 721 days. FOWLS. Nu Method of | Duration of — Number.; Dose. Taoedlatiol. | Life. Result. 2 22 2 | 474 10:0 mg. Intrap. K. 77 days | No tuberculosis. 9 476 47:0 mg.| Intramus- | D. 14 ,, No tuberculosis. cular. 2 2A : 159 : Guinea-pigs.—T wo guinea-pigs were inoculated intraperitoneally with 1:0 mg. and 0-1 mg. respectively, the former dying in 12 days, the latter in 23 days with general tuberculosis. Two guinea-pigs were inoculated subcutaneously, one with 1:0 mg. the other with 0:1 mg. and both succumbed to general tuberculosis in 46 days and 60 days respectively. _Fowls.—Fow] 474 was inoculated intraperitoneally with 10 mg. of culture, and when killed in 77 days was perfectly healthy. CALF 510. Virus Z III. Subcutaneous inoculation with culture derived from the original material. Dose—50 milligrammes. Date of Inoculation—February 7, 1907. [Age about 9 weeks.] Killed when in good general health—May 14, 1907. [96 days after inoculation.] Clinical History. There was no sign of ill-health during the experi- ment. Temperature. The temperature was slightly raised during the first three weeks, highest point 39-9, after which it fell and remained normal until death. ' Tuberculin Test. April 30, 1907, 82 days after the experiment began. Dose of tuberculin (Bovine), 2;0cc. Rise of tem- perature, 1°5° C. Weights. Kilos. February 7, 1907 soe aids ee» 78°90 May 14, 1907 ... ans was es 10611 Total gain in weight.—27°21 kilos. POST-MORTEM EXAMINATION OF CALF 510. General condition good. Local Lesion—At the seat of inoculation on the right side of the neck there was a prominent fluctu- ating tumour measuring 15 by 10 by 9cm., and weighing 622 grammes. On section it was a cyst, tense with yellow purulent fluid containing small caseous flakes. The walls were lined by slightly roughened and pitted grey granulation tissue in which were a few gritty projecting points, and they were crossed by smooth fibrous cords up to 8mm. in thickness. Prescapular Glands.—The right prescapular gland measured 6°5 by 3 by 1-4cm. On section through the middle it appeared normal. A further superficial slice of cortex revealed two gritty foci and a small collection of caseo-calcareous tubercles the size of a wheat-grain. The left prescapular gland measured 5:2 by 2°7 by l-lem. On section it was normal. Prepectoral Glands.—Normal. Cervical Glands.—The right inferior cervical gland felt firm, and measured 1°3 by 1°8 by1lem. Onsection the greater part of the gland was filled with a fine caseo-calcareous network, and there was also an irregular caseous patch with calcareous streaks, mainly firm but softening at one point near the cortex. Other cervical glands were normal. Spleen.—The spleen weighed 339 grammes, and on section was normal, All other organs and glands were normal. Microscopical examination of a smear preparation from the emulsion of cervical gland showed a few tubercle bacilli. CALF 516. Virus ZIII. Subcutaneous inoculation with culture derived from the original material. Dose—50 milligrammes. Date of Inoculation—February 7, 1907. [Age about 9 weeks.] Killed when in good general health—June 10, 1907. [123 days after inoculation. ] Temperature. Slightly raised during earlier part of experiment ; highest point 39:7; it quickly fell and remained normal until death. Tuberculin Test. April 30, 1907, 81 days after the experiment began. - Dose of tuberculin, 2‘0cc. (Bovine). Rise of tem- perature, 1:1° C. Weights. Kilos. February 7, 1907 nee at we 56°23 June 10, 1907 ... tae sea 64:85 Total gain in weight—8'52 kilos. 34684 POST-MORTEM EXAMINATION OF CALF 516. General condition good. Local Lesion.—At the seat of inoculation in the subcutaneous tissues on the right side of the neck there was a tumour measuring 8 by 6 by 4cm. On section it was a moderately thick-walled fibrous cyst filled with mucoid caseo-pus and serous fluid; the internal walls were roughened and lined by grey granulation tissue, which was beset with gritty foci. Prescapular Glands.—The right prescapular gland measured 5 by 2'4 by lcm. On section it contained just beneath the capsule a small collection of soft slightly gritty caseous nodules up to a millet eeed in size, and an occasional opaque focus in the rest of the gland. 2B 159 3 Guinea-pigs.—T wo guinea-pigs were inoculated intraperitoneally with 1-0 mg. and 0-1 mg, respectively, the former dying in 12 days, the latter in 23 days with general tuberculosis. Two guinea-pigs were inoculated subcutaneously, one with 1:0 mg. the other with 0-1 mg. and both succumbed to general tuberculosis in 46 days and 60 days respectively. Fowls.—Fowl 474 was inoculated intraperitoneally with 10 mg. of culture, and when killed in 77 days was perfectly healthy. CALF 510. Virus ZIII. Subcutaneous inoculation with culture derived from the original material. Dose—50 milligrammes, Date of Inoculation—February 7, 1907. [Age about 9 weeks.] Killed when in good general health—May 14, 1907. [96 days after inoculation.] Clinical History. There was no sign of ill-health during the experi- ment. Temperature. The temperature was slightly raised during the first three weeks, highest point 39°9, after which it fell and remained normal until death. ' Tuberculin Test. April 30, 1907, 82 days after the experiment began, Dose of tuberculin (Bovine), 2:0cc. Rise of tem- perature, 1:5° C. Weights. Kilos. February 7, 1907 oa sive oo» 78°90 May 14, 1907 ... oe ous ee 106711 Total gain in weight.—27°21 kilos. POST-MORTEM EXAMINATION OF CALF 510. General condition good. Local Lesion.—At the seat of inoculation on the tight side of the neck there was a prominent fluctu- ating tumour measuring 15 by 10 by 9cm., and weighing 622 grammes. On section it was a cyst, tense with yellow purulent fluid containing small caseous flakes. The walls were lined by slightly roughened and pitted grey granulation tissue in which were a few gritty projecting points, and they were crossed by smooth fibrous cords up to 8mm. in thickness. Prescapular Glands.—The right prescapular gland measured 6°5 by 3 by J-4cm. On section through the middle it appeared normal. A further superficial slice of cortex revealed two gritty foci and a small collection of caseo-calcareous tubercles the size of a wheat-grain. The left prescapular gland measured 5:2 by 2:7 by 1-1em. On section it was normal. Prepectoral Glands.—Normal. Cervical Glands.—The right inferior cervical gland felt firm, and measured 1°3 by 18 by1lcm. Onsection the greater part of the gland was filled with a fine caseo-calcareous network, and there was also an irregular caseous patch with calcareous streaks, mainly firm but softening at one point near the cortex. Other cervical glands were normal. Spleen.—The spleen weighed 339 grammes, and on section was normal, All other organs and glands were normal. Microscopical examination of a smear preparation from the emulsion of cervical gland showed a few tubercle bacilli. CALF 516. Virus Z III. Subcutaneous inoculation with culture derived from the original material. Dose—50 milligrammes. Date of Inoculation—February 7, 1907. [Age about 9 weeks.] Killed when in good general health—June 10, 1907. [123 days after inoculation. ] Temperature. Slightly raised during earlier part of experiment ; highest point 39:7; it quickly fell and remained normal until death. Tuberculin Test. April 30, 1907, 81 days after the experiment began. - Dose of tuberculin, 2°0cc. (Bovine). Rise of tem- perature, 1°1°C. Weights. Kilos. February 7, 1907 site se we 56°23 June 10, 1907 ... aie 06 64:85 Total gain in weight—8°52 kilos. 34684 POST-MORTEM EXAMINATION OF CALF 516. General condition good. Local Lesion.—At the seat of inoculation in the subcutaneous tissues on the right side of the neck there was a tumour measuring 8 by 6 by 4cem. On section it was a moderately thick-walled fibrous cyst filled with mucoid caseo-pus and serous fluid; the internal walls were roughened and lined by grey granulation tissue, which was beset with gritty foci. Prescapular Glands.—The right prescapular gland measured 5 by 2:4 by 1cm. On section it contained just beneath the capsule a small collection of soft slightly gritty caseous nodules up to a millet seed in size, and an occasional opaque focus in the rest of the gland. 2B 160 The left prescapular gland measured 5 by 2:4 by 12cm. On section it was normal. Prepectoral Glands.——Normal. Cervical Glands.—The right inferior cervical gland measured 1:8 by 1:3 by 1 cm., and on section resembled the right prescapular gland. In the cortex of the right superior cervical gland there were two irregular soft caseous gritty tubercles. Lungs.—The lungs showed on the surface an occa- sional congested lobule without any sign of tuberculosis. On section there were numerous congested patches slightly oedematous, but air-containing. Lymphatic Glands.—In an anterior mesenteric gland there was a single caseous gritty focus. There were two opaque foci in the left submazillary gland. There was no sign of tuberculosis in any other organ or gland. Microscopical examination of a smear preparation of the emulsion of right inferior cervical gland showed numerous tubercle bacilli, PIG 170. Virus Z III. Fed with culture derived from the original material. Dose—100 milligrammes. Date of Feeding—February 8, 1907. [Age about 10 weoks.] Killed when in good general health—June 7, 1907. Temperature. Normal throughout. Tuberculin Test. April 30, 1907, 81 days after the experiment began. Dose of tuberculin (Bovine) 2:0 cc. Rise of tem- perature 1:0° C, Weights. Kilos. . February 8, 1907 “es nee 10°43 June 7,1907 ... ca on 32°19 Total gain in weight.—21°76 kilos. POST-MORTEM EXAMINATION OF PIG 170. General condition good. Tongue.—Normal. Pharynx.—Normal. Tonsils—In each tonsil there were a few soft opaque yellow foci (? secretion), no tubercle bacilli found. Submaaillary Glands.—The submaxillary glands on the right side, anterior to the salivary gland, the largest measuring 2 by 8 cm. in thickness, contained several softening caseous nodules with fine gritty streaks replacing a considerable part of the gland substance. In the rest of the gland substance there were discrete caseo-calcareous tubercles. The nodules and tubercles were easily picked out. One of the corresponding glands on the left side was enlarged and similarly beset with nodules; the others showed a few discrete yellowish tubercles the size of rape seed. Cervical G'lands.—Several smaller glands on each side of the neck posterior to the submaxillary salivary glands were normal. Parotid Glands.—Normal. [119 days after the experiment began.] Abdomen. Intestines.—Normal. Mesenteric Glands.—The mesenteric glands all con- tained discrete softening caseous slightly gritty nodules, the majority the size of a millet seed, but ranging up to a hemp seed. Coeliac Glands.—Two showed a few opaque yellow gritty foci. A small gland attached to the pancreas showed similar foci. Spleen.—Normal. Liver.—On the surface of the liver there were two grey points. On section the substance was normal. Portal Glands.—In one portal gland there was a yellowish gritty submiliary tubercle Kidneys.—Normal. Suprarenal Bodies.—Normal. Thorax. Lungs.—On the surface of the right lung just beneath the pleura there were two grey tubercles, one opaque in the centre. On the surface of the left lung there were also two. On section through the sub- stance, a single yellowish gritty tubercle was found in the left lung. Bronchial Glands.—Normal All other organs and lymphatic glands were normal. Microscopical Examinations. (Smear Preparations ) Tonsil (foci).—No tubercle bacilli. : Mesenteric Gland (nodule).—No tubercle bacilli. Coeliac Gland (focus).—No tubercle bacilli. Liver (focus).—No tubercle bacilli, 161 PIG 174. Virus Z IL. Subcutaneous inoculation with culture derived from the original material. Dose—50 milligrammes. Date of Inoculation—February 8, 1907. [Age about 10 weeks. ] Died—May 30, 1907. [111 days after inoculation.] Temperature. The temperature was practically normal through- out (39°6 highest point). Tuberculin Test. April 30, 1907. [81 days after the experiment began.] Dose of tuberculin (Bovine), 2:0 cc. No reaction. Weights. Kilos. February 8, 1907 aise ie ee 13°60 May 30,1907 ... se ts ee = 37:18 Total gain in weight.—23'58 kilos. t POST-MORTEM EXAMINATION OF PIG 174. | Local Lesion.—In the subcutaneous tissues of the abdomen to the left of the middle line there was a rounded slightly prominent tumour, 3 cm. in diameter. On section it was found to be a cyst with smooth internal walls filled with yellow caseo-pus. In the subcutaneous tissues around there were some scattered discrete fibro-caseous tubercles. Inguinal G'lands.—One left inguinal gland measured 2:3 by 1°5 cm. and consisted almost throughout of pinkish caseo-necrotic tissue slightly gritty in parts. Four smaller glands were normal. The right inguinal glands were normal. Precrural Glands.—The left precrural gland was two-thirds replaced by pinkish caseo-necrotic sub- stance; the other third consisted of soft yellow slightly gritty material. The right precrural gland was normal. Diaphragm.—On the peritoneal surface of the diaphragm on both sides of the central tendon there were several flattened grey translucent nodules, some slightly opaque in the centre and about 1 mm. in diameter. On the pleural surface on the left side there were two similar nodules. Lungs.—The lungs were congested and oedematous but floated in water. Just beneath the pleura on the surface were scattered minute glassy foci, a few of which showed minute opaque centres. 5 Bronchial Glands.—Normal. Ventral Mediastinal Glands.— The left ventral mediastinal gland was enlarged, congested, and partly replaced by a caseo-necrotic slightly gritéy nodule. The right gland was normal. Spleen.—Normal. Liver.—The convex surface of the liver was covered with a lace-like network of fine greyish white fibres which could be picked-up and stripped away. The substance of the liver was deeply congested. Kidneys.—The pelvis of the left kidney was dilated and the cortex contained two cysts filled with fluid. The kidneys showed no tuberculous lesions. Coeliac Glands.—Normal. Iliac Glands.—In a left iliac gland at one extremity the substance was a little firm and showed some opaque yellow streaks. Brain.—Normal. There was no sign of tuberculosis elsewhere in the body. Microscopical Examinations. (Smear Preparations.) Lung (foci).—No tubercle bacilli. E. of Left Precrural Gland—A few tubercle bacilli. Iliac Gland (foci).—One doubtful tubercle bacillus. Liver (substance).—No tubercle bacilli. 34684 2B2 162 VIRUS Z IV (Rhesus Monkey). CULTURE INOCULATIONS. LUNG. May 4, 1907. The culture used was derived from the original material. The 13th generation ; 22 days old, and had been in cultivation a period of 463 days. GUINEA-PIGS. RABBITS. Method of| Duration Method of| Duration Number.} Dose. | Inocula- of Result. Number.) Dose. Inocula- of Result. tion. Life. tion. Life. 2468 | 1:0 mg.| Intrap. | D.25 days| G. T. 1326 |10°0mg.) Intrav. | D. 22days| G.T. 2467 | 1:0 mg.| Subcut. |D. 108, |G. T. 1325 | 1:0 mg.} Intrav. |K.122 ,, elven 1328 |30-0mg.| Subent. |K.122 ,, |Local’ T. and case- ous no- dules in lungs. 1327 | 100mg.) Subcut. | K.122 ,, | Local T. and a single nodule in the lung. VIRUS Z IV (Rhesus Monkey). Original material—The tuberculous material was obtained from an adult Macacus Rhesus which had been, specially caught in the Indian jungle for the Royal Commission on Tuberculosis. noticed to have a cough and was therefore killed on J anuary 26, 1906. Post-mortem Examination. — The lungs showed apical and marginal emphysema and were marked on the surface by numerous scars. The lung sub- stance was filled with hard irregular nodules ; these on section were caseous and many of them were softened in the centre, forming cavities filled with caseo-pus. In the right lung there was a large air- containing cavity with fibrous walls lined by caseo- pus. Each of two bronchial glands contained a caseous nodule. In the spleen there was a caseous nodule and one in the left kidney. The liver showed sparsely scattered miliary tubercles. In smear pre- parations from the spleen, lung and bronchial gland, long beaded tubercle bacilli were found. CULTURES. A culture was isolated from the lung, and this grew luxuriantly on the differential media, forming on glycerin agar and glycerinaied potato rich yellow wrinkled and warty layers. INOCULATION EXPERIMENTS. (2) With original material from Rhesus Monkey. No inoculation experiments were performed with the original material. (6) With culture from this material, Rabbits—Rabbit 1326 was inoculated intravenously with 10 mg. of culture and died in 22 days of general tuberculosis. The lungs did not collapse normally, and were closely filled with grey tubercles with yellow centres. Mediastinal glands partly caseous. Moderately numerous grey tubercles were visible in the On arrival it was cortices of the kidneys. spleen or liver. Rabbit 1325 received 1:0 mg. intravenously and when killed after 122 days had gained in weight and showed slight general tuberculosis. The lungs were crepitant, and on the surfaces beneath the pleura were minute translucent tubercles with opaque centres. The surfaces of the kidneys were pitted and showed discrete translucent nodules from which soft caseous streaks passed through the pyramids ; the pelvis of the right contained a little caseo-pus. In the tissue beneath each nipple there was soft yellow caseous substance which microscopically showed moderately numerous tubercle bacilli. No disease elsewhere. Rabbits 1327 and 1328 were inoculated subcu- taneously, the former with 10 mg. the Jatter with 30 mg., and were killed after 122 days, Rabbit 1327 showed a cystic local lesion, a soft caseous nodule in ‘an axillary gland and a single encapsuled caseous miliary tubercle in the right lung. Rabbit 1328 had a larger caseo-purulent local cyst, but the axillary glands were normal. The lungs contained, mainly on the surface beneath the pleura, moderately numerous irregular flat translucent nodules caseating in the centre ; the nodules were confluent at the anterior and posterior extremities and there were caseating marginal nodules. With the exception of a soft caseous nodule in a bronchial gland there was no tuberculosis else- where. No foci could be seen in the Guinea-pigs. —Guinea-pig 2468 died of general tuberculosis in 25 days after the intraperitoneal inoculation of 1:0 mg. " Guinea-pig 2467 was inocu- lated subcutaneously with 1:0 mg. and died in 108 days; the tuberculosis was of the chronic type and the lymphatic glands were but little enlarged and showed only slight caseation, except for those nearest to the local lesion. 163 I VIRUS ZV (Chimpanzee), CULTURE INOCULATIONS. MESENTERIC GLAND. May 4, 1907. Inoculated with culture derived from the original maierial. The culture used was the 10th generation, a total period of 341 days, GUINEA-PIGS, 23 days old, and had been in cultivation SEPTEMBER 24, 1908. Inoculated with culture derived from the original material. The culture used was the 4th generation, 14 days old, and had been in cultivation a total period of 850 days. Method of s Number.) Dose. Inocula- | Duration of Result. : ti Life. ion. 480 10-0 mg. | Intrav. | K.77 days | Few tuber- cles in lungs. 482 10°0 mg. | Intrav. | K.46 _,, Minute tu- bercles in lungs. 478 | 100:0 mg. | Intramusc.| K. 84 _,, Local T. Tubercle in liver, Inoculated with an emulsion from the lung of Fowl 482. NovemBer 10, 1908. 4 , Rabbit 1918 Method of : Animal. Tnocula- Pes of Result. : e. tion. Intrav. | K. 58 days |2 caseous nodules in lungs. Subcut. | K.41 ,, | Harly G. T. ‘ Guinea-pig 3347 RABBITS. Method of : Method of z Number.) Dose. | Inocula- | Duration of | pou, Number] Dose. | Inocula- | Duration of) posit, tion Life. ; Life. tion. 2466 )1:0 mg. | Intrap. | D. 22 days| G. T. 1322 |10°0mg.| intrav. | D. 19 days| Acute T. 2465 |1:0mg. | Subeut. | D.111_,, Ghpaae 1321 10mg.) Intrav. | K.122 ,, | Chronic Ls G. T. 1324 |30°Omg.| Subcut. | K.122 ,, | Local T. 4 caseous nodules in the lungs. 1323 |10°Omg.} Subcut. | K.122 ,, | Local T. only. II. FOWLS. 164 VIRUS Z V (Chimpanzee). station. Original material—This chimpanzee was acquired for experimental purposes, and died at the isolation: The post-mortem examination was made on May 28, 1906, and revealed acute miliary tuberculosis, the: lungs and kidneys being closely filled with grey tubercles. thickened bases and serpiginous outlines ; in the large intestine there was one. In the small intestine there were four ulcers with The mesenteric glands were adherent to each other ; they were enlarged and to a great extent replaced by softening caseous substance. - CULTURE. A culture was isolated directly from a mesenteric gland. On the various differential media the culture produced characteristic luxuriant pigmented growths, resembling in this respect a human tubercle bacillus of Group IT. INOCULATION EXPERIMENTS. (a) With the original material from Chimpanzee. Guinea-pigs. — Two guinea-pigs were inoculated intraperitoneally, Guinea-pig 2050 with an emulsion of the lung, and Guinea-pig 2051 with an emulsion of the mesenteric gland. Guinea-pig 2050 died in nine days of a subacute non-tuberculous infection. Guinea- pig 2051 died in 26 days of general tuberculosis. (6) With culture from mesenteric gland of Chimpanzee. Rabbits.—Rabbit 1322 was inoculated intravenously with 10 mg., and died in 19 days. The lungs did not collapse normally, and were closely filled with minute grey translucent tubercles. Numerous grey foci were visible in the cortices of the kidneys. None could be seen in the spleen or the liver. Tubercle bacilli were sparse in smear preparations made from the spleen, liver, lung, and kidneys. Rabbit 1321 was killed 122 days after the éntra- venous inoculation of 1:0mg.; in the lungs there were two minute transparent tubercles with opaque centres. The kidneys contained in the cortices a few caseous nodules ; in the medulla of each kidney there were about fifteen yellow caseous miliary tubercles, a few gritty. The right elbow joint and the fifth, sixth, and seventh costo-vertebral-articulatioris contained soft caseous substance. No tuberculosis elsewhere. Two rabbits were inoculated subcutaneously and were killed in good health 122 days later. Rabbit 1323 received 10 mg. and showed only a soft caseous nodule in the skin, over which was a small ulcer. Rabbit 1324, inoculated with 30 mg., developed a cystic local lesion and four caseating nodules in the lungs up to about a hemp seed in size. Guinea-pigs. — Two guinea-pigs were inoculated No. 2466 intraperitoneally with 1°0 mg. and No. 2465 subcutaneously with 1-0 mg. Guinea-pig 2466 died in 22 days with general tuberculosis. Guinea-pig 2465 succumbed to chronic general tuberculosis 111 days after inoculation; the spleen was enlarged and contained scattered calcareous granules. The right kidney contained a grey nodule in the cortex extending through the medulla to the pelvis where its apex was yellow and soft. The liver, not enlarged, contained a network of grey foci. In the lungs there were numerous discrete tubercles, the majority perfectly translucent. The bronchial, lumbar, iliac and portal glands contained calcareous granules easily picked out. Fowls.—Fowls 480 and 482 were each inoculated intravenously with 10mg. of culture. Fowl! 480. was killed after 77 days, and Fowl 482 after 46 days ; both showed a few minute tubercles in the lungs, and no disease elsewhere. Fowl 478, inoculated intramuscu- larly with 100mg., was killed 84 days later, and showed local tuberculosis only. VIRUS Z VI (Cat). CULTURE INOCULATIONS. MESENTERIC GLAND. I,.—SePTEMBER 10, 1907. Inoculated with the culture derived from the original material. The culture used was the 5th generation, 21 days old, and had been in cultivation a total period of 105 days. GUINEA-PIGS. ile Method of | Duration Method of | Durati Number. | Dose. Inoculation. | vf Life. Result. Number. | Dose. Tnosulatian. of ‘Life. Result. | 2614 | Ol mg.| Intrap. D. 20 days| G. T. 1456 | 1:0 mg. Intrav. D. 17 days | G. T. 2613 | O-1mg.} Subcut. D.60 , | GT. 1452 | O-lmg.| Intrayv. D.19 ,, | GT. 1453 [ 0:1mg.| Intrav. D.20 ,, | GT. 1455 |10-Omg.| Subeut. | Killed (ill)} @. T. 71 days. 1454 | 1:0mg.| Subcut. | D. 44 days| G. T. 165 VIRUS Z VI (Cat)—continued. II. INOCULATIONS WITH EMULSION OF ORIGINAL MATERIAL, MESENTERIC GLAND. May 29, 1907. Method of | Duration Number, | Tnoenlation, | of Lite ae Rabbit 1356 Intrap. D. 40 days | G. T. Rabbit 1355 Subcut. K. 58 ,, Early G. T. Guinea-pig 2507 | Subcut. D.60 ,, G. T. Guinea-pig 2508 Intrap. D. 98 =, G. T. Emulsion of Lung of Rabbit 1355. JULY 26, 1907. Method of | Duration Number. Inoculation of Life. Result, Fowl! 290 Intrap. K. 129 days | Slight pe ri- toneal tuber- culosis. Fowl 292 Intramus- | K.129 ,, | Local tubercu- cular. losis only. VIRUS Z VI (Cat). Original material—The specimen received was the enlarged mesenteric gland from a case of casual tuberculosis in a cat sent by Sir John McFadyean, May 28, 1907. The gland measured 4°6 by 3°5 by 25cm. ‘On section it was found to consist of three parts fused together, and was entirely composed of yellow caseous substance softening in parts. CULTURE. A culture was isolated direct from the mesenteric gland. Its characters were those of a bovine tubercle bacillus belonging to Class II. On glycerin serum it produced a moderately thick creamy layer, the surface of which was ribbed ; on glycerin-agar after ten weeks there was a thin grey translucent layer, which did not increase subsequently; the final growth on glycerinated potato consisted of numerous minute white raised colonies. On glycerin gelatin in ten weeks the surface was three-quarters covered by a thin transparent pellicle dotted over with greyish white foci. INOCULATION EXPERIMENTS. (a) With the original material from the Cat. Rabbiis—Rabbit 1356 was inoculated intraperi- ‘toneally with an emulsion of the mesenteric gland, and died in 40’days of general tuberculosis. The peritoneum and mesentery were studded with caseous nodules and the omentum was thickened. The spleen and kidneys contained moderately numerous tubercles -and the lungs were closely filled with caseous tubercles. Rabbit 1355, inoculated subcutaneously, was killed after 58 days, and showed early general tuberculosis ; the lungs contained numerous discrete caseating nodules, Smear preparations showed very numerous tubercle bacilli. Guinea-pigs.—T wo guinea-pigs were inoculated, one intraperitoneally, the other subcutaneously. Both died of general tuberculosis in 98 and 60 days respectively. Fowls. —'Two fowls, Nos. 290 and 292, were inoculated, the former intraperitoneally, the latter intramuscularly, with an emulsion made from the lung of Rabbit 1355 (See above). Fowl 290, killed 129 days later, showed slight peritoneal tuberculosis only. Fowl 292 also killed after 129 days, was healthy except for slight tuber- culosis at the seat of inoculation. (0) With culture from Cat’s mesenteric gland. Rabbits—Three rabbits 1456, 1452, and 1453, were inoculated intravenously, and died of general tubercu- losis. Rabbit 1456 received 1:0 mg. and died in 17 days. Rabbits 1452 and 1453 received 0:1 mg., and died in 19 and 20 days respectively. Two rabbits were inocu- lated subcutaneously, No. 1454 with 1:0 mg. and No. 1455 with 10 mg.; the former died of general tuber- culosis after-44 days, the latter after 71 days. Guinea-pigs.—Guinea-pig 2614 died in 20 days after the intraperitoneal inoculation of 0:1 mg., and Guinea- pig 2613 in 60 days after the subcutaneous inoculation of 0-1 mg., both of general tuberculosis. 167 INVESTIGATION OF AVIAN TUBERCLE BACILLI OBTAINED FROM BIRDS AND SWINE. BY F. GRIFFITH, M.B. 34684 ae 168 INVESTIGATION OF AVIAN TUBERCLE BACILLI OBTAINED FROM BIRDS AND SWINE. TABLE OF CONTENTS. PAGE Introduction 170 Summary of Results ee we 170 Table of Origins of the Avian Vedas cued teers Beds ss ons we = LTA (For the Table of Origins of the Avian Viruses obtained from Swine, see the Report on Swine Tuberculosis, Volume III.) Cultural Characters of the Avian Tubercle Bacillus... 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A II. Subcutaneous Inoculation of culture derived from the original material. Dose—50-0 milligrammes. Date of Inoculation—May 22, 1906. [Age 5 weeks.] Killed when in good health—August 31, 1906. Clinical Notes. Eighteen days after inoculation on the left side of the neck there was a firm subcutaneous thickening measuring 63 by 5 cm. by about 1 cm. in greatest thickness. On the 35th day the thickening had diminished a Temperature. , Mey JUNE [101 days after inoculation. ] little in extent. The left prescapular was probably normal in size ; it was 5 cm. long. Subsequently the local thickening was replaced by a small oval tumour, and the prescapular gland became definitely enlarged. The calf remained in good health during the experiment. (Chart to July 10.) JOLY From July 10 to August 31 the temperature remained normal. Weights. Kilos. May 22 tas ave ils 28°57 July 17 wee ius ais aes 47°16 August 31... se ae ive 61:22 Total gain of weight during eaperiment.—32°60 kilos. Average rate of gain per week.—2'2 kilos. Local Lesion.—At the seat of inoculation, on the left side of the neck, was an oval tumour measuring 5 by 4°5 by 2 cm. Left Prescapular Gland.—The left prescapular gland measured 6:3 by 3°8 by 25 cm. On sectioa rather more than half tke cortex was replaced by grey trans- lucent necrotic tissue with opaque gritty foci, and about half the medulla was filled up with a similar mass. The Right Prescapular, Prepectoral and Agillary Glands were normal. Thorax. Pleura.—Normal. Lungs.—Scattere1 throughout the lungs were minute congested foci. In addition, in the right middle lobe there were six grey tubercles with slightly opaque centres ; the largest, 1°5 mm. in diameter, on section contained a minute calcareous centre. Thoracic Glands.—The bronchial glands on each side contained in their cortices scattered irregular yellowish gritty tubercles up to 1:5 mm. in diameter. The long mediastinal gland contained similar tubercles. One of the other mediastinal glands contained a tubercle. Heart.—Normal. 34684 Abdomen. Omentum and Peritoneum.—Normal. Spleen and Liver—Normal. Portal Glands—Three portal glands contained a few yellow gritty foci. Kidneys.—Normal. Suprarenals.—The right contained in the cortex one pinhead-sized yellow caseo-calcareous tubercle. The left was normal. Renal, Lumbar, and Iliac Glands.— Normal. Alimentary Tract. Tongue, Pharynx, Palate, Tonsils.—Normal. Pharyngeal, Submawillary, Hyoid, and Parotid Glands.—Normal. Stomach.—Normal. Intestines.—1n the first foot of the small intestine there were eight soft yellow tubercles up to a pinhead in size. In the rest of the small intestine three were counted. : The large intestine was normal, Mesenteric Glands.—Many of the mesenteric glands showed punctiform hemorrhages, and in some of these close beneath the capsule could be seen very small irregular yellow foci. In one gland there was a distinctly yellow necrotic patch about the size of a wheat grain. Many of these small foci had a yellow periphery with a transparent centre, and they cracked on pressure. Tleo-colic and Colic Glands.—Normal. Testes. —Normal. 2T2 292 Various Lymphatic Glands. Precrural, Popliteal, Gluteal, Ischiatic, Pudic.— Normal. Microscopical Examination. Emulsion of Mediastinal Gland.—No tubercle bacilli seen. Tubercle from Lung.—No tubercle bacilli seen. Focus from Mesenteric Gland.—No tubercle bacilli seen. : Caseous patch from Mesenteric Gland.—No tubercle bacilli seen. Tubercle from Small Intestine.—Four tubercle bacilli seen. Lubercle from Suprarenal—Numerous bacilli. tubercle GOAT 38. Virus A II. Fed once with culture derived from the original material. Dose—100-0 milligrammes. Date of Feeding—March 1, 1907. Killed when in good health—June 19, 1907. Clinical History. The animal showed no sign of ill health during the experiment. Temperature. The temperature rose to 401° C. on the 20th day and fell to 39:2° C. three days later; subsequently it varied between 38:9° C. and 39°6° C. Tuberculin Tests. April 9, 1907. Dose 2:0 ce. of bovine tuberculin. [39 days after the experiment began.] Positive re- action. Rise of temperature 2°1° C. April 30,1907. Dose 4:0 cc. of avian tuberculin. [60 days after the experiment began.] Positive re- action. Rise of temperature 1'8° C. June 13,1907. Dose 2:0 cc. of bovine tuberculin. [104 days after the experiment began.] Positive re- action. Rise of temperature 1:2° C. POST-MORTEM EXAMINATION. General condition good. Tongue.—Normal. Tonsils.—Normal. Larynx and Pharyne.—Normal. Submaxillary Glands.—Normal. Pharyngeal Glands.—Normal. Cervical Glands.—Normal. Abdomen. Peritoneum and Omentum.—Normal. Intestines.—In the small intestine there were two soft caseous slightly gritty nodules, the largest the size of a millet seed. The large intestine was normal. Coeliac Glands.—The glands situated dorsally and to the left of the cardiac orifice of the stomach con- tained a moderate number of irregular yellowish [Age about 10 months.] [110 days after feeding. ] nodules and small patches composed of soft caseous slightly gritty substance. Mesenteric Glands——The mesenteric glands were normal with the exception of one terminal gland which contained in the cortex soft yellow caseous slightly gritty nodules easily shelled out from the gland substance. Spleen.—The spleen appeared normal. Liver.—Normal. Portal Glands —Normal. Kidneys.—Normal. Suprarenals.—Normal. Lumbar, Iliac, and Renal Glands.—Normal. Thoras. Pleura.—Normal. Lungs.—In the right posterior lobe there was one small firm’ grey patch and in the rest of the lung an occasional grey nodule which did not appear to be tuberculous. Bronchial and Mediastinal Glands.—Normal. Heart.—Normal. Various Lymphatic Glands. Parotid, Prescapular, Axillary, Precrural, Popliteat, and Gluteal Glands.—Normal. Microscopical Examination. (Smear Preparations.) Lung (grey patch).—No tubercle bacilli seen. Coeliac Gland (gland near cardiac orifice of stomach). —No tubercle bacilli seen. Mesenteric Gland (emulsion).—Two tubercle bacilli seen. Small Intestine (nodule No. I).— One tubercle bacillus seen. Small Intestine (nodule No. II).—? One tubercle bacillus seen. 293 CHIMPANZEE 20. Virus A II. Subcutaneous inoculation of culture derived from the original material. Dose—50-0 milligrammes. Date of Inoculation—May 22, 1906. Died—March 25, 1908. Clinical Notes. During the three weeks immediately following the inoculation the animal seemed at times a little unwell, and its appetite was not so good as usual. On the 18th day there was a slightly raised firm swelling at the seat of inoculation measuring 5 by 4 cm. An axillary gland on the right side appeared to be enlarged. On the 35th day after inoculation the local swelling was fluctuating and measured about 5 by 4.cm.; the skin over it was thinned and reddened and showed two small ulcers discharging yellowish pus. ‘The animal was well. Subsequently the discharge ceased and the swelling diminished in size being finally represented only by a small raised thickening. The animal subsequently showed no sign of ill [673 days after inoculation. ] health until shortly before its death when it developed diarrhoea. Tuberculin Test. February 8, 1907. began. ] reaction. [262 days after the experiment Dose of tuberculin (bovine) 1:0 cc. No POST-MORTEM EXAMINATION. General condition fair. Local Lesion —At the seat of inoculation in the skin of the back there were four white linear scars the largest 12°5 mm. in length, and the skin seemed puckered. All organs and glands appeared normal and there was no sign of tuberculosis. 294 VIRUS A III. (Lung of a Crane). CULTURE INOCULATIONS. I—Juvxy 14, 1906. The strain was derived from the original material, and had been in cultivation a total period of 108 days. The culture used was the 4th generation, 22 days old. RABBITS. GUINEA-PIGS. Number.|] Method.| Dose. al of Result. Number. | Method.}| Dose DE of Result. 990 Intrav.| 0-1 mg.| D. 33 days | G.T. 2145 | Intrap.| 1-0 mg. | K. 184 days | Local T. 992 Intrap.{ O1mg.} D. 39 ,, | G.T. 2146 | Intrap.} Ol mg.;K.184 ,, | Local T. 988 | Subcut.} 10°0 mg.; D.157 ,, | Chronic T. 2143 | Subcut.| 10°0 mg.| K.184 ,, | Local T. 989 |Subcut.| 100mg. D. 93, { ae 2144 | Subcut.| 10-0 mg.|K.184 ,, | Local T. CALF 520. Virus A III. Subcutaneous inoculation of culture derived from the original material. Doge—100-0 milligrammes. Date of Inoculation—February 6, 1907. [Age: 9 months. | Killed when in good health—June 8, 1907. [122 days after inoculation.] Temperature. The temperature remained normal throughout. Tuberculin Tests. May 7, 1907. Dose 3:0 cc. bovine tuberculin. [90 days after inoculation.] Negative. No rise of temperature. June 6, 1907. Dose 5:0 cc. bovine tuberculin. [120 days after inoculation.] Negative. No rise of temperature. Weights. Kilos. February 6, 1907 ... me 21227 June 8, 1907 oe 5 oes 222°27 Total gain of weight.—10 kilos. POST-MORTEM EXAMINATION. General condition good. Local Lesion. —The local lesion in the subcutaneous tissue on the left side of the neck measured 8°5 by 6 by 4cm. On section it consisted of a fibrous cyst lined with greyish granulation tissue and filled with a mucoid caseo-purulent substance, Left Prescapular Gland.—The left prescapular gland measured 7°6 by 3°5 by 1:°7cm. On section it con- tained a cyst the size of a sparrow’s egg filled with yellow caseo-pus together with a firmer mass of caseous substance beneath the capsule. There was also a small caseo-purulent nodule and scattered minute soft opaque foci. Right Prescapular Gland.—The right prescapular gland measured 8 by 3:7 by 21cm. It was swollen as the result of a recent tuberculin inoculation. On section it was normal, Prepectoral, Cervical, and Axillary Glands.—Normal. Thoraz. Pleura.—Normal, Lungs.—Normal. Thoracic Glands.—In the long mediastinal gland there were in the cortex a moderate number of minute discrete irregular soft caseous foci. In a second mediastinal gland and in a bronchial gland there were one or two similar foci. Heart.—Normal. Abdomen. Peritoneum and Omentum.—Normal, Intestines.—In the small intestine there were three small greyish elevations of mucous membrane each with a minute yellow purulent centre. Mesenteric Glands.—Many of the mesenteric glands contained each a single soft caseous tubercle the size of a millet seed easily shelling out of the gland sub- stance. Spleen.—Normal. 2. RATS. “ous. ; Number. | Method mg. Number. | Method.| Dose. D al of Result. 7, 1907. 2369 | Intrap: 108 Intrap. | 500 mg.| D. 24 days| Numerous T.B. n the skin in organs. 29368 a axillary 109 | Intrap.| 100 mg.| D. 285 ,, Ditto. erculosis 2370 Intrap 110 Intrap. | 10°0 mg.| D. 20_,, Ditto. 2367 Intrap 111 Intrap.| 1:0 mg.| D. 376 _,, Omental nodules and afew T.B. = ——s iu organs. | 112 |Subcut.| 500mg.; D. 8 ,, | T.B. in organs. | 113 |Subcut.| 100mg.| D. 18 ,, . Ditto. i 114 | Subcut.| 10-0 mg.| D.290 ,, | Slight local T. Few T.B. in organs. Number: 115 |Subcut.} 1:0 mg.| K.278 ,, | NolocalT. T.B. in spleen. 116 117 RABBITS. Duration of 118 1.) Dose. Life. Result. | 34684 1:0 mg. 5:0 mg, K.211 ,, K. 464 ,, bacilli. 1:0 mg. | D. 19 days | G.T. Chronic T. (intra-caecal). Slight local T. Fesruary 25, 1907. CAT 94. Fed with organs from Rabbit 1236. 107 days. P.M.—A few foci in a Peyer’s patch containing tubercle No tuberculosis elsewhere. Killed : June 12, 1907. 2 TF © i i ' : j ; fe : Rey . a ae aS 295 Liver.—In the liver there was a single minute grey ‘translucent tubercle. Kidneys.—Normal. Suprarenal Bodies.—Normal. Renal, Lumbar, and Iliac Glands.~—Normal. Tongue, Tonsils, Larynx, and Pharynz —Normal., Various Lymphatie Glands. Precrural, Coeliac, Popliteal, Gluteal, Pudic, Sub- maxillary, and Pharyngeal Glands.—Normal,' aa Microscopical Examinations. (Smear Preparations.) Local Lesion.—Tubercle bacilli very numerous, Prescapular Gland (left). — Tubercle bacilli numerous. Liver (tubercle).—One tubercle bacillus seen. Mediastinal Gland (tubercle).— A few tubercle bacilli seen. Mesenteric Gland (nodule).—Moderately numerous tubercle bacilli. Small Intestine (purulent focus).—No tubercle bacilli seen. GOAT 40. Virus A III. Subcutaneous inoculation of culture derived from the original material. Dose—100-0 milligrammes. Date of Inoculation—February 6, 1907. [Age 10 months.] Killed—June 14, 1907. [128 days after inoculation.] Clinical History. On the left side of the neck, 17 days after inocula- tion, there was an ill-defined tumour about 8-2 by 5:1 em. The centre of the tumour fluctuated and the “surrounding swelling extended on to the front of the neck. Subsequently the tumour became more promi- nent and fluctuant and finally ulcerated. Both prescapular glands became enlarged, the ulcer having extended on to the right side of the neck. Subse- quently the ulcer slowly healed. At no time was there any obvious sign of ill-health. Temperature. MARCH ’ FEBRUARY / u APRN The temperature during the first two months is shown above. From April 10 to June 14 the temperature was irregular and somewhat raised, the maximum range of variation being from 38°7° OC. to 40°6° C. e Tuberculin Tests. April 9, 1907. Dose 2:0 cc. of bovine tuberculin. [62 days after inoculation.] Rise of temperature 0°6° C., from 40°7° to 41-3° C. April 30,1907. Dose 4:0 cc. of avian tuberculin. [83 days after inoculation.] Rise of temperature 1:2° C., from 40°2° to 41-4° C. F June 13, 1907. Dose 2:0 cc. of bovine tuberculin. [127 days after inoculation.] Rise of temperature 13° C., from 39°9° to 41°2° ©. Weight when killed—12.24 kilos. POST-MORTEM EXAMINATION, General condition fair. Local Lesion.—At the seat of inoculation on the left side of the neck there were in the skin two small ulcers and one linear ulcer covered with scabs, these being all that remained of the large deep ulcer which developed soon after inoculation. In the subcutaneous tissue and in the muscle there was a lobulated tumour measuring about 8 by 6 cm., which consisted of cica- trical tissue and muscle closely beset with caseous nodules varying in size from a millet seed to a 9 aol sparrow’s egg, the majority being small. The largest consisted of grey translucent capsules filled with yellow caseo-pus ; the smallest were caseous and often slightly gritty. Left Prescapular Gland.—The left prescapular gland was enlarged and rounded and about the size of a duck’s egg measuring 7 by 5 by 4 cm. On section it consisted throughout of caseo-necrotic tissue with fine gritty streaks; it was pinkish in parts and rather friable. The capsule was thickened. Lesion on the right side of the Neck.—On the right side of neck almost in middle there was a small subcu- taneous tumour, the size of a sparrow’s egg which on section consisted of a fibrous-walled cyst lined with granulation tissue and filled with mucoid caseo-pus. There were some discrete soft caseous tubercles in the muscle between the two tumours. Fight Prescapular Gland.—The right prescapular gland measured 5 by 2°8 by 14cm. On section it was oedematous and contained at one extremity a caseous slightly gritty nodule measuring about 6 mm. in diameter with several opaque yellow foci in the tissue around it. Prepectoral Glands.—The left reniform prepectoral gland was oedematous. The left round gland was normal. In the right reniform prepectoral gland: there was a caseo-calcareous submiliary tubercle. Cervical Glands.—The left inferior cervical gland was enlarged and measured 1'2 cm. in diameter. On section it was oedematous and contained one caseous miliary tubercle and two opaque foci. The right inferior cervical gland was normal. A left middle cervical gland contained several dis- crete caseous tubercles the size of hemp seed. Therax, Pleura —Normal. Lungs.—The lungs were crepitant.. In the right anterior lobe there were several firm reddish grey lobules ; these on section contained yellow caseo- purulent nodules which shelled out, leaving cavities. In the right tongue sbaped piece the consolidation was similar but more extensive, the lobules coalescing and containing more numerous purulent nodules. In the right posterior lobe there were scattered discrete grey nodules up to 6 1am. in diameter; they were grey and translucent with caseo-calcareous centres. The left lung was affected similarly to the right, the tongue shaped piece being also more extensively affected than the rest of the lobe. Dorsal Mediastinal Glands.—These glands contained discrete caseous gritty tubercles varying up to a millet seed, the gland being not much enlarged. Bronchial Glands.—The bronchial glands were small and contained occasional minute calcareous tubercles. Heart.—Normal. Abdomen. Peritoneum and Omentum.—Normal. Diaphragm.—Normal. Stomach.—In the wall of the stomach just beneath the peritoneum there was a caseous tubercle the size of arape seed. Beneath the mucous membrane and projecting slightly there was a second nodule the size of amillet seed, grey with a yellowish purulent centre. 96 Intestines—On the mucous surface of the sniall intestine there were seven scattered opaque tubercles varying from a point up to nearly a millet seed ; one in a Peyer's patch was ulcerated. . In the large intestine on the surface there were three soft caseous miliary tubercles. Mesenteric Glands.—The cortices of the mesenteric glands were rather closely filled with irregular cagseous gritty nodules; they affected the superficial part of the cortex which from the surface seemed to be about half replaced. On section they were easily picked out leaving cavities in the gland ; some covered a consider- able area up to 1 cm. or more in superficial diameter. Pancreatic Glands.—Two glands in the pancreas contained in the cortices caseous gritty tubercles the size of rape seed. Spleen.—The spleen appeared normal. Liver—Except for one small irregular collection of opaque foci the liver appeared normal. Portal Glands.—The portal glands were beset with discrete opaque nodules. The larger were caseous and gritty, easily shelling out, resembling the mesen- teric glands. The small (the majority) were merely calcareous grains also easily shelling out. Kidneys.—The left kidney contained one opaque miliary tubercle. The right was normal. Suprarenal Bodies.—Normal. Lumbar Glands.—In the lumbar glands there was an occasional opaque tubercle. Iliac Glands.—The iliac glands contained in the cortices discrete caseo-calcareous submiliary tubercles. Pharynz, Larynx, Tongue, Tonsils, and Trachea.— Normal. Various Lymphatic Glands. Precrural Glands.—Normal. Popliteal Glands.—Normal. Submazillary Glands.—The right contained a cal- careous grain ; the left a caseous tubercle. Parotid Glands.—There was a calcareous grain in each. Left Posterior Pharyngeal Gland.—The gland was considerably enlarged, being nearly twice the size of the right, and contained in the cortex caseo-caleareous tubercles and nodules replacing a considerable part of the gland substance ; they were all easily picked out. Right Posterior Pharyngeal Gland.—This contained several calcareous grains the size of rape seed, easily picked out from the gland substance. Microscopical Examination. (Smear Preparations.) Left Local Lesion showed numerous tubercle bacilli. Right Local Lesion showed moderately numerous tubercle bacilli. Left Prescapular Gland showed numerous tubercle. bacilli. Lung (emulsion).—T wo tubercle bacilli seen. Portal Gland showed a few tubercle bacilli. Mesenteric Gland showed a few tubercle bacilli. Large Intestine showed a few tubercle bacilli. Small Intestine showed a few tubercle bacilli. Stomach.—No tubercle bacilli seen. 297 CAT 92. Virus A III. Subcutaneous inoculation with culture derived from the original material. Dose—10-0 milligrammes. Date of Inoculation—February 6, 1907. Died—March 27, 1907. [49 days after inoculation. ] Weights. February 6, 1 907 er'3090 March 27, 1907 1140 Loss in weight.—950 grammes. POST-MORTEM EXAMINATION. Local: Lesion —In the skin of the back to the right of the middle line there was a small circular ulcer 6 mw. in diameter, communicating with a smooth walled cavity in the subcutaneous tissues which con- tained a little turbid serum. Azillary Glands.—The right axillary gland was enlarged and about half replaced by firm caseous sub- stance. A second gland on the ribs was enlarged and beset with caseous foci. “The left axillary glands were normal. Inguinal Glands.—Normal. Thorax. Lungs.—On the surface of the lungs there were some small grey patches from which on section a little pus exuded. The lungs were otherwise normal. Bronchial Glands.—Normal. Abdomen. On the mesentery there was a number of flattened glassy bodies with white streaks in the centre. Other organs and lymphatic glands showed no tuberculosis. Microscopical Examinations. (Smear Preparations.) Aaillary Gland.—Very numerous tubercle bacilli. Lung (substance).—No tubercle bacilli. Liver (substance) —No tubercle’bacilli. Spleen (substance).—No tubercle bacilli. Kidney (substance).—-No tubercle bacilli CAT 94. Virus A IIL. Fed once with the organs of Rabbit 1236 [Inoculated intravenously with 1:0 mg. of culture derived from the original material. | Date of Feeding—February 25, 1907. Killed when in good. health—June 12, 1907. [107 days after the experiment began.] Weights. grammes. February 25, 1907 3400 June 12, 1907 ... 3120 Loss.—280 grammes. POST-MORTEM EXAMINATION. General condition good. Intestines. — The Peyer’s patches showed some opaque foci. Mesenteric G'lands.~The mesenteric glands were not enlarged and showed no opacities. é No tuberculous lesion could be seen in any other organ or lymphatic gland. Microscopical Examinations. (Smear Preparations.) Peyer's patch.—Four tubercle bacilli seen. Mesenteric Gland I.—A few tubercle bacilli seen. Mesenteric Gland II.—Two tubercle bacilli seen. 34684 = ‘snyTIOeq epo1eqny aoe “ ‘i 7 ‘ aurlAog B Suljquieses my ut er par txewua SBA OINYTNO SID, 5 Ls td | “Sur To | ymoqng) gqqTt ‘L'O| “ 6¢’a | ‘Suro.t | ynoqng| gett a ‘L'9| “ 8o°a | ‘800.01 | ‘sMoqng) LETT - Lp) “ sr | ‘BM T.0 | ynoqng) ¢98z ‘LD “ 91-qd | Ba p.o | dequy} gett ‘L'p| ‘ tb 'a | B01 |ymoqng} gore Teal | © gpg | Buoy | denoy|) ror “uaods worj AvaTIS UI HES ‘LD ““¢t-q | ‘8a f.9 | ‘deaquy 79S * 02 'a ‘3M .9 | ‘avizUy 6SIT "H'L 921g, “stsopnoreqny ‘Lemoy< |- jo usis ON — ‘Wd ‘LO | skep zg‘ | ‘Smo | deny | 99%% skep 21 | “ur .T | “Aezz0T | OTL ‘sXep 68 * PATH ‘BU (.T 2 08 ‘ “OFUT . : . . “OFUT, . . . oe q[nsey jo woryeng esoq | ponzeyy | “soqurn yy q[usey jo woneing esoq |'pojeyy | ‘soquny “OAT TMOT “SDId-VONIND ‘SLIGGVa ‘pro sfep 1% wey ‘9061 ‘ ‘9g syaquies9q, «WO pasn Sv OIN}[ND Jo MoNedeues YIP eq, ‘“Bep Tg “WoOIqeA -Y[Nd Jo NORM [e}OT, “QGP JLB Fo purls aepndeosaad wiosz poatsep uresyg = . co TAUALTNAIs a> NA ‘Loss | “ FOL | Sar 0.01 | moqng) 896 ‘Lo prouniays Gaagemaguy. |S) ~ OMe a eeraor | eas) 186 “Leo |] “ gst] “S01 | ymoqng| OFTS A O18 “so[N pou snoeied peach : ‘ ; : i Roel ¢ ‘ ee any ‘pourequ00 pars “y a oe, "te ad) ope | tener) S06 Loot] “ Zgr-M| 8m 0.01 | ynoqng| 6@1S deosoid 4je, © uote : 2 ‘ . i : . a eae on aus oruorgg| ‘ TLL] ‘Barg.7 | ‘deyuy | 796 L 120°T 6y ‘a| ‘3uy.0 | deauy] gelg L‘9 “ cea ‘Sur OT “ABIBOT FOL pue Ce - 09880 — Wd ‘LD skep 9% ‘a ‘Su 1-0 “AeI{Uy £96 “, [e00'J skep 081 “N ‘Sa O-T ‘deayuy LElLs “BAeP BOT ‘trol “ 29°q@ | ‘Suro.or | -aeagaT 90T “9Q06T ‘OZ 2990990 : POTIEH ae ‘enn ae ; ; . “Bur (.0G : 280q “qpns{y ieaeiaarn ‘880q_) =| (poqyey | ‘soqmnyy “4 nse yy z ‘asoq] |" poqqey | “Asquinyy L‘p| ssep ce‘a | 80r0.0¢ | “aetur | «= 80T iesnernnene Fo Dopernd Jo wo1yeang ai ‘9cF ATVO “SLITGV a “SDId-VHNIND “qusay yo een ‘es0q =| ‘pomp | “soquinyy “STMOU "PIO shep €% ¢ ‘mone10ues YP 4} SBA pasn eINg[Nd oq, shep ‘plo sup zz ‘pasn sv oingjno Jo woNvseued y4F egy, | BET JO pomed jej07 & UOTZeAT}[NO UI Ueeq peY pues ‘TeIIOjVUL [VULTIIO OY} WOI} POATJop BEA UTCI4S OUT, ‘9061 ‘3g IsapnyY—IT ‘shep 68 ‘WOT]VATY[NO JO WOI4eINp \ejoJ, “[eIExeM yeuIM0 MoI, paaltop urer4g ‘9061 ‘p ATAL—T ‘SNOILVINOONI HHOLTAO “(queseoyd @ JO JOATT) “AI V SOUIA GUINEA-PIGS. Duration of Hed, Life. Dose. Result. rap.| 1:0 mg. | D. 45 days | Subacute avian T. cut.| 10mg. | D.80 ,, | Local T. 5. Material Inoculated. lof Calf 494, duration of local lesion. E. of a E. of right local lesion. E. of left prescapular gland. E. of right prescapular gland. GUINEA-PIGS. Number. | Method.| Dose. ee of Result. 2903 | Subcut. | 50°0 mg. | D. 118 days} Local T. 2904 | Subcut.| 100mg.) D.120 ,, | Local T. pat Result. ays | No T. y» {No T. 4) | Local T. ” Local T. | 2 U* 299 CALF 456, Virus A IV. Subcutaneous inoculation of culture derived from the original material Dose—50-0 milligrammes, Date of Inoculation—J uly 4, 1906. Weight at Inoculation—59-86 kilos. Clinical Notes. _ Asmall circular swelling developed at the seat of inoculation on the left side of the neck, and the left cde age erelg Ny slightly enlarged. The calf Temained in good health and srew ll i the period of the experimen - vores Temperature, On the third day after inoculation the temperature Tose to 39°7°C. It then remained normal until the 12th day when it rose to 39:9° GC. On the 14th day it was again normal (38°9° C.) and remained normal subsequently. Tuberculin Test.—The calf was not tested subsequent to inoculation. Weights. Kilos. July 4,1906 ... 59°86 July 17,1906 ... 66°21 October 20, 1906 97°95 Total gain of weight—38-09 kilos. Average rate of gain per week—2°53 kilos, POST-MORTEM EXAMINATION. General condition very good. Local Lesion.—In the subcutaneous tissues on the left side of the neck there was a flattened oval tumour measuring 7 by 5°7 by 1'3cm. On section it consisted of grey translucent tissue with a caseo-calcareous network. It was adherent to skin and muscle, but did not infiltrate either. Left Prescapular Gland.—The left prescapular gland measured 6°3 by 3:2 cm. On section it showed four nodules ; one about 1°3 cm. in diameter consisted of fine translucent tissue with a close caseous network and an occasional gritty streak. The others consisted [Age about 13 weeks. ] Killed when in good health—Octoher 20, 1906. [108 da ye after inoculation. ] of collections of caseo-calcareous grains. The rest of the gland substance appeared quite normal. Right Prescapular Gland.—Normal. Prepectoral and Axillary Glands —Normal. Thorac. Pleura and Heart.—Normal. Thoracic Glands. —Normal. Lungs.—The lungs showed in the margins of the anterior lobes a few small dark collapsed areas; otherwise they were normal. al bdomen. Omentum and Peritoneum.—Normal. Spleen, Liver, Portal Glands—Normal. Kidneys and Suprarenals,—Normal. Renal, Lumbar, and Iliac Glands.—Normal. Alimentary Tract. Tongue. Pharynz, Palate, Tonsils.—Normal. Submawillary, Pharyngeal, Parotid, and Hyoid Glands.—Normal. Intestines.—Normal. Mesenteric Glands.—Normal. Testes.—Normal. Various Lymphatic Glands. Precrural, Popliteal, Gluteal, Ischiatic, Pudic.— Normal. Microscopic Examination. Prescapular Gland—Moderate number of tubercle bacilli. CALF 494. Virus AIV. Subcutaneous inoculation of culture derived from the original material. Dose—50:0 milligrammes into left side of neck. Date of Inoculation—January 29, 1907. Weight—when killed, 187-29 kilos. 1:0 milligramme into right side of neck. [Age about 18 weeks. ] Killed when in good health—July 2, 1907. [154 days after inoctlation.] Temperature. On the 13th day after inoculation the temperature Tose to 39°8° C. and reached a maximum of 40°8° C. on the following day. The temperature remained high for thirteen days ; during the remaiving period of the experiment it was normal. Zuberculin Tests. April 9, 1907. Dose 2:0 ce. of bovine tuberculin. [70 days after inoculation.] Rise of temperature 0: io 8° ©. April 30, 1907. Dose 4:0 cc. of avian tuberculin [91 days after inoculation.] Rise of temperature 14° 0. June 13, 1907. Dose 4:0 ce. bovine tuberculin. [185 days after inoculation.] Negative reaction. No rise of temperature POST-MORTEM EXAMINATION. Condition good. Local Lesions (I. On the left side of the neck),— In the subcutaneous tissue on the left side of the neck 300 there was a rounded prominent tumour about the size of a hen’s egg. On section it consisted of a fibrous walled cyst filled with yellow muco-purulent slightly gritty substance together with a tew firmer caseo- necrotic masses; there were several soft caseous tubercles in and between the muscle fibres. (II. On the right side of the neck).—On the right side of the neck under the skin there was a tumour about the size of a thrush’s egg, which on section was a cyst filled with muco-purulent gritty substance. Left Prescapular Gland.—The left prescapular gland measured 8:1 by 4 by 2°5 cm. On section there was a softening caseo-necrotic gritty mass occupying about half the gland; there was a second smaller similar nodule separated from the former by an isthmus of normal gland substance. Right Prescapular Gland—The right prescapular gland measured 7 by 3 by 1:2 cm. It contained in the cortex just beneath the capsule three soft caseous gritty nodules up to a pea in size; the rest of the gland was normal. Prepectoral, Cervical, and Axillary Glands on both sides were normal. Thora’. Pleura.—Normal. Lungs.—Normal. Bronchial and Mediastinal Glands.—Normal. Pericardium and Heart.—Normal. Abdomen. Peritoneum and Omentum.—Normal. Intestines.—Normal. Mesenteric Glands.—Normal. Spleen.—Normal. Liver.—In the liver just beneath the capsule there was a single grey miliary tubercle with a calcareous centre which ‘microscopically showed moderately numerous tubercle bacilli. Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Renal, Lumbar, and Iliac Glands.—Normal. Tongue, Larynx, Pharynx, and Tonsils—Normal. Various Lymphatic Glands. Coeliac, Precrural, Gluteal, Popliteal, Parotid, Submacillary and Retropharyngeal Glands.—Normal. Microscopical Examination. (Smear Preparations. ) Left Local Lesion.—Numerous scattered tubercle bacilli and a few small clumps. Right Prescapular Gland.—Moderately numerous tubercle bacilli. Right Local Lesion.—A few scattered tubercle bacilli. Liver Tubercle—Moderately numerous bacilli. tubercle Animals Inoculated. CALF 568. Virus A IV. Subcutaneous inoculation with culture derived from the Prescapular Gland of Calf 494. Dose—100-0 milligrammes. Date of Inoculation—November 9, 1907. Killed when in good health—May 8, 1908. Clinical History. There was no sign of ill-health during the experi- ment. Temperature. It rose to 39°7° on the 11th day and remained at this level until the 21st day when it reached 40-1°. After the 29th day it remained normal. Tuberculin Test. March 13, 1908. [125 days after the experiment began.] Dose of tuberculin (bovine) 20 cc. No re- action. Weights. Kilos. November 9, 1907 85:25 May 8, 1908 196°81 Total gain in weight.—111-56 kilos. POST-MORTEM EXAMINATION. Generai condition good. [Age about 20 weeks.] [181 days after inoculation. } Local Lesion—At the seat of inoculation in the muscle on the left side of the neck there was a roughly pear-shaped tumour measuring 8 by 6 by 35 cm. Ii was cystic and on section was filled with yellow caseo- pus. Left Preseapular Gland.—It measured 6:2 by 2°5 by 18cm. On section it contained two hard nodules measuring about 1 and 2 cm. in diameter, and com- posed of caseo-calcareous tissue which fairly readily shelled out leaving in the gland substance cavities with somewhat congested walls. Right Prescapular Gland.—It measured 5:5 {by 2°5 by 1:4 cm. and on section was normal, Lungs.—In the lung there was a nodule the size of a millet seed composed of soft grey translucent tissue. There was no tuberculosis elsewhere, Microscopical Examination. (Smear Preparations.) Local Lesion.—Numerous tubercle bacilli. Prescapular Gland.— Numerous tubercle bacilli. Lung (nodule).—No tubercle bacilli. 301 VIRUS A V. (Spleen of a Pheasant). CULTURE INOCULATIONS. JuLy 4, 1906. Strain derived from original material. Total duration of cultivation, 88 days. The 4th generation of culture was used, 22 days old. CALF 458. Subcutaneous. Dose : 50:0 mg. Killed : October 15, 1906. 103 days. P.M.—Caseo-calcareous and fibroid tumour ut seat of inoculation. The left prescapular gland con- tained two small caseo- calcareous nodules. There was no tuberculosis else- where. RABBITS. GUINEA-PIGS. Number. |Method.| Dose. pul ae os Result. Number. | Method.} Dose. aaa of Result. ‘ | ; 958 | Intrav. | 0°01 mg. | D. 145 days Chronic 212% | Intrap. | 1:0 mg. | K. 18% days | Local T. G.T. 959 | Intrap.| 1:0 mg. | D. 28 ,, | Acute T. 2128 | Intrap. | 0-01 mg.|K.182 ,, |NoT. 960 | Intrap. | 0°01 mg. | K. 198 _,, ae 2125 |Subcut.| 10:0 mg.|D 127 ,, orb, z ; : .| D. 52 ., | HarlyG.T. joint. Poe enon | rae — 2126 |Subcut.| 10°0 mg.| K.182 ,, |NoT. 957 Subcut. | 10°0 mg. | K.168 ,, | Chronic G.T. v 2x 34684 302 CALF 458. Virus A V. Subcutaneous inoculation of culture derived from the original material. Dose—50°0 milligrammes. Date of Inoculation—July 4, 1906. Weight—51°69 kilos. [Age about 10 weeks.] Killed when in good general health—October 15, 1906. [103 days after inoculation.] Clinical Notes. A small tumour developed at the seat of inoculation on the left side of the neck and the left prescapular gland became slightly enlarged. On October 12, 100 days after inoculation, the calf was found to be unable to stand on its forelegs. It eet to be in good health, and its appetite was goo The animal was killed three days later. Temperature. On the 12th day after inoculation the temperature rose, reaching a maximum (40°2° C.) on the 14th day. It remained high for nearly two weeks, returning to the normal on the 25th day. During the remaining period of the experiment it was approximately normal. Tuberculin Test. The calf was not tested subsequent to inoculation. Weights. Kilos. July 4, 1906... ee sie we 51°69 July 17, 1906 at wae, SD October 15,1906... ie ai 87.07 Lotal gain in weight.—35'38 kilos. Average rate of gain per week.—2'6 kilos. POST-MORTEM EXAMINATION. General condition good. Local Lesion.—In the subcutaneous tissues on the left side of the neck there was a slightly prominent yumour 6°3 by 4°5 by 1:2.cm. On section it consisted of pinkish translucent tissue containing caseo-calcareous granules and small irregular caseous nodules. Left Prescapular Gland.—The left prescapular gland was 5 cm. in length and contained (one at each extremity) two nodules, one 0°6 cm. the other 1:2 cm. in diameter, consisting of caseo-calcareous granules. Right Prescapular Gland, Prepectoral, and Axillary Glands.—Nor al. Thorax. Pleura and Heart.—Normal. Lungs and Thoracic Glands.—Normal. Abdomen. Peritoneum and Omentum.—Normal. Spleen.—Normal. Liver, Gall Bladder, Portal Glands.—Normal. Kidneys and Suprarenals.—Normal, Renal, Lumbar, and Iliac G'lands.—Normal. Alimentary Tract. Tongue, Pharynz, Palate, Tonsils.—Normal. Pharyngeal, Submazillary, Hyoid, and Parotid Glands.—Normal. Intestines.—Normal. Mesenteric Glands.—Normal. Joints—The joints were carefully examined, and were quite normal in appearance. Various Lymphatic Glands. Precrural, Popliteal, Gluteal, Ischiatic, Pudie.— Normal. Microscopic Examination. Prescapular Gland.—Tubercle bacilli in moderate numbers. 303 VIRUS A VI. (Spleen of a Pheasant). CULTURE INOCULATIONS. SEPTEMBER 21, 1907, The strain was derived from the original material, and had been in cultivation a total period of 529 days. The culture used was the 6th generation, 21 days old. RABBITS. PARROTS. Number. | Method.| Dose. Parsee a Result. Number. | Method.| Dose. ee . Rent | 1484 | Intrav.| 1:0mg.| D. 12 days, Acute T, Subcut. | 10-0 mg.} D. 95 days |G.T 1483 | Intrav.| 0-1mg.| D. 43 ,, | GT. Fed | 10mg.{ D.76 , [GT | 1482 | Intrav.|0-01 mg.} D. 40 ,, | GT 1485 | Intrap.| 10mg.]D. 59 ,, | GT. 1486 | Intrap.| O1mg.| D. 8 ,, |G.T. 1488 | Subcut.| 500 mg.| D. 52 ,, | GT. 1487 | Subeut.| 01 mg. | D.267 ., | Chronic G.T. GUINEA-PIGS. FOWLS. i Duration of Number.| Method.) Dose. ee of! Result. Number. | Method} Dose. Life. Result. 2625 | Intrap.} 1:0 mg. | D. 56 days | Subacute 814 | Intrap.| 0:01 mg.| D. 45 days} G.T. avian T. 2624 | Intrap.| 0-1 mg. | D. 57 ,, | Subacute 312 | Intrap.| 0-1 mg.| D.207 ,, | G.T. avian T. a : 1 T. sub.). GRAS: | Sobek | MeO mg, | eee oy. yee 310 |Subcat.|1-0 mg.|D. 79 ,, | GT. 2622 | Subcut.| 50-0 mg.} K.171 ,, | Local T. 2X2 34684 ‘ ' ~ ° : ‘ j ‘ Mo - ti \ , 2 rb 1 i é : ! a . ‘ ‘ 4 , 1907. local gland seous hinute rs and Interic sand Fusely h the retion e also liver, bus in bercu- work, CAT 96. Intraperitoneal. Dose: 1-0 mg. Died : May 20, 1907. 69 days. P.M.— Omentum thickened and retracted ; composed of grey tissue with a caseous net- Patches of opaque foci on the surfaces of the lungs. No tuberculosis elsewhere. Duration of Life. Result. K. 153 days | Local T. K.153 ,, | No T. K.153,, Local T. K. 153 ,, | Local T. RATS. Duration of Number. |Result. Number. | Method Dose. ate Result. trap. | 20°0 mg.| K. 82 days|}G.T. Nume- 1278 Ir. 127 | Intrap g Lae in organs. ee 125 | Intrap.| 1:0 mg.| K.177_, Nees! Ds -B. in re organs. ; ‘lL mg.| K.173 ,, |NoT. T.B. sisi |e ae | | ee inliverand spleen. 1282 iT. : . | 0°01 .| K. 574 ,, Local T. tee ee ane T.B. in lung and spleen. .| 20:0 mg.| K.244 ,, |No'T. T.B 128 | Subcut 4 : in spleen. 2" 34684 305 VIRUS A VII. (Spleen of a Fowl). CULTURE INOCULATIONS. I.—January 24, 1908. The strain was derived from the original material, and had been in cultivation a total period of 430 days. The culture used was the 8th generation, 21 days old, ~ PONY 10. GOAT 54. RABBITS. Subcutaneous. Subcutaneous. D ; oo Dose: 100-0 mg. Dose : 50:0 mg. Number. | Method.| Dose. stl Or |) Result. Died : March 2, 1908. Killed : May 22, 1908 38 days. 119 days. 1608 Intrav. | 1:0 mg. | D. 47 days | G.T. ae P.M.—Cyst with caseo: “8 P.M.—Fibrous walled necrotic abetents at seat 1602 | Intrav.| Olmg. | D.33 ,, G.T. ecystatseatofinoculation; 4 in ocilation. | (Caseo- i ae ae necrotic nodules in adja- Death from septic peri- cenuglands, Enonemesen- Seaianadceie teric gland numerous PY softened caseous gritty M-rca 11, 1908. collections were seen. A left iliac gland contained PIG 246. four yellow tubercles. Fed with emulsion of lung, liver, spleen, and kidneys from Rabbit 1603. GUINEA-PIGS. Killed : June 12, 1908. : : 93 days. Material Duration of Inoculated. Number. Methad. Life. - Result. P.M.—All the mesen- teric and colic glands contained caseous gritty E. of local 3190 Subcut. | K. 103 days Local T. nodules, some confluent. lesion. On the surface of the E. of pre- 3189 Subcut | K. 103 a Local T. jungs there were three or scapular four translucent tubercles gland. and a group of minute E. of cervical | 3192 |Subcut.| K.103 ,, |Healthy.) foci, There was no gland. tuberculosis elsewhere. E. of mesen- | 3191 |Subcut./ K.103 ,, | Healthy. teric gland. GUINEA-PIGS. Material Duration of | Teamulnted, Number. Method. Life. Result. E. of mesen 3198 | Intrap. | K. £03 days | Local T. teric gland (a). 3197 | Subcut.| K.103 ,, | LocalT. E. of mesen- teric gland | 3199 | Intrap.|K.103 ,, |Healthy. (2). 306 PIG 176. Virus A VII. Fed once with culture derived from the original material. Dose—50-0 milligrammes. Date of Feeding—March 12, 1907. [Age 8 weeks.] Killed when in good health—January 21, 1908. [315 days after the experiment began.] Clinical History. There was no ill-health during the sign of experiment, Temperature. Normal throughout. Tuberculin Tests. (93 days after the experiment Rise of tempera- June 13, 1907. began.] Dose of tuberculin 4:0 cc. ture 1-3° C. September 18, 1907. ment began.] Dose of tuberculin 2:0 cc. temperature 2°3° C. October 29, 1907. began.] Dose of tuberculin 4:0 ce. [190 days after the experi- Rise of [231 days after the experiment Rise of tempera- ture 0°8° C. Weights. Kilos. March 12, 1907 ane is ads 9:97 January 21,1908 ... wes aes 65°75 Total gain in weight—55°78 kilos. POST-MORTEM EXAMINATION. General condition good. Submazillary Glands.—In the largest submaxillary gland on each side there were two calcareous tubercles. the size of rape seed, just beneath the capsule. There- was also a gritty granule in the left gland. Intestines.—Normal. Mesenteric Glands.—The majority of the mesenteric glands contained irregular caseo-calcareous nodules easily shelling out of the substance ; several of the glands were considerably replaced. The remaining glands were normal. Colic Glands.—Many of the colic glands contained’ calcareous granules. Portal Glands.—Normal] in size. In each of two- there was a single minute calcareous granule. Iliac Glands.—The left iliac gland contained about six irregular soft caseous and calcareous nodules up to a hemp seed in size. The central gland contained three similar nodules. All organs and other lymphatic glands were normal. Microscopical Examinations. (Smear Preparations.) Portal Gland (focus).—One beaded tubercle bacillus.. Mesenteric Gland.—Moderately numerous tubercle bacilli. There were no tubercle bacilli seen in smears from the lung, liver, spleen, kidney, and iliac gland. No growth was obtained in cultures made from the mesenteric, submaxillary, and iliac glands. PIG 178. Virus A VII. Fed once with culture derived from the original material. Dose—100°0 milligrammes. Date of Feeding—March 12, 1907. [Age 8 weeks.] Killed when in good health—June 21, 1907. [101 days after feeding.] Clinical History. The animal remained in good health throughout the experiment. Temperature. Approximately normal throughout. Tuberculin Test. 93 days after feeding. Reaction negative. ; Weights. Kilos. March 12, 1907 ‘ise itis san 9°97 , June 21, 1907 se ‘igs ave 17°68 Total gain of weight—T-71 kilos. POST-MORTEM EXAMINATION. General Condition.—Animal appeared well; it was fat and in good condition. Tongue, Larynx, Pharyna and Trachea.—Normal. Tonsils.—In each tonsil there were several small collections of soft yellowish substance. (? secretion.) Submasillary Glands.—Normal. Cervical Glands.—Normal. Intestines —The large and small intestines, except for slight congestion, appeared normal. Mesenteric Glands.—Many of the mesenteric glands showed fine yellowish markings between the lobules, an appearance sometimes present in normal glands of the pig. In addition there were also two small linear caseous gritty nodules up to nearly 2 mm. in length. Spleen.—Normal. Liver.—Normal. Portal Glands—In the portal glands there were: streaks and foci similar to those in the mesenteric glands. Kidneys.—Normal. Suprarenal Bodies.—Normal. Lumbar and Iliac Glands.—Normal. Thorax. Lungs.—The lungs were irregular and patchily con- gested, and a little oedematous , 307 Heart.—Normal. Bronchial and Mesenteric Glands.—These appeared normal. Various Lymphatic Glands. Parotid, Prescapular, Precrural, Ingui 1m teal Glands.—Normal. anon and Ropts Microscopical Examinations. (Smear Preparations.) Spleen (substance).—Tubercle bacilli moderately numerous. Liver (substance).—Tubercle bacilli present in large numbers. Kidney (substance).—Tuberele bacilli scanty. Lung (substance).—Tubercle bacilli in clumps. Bronchial Gland (substance).—Very numerous tubercle bacilli. Portal Glands (streaks).—Numerous tubercle bacilli, Coeliac Gland (streaks).—Numerous tubercle bacilli. Mesenteric Gland (streaks).—Swarms of tubercle bacilli. Mesenteric Gland (nodule).—Swarms of tubercle bacilli. Intestine (scraping).—Very numerous tubercle bacilli. Tonsil (pus).—A moderate number of tubercle bacilli. PIG 180. Virus A VII. Subcutaneous inoculation of culture derived from the original material. Dose—50°0 milligrammes. Date of Inoculation—March 12, 1907. [Age 8 weeks.] Killed when in good health—June 25, 1907. [105 days after inoculation.] Clinical History. During the whole experiment the animal showed no -symptoms of ill-health. Temperature. Approximately normal throughout. Tuberculin Test. 93 days after inoculation. Reaction negative. Weights. Kilos. March 12, 1907 ve ‘ins wait 7:25 June 25, 1907 eee ats 13°24 Total gain in weight—5'99 kilos. . POST-MORTEM EXAMINATION. General condition.—The animal was in fair condition but bad not grown much. “Local Lesion.—In the subcutaneous tissue at the seat of inoculation in the abduminal wall there. was a circular nodule 3cm. in diameter by 1:3cm. in thickness. On section it was a fibrous cyst with smooth internal walls and filled with bright yellow pus. Inguinal Glands.—The left inguinal gland was en- larged and contained at one extremity a collection of caseous slightly gritty nodules softening at their peri- phery and capable of being picked out though not* very easily. They replaced about balf of the gland. A second gland showed foci of congestion only. Left Sternal Gland.—The left sternal gland was en- larged and showed at one extremity some discrete irregular caseous and gritty nodules mostly about the size of rape seed. Right Inguinal and Right Sternal Glands.—Normal. ~ Precrural Glands.—Normal. Agillary Glands—Normal. Thorax. Lungs.—The lungs were pink in colour and col- lapsed well. The substance was beset with small congested foci; one minute translucent focus was -also seen. Bronchial Glands—Normal. Mediastinal Glands.—Normal. Abdomen. Intestines.—The intestines appeared normal. Mesenteric Glands.—In the cortices of some of the mesenteric glands there were numerous yellowish softening caseous slightly gritty streaks and irregular foci; other glands were enlarged and firm, containing either a network of waxy caseous tissue, or diffusely caseous nodules opaque and softened in parts and not easily separated from the surrounding gland substance. Spleen.—Normal. Liver.—Normal. Portal Glands.—Normal. Kidneys.—On the surfaces of the kidneys just beneath thea capsules an occasional grey point could be seen. Suprarenal Bodies.—Normal. Lumbar and Iliac Lymphatic Glands.—Normal. Tongue, Larynx, and Pharyna.—Normal. Tonsils.—In the tonsils there were yellowish nodules of secretion. Various Lymphatic Glands. Submazillary, Parotid, Pharyngeal, Cervical, Pres- capular, and Popliteal Glands.—Normal. Microscopical Examinations. (Smear Preparations.) Lung (substance).—A few tubercle bacilli. Spleen (substance).—A few tubercle bacilli. Liver (substance).—More numerous tubercle bacilli. Kidney (substance).—A few tubercle bacilli. Local Lesion.—Moderately numerous tubercle bacilli. _ Inguinal..Gland (emulsion).—Swarms of tubercle bacilli. Mesenteric Gland (substance).—Large numbers of tubercle bacilli. Intestine (scraping). — Very numerous tubercle bacilli Stomach (scraping).—No tubercle bacilli seen. Tonsil (secretion).—A few tubercle baciili. 308 PIG 182. Virus A VII. Intraperitoneal inoculation of culture derived from the original material Dose—10-0 milligrammes. Date of Inoculation—March 12, 1907. [Age 8 weeks.] Killed in good health—June 26, 1907. [106 days after inoculation.] Clinical History. There were no signs of illness during the experiment. Temperature. Approximately normal throughout Tuberculin. Test. 93 days after inoculation. Reaction negative. Weights. Kilos. March 12, 1907 10°90 June 26, 1907 18°56 Total gain in weight.—7-66 kilos. POST-MORTEM EXAMINATION. General condition good. Abdomen. Parietal Peritoneum.—The parietal peritoneum was not affected. Omentum.—The omentum was not thickened or retracted, but it showed moderately numerous discrete translucent streaks and small patches. The majority of these were flat without any thickness. but there were scattered firm nodules, mainly in the free margin, which attained the size of a wheat grain, and showed central opacities consisting either of soft yellow slightly gritty substance or irregular calcareous grains. Meso-colon.—On the mesentery of the large intestine there were sparsely scattered nodules up to7 by 5 mm., sometimes composed wholly of translucent tissue and only occasionally showing yellow gritty foci in the centre. Mesentery.—The central part of the mesentery appeared normal, but near the mesenteric glands it showed radial translucent thickenings beset with opaque foci. Intestines.—Small intestine. the small intestine was pale. appeared normal. Coeliac Glands.—The glands situated dorsally to the stomach, near the cardiac orifice,appeared enlarged, but showed no obvious tuberculous change. Mesenteric Glands—The mesenteric glands were uniformly enlarged ; on section in some the substance appeared normal, but in the majority could be seen numerous minute yellowish streaks and foci ; in others The epithelium lining The large intestine these had run together and formed small irregular a caseous slightly gritty nodules. Spleen.—The lymphatic fringes on the spleen were a little enlarged ; in the substance greyish translucent foci were visible. , Liver.—The liver appeared normal. Portal Glands—The portal glands were enlarged but showed no opacities. Kidneys.—In the cortices of the kidneys there were numerous irregular grey translucent foci. Suprarenal Bedies.—These appeared normal. Lumbar Lymphatic Glands.—These glands were a little enlarged. Iliac Lymphatic Glands.—Normal. Diaphragm.—On the peritoneal surface of the diaphragm there was a yellowish calcareous submiliary tubercle. Thorax. Lungs.—On the surface of the right caudal lobe just. beneath the pleura there was a grey miliary tubercle. In the rest of the lung there was an occasional translucent focus. Thoracic Glands.—The bronchial glands appeared normal. The Dorsal Mediastinal Glands were normal. The Ventral Mediastinal Glands were enlarged and firm, and showed a few sparsely scattered yellowish calcareous grains. Various Lymphatic Glands. Submasllary, Parotid, Cervical, Prescapular, Axillary, Precrural, Inguinal, and Popliteal Glands. —Normal. Microscopical Examinations. (Smear Preparations.) Spleen (substance).—Tubercle bacilli moderately numerous. Liver (substance).—Tubercle bacilli numerous. Kidney (substance}.—No tubercle bacilli seen. Small Intestine (scraping).—Tubercle bacilli in large numbers. Large Intestine (scraping).— Tubercle bacilli numerous. Mesenteric Gland (substance).—Swarming with tubercle bacilli. Mesenteric Gland (nodule).—Swarming with tubercle. bacilli. Portal Gland (substauce).—Moderately numerous tubercle bacilli. Coeliac Gland (substauce)-—Swarming with tubercle bacilli. 309 CAT 96. Virus A VII. Intraperitoneal inoculation of culture from the original material. Dose—1°0 milligramme. Date—March 12, 1907. Died—May 20,1907. [69 days after inoculation.] Weights. grammes. March 12, 1907 1650 May 20, 1907 ... 1300 Loss.—350 grammes, POST-MORTEM EXAMINATION. Omentum.—The omentum was thickened, retracted and adherent to the spleen ; it consisted of firm trans- lucent tissue with a caseous network. The Mesentery and Parietal Peritoneum were normal. Lungs.—The lungs were oedematous and showed on the surfaces numerous greyish white patches, small ole irregular and made up of collections of opaque oci. Other organs and glands appeared normal. Microscopical Examinations. One tubercle bacillus was seen in a smear made from a caseous patch of the lung ; none in the spleen, liver, and kidney. PONY 10. Virus A VII. Subcutaneous inoculation of culture derived from the original material. Dose—100-0 milligrammes. Date of Inoculation—January 24, 1908. Died—March 2, 1908. Clinical History. The animal appeared well until a few days before a death. It then became quiet and was always lying own. Death was due to acute septic peritonitis, the cause of which could not be discovered. A small raised tumour had developed at the seat of inoculation but the skin remained unbroken. Temperature. Normal. Weights. Kilos. January 24, 1908 103-40 March 2, 1908 84°35 Total loss in weight.—19-05 kilos. POST-MORTEM EXAMINATION. Local Lesion—At the seat of inoculation on the left side of the neck there was a circular tumour 4 by 1°5 cm. which on section was a fibrous walled cyst with a central cavity containing reddish necrotic masses and serous fluid. The skin over it had not broken but it was thinned and dead. Left Prescapular Glands.—In the left group of prescapular glands there was one about 1°2 cm. in greatest length which was about half replaced by firm bright yellow necrotic substance ; in a second there was a small necrotic patch. All the other glands were normal. Left Cervical Glands.—A left cervical gland con- tained a few minute calcareous grains. Thoraz. Lungs.—In the lungs there were sparsely scattered nodules, similar to those in the liver, which were distinctly palpable and varied up tc nearly a millet [Age about 1 year.] [38 days after inoculation. ] seed in size. crepitant. Bronchial Glands.—Normal. The lung parenchyma was pink and Abdomen. Peritoneal Cavity—The peritoneal cavity was filled with turbid fluid and there was a deposit of yellowish fibrin on the intestines which were glued together. The mesentery was contracted. Spleen.—Normal. Liver.—In the liver there were moderately numerous nodules up to nearly a millet seed in size. They were very irregularly distributed and about half the right lobe was unaffected. They were composed of soft grey tissue with a yellowish tint. Kidneys.—Normal. Intestines.—The walls of the intestines were thickened with inflammatory exudation. There was no sign of any perforation. The Mesenteric, Colic, Rectal and Ventral Mediastinal Glands were enlarged and contained necrotic patckes becoming purulent. Other lymphatic glands were normal. Microscopical Examinations. (Smear Preparations.) Local Lesion.—Very numerous tubercle bacilli. Left Prescapular.—Very numerous tubercle bacilli. Left Cervical.— One tubercle bacillus: other organisms present. Peritoneal Fluid.— Swarms of organisms. Liver Nodules.—Blue-staining organisms. Nore.—The tuberculosis was limited to the seat of inoculation and the nearest lymphatic glands. Death was due to acute septic peritonitis and pyaemia. blue - staining GOAT 54. Virus A VII. Subcutaneous inoculation of culture derived from the original material. Dose—50'0 milligrammes. Date of Inoculation—January 24, 1908. [Age 5 months. ] Killed when in good health—May 22, 1908. [119 days after inoculation. ] Clinical History. The animal showed no sign of ill-health during the experiment. 34684 Temperature. Normal throughout. 2Y 310 Tuberculin Tests. [49 days after the experiment Mareh 13, 1908. Rise began.] Dose of tuberculin (Bovine), 2°0 cc. of temperature, 1:A° C. May 13, 1908. [110 days after the experiment began.] Dose of tuberculin, 2:0 cc. (Avian). Rise of temperature, 2°2° C. POST-MORTEM EXAMINATION. General condition good. Local Lesion.—In the subcutaneous tissue on the left side of the neck there was a prominent rounded tumour about 7 cm. in diameter. In the underlying muscles there were numerous small nodules filled with caseous substance. On section the tumour consisted of a thick fibrous walled cyst filled with softening caseo-necrotic substance. Prescapular Glands—The left prescapular gland measured 55 by 3 by 2cm. On section it contained two softening caseo-necrotic nodules almost replacing the gland substance, the largest measuring 3 by 2°5 cm. The right prescapular gland measured 3:6 by 1:7 by 07cm. On section it was normal. Prepectoral Glands.—Normal. Cervical Glands.—A left cervical gland, a little larger than a sparrow’s egg, was replaced by pinkish softening caseo-necrotic substance. A second gland contained a caseous gritty nodule 6 mm. in diameter. In a third smaller gland there were several small caseo-calcareous nodules. Axillary Glands.—Normal. Thorax. Pleura.—Normal. Lungs.—Normal. Bronchial and Mediastinal Glands.—Normal. Heart.—Normal. Abdomen. Omentum and Peritoneum.—Normal. Intestines —Normal. Mesenteric Glands.—In one mesenteric gland there were numerous softened yellow collections of caseous gritty substance. Spleen.—Normal. Liver.—Normal. Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Iliac Glands.—A left iliac gland contained four yellow tubercles. Renal, Coeliac, and Lumbar Gtanas.—Normal. Tongue, Tonsils, Larynx, and Pharynx.—Normal. Various Lymphatic Glands. Parotid, Submazillary, Retro-pharyngeal, Precrural, and Popliteal Glands.—Normal. Microscopical Examinations. (Smear Preparations.) Local Lesion—Numerous tubercle bacilli. Prescapular Gland.—Numerous tubercle bacilli. Lung.—No tubercle bacilli. Liver.—No tuberele bacilli. Left Iliac Gland.—No tubercle bacilli. Mesenteric Gland.—A few scattered tubercle bacilli. PIG 246. Virus A VII. Fed with the organs of Rabbit 1603 (which had been inoculated intraveneously with 1:0 mg. of culture of the original material). Date of Feeding—March 11, 1908. Killed in good health—June 12, 1908. Temperature. Normal throughout. Tuberculin Pest. May 13, 1908. began.] Dose of tuberculin, 2°0 cc. {63 days after the experiment No reaction. Weights. Kilos. March 11, 1908 27°66 June 12, 1908 43-08 Gain in weight.—15:42 kilos. POST-MORTEM EXAMINATION. Condition good. Tongue, Tonsils, Pharyna, Submasillary Glands and Intestenes.—Normal. Mesenteric Glands.—All the mesenteric glands con- tained yellow caseous nodules with a few gritty streaks. Most of the nodules were firm except round the margins which were softening a little, and which could be separated from the surrounding gland tissue. [Age 15 weeks.] [93 days after feeding. ] In several of the glands the nodules were closely placed and confluent replacing the majority of the superficial part of the cortices of the gland. Colic Glands.—They resembled the mesenteric. Lungs.—On the surface of the lungs there was a small collection of transparent foci and three or four minute transparent tabercles. All other organs and glands were normal. Animal Inoculations. Three guinea-pigs were inoculated with emulsions of the mesenteric glands ; two developed local tuber- culosis only, the third remained healthy. Microscopical Examinations. (Smear Preparations.) Lung (substance).—No tubercle bacilli. Liver (substance).—No tubercle bacilli. Spleen (substance).—No tubercle bacilli. Intestine (scraping).—No tubercle bacilii. Lung (tubercle).—No tubercle bacilli. Mesenteric Gland.—Numerous tubercle bacilli. a total 4 1 | | ol 524 | | G 3 ae UINEA-PIGS. RATS. Dose : 200°0 | Method.| D Duration of : Died: June 27, rae Life. Result. Number. | Method.| Dose. Rue of) Result. 30 days. Int Pa ntrap. . ‘ Pal —The Jon Tap. |1000 mg.) D. 29 days ! aaa 132 | Intrap.| 200-0 | D. 131 days |G.T. Nume- almost entirely I : ian T. mg. rous T.B. ee cae D eee She 133 | Intrap. | 1:0 mg.| K. 497 ae owed patches o ? ntrap. ‘O mg. : » |Local T. tion everywhere. Intrap. | 10mg. | D. 52. ,, pena severe. A: were numerous in : at few T.B principal organs, Sabot | LOO Dmg Ky Sy [eae in in organs. small intestine, ant skin, the glands examin¢ eens 4 —_ a dd a FOWLS. Number. |Method.| Dose, | Duration of) Resut Mate g : Life. esult. Inocu 252 |Subcut.| 1:0 mg. | K. 254 days | G. T. E. of lu 250 |Subcut.} 0°01 mg. | K. 254 ,, |G. T. BE. of lun 248 |Subcut.| 0:0001 | D.190 ,, | G.T. mg. Pe 246 |Subcut.| 0:000001 | K.254 ,, |G.T. mg. 244 | Subcut. | 0:0000001|K.254 ,, |Slight & mg. G. T. iy na NEA-PIGS. eS Gg ose. pos st Result. | ; fe a ee 0 mg. | K. 163 days | Local Ts Omg.| D. 61 -, | Local T. 0 mg.| K.163 ,, Local T. 34884 2 Y* 311 VIRUS A VIII. (Spleen of a Fowl). CULTORE INOCULATIONS. IYI.—Aprin 7, 1908. The strain was derived from the original material, and had been in cultivation a total period of 501 days. The culture used was the 8th generation, 22 days old. RABBITS. PIG 248. PARROT 18. : a Intravenous. Fed: About 20-0 mg. Number Method. Dose. ve 2 Result. | Dose: 50:0 mg. Died 113 days. Died : May 24, 1908. ee > : replace by caseo- 19717 | Intrav.| 0:00001 | D. 100 days | C ue oni ; 47 days. a - ia y aan ae a. P.M.—Acute tuber- Caseous a ae and a is. foci in the liver; 1721 | Intrav. |0-0000001} D.198 ,, | Chronic T. culosis eee mee The culture had evi- dently been inhaled. GUINEA-PIGS. FOWLS. : | away | Number.| Method.| Dose. | a at Result. Number.| Method., Dose. | Ravel of! Result. 3141 | Intrap.| 1:0 mg. | D. 50 days \ é hacuie 376 | Intrav. ae K. 94 days | G.T. severe. avian T. ' 1| K.94 ,, | G.T. mode- 3142 | Intrap.| 10mg. | D.36 ,, | 3878 | Intrav. | 0 ee rece severe. 3880 | Intrav. |0:0000001/ K.94 ,, | G.T. slight. mg. 2Y 2 34684 312 CALF 524. Virus A VIII. Intravenous inoculation of culture derived from the original maierial. Dose—200°0 milligrammes. Date of Inoculation—May 28, 1907. [Age 14 weeks.] Died—June 27, 1907. [80 days after inoculation. ] Temperature . MY 1907 JUNE Weights. Kilos. May 28, 1907 ... 31°75 June 27, 1907 26°75 Total loss in weight.—5-0 kilos. POST-MORTEM EXAMINATION. Condition poor. Local Lesion.—Along the vein of the ear there was some thickening surrounded by a collection of muco- pus, and in the corresponding parotid gland there were a few caseous patches. Thoras. Pleura.—The pleura appeared normal. Lungs ——The anterior, the middle, and a consider- able part of the ventral portions of the posterior lokes were dark reddish grey and airless. In the remaining portions of the posterior lobes con- taining air, there were scattered congested lobules. The dark red anterior lobes were mottled with numerous yellowish patches and small grey nodules. In some parts the caseous patches were extensive, replacing a number of lobules; similarly in the rest of the solid areas there was extensive caseation of the pneumonic parts. Muco-pus could be expressed from the bronchioles, and the larger bronchi algo contained pus. Thoracic Glands—The bronchial and mediastinal glands did not appear enlarged ; they were congested and oedematous but showed no caseation except for two small yellowish tubercles. The sternal glands and the glands at the entrance to the thorax were congested. Heart and Pericardium.—Normal. Abdomen. Peritoneum, Omentum, and Diaphragm —Normal. Antestines.—Normal. Mesenteric Glands.—Normal. Spleen.—The spleen weighed 254 grammes ; it was somewhat enlarged and swollen, and on section the sub- stance was dark red with a tendency to be diffluent. Liver.—The surface of the liver appeared normal ; on section grey points and streaks could be seen, probably interlobular connective tissue. Portal Glands.—In a portal gland there was an undefined small yellowish patch. Kidneys.—The cortices of both kidneys were pale Suprarenal Bodies.—Normal. Lumbar, Renal, and Iliac Lymphatic Glands —T hese glands were uniformly enlarged and oedematous. Left Prescapular, Prepectoral, Inferior Cervical, and Axillary Glands—All were congested, several being dark red. Left Superior Cervical and Submasillary Glands.— These glands were a little oedematous. The corre- sponding glands on the right side resembled those on the left side. Precrural, Popliteal, Ischiatic, and Gluteal Glands. —These appeared normal. In none of the lymphatic glands, except the parotid, was there any caseation. Microscopical Examinations. (Smear Preparations } Lung (substance).—Numerous tubersle bacilli. Liver (substance). — Very numerous scattered tubercle bacilli and some smali clumps. Spleen (substance), — Very numerous tubercle bacilli. Kidney (substance).—Moderately numerous tubercle bacilli. Popliteal Gland bacilli, Mesenteric Gland tubercle bacilli, Small Intestine (scraping).—Very numerous tubercle bacilli. (substance).—Numerous tubercle (substance). — Very numerous 313 CALF 544. Virus A VIII, Intravenous inoculation of culture derived from the original material Dose—10-0 milligrammes. Date of Inoculation—May 28, 1907. [Age 64 weeks.] . Died—June 30, 1907. [33 days after inoculation.] Temperature. WAY 190] JUNE ki Weights. Kilos. May 28, 1907 re 35°38 June 30, 1907 we we 31:29 Total loss in weight.—4:09 kilos. POST-MORTEM EXAMINATION. Condition poor. Thorax. Plewra.—Normal. The pleural cavities contained a small amount of ‘fluid. Lungs. — The left anterior and antero - inferior portions of the caudal lobes were dark red, solid and ‘airless. They showed a few collections of ‘small greyish patches. The rest of the caudal lobe was beset with pneumonic nodules. The corresponding parts of the right lung, including the middle lobe, were in a similar condition of red hepatisation, but in -the anterior lobe the substance was closely beset with greyish early caseous patches. Thoracic Glands—The bronchial and mediastinal glands were congested and oedematous. Pericardium and Heart.—Normal. Abdomen. Peritoneum and Omentum.—Normal. Intestines. —The intestines appeared normal. Mesenteric Glands.—These glands were not enlarged :and appeared normal. Spleen.—The spleen weighed 50Y grammes ; it was considerably enlarged and firm in consistency. On section the splenic pulp was dark red, but otherwise normal in appearance. Liver.—The interlobular network of the liver was prominent. Portal Glands.—In the portal glands there were some scattered small discrete slightly yellow patches. Kidneys.—The kidneys appeared normal. On section the pelves were oedematous. Tongue, Tonsils, Larynx, and Pharynz.—Normal. The Subcutaneous Tissues of the body and the extra- peritoneal tissue were oedematous. Various Lymphatic Glands. Parotid, Submasillary, Cervical, Prescapulur, Pre- pectoral, Axillary, Precrural, Popliteal, Gluteal, Ischiatic, Coeliac, Lumbar, and Iliac Glands.—All were odematous with varying amounts of congestion, but showed no caseation. Microscopical Examinations. (Smear Preparations.) Liver (substance).—Tubercle bacilli numerous. Lung (substance).—Tubercle bacilli numerous, but not so numerous as in the liver. Spleen (substance).—Tubercle bacilli numerous, but not so numerous as in the liver. Intestine (scraping).—Tubercle bacilli scanty. 314 DOG 152 (Puppy). Virus A VIII. Intravenous inoculation of culture derived from the original material. Dose—1‘0 milligramme. Date—May 28, 1907. Killed—December 8, 1907. Weight—3200 grammes. Tuberculin Tests. September 18, 1907. [113 days after the experi- ment began.] Dose of tuberculin (bovine) 4:0 cc. No reaction. October 29, 1907. [154 days after the experiment began.] Dose of tuberculin (avian) 4:0 cc. No re- action. [194 days after inoculation. ] POST-MORTEM EXAMINATION. Carcass in good condition. Liver.—In the liver there were two small grey nodules composed of soft translucent tissue. There was no sign of tuberculosis elsewhere. Microscopical Examination of smears made from the lungs, liver, and spleen showed no tubercle bacilli. DOG 154 (Puppy). Virus A VIIL. Intravenous inoculation of culture derived from the original material. Dose—10-0 milligrammes. Date—May 28, 1907. Killed—November 15, 1907. [171 days after inoculation.] Weight—Initial : 3530 grammes. Final: 10,400 grammes. Tuberculin Tests. September 18, 1907. [113 days after the experi- ment begau.] Dose of tuberculin (bovine) 4:0 cc. No reaction. October 29, 1907. [154 days after the experiment began.] Dose of tuberculin (avian) 4°0 cc. No re- action. POST-MORTEM EXAMINATION. Carcass in good condition. The animal was perfectly healthy and showed no trace of the inoculation. PONY 2 (Aged). Virus A VIII. Subcutaneous inoculation with culture derived from the original material. Dose—50°0 milligrammes. Date of Inoculation—August 15, 1907. Killed when in good health—January 20, 1908. Clinical History. A small rounded prominent tumour was formed by the inoculation of 10 ce. of emulsion. This flattened out and 24 hours later was asoft flattened but slightly raised tumour 8 by 10cm. The tumour became more prominent, tense, and fluctuating. A small hole formed six days after inoculation. The ulcer became larger and discharged. Temperature. Normal. Tuberculin Test. October 29,1907. [75 days after the experiment began.] Dose of tuberculin (avian) 4:0 cc. Rise of temperature 2°3° C. Weights. Kilos. August 15, 1907 334-68 January 20,1908 ... 365-62 Total gain in weight.—30 84 kilos. POST-MORTEM EXAMINATION. General condition good. Local Lesion.—In the skin at the seat of inocula- tion on the left side of the neck there was an ulcer covered with red granulations and measuring 2:0 by 05 cm.; the edges were smooth and turned in. On section through it there was found in the deeper [158 days after the experiment began. ] layers of the skin a number of yellowish fairly firm caseous nodules. Prescapular Gilands.— Left prescapular glands. Two of the glands in the prescapular group were firm, the largest the size of a sparrow’s egg ; this contained a firm yellowish waxy caseous nodule easily shelling out and replacing nearly half the substance; the second gland was smaller but similarly affected. The right prescapular glands were normal. Cervical Glands.—Normal. Thorac. Lungs.—In the lungs there was a firm grey trans- lucent patch 1 cm. in length beset with opaque foci (nodule I). There was a second circular nodule 1-2 cm. in diameter slightly raised on the pleural surface which on section was perfectly translucent. There was a third small nodule slightly opaque in the centre. Abdomen. In the omentum there was a firm fibrous nodule containing in the centre brownish yellow calcareous substance. , There was a mass of yellowish grey necrotic sub- stance around and infiltrating the vessels in one part of the mesentery (parasitic). Liver.—The liver contained, scattered deeply in its substance, four or five collections of calcareous granules enclosed in firm fibrous capsules resembling those seen in the other ponies. All other organs and glands showed no tuberculosis. 315 Microscopical Examinations. (Smear Preparations. ) Prescapular G'land.—Moderately numerous tubercle bacilli. Lung (nodule I).—One tubercle bacillus seen. Liver.—No tubercle bacilli. Mesentery.—No tubercle bacilli. Cultures. No growth was obtained in cultures made from two prescapular glands and a lung nodule. PONY 4 (Aged). Virus A VIII. Fed once with culture derived from the original material. Dose—100°0 milligrammes. Date of Feeding—August 15, 1907. Killed when in good health—January 22, 1908. [160 days after the experiment began. ] Clinical History. There was no sign of ill-health during the experiment. Temperature. Normal. Tuberculin Test. October 29, 1907. [75 days after the experiment began.] Dose of tuberculin (avian) 4:0 cc. Rise of temperature 2°0° C. Weights. kilos. August 15, 1907 21264 January 22, 1908 ... 235°36 Total gain in weight.—22°72 kilos. POST-MORTEM EXAMINATION. General condition good. Intestines.—Beneath the mucous membrane of the small intestine two minute opaque foci were seen. Large Intestine—Normal. Mesenteric Glands.—There was an opaque focus in one mesenteric gland. Mesentery—In the mesentery between the layers there were about 20 hard irregular nodules (parasitic) up to 1 cm. in diameter. On section they showed a firm dirty yellow shell containing soft yellow necrotic substance in the centre. There was slight thickening and necrosis of a mesenteric artery. In conditions resembling these in other ponies small worms have been found. Liver—There were a few scattered calcareous nodules within grey capsules similar to those seen in other adult ponies. All other organs and glands were normal. icroscopical Examinations. (Smear Preparations.) Liver.—No tubercle bacilli. Intestine (opaque focus).—No tubercle bacilli. Intestine (gritty nodule).—No tubercle bacilli. Mesentery (nodules).—No tubercle bacilli. Mesenteric Gland (opaque foci).—No tubercle bacilli. PIG 248. Virus A VIII. Intravenous inoculation with culture derived from the original material. Dose—50:0 milligrammes. Date of Inoculation—April 7, 1908. Died—May 24, 1908. Clinical History. The animal quickly became ill, and emaciated rapidly. It was pale and had an infrequent hoarse cough, Tuberculin Test. May 18, 1908. [36 days after inoculation.] Dose of tuberculin (avian) 2:0 cc. No reaction. Weights. kilos. April 7, 1908 is ae 46°25 May 24, 1908 x0 31-74 Loss in weight.—14°41 kilos. POST-MORTEM EXAMINATION. Body.—Emaciated. Lungs.—The lungs filled the pleural cavities. The extremities of the anterior lobes were reddish grey and solid. The rest of the lungs was oedematous and emphysematous, but not congested. [47 days after inoculation.] [Age 19 weeks. ] Bronchial Glands.—They were a little firmer than normal ; the right showed an opaque yellowish focus. Spleen.—The spleen was enlarged, and the pulp was firm and dark red in colour. Liver.—Norwal. Portal Glands.—These glands were firm, and showed scattered streaks of soft yellowish necrotic substance. Intestines.—There were patches of congestion in the mucous membranes. : There was no sign of tuberculosis elsewhere. Microscopical Examinatiens (Smear Preparations. ) Lung (substance) Liver (do.) | samen tubercle bacilli. Spleen do.) Kidney (substance}.—A few tubercle bacilli. Intestine (Peyer's patch) Numerous tubercle Mesenteric Gland eae bacilli. sonk 48. 44, RABBITS. N ous. Subcutaneous. = umber. | Method. 0 mg. Dose : 50:0 mg. Number. | Method.| Dose. ti eae of Result. | mber 10, Killed when very ill: a 162 Intrav. September 30, 1907. (Puppy). eT 33 disse 1887 | Intrav.| 0-1 mg. | D. 26 days} G.T. were some P.M.—Small local ulcer. 13885 | Intrav. | 0-01 oid 164 Intrav. |, tubercles Gritty caseo - necrotic sii Hi | Aes TE a eo loculation ; masses in left prescapular 1384 | Tntrav. | 0-001mg.| D. 78 ,, |G. ~, bulargland gland. The lungs con- by caseo- tained a moderate number, A een Ae ee eee es. The of translucent nodules, bd three some with gritty centres. cles, and The liver showed a num- all caseous ber of yellowish necrotic nthe ileo- foci. “There were three minute foci in one kidney. GUINEA-PIGS. The Peyer’s patches were aia ae eee Number. | Method.| Dose. ae uf Result. (T.B. swarming). The hee ; mesenteric glands were en- larged and caseo-necrotic. 2556 | Intrap.| 1:0 mg. | D. 64 days | Subacute There were caseous avian T. patches in the thoracic 2555 | Intrap.| Ol mg. | K. 202 ,, | Local T. and portal glands. T.B. were seen in a smear from 2557 | Subcut.) 10-0 mg.) K. 202 ,, |NoT. the spleen. MONKEY 188. GUINEA-PIGS. MICE. Intravenous. ; | as Dose : 50:0 mg. xthod.| Dose. ee of Result. Number. | Method.| Dose. ae of Result, Died : Dec. 15, 1907. ; 18} days. beut. | 50°0 mg. | D. 106 days | Local T. 103 | Intrap. | 10:0 mg.| K. 260 days og a ent, | BOOing, | De 99 | Local T. 101 | Intrap. 1:0 mg.| D. 43, = 3 % 28 100 | Intrap.| 0-1 mg.| D.191_,, go a a 99 | Intrap. | 0:01 mg.| K. 350 _,, _ + op : 105 Subcut. | 50:0 mg.} D.105_,, 5, é 106 | Subeut. | 500 mg.) D. 128, Z 3 104 Subcut.| 10:0 mg.| K.127 ,, Pa is 102 | Subcut.| 1:0 mg.| K.350 ,, sl i f umber. | Method.| Dose. ee Result. 384 Intrap.| 0-1 mg. | D. 50 days G. T. 382 | Intrap.| 0-01mg.| D.46 , | G.T. 390 Intrap.| 00001 | K.50 , Sli gh t mg. G. T. 388 | Intrap.| 0:00001 | K.50 ., |Slight 2 mg. G. 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Fed once with culture derived from the original material. Dose—50:0 milligrammes. Date of. Feeding—July 9, 1907. [Age 9 weeks, ] Killed when in good health—November 28, 1907. Temperature. Normal. -Tuberculin Tests, September 18, 1907. [71 days after the experiment began.] Dose of tuberculin 2-0 cc, Negative re- action. October 29, 1907. [112 days after the experiment began.] Dose of tuberculin 4:0 ce. Positive re. action. Rise of temperature 2°1° C. Weights. kilos. July 10,1907 ... 14:06 November 28, 1907 41:72 Zotal gain in weight.—27°66 kilos. POST-MORTEM EXAMINATION, General condition good. Tongue, Tonsils, Larynx, and Pharynx.—Normal. Parotid, Submaxillary, Pharyngeal, and Cervical Glands.—Normal. Abdomen. Omentum and Peritoneum.—Normal. Intestines.—Normal. Mesenteric Glands.—The mesenteric glands were [142 days after feeding.] slightly enlarged, almost all contained caseous gritty nodules and masses replacing in many cases the greater part of the gland substance ; these nodules easily shelled out leaving smooth walled cavities. Colic Glands.—Several of the small colic glands contained nodules similar to those in the mesenteric glands. Spleen.—Normal. Liver and Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Renal, Coeliac, Lumbar, and Iliac Glands.—Normal. Thorax. Pleura.—Normal. Lungs.—Normal. Bronchial and Mediastinal Glands —Normal. Heart.—Normal. Other Lymphatic Glands. Prescapular, Precrural, and Inguinal Glands.— Normal. Smear preparations from the Lung, Spleen, Liver and Kidney showed no tubercle bacilli. Mesenteric Gland showed moderately numerous tubercle bacilli. PIG 198. Virus A X. Fed once with culture derived from the original material. Dose—50-0 milligrammes. Date of Feeding—July 9, 1907. Killed when in good health—October 28, 1907. [Age 9 weeks. ] Temperature. Normal. Tuberculin Test. September 18,1907. [71 days after the experiment began.] Dose of tuberculin 2:0 cc. No reaction. . siti Weights kiloa, 13°60 July 10, 1907 ... 59-44 October 28, 1907 Gain in weight.—25°84 kilos. POST-MORTEM BXAMINATION. General condition good. Tongue, Tonsils, and Pharynz.—Normal. Submazillary Glands—In one right submaxillary gland there was a soft caseous tubercle. Pharyngeal Glands.—Normal. Cervical Glands.—Normal. Abdomen. Peritoneum and Omentum.—Normal. 34684 [111 days after feeding. ] Intestines.—Normal. Gastric Glands.—Normal. Mesenteric Glands—The mesenteric glands were enlarged and almost all were filled and practically re- placed by soft caseous gritty masses which easily shelled out leaving smooth walled cavities. Spleen.—Normal. Liver.—Normal. Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Renal, Coeliac, Lumbar, and Iliac Glands.—Normal. \ Thorax. Pleura.—Normal. Lungs.—On the surfaces of the lungs just beneath the pleura there were several grey completely trans- lucent nodules up to nearly a hemp seed in size (? parasitic) ; there was also a small grey patch around a bronchiole which on section contained several round worms. Bronchial Glands.—In one bronchial gland there- was a minute opaque focus. The rest were normal. 3A 322 Medrastinal Glands.—Normal. Heart.—Normal. Various Lymphatic Glands. Parotid, Prescapular, Prepectoral, Axillary, Pre- crural, Ischiatic, Gluteal, and Popliteal Glands.— Normal. Smear preparations from the Lung (substance and nodule), Spleen, Liver, Kidney, and Intestinal Mucous Membrane showed no tubercle bacilli. Mesenteric Gland a few scattered tubercle bacilli. PIG 200. Virus A X. Fed once with culture derived from the original material. Dose—50-0 milligrammes. Date of Feeding—July 9, 1907. [Age 9 weeks.] Killed when in good health—November 12, 1907. Temperature. Normal. Tuberculin Tests. September 18,1907. [71 days after the experiment began.] Dose of tuberculin, 2:0 cc. No reaction. October 29, 1907. [112 days after the experiment began.] Dose of tuberculin, 4:0 cc. Positive reaction. Rise of temperature, 2°2° C. Weights. kilos. July 10, 1907 ... a sa we =: 16°78 November 12,1907 ... 28°11 Gain in weight.—11°33 kilos. POST-MORTEM EXAMINATION. General condition good. Tongue, Tonsils, and Pharynz.—Normal. Submaxillary Glands.—Normal. Pharyngeal Glands.—Normal. Cervical Glands.—Normal. Abdomen. Peritoneum and Omentum.—Normal. 4£ntestines.—Normal. Mesenteric Glands.—The mesenteric glands were not obviously enlarged, but were closely beset throughout the whole chain with yellow caseous gritty nodules ; these nodules easily shelled out ; they replaced the greater part of the gland substance. Spleen.—Normal. [126 days after feeding. ] Liver —Normal. Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Renal, Coeliac, Lumbar, and Iliac Glands.—Normal. Thorax. Pleura.—Normal. Lungs.—The lungs showed a few congested patches on the dorsal surfaces, and just beneath the pleura there was a single soft grey nodule. Bronchial and Mediastinal Glands.—Normal. Heart.—Normal. Various Lymphatic Glands. Parotid, Prescapular, Prepectoral, Amillary, Pre- crural, Ischiatic, Popliteal, and Gluteal Glands.— Normal. Microscopical Examination. (Smear Preparations.) Lung «. agi Spleen... | Liver ... Kidney Peyer's Patch | Mesenteric Gland.—Moderately numerous tubercle bacilli. - fae tubercle bacilli. GOAT 42. Virus A X. Subcutaneous inoculation of culture derived from the original material. Dose—50°0 milligrammes. Date of Inoculation—July 9, 1907. Killed when in good health—October 15, 1907. Temperature. The temperature rose sharply on the fourth day after inoculation to 41°6° C. and slowly fell until the ninth day. It subsequently varied between 39° and 39-8° for several weeks until it became steady and normal. [Age 9 months. ] [98 days after inoculation.] Tuberculin Test. September 18, 1907. began.] Dose, 2:0 cc. of tuberculin. negative. [71 days after the experiment Reaction 323 Weights. kilos, July 9,1907 ... a st oat 26-30 October 15, 1907 viet oe = 27°66 Gain in weight.—1°36 kilos. POST-MORTEM EXAMINATION. General condition good. Local Lesion.—On the left side of the neck, situated in the muscle, there was a somewhat pear-shaped tumour 8 cm. long and 5 em. at its widest diameter. On section it consisted of a firm translucent fibrous wall enclosing pinkish caseo-necrotic substance with gritty streaks ; in the centre there was a cavity con- taining turbid fluid. The tumour was situated near the gland, and in the neighbouring tissue between them were discrete caseo- calcareous tubercles and nodules up to a wheat grain in size. Prescapular G'lands.—The left prescapular gland consisting of two parts joined by a narrow neck each measuring 2°8 by 22 cm. On section it contained pinkish caseo-necrotic masses with calcareous streaks replacing more than half the substance. The right prescapular glands measured each 1°5 by 16cm. On section they were normal. Prepectoral G'lands.—Normal. Cervical Glands.—Normal. Thorax. Pleuwra.—Normal. Lungs.—At the extremities of the caudal lobes of both lungs there were areas beset with small, and for the most part ill-defined grey nodules, many of which were yellowish in the centre ; they were in the depth of the lung, but could be felt on palpation beneath the pleura. They were produced by Strongylus Filaria ; otherwise the lungs were normal. Bronchial and Mediastinal Glands.—Normal. Heart.—Normal. Abdomen. Omentun.—In the omentum, attached by a narrow pedicle, there was a reddish nodule 1 by ‘5 cm., which on section contained yellowish necrotic substance. _ Peritoneum.—On the parietal peritoneum on the left side there were three grey nodules the size of rape seeds. Diaphragm.—Normal. Small Intestine-—Normal. Large Intestine.—In the large intestine beneath the mucous membrane there was a caseo-purulent nodule the size of a millet seed. Gastric Glands.—Normal. Mesenteric Glands.—The majority of the mesenteric glands contained in the cortices irregular calcareous granules, in many places forming a network which in all cases was easily picked out from the substance. Colic Glands.—Many of the colic glands contained nodules resembling those in the mesenteric glands. Spleen.—Normal. Liver.—Normal. Portal Glands.—Normal. Kidneys.—In each kidney there was a single grey focus, Zestes.—In a testis there were several wedge-shaped streaks of whitish gritty substance. Tongue, Tonsils, Larynx, and Pharynz,—Normal. Various Lymphatic Glands. Parotid, Submawillary, Posterior pharyngeal, Pre- crural, Renal, Coeliac, Lumbar, Iliac, and Popliteal Glands.—Normal. Microscopical Examinations. (Smear Preparations.) Local Lesion.—Very numerous tubercle bacilli. Left Prescapular Gland.—Moderately numerous tubercle bacilli. _ Lung (nodule).—No tubercle bacilli. Kidney focus.—No tubercle bacilli. Omental Nodule.—A few tubercle bacilli. Mesenteric Gland.—Two tubercle bacilli seen. Intestine (scraping from).—No tubercle bacilli. Testis (nodule).—A few tubercle bacilli. GOAT 44. Virus A X. Subcutaneous inoculation of culture derived from the original material. Dose—10-0 milligrammes. Date of Inoculation—July 9, 1907. [Age 7 months. ] Killed when in good health—December 10, 1907. [154 days after inoculation.] Temperature. After August 16, 1907, the temperature remained normal except for a short rise in November. 34684 3A 2 324 Tuberculin Tests. September 18,1907. [71 days after the experiment began.| Dose 2:0 cc. of tuberculin. No reaction. October 29, 1907. [112 days after inoculation] Dose of tuberculin 4:0 cc. Positive reaction. Rise of temperature 2:7° C. Weights. kilos. July 9, 1907... An 21°77 - December 10, 1907 ... 25°40 Gain in weight.—3°63 kilos. POST-MORTEM EXAMINATION. General condition good. Local Lesion —At the seat of inoculation on the left side of the neck, in the subcutaneous tissue and in the muscle, there were scattered irregular caseo- calcareous tubercles, 2 few elongated but none larger than millet seeds. Prescapular Glands.—The -left prescapular gland was enlarged measuring 4 by 2°3 by 22cm. Almost half the gland was replaced by a mass of pinkish caseo-necrotic tissue easily shelling out. In the other half there was a pea-sized nodule of similar substance. The right prescapular gland measured 2°9 by 1:2 by 0'7 cm. On section it was normal. Cervical Glands.—In the cortex of the left superior cervical gland just beneath the capsule were scattered caseo-calcareous granules. Other cervical glands normal. Agillary Glands.—Normal. Thorax. Pleura.—Normal. Lungs.—At the caudal extremities of each posterior lobe both in the centre and in the margins, were several firm grey patches, translucent around the peri- phery, becoming slightly opaque and yellowish tewards the centre; the patches measured up to 3 cm. in diameter and scattered around them were smaller nodules similar in structure. The anterior lobes showed an occasional firm grey nodule. These nodules were due to parasites (Strongylus Filaria) which were easily found on microscopical examination. Mediastinal Glands.—Normal. Bronchial Glands.—Normal. Heart.—Normal. Abdomen. % Peritoneum and Omentum.—Normal. Intestines.—Normal. Mesenteric Glands.—Normal. Tleo-Colic Glands.—In these glands just beneath the capsule there were several small patches composed of yellowish caseous gritty collections. Spleen.—Normal. Liver.—In the liver just beneath the surface there were three translucent miliary tubercles with opaque gritty centres ; no more were seen on section. Portal Glands.—Normal. Kidneys.—Normal. Renal, Coeliac, Lumbar, and Iliac Glands.—Normal, Tongue, Tonsils, Larynx, and Pharynx.—Normal. Uterus, Ovaries, Mammary, and Supramammary Glands.— Normal. Submazillary, Precrural, and Popliteal Glands.— Normal. Microscopical Examinations. (Smear Preparations.) Prescapular Gland numerous tubercle bacilli. Lung (nodule).—A few tubercle bacilli. Lung (nodule ; fresh specimen).—Motile worms. Liver (tubercle).—No tubercle bacilli. (emulsion). — Moderately GOAT 48. Virus A X. Subcutaneous inoculation of culture obtained from the original material. Dose—50°0 milligrammes. Date of Inoculation—July 9, 1907. [Age about 5 months. ] Killed when very ill—September 30, 1907. [83 days after inoculation.] Temperature. The temperature rose to 41°C. on the 5th day. It subsequently varied between 39°1° and 40°5° with a gradual downward inclination. Weights. kilos. July 9, 1907 ... ais 12:25 September 30, 1907... 8°39 Loss on initial weight.—3°86 kilos. POST-MORTEM EXAMINATION, General condition poor. Local Lesion —At the seat of inoculation on the left side of neck there was an oval ulcer measuring 5 by 4 cm., the base of which was formed by the muscle ; the floor was about two-thirds clean, while one-third was covered with pus coming from under the margin. The rest of the margin was thin and very little undermined. Left Prescapular Gland.—The left prescapular gland measured 6 by 4 by 2cm. On section the substance was almost replaced by two masses of caseo-necrotic slightly gritty substance. There was some oedema around the left prescapular gland, and several flattened firm fibro-caseous tubercles up to 3 mm. in diameter. Right Prescapular Gland.—This gland measured 2-7 by 1:5 by 0°6 cm. ; on section it was normal. The tissues around it were slightly oedematous. Prepectoral Glands.—Normal. Cervical Glands.—Normal. Thorax. Pleura.—Normal. Lungs.—The lungs did not quite collapse, they were oedematous and slightly congested ; they showed on the surfaces numerous irregular translucent nodules, the majority with opaque yellowish, occasionally gritty centres; they were distinctly palpable. On section through the substance, they were distributed throughout but in fewer numbers. Many were caused by the Strongylus Filaria. 32 Thoracic Lymphatic Glands. The Bronchial and Dorsal Mediastinal Glands were not obviously enlarged, and contained a moderate number of irregular, discrete, caseous patches, similar to those in the portal glands. Heart.—Normal. Diaphragm.—Normal. Abdomen. Omentum.—In the omentum there were two flat translucent tubercles with opaque centres up to 2 mm. in diameter. Peritoneum.—Normal. Intestines.—The Peyer’s patches of the small intes- tine were closely beset with irregular sharply marked pits varying up to 3 and 4 mm. roughly ; the margins of the pits were slightly mora opaque than the rest of the patch and yellowish in colour. Large Intestine.—Normal. Gastric Glands.—They tubercles. Mesenteric Glands.—These glands were enlarged and firm, and the majority of them had their cortices almost completely replaced by firm caseo-necrotic substance, in many parts of which could be seen a network of congested capillaries. showed small caseous - uv Spleen.—Normal. Liver.—The surface of liver presents a greyish mottled appearance from the presence of numerous irregular grey and yellowish necrotic foci. Onsection through the substance they were much less numerous. Portal Glands.—The portal glands contained in the cortices small irregular discrete yellowish caseous patches. Kidneys.—In the cortex of left kidney there were three minute greyish white foci. Right Kidney.—Normal. Suprarenal Bodies.—Normal. Renal, Coeliac, Lumbar, and Iliac Glands.—Normal. Tongue, Tonsils, Larynx, and Pharynx.—Normal. Parotid, Submazillary, Pharyngeal, Precrural, Popliteal, Gluteal, and Ischiatic Glands.—Normal. Microscopical Examinations. (Smear Preparations.) Lung (substance) showed moderately numerous tubercle bacilli with a few small clumps. Spleen (substance) Liver ( do. ) Omentum (nodule) showed a few tubercle bacilli. Peyer's patch showed swarms of tubercle bacilli. Some scattered tubercle bacilli, but chiefly in small clumps. DOG 162 (Puppy). Virus A X. Intravenous inoculation with culture derived from the original material. Dose—50°0 milligrammes. Date of Inoculation—July 9, 1907. Killed—December 2, 1907. Tuberculin Tests. The animal was tested 71 days and 112 days after ‘inoculation and gave no reaction. The dose on the ‘first occasion was 4°0 cc. of bovine tuberculin ; on the -second 4:0 ce. of avian tuberculin. Weights. grammes. July 9, 1907 ... sie vee ses 4850 December 2,1907 ... oe sit 9400 Gain in weight.—4550 grammes. [146 days after inoculation. ] POST-MORTEM EXAMINATION. General condition good. Lungs.—In the lung there were two translucent foci of doubtful nature. Kidneys.—In one kidney there were two small grey nodules with opaque centres. All other organs and glands were quite healthy. No tubercle bacilli could be found in smears made from the substance of the lung, liver, and spleen. DOG 164 (Adult). Virus A X. Intravenous inoculation with culture derived from the original material. Dose—50°0 milligrammes. Date of Inoculation—June 9, 1907. Weight—10,270 grammes. Killed—November 28, 1908. [142 days after inoculation. ] Tuberculin Tests. The animal was tested 101 days after inoculation -and gave no reaction. It was retested 142 days after ‘inoculation and gave a rise of temperature of 1 C. “The dose on the first occasion was 4°0 cc. of bovine tuberculin ; on the second 4:0 cc. of avian tuberculin. POST-MORTEM EXAMINATION. General condition good. Lungs.—Normal. Spleen.—In the spleen there were two nodules, one projecting, the size of a small pea; on section they 326 consisted of soft grey translucent tissue (not tuber- culous). Kidneys.—In the cortices of the kidneys there were a few scattered grey nodules with, in a few cases, opaque centres the largest the size of a rape seed. (Non-tuberculous nodules often found in the kidneys of dogs.) All other organs and glands appeared normal. Microscopical Examinations. (Smear Preparations.) Spleen (nodule).—No tubercle bacilli. Kidney (nodule),—No tubercle bacilli. Cultures. Cultures were sown from the liver, kidney, and spleen substance. In each case moderately numerous. typical avian colonies developed. MONKEY 188. Virus A X. Intravenous inoculation of culture from the original material. Dose—50°0 milligrammes. Date—November 27, 1907. Died—December 15,1907. [18 days after inoculation. ] POST MORTEM EXAMINATION. Lungs.—Thelungscollapsed normally ; they were con- gested and slightly oedematous and on close inspection a few minute grey foci up to 1:0 mm. could be seen. Bronchial Glands.—They were oedematous ; other- wise normal. Spleen.—The spleen was swollen measuring 6 by 4 by 1:5 cm. On section the malpighian bodies were enlarged ; no definite grey points could be seen. Liver.—The substance showed a fine grey network outlining the lobules and in patches apparently made up of minute grey points. Kidneys.—Tke cortices were pale. Other organs and glands normal. Microscopical Examinations. Numerous tubercle bacilli were demonstrated in smears made from the lung, liver, spleen, and bronchial gland ; a few in the kidney. MONKEY 190. Virus A X. Intravenous and intraperitoneal inoculation of culture from the original material. Dose—1:0 milligramme. Date—November 27, 1907. Died—December 23, 1907. POST-MORTEM EXAMINATION. Lungs.—The lungs were oedematous and the left was congested. No tubercles could be seen. Bronchial Glands.—The bronchial glands were en- larged and oedematous ; they showed in the cortices ‘numerous opaque yellowish foci. Abdomen. Omentum.—The omentum was not retracted, but was uniformly thickened with greyish opaque tubercles lying chiefly along the blood vessels, varying up to a millet seed in size. The spaces between the vessels were covered with minute grey tubercles. Mesentery.—On the mesentery there were similar tubercles but not so numerous. Spleen.—The spleen measured 6 by 3°5 by 1°5 cm. (0:5 intravenously and 0°5 intraperitoneally. ] [26 days after inoculation. ] It was soft and on section the pulp was closely packed with minute grey points. Liver.—Normal. Portal Glands.—They were oedematous but showed no sign of caseation. Kidneys.—Normal. Abdominal Glands.—They were oedematous but showed no sign of caseation. Microscopical Examinations. (Smear Preparations.) Lung (substance).—No T.B. Spleen (substance ).—Moderately numerous T.B. Liver (substance).—A few T.B. Bronchial Gland.—~Numerous T.B. 3 2 7 MONKEY 196. Virus A X. Intravenous inoculation of culture from the original material. Dose—10°0 milligrammes. Date—November 27, 1907. Died— December 26, 1907. POST-MORTEM EXAMINATION. Eungs.—The lungs felt slightly firm and were a little oedematous. The right was slightly congested and both showed dark pigmentary mottling. Spleen.—The spleen was slightly enlarged. On section the malpighian bodies were prominent but -otherwise the pulp was normal in appearance. Liver.—The substance was pale. The surface beneath the capsule was normal, but on section numerous rather indefinite grey points could be seen through a hand lens. Kidneys.—They were pale but otherwise normal. [29 days after inoculation.] Lymphatic Glands.—They appeared enlarged and were oedematous but showed no sign of caseation, Microscopical Examinations. (Smear Preparations.) Lung (substance).—Numerous tubercle bacilli. Liver (substance).— Very numerous tubercle bacilli. Spleen (substance).— Very numerous tubercle bacilli. Kidney (substance).—A few tubercle bacilli. Mesenteric Gland.—Very numerous tubercle bacilli. MONKEY 198. Virus A X. Intravenous inoculation of culture from the original material. Dose—100-0 milligrammes. Date—November 27, 1907. Died—December 25, 1907. POST-MORTEM EXAMINATION. Lungs.—The lungs were pink and crepitant ; they showed a few patches of congestion and emphysema, but no foci were visible. Spleen.—The spleen was swollen, measuring 6°5 by 35 by 15 cm. The pulp was firm, and on section showed a faint grey tracing, and a moderate number of irregular small necrotic areas. Liver.—The liver was pale, and in the darker areas could just be seen numerous minute grey points. Kidneys.—Normal. Suprarenal Bodies.—Normal. [28 days after the inoculation.] Lymphatic Glands. All the lymphatic glands were enlarged and oedematous, and showed grey points which could be seen best in those that were congested. Microscopical Examinations. (Smear Preparations.) Spleen (substance).—A mass of tubercle bacilli. Liver (substance).—A mass of tubercle bacilli. Mesenteric Gland.—A mass of tubercle bacilli. Lung (substance).—Numerous tubercle bacilli. Cidney (substance).—Numerous tubercle bacilli. PIG 260. Virus A X. Intravenous inoculation with culture derived from the original material. Dose—50°0 milligrammes. Date of Inoculation—May 21, 1908. Clinical History. The animal quickly became ill with laboured breathing, and rapidly lost flesh. ‘ Temperature. On the 12th day after inoculation the temperature rose to 40°9° C. ‘It remained high (between 39:9 C. and 41-0° C.) for 34 days; it then fell rapidly, death ensuing within a few days. Weights. Kilos. May 21, 1908 igs as sa 29°48 July 11, 1908 os fi ae 16°77 Loss in weight.—12°71 kilos. POST-MORTEM EXAMINATION. Lungs.—The lungs were firm, and did not collapse normally. They were not congested, but were [Age 15 weeks.] Died—July 11, 1908. [51 days after inoculation.] oedematous and emphysematous. ‘There were a few scattered reddish-grey patches. Spleen.—The spleen was slightly swollen. Liver.—Normal in appearance. Kidneys.—Normal in appearance. Intestines,—They were thin and congested. Lymphatic Glands.—The lymphatic glands showed occasionally very indefinite, small yellowish grey patches. Microscopical Examinations. (Smear Preparations.) Lung (substance).—Very numerous tubercle bacilli. Spleen (substance).—Swarms of tubercle bacilli. Liver (substance).—Very numerous tubercle bacilli. Intestine (scraping).—Numerous tubercle bacilli. Mesenteric Gland.—Swarms of tubercle bacilli. Precrural Gland.—Numerous tubercle bacilli. 328 PIG 262. Virus A X. Intravenous inoculation with culture derived from the original material. Dose—10:0 milligrammes. Date of Inoculation—May 21, 1908. Died—July 30, 1908. [Age 15 weeks. ] [70 days after inoculation.] - Temperature. Weights. Kilos. May 21, 1908 30°38 July 30, 1908 20°86 Loss of weight.—9-52 kilos. POST-MORTEM EXAMINATION. Carcass emaciated. Thorax. Lungs.—The lungs were voluminous and cut some- what firmly. The anterior inferior extremities were reddish grey, and solid ; the rest of the lungs showed on section patchy congestion and slight oedema. The bronchial glands were congested. Abdomen. Intestines.—Normal. Spleen.—The spleen weighed 140 grammes, and on section the pulp was a little firm and red. Liver.—The liver was a pale yellowish colour. Portal Glands.—In a portal gland there was a small: soft, yellowish necrotic nodule. Kidneys.—Normal. Tonsils.—In the tonsils there were a few soft yellow miliary nodules, and in one a small ulcer was seen. The majority of the lymphatic glands were con- gested, but showed no definite foci. Other organs and glands appeared normal. Microscopical Examinations. (Smear Preparations.) Lung (substance).—Swarms of tubercle bacilli. Spleen (substance).—Swarms of tubercle bacilli. Liver (substance).—Swarms of tubercle bacilli. Mesenteric Gland.—Swarms of tubercle bacilli. Cidney (substance).— Very numerous tubercle bacilli. Inguinal Gland.—Very numerous tubercle bacilli. Intestine tubercle - bacilli. (scraping).—Very numerous PIG 264. Virus A X. Subcutaneous inoculation with culture derived from the original material. Dose—50°0 milligrammes. Date of Inoculation—May 21, 1908. Killed—September 3, 1908. Clinical History. There was no sign of ill-health during the experiment. Weights. Kilos. May 21, 1908 ... ae obs eee = 31°75 September 3, 1908... tee we = 43:54 Gain in weight —11:79 kilos. POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion.—At the seat of inoculation there was a tumour about the size of a hen’s egg in the sub- cutaneous tissue, over which the skin was intact. On [Age 15 weeks. ] {105 days after inoculation. ] section it consisted of a thin-walled cyst filled with soft yellow caseous substance. Inguinal Glands.—A left inguinal gland contained a nodule about the size of a sparrow’s egg composed of soft caseous substance. In a right inguinal gland there were three or four soft caseous slight gritty tubercles. Kidneys——In the cortices there was a moderate number of translucent miliary nodules of doubtful nature (no T.B.). There was no tuberculosis elsewhere. Microscopical Examinations.—Smear preparations from the lung, liver, spleen, intestine, and mesenteric gland showed no tubercle bacilli. PONY 382 (Aged). Virus A X. Intravenous inoculation of culture derived from the original material. Dose—10°0 milligrammes., Date of Inoculation—May 21, 1908. Died—June 5, 1908. Clinical History. The animal began to breathe quickly within a few [15 days after inoculation.] days after the inoculation. The frequency increased and the animal went down, June 4, breathing violently. © ; 329 Temperature. The temperature was at 40° C. the day after the injection and remained high until the animal's death : the highest point reached was 40°5° O. Weights. Kilos. May 21, 1908 wee 273-00 June 5, 1908 see 259°39 Total loss of weight.—13'61 kilos. POST-MORTEM EXAMINATION. General condition fair. Thorax. Pleura.—Normal. Lungs.—The lungs were voluminous, heavy, and water-logged. The parenchyma was deeply congested as well as oedematous, especially the anterior: inferior portions which sank in water, Mediastinal and Bronchial Glands.—Normal. Heart.—Normal. Abdomen Omentum and Peritoneum.—Normal. Intestines.—Normal. Mesenteric Glands.—Normal. Colic Glands.—Normal. Spleen —The spleen was swollen and dark red in colour. It weighed 1:47 kilogrammes. Liver.—The liver substance was pale. Portal Glands.—Normal. Kidneys.—In the cortices of the kidneys there were scattered hemorrhagic foci. Suprarenal Bodies.—Normal. Renal, Lumbar, Coeliac, and Iliac Glands.—Normal. Longue, Tonsils, Laryna, and Pharynx.—Normal. Parotid, Submazillary, Retro-pharyngeal, Cervical, Prescapular, Prepectoral, Agillary, Precrural, Pudic, and Popliteal Glands were dark and congested. Microscopical Examinations. (Smear Preparations.) Lung showed moderately numerous tubercle bacilli. Liver showed a few tubercle bacilli. Spleen showed a few tubercle bacilli. Kidney showed a few tubercle bacilli. Prescapular Gland showed a few tubercle bacilli. Mesenteric Gland showed a few tubercle bacilli. CALF 546. Virus A X. Intravenous inoculation of culture derived from the original material. Dose—50-0 milligrammes. Date of Inoculation—May 30, 1907. [Age about 12 weeks, ] Died—July 1, 1907. [32 days after inoculation.] Temperature. MAY 1907 JUNE 39 38 Weights. Kilos. 55°76 May 30, 1907 ... pe July 1, 1907 ae Total loss in weight.—4°97 kilos. POST-MORTEM EXAMINATION. Condition poor. Thorax. Pleura.—Normal. Lungs.—In the cephalic lobes, the right middle and 34684 JULY: the anterior portions of the caudal lobes, there were large dark red areas of consolidation, which, however, did not completely replace those parts of the lungs, since they still floatedin water. The rest of the lung tissue was moderately congested and oedematous. Bronchial and Mediastinal Glands.—These glands were congested and somewhat oedematous but showed no caseation. Pericardium. — The pericardium was a little in- jected. : mat The tissues of the posterior mediastinum were oedematous. 3B 330 Abdomen. Peritoneum and Omentum.—Normal. Intestines.—Normal. Mesenteric Glands.—These glands appeared normal. Spleen.—The spleen was greatly enlarged and weighed 907 grammes. On section the substance was dark red and contained two greyish nodules, becoming yellow towards the centre and about 6 mm. in ‘diameter. Liver.—The surface of the liver showed the inter- lobular network well marked, and on section the substance was congested. Portal Glands.—These glands appeared normal. Kidneys——On the surface of the kidneys beneath the capsule there a few foci of congestion. ~ Suprarenal Bodies.—These appeared normal. Coeliac, Renal, and Lumbar Glands.—All were a little enlarged and congested. Tongue, Pharynz, Larynx, and Tonsils.—Normal., Various Lymphatic Glands. Precrural Glands.—Normal. Gluteal and Popliteal G'lands.—These glands were oedematous. Left Prescapular Gland.—This gland was enlarged and oedematous from a recent inoculation of tuberculin. Many of the remaining lymphatic glands showed some amount of congestion but no caseation. Microscopical Examinations, (Smear Preparations.) Lung (substance) showed numerous scattered tubercle bacilli and a few small clumps. Mediastinal Gland (substance) showed numerous tubercle bacilli. Spleen (pulp). — Moderately numerous tubercle bacilli. Liver (substance).—Numerous tubercle bacilli. Scraping from Intestine showed a few tubercle bacilli. GOAT 62. Fed once with culture derived from the original material. teat.) Dose—60 milligrammes. Virus A X. (Dose given with milk through a rubber Date of Feeding—January 28, 1909. [Age 5 days.] Killed when in good general health—March 25, 1909, [56 days after inoculation.] Weights. Kilos. January 29, 1909 ‘ sta we = 805 March 25, 1909.., a ere v =: 6°30 Gain in weight,—3°25 kilos. POST-MORTEM EXAMINATION. Carcass in fair condition. Tonsils—Each tonsil showed a few caseous streaks and foci. Parotid Glands.—The right gland was enlarged and contained several soft caseous areas beneath the cortex. The left was normal. Cervical Glands.—The right superior cervical gland showed small caseous patches and foci in the cortex. The other right cervical glands and the left were normal. Submacillary Glands.—The submaxillary gland on each side was a little enlarged, and contained nodules formed of a caseous network in grey translucent tissue. Posterior-pharyngeal Glands.—The right gland was about the size of a robin’s egg, twice the size of that on the other side. It was firm, and on section was about half replaced by caseous tissue softening in parts. The gland on the left side contained caseous patches. Abdomen. Intestines.—The first Peyer’s patch was much thick- ened and closely beset with serpiginous ulcers ; beneath the raised portions at the margins of the ulcers were numerous opaque caseous foci, and the floors were lined by easily detached soft caseo-necrotic substance. All the other Peyer’s patches were similarly affected, and in many instances the base of the ulcer was formed merely by peritoneum. The margins of the ulcers were congested. The solitary follicles were prominent, congested, and showed at the top of each a small punched-out ulcer. The large intestine appeared normal. Mesenteric Glands.—The mesenteric glands were enlarged and firm, and on section they consisted of firm grey tissue beset with caseous areas, and many of the glands were considerably replaced by caseous tissue. Ileo-colic Gland.—The ileo-colic gland, about the size of a pigeon’s egg, showed a narrow margin of the cortex replaced by caseous tissue. Spleen.—Normal. Liver.—In the liver there were sparsely scattered minute irregular yellowish-grey foci. Portal Glands—In the cortex of one portal gland there were a few irregular caseous patches and foci. Kidneys.—Normal. Thorax. Lungs.—The lungs appeared normal except for an occasional minute translucent focus. All other organs and glands appeared normal. Microscopical Examinations. (Smear preparations.) Intestine (scraping).—Numerous tubercle bacilli. Mesenteric Gland.—Moderately numerous tubercle bacilli. Portal Gland Gastric Gland Lung (substance) } Liver ; nae wer ; i tubercle bacilli. i A few scattered tubercle bacilli ~ Spleen 5 Kidney is 331 GOAT 64. Virus A X. Fed once with culture derived from the original material. Dose—60 milligrammes. (Dose given with milk through a rubber teat.) Date of Feeding—January 28, 1909. [Age 5 days. ] Killed when in good general health—March 25,1909. [56 days after inoculation.] Weights. Kilos. January 29,1909... ae sus 3°05 March 25, 1909 tes as wt 5°40 Gain in weight.—2°35 kilos. *POST-MORTEM EXAMINATION. General condition. —Fair. Tonsils—The crypts of each tonsil were lined by soft caseous substance. Submawillary Glands.—The left submaxillary gland contained caseous foci. The right was normal. Cervical Glands.—Normal. Posterior Pharyngeal Glands. — Each posterior pharyngeal gland contained small softening caseous patches which in the left were confluent and formed caseous areas. Abdomen. Iniestines—The long Peyer’s patch was greatly thickened and closely filled with narrow ulcerated clefts running round the gut. At the distal half they became wider and serpiginous. The floors of the ulcers were lined by yellowish softening caseo-necrotic tissue. All the other Peyer’s patches were similarly extensively ulcerated, and in many cases the ulceration had extended down to the peritoneum. Mesenteric Glands—The mesenteric glands were enlarged, one to nearly the size of a hen's egg. The gland substance was firm and beset with caseous patches, most marked just beneath the capsule, and with caseous foci. Lleo-Colic Glands.—One ileo-colic gland was enlarged and the cortex contained a network of caseous tissue while the rest of the gland was beset with caseous foci. Spleen.—Normal. Liver.—In the liver substance there were numerous minute grey points just visible. Portal Glands.—The portal glands were a little firm and showed irregular opaque caseous foci in the cortices, Kidneys.—In the cortices of the kidneys just beneath. the surface could be seen a moderate number of ill- defined grey foci. Thorax. Lungs.—In the lungs could be seen sparsely scattered grey points of a doubtful nature. Mediastinal Gland.—In the largest mediastinal gland a few opaque points could be seen. All other organs and glands appeared normal. Microscopical Examinations. (Smear preparations.) Tonsil (caseous substance).—A moderate number of tubercle bacilli. Intestine (scraping ).—Numerous tubercle bacilli, Lung (substance).—No tubercle bacilli. Spleen (substance).—No tubercle bacilli. Liver (substance).—No tubercle bacilli. Kidney (substance).—No tubercle bacilli. CALF 548. Virus A X. Intravenous inoculation of culture derived from the original material. Dose—100°0 milligrammes. Date of Inoculation—May 30, 1907. [Age about 10 weeks.] Killed when dying—July 9, 1907, [40 days after inoculation.] Temperature. MAY 1907] JUNE Weights. ; Kilos. May 30,1907 ... a. se wns 53106 July 9,1907 cee 89°45 Total loss in weight —13°61 kilos. 34684 JUN POST-MORTEM EXAMINATION. Condition poor. Thorax. Pleura.—Normal. » 332 Lungs.—The lungs were not severely affected ; they did not collapse quite normally and were oedematous. They showed, scattered throughout, dark red and reddish grey areas consisting of lobules in various stages of pneumonia ; the dark red patches were com- pletely airless; the reddish grey showed much oedema, but did not sink in water. Bronchial and Mediastinal Glands.—Normal. Abdomen. Peritoneum and Omentum.—Normal. Intestines—The intestines appeared normal. Mesenteric Glands.—These glands appeared normal. Spleen.—The spleen was enlarged, and the pulp was ‘soft and dark red. Liver.—The surface of the liver presented a reddish mottled appearance, and the interlobular network was well marked. On section through the substance, parts were reddish grey and smooth, while there were scattered areas of a reddish yellow colour, the cut surfaces of which were finely granular. Portal Glands.—These glands appeared normal, Kidneys.—The kidneys were pale, and just beneath the capsule could be seen rather indefinite irregular paler grey foci. Suprarenal Bodies.—They appeared normal. Coeliac, Renal, Lumbar, and Iliac Glands.—Normal. Tongue, Pharynx, Larynx and Tonsils.—Normal. Various Lymphatic Glands. Submasillary, Parotid, Pharyngeal, Cervical, Pres- capular, Prepectoral, Precrural, Pudic, Giluteal and Popliteal Glands all appeared normal. Microscopical Examinations. (Smear Preparations.) Lung (substance).—Numerous tubercle bacilli. Spleen (substance).—Numerous tubercle bacilli. Lnver (substance).— Very numerous tubercle bacilli, scattered and in clumps. Kidney (substance).—Moderately numerous tubercle bacilli. Mesenteric Gland (substance).—Tubercle bacilli, not quite so numerous as in the liver. Intestine (scraping).—Moderately numerous tubercle bacilli, CALF 550. Virus A X, Intravenous inoculation of emulsion made with the spleen of Calf 548 (inoculated with culture of the original material). Dose—20:0 cubic centimetres, containing numerous tubercle bacilli. Date of Inoculation—July 9, 1907. Died—August 14, 1907. [Age about 8 weeks.] [86 days after inoculation.] Temperature. JULY 190] AuGusT Weights. Kilos. July 9, 1907 46°25 August 14, 1907 31:74 Total loss of weight.—14°51 kilos. POST-MORTEM EXAMINATION, General condition poor. Thorax. Pleura.—The pleura appeared normal, Lungs.—The lungs did not quite collapse ; the ven- tral portions of the anterior and posterior lobes con- tained dark red solid airless areas. The rest of the lungs was congested and oedematous containing. i Thoracic Glands.—The bronchial and mediastinal glands were congested and a little oedematous but showed no caseation. Pericardium and Heart.—Normal. Abdomen. Peritoneum and Omentum.—Normal. but air 333 Intestine.—The upper part of the small intestine showed some patchy congestion of the mucous membrane. , ‘Spleen.—The spleen was enlarged and swollen and dark red in colour. Liver.—The substance of the liver was speckled with very numerous minute grey and slightly yellowish foci. Portal Glands.—In the portal glands there were scattered discrete irregular yellowish necrotic patches. Kidneys.—The kidneys appeared normal except for oedema of the pelves. Suprarenal Bodies.—These appeared normal. The Pharynz and the base of the Tongue were deeply congested, (?) hypostatic. Tonsils.—Normal. Various Lymphatic Glands. Posterior Pharyngeal Glands.—These were deeply congested. Parotid, Submazillary, Prescapulur, Cervical, Pre- pectoral, Axillary, ' Precrural, Popliteal, Gluteal, Ischiatic, Coeliac, Mesenteric, Colic, Lumbar, and Iliac Lymphatic Glands——All showed varying amounts of congestion and oedema, Microscopical Examinations. (Smear Preparations.) Lung (substance) showed numerous tubercle bacilli. Spleen (substance) showed fairly numerous tubercle bacilli. Liver (substance) showed numerous tubercle bacilli. Kidney (substance) showed a few tubercle bacilli. -Mediastinal Gland (substance) showed numerous tubercle bacilli. Intestine (scraping) showed a moderate number of tubercle bacilli. CALF 554. Intravenous inoculation of emulsion made with the original material). Virus A X, the spleen of Calf 548 (inoculated with culture of Dose—10-0 cubic centimetres, containing numerous tubercle bacilli. Date of Inoculation—July 9, 1907. [Age about 7 weeks.] Died—August 13, 1907. [35 days after inoculation.] Temperature. JY 1407 Wl Weights. Kilos. 41:27 i ain July 9, 1907 31-99 August 13, 1907 Total loss in weight.—6'35 kilos. POST-MORTEM EXAMINATION ‘Condition poor. Thorax. Pleura.—Normal. Lungs.—The lungs did not collapse normally ; the whole of the anterior lobes and the right middle lobe were dark red, solid, and airless with an indication of a fine greyish mottling. Parts of the anterior portions of the caudal lobes were similarly consolidated, but the rest, the majority, was congested and oedematous, though still air containing. Bronchial and Mediastinal Glands—These glands were oedematous, many of them congested, but none showing any caseation. Pericardium and Heart.—Normal. 334 Abdomen. Peritoneum, Omentum, and Diaphragm.—Normal. Intestines —The intestines appeared normal. Spleen—The spleen was enlarged, firm, and the substance dark red, otherwise normal in appearance. Liver—The substance of the liver was speckled with very minute irregular greyish foci. Kidneys—The kidneys were somewhat dark in colour and showed in the cortices just beneath the capsule hemorrhagic foci ; on section the pelves were oedematous. Suprarenal Bodies ——Normal. The upper part of the Laryna, the Epiglottis, and the Pharynx were congested. : The tissues about the body generally were oede- matous. Various Lymphatic Glands. Parotid, Submazillary, Cervical, Prescapular, Pre- pectoral, Axillary, Precrural, Popliteal, Ischiatic, Gluteal, Coeliac, Lumbar, Iliac, and Mesenteric Glands. —AlI were oedematous, and many of them congested, but in none was there any caseation, Microscopical Examinations. (Smear Preparations.) Lung (substance) iss toe Spleen (substance) ... an | Liver (substance) wile te Mediastinal Gland (substance) | Intestine, small (scraping) ... Kidney (substance).—A moderate number of tubercle Showed numerous tubercle bacilli. bacilli. CALF 562. Virus A X. Intravenous inoculation of an emulsion made from the spleen of Calf 550. Dose—10:0 cubic centimetres. Date of Inoculation—August 14, 1907. [Age about 9 weeks. ] Died—September 15, 1907. [32 days after inoculation. | Temperature. The temperature rose to 39°7° on the 15th day, and remained about that point until the 28th day, when it reached 40°3° C, It dropped two days later and re- mained normal until death. Weights. Kilos. August 14,1907... Be ae 85°83 September 15, 1907 ... 28°57 Total loss in weight.—7 26 kilos. POST-MORTEM EXAMINATION. General condition poor. Thorax. Pleura.—Normal. Lungs.—The lungs collapsed fairly well, but were deeply congested and oedematous, especially in the anterior lobes and the anterior thirds of the posterior lobes. No part, however, was completely airless. Bronchial and Mediastinal Glands.—Normal. Pericardium and Heart.—Normal. Abdomen. Peritoneum and Omentum.—Normal. Intestines —The mucous membrane of the intestine was irregularly congested. Mesenteric Glands.—Normal in appearance. Spleen. — The spleen weighed 141 grammes; on section it appeared normal. Liver.—Normal. Portal Glands.—Normal, Kidneys.—The kidneys were rather dark in colour, otherwise normal. Suprarenal Bodies.—Normal. Renal, Lumbar, and Iliac Glands.—Normal. Pharynz.—The pharynx was slightly congested Tongue, Laryna, and Tonsils—Normal, The Prescapular, Axillary, Cervical, and Pharyngeat Lymphatic Glands were congested. Various Lymphatic Glands. Coeliac, Precrural, Popliteal, Gluteal, and Ischiatic Glands.—Normal. e Microscopical Examinations. (Smear Preparations.) Lung (substance).—Moderately numerous tubercle bacilli. Spleen (substance).—Numerous tubercle bacilli. Liver (substance).—Moderately numerous tubercle. bacilli. Kidney (substance).—A few tubercle bacilli. Mesenteric Gland (substance).— Very numerous tubercle bacilli with a number of small clumps, Intestine (scraping).—Numerous tubercle bacilli as- in the mesenteric gland. 5 CALF 560. Virus A X. {ntravenous inoculation of an emulsion made from the liver of Calf 554. Dose—9°0 cubic centimetres. Date of Inoculation—August 14, 1907. [Age 9 weeks.] Died—September 23, 1907. [40 days after inoculation. ] Temperature. wd "Us 38 Weights. Kilos. August 14, 1907 Pee 8 .. 44°90 September 23,1907 ... ae .» 33°56 Total loss in weight.—11:34 kilos. POST-MORTEM EXAMINATION. General condition poor. 2 Thorax. Pleura.—Normal. Lungs.—The anterior lobes of both lungs and more than half of the left posterior lobe were dark reddish grey and airless. On section the solid areas presented a greyish mottled appearance and minute drops of pus could be expressed ; the rest of the lung tissue was crepitant, containing only a few consolidated patches. Bronchial and Mediastinal Glands.—These glands were congested and oedematous, but showed no casea- tion. Pericardium and Heart.—Normal. Abdomen. Peritoneum and Omentum.—Normal. Intestines. —Normal. Mesenteric Glands.—The mesenteric glands were oedematous but otherwise normal. Spleen.—The spleen weighed 196 grammes. section the splenic pulp appeared normal. Liver.—The liver was normal. Portal Glands.—Nprmal. Kidneys.—The kidneys were a little pale and showed minute subcapsular hemorrhages The pelves were oedematous. Suprarenal Bodies.—Normal. Renal, Coeliac, Lumbar, and Iliac Lymphatic Glands. —Normal. Tongue, Lurynx, Pharynx, and Tonsils——Normal. Various Lymphatic Glands. Parotid, Submazillary, Cervical, Prescapular, Pre- pectoral, Axillary, Precrural, Popliteal, and Gluteal Lymphatic Glands.—These glands were oedematous, especially those in the anterior part of body, and some showed also minute foci of congestion. Microscopical Examinations. (Smear Preparations.) Lung (substance).—Showed very numerous tubercle bacilli. Liver (substance). — Showed numerous tubercle bacilli, some in clumps. Spleen (substance).—Showed moderately numerous tubercle bacilli. Kidney (substance).—Showed moderately numerous tubercle bacilli. Prescapular Gland (substance).—Showed a few tubercle bacilli. Mesenteric Gland (substance).—Showed numerous tubercle bacilli. 336 CALF 558. Virus A X. Intravenous inoculation with an emulsion made from the liver of Calf 562. Dose—5:0 cubic centimetres. Date of Inoculation—September 16, 1907. [Age about 14 weeks. ] Died—October 31, 1907. [45 days after inoculation. ] Temperature.. The temperature rose to 398° C. on the 35th day and reached 40:1° on the 38th day. On the 42nd day it fell again to normal. Weights. Kilos. September 16, 1907 34:00 October 31, 1907 85°37 Gain of weight.—1:37 kilos. POST-MORTEM EXAMINATION. General condition poor. Thorax. Plewra.—Normal. Lungs.—The lungs were rather heavy, not collapsing normally ; the anterior lobes of both lungs were toa great extent reddish grey and airless. The anterior portion of the left caudal lobe was also pneumonic, and in the substance of the rest of the posterior lobes there were scattered pneumonic lobules most numerous on the left.side. The pneumonic parts were in a con- dition of red hepatisation without any necrosis. Bronchial and Mediastinal Glands—They were enlarged and firm, and on section the cortices were replaced by a definite yellowish waxy caseous network. Heart and Pericardium.—Normal, Abdomen. Peritoneum and Omentum.—Normal. Intestines —Normal. Mesenteric Glands.—These were oedematous but showed no caseation. é Spleen.—The spleen weighed 252 grammes. The sub- stance was firm and showed some sparsely scattered ill- defined greyish necrotic patches, and also some promi- nence of the malpighian bodies, many of which appeared slightly necrotic in the centre. Liver.—On the surface of the liver there was an occasional minute greyish white focus. On section the substance, beyond slight congestion, appeared normal. Portal Glands.—The portal glands were moderately enlarged. On section the cortices were firm, and to a great extent consisted of greyish necrotic tissue with a yellowish tint, in parts in the form of a network, in others as uniform nodules. Kidneys——The pelves of the kidneys were oede- matous; otherwise the kidneys were normal in appearance. Suprarenal Bodies.—The cortices of the suprarenal bodies were closely beset with minute grey foci. Renal, Coeliac, Lumbar, Iliac, Parotid, Submazillary, Pharyngeal, Cervical, Prescapular, Prepectoral, Pre- crural, Popliteal, Gluteal and Ischiatic Glands showed varying amounts of oedema, but no caseation. Microscopical Examinations. (Smear Preparations.) Lung (substance) «.. Spleen (necrotic patch) Liver (substance).—Moderately numerous tubercle bacilli. Kidney (substauce).—A few tubercle bacilli. Bronchial Gland (substance) {Numerous tubercle bacilli, Portal Gland (substance) ... vey tubercle bacilli Small Intestine (scraping) ... eo CALF 564. Virus A X. Intravenous inoculation with an emulsion made from the liver of Calf 560. Dose—9-0 cubic centimetres, Date of Inoculation—September 23, 1907. Died—November 7, 1907. Temperature. The highest point reached was 4U:1° C. on the 25th day. It remained about that point until the 32nd day, when it fell and subsequently remained about normal. Weights. Kilos. September 23, 1907 ... aes ve» = 50°80 November 7, 1907... gee vee = 46°72 Zotal loss of weight.—4:08 kilos. POST-MORTEM : EXAMINATION. General condition poor. Thorax. Pleura.—Normal. Lungs.—The anterior lobes of the lungs were deeply congested and several portions were quite airless. The anterior portions of the caudal lobes resembled the anterior lobes, whilst the remaining portions of the posterior lobes were moderately congested and very oedematous. [Age about 15 weeks.} [45 days after inoculation. ] Thoracie Lymphatic Glands——The bronchial and mediastinal glands were congested and firm and on section the cortices were closely beset with yellow foci. Heart.—Normal. Abdomen. Peritoneum and Omentum.—Normal. Intestines.—Normal. Mesenteric Glands.—They appeared normal. Spleen.—The spleen weighed 336 grammes ; the sub- stance was firm and on section there were several small’ ill-defined yellowish grey necrotic patches. Liver.—The liver was congested but otherwise normal. Portal Glands.—The portal glands were moderately enlarged and the cortices were firm and beset with. yellowish streaks and foci forming a network. Kidneys.—-The cortices of the kidneys were speckled’ with hemorrhagic points and the tissue in the pelves- was oedematous, 337 Suprarenal Bodies.—Normal. Renal, Coeliac, Lumbar, and Iliac Glands.—Normal. Tongue, Tonsils, Larynx.and Pharyna.—Normal. Various Lymphatic Glands. Parotid, Submazillary, Pharyngeal, Prescapular, Cervical, Prepectoral, Axillary, Precrural, Popliteal, Gluteal and Ischiatic Glands.— Oedematous but otherwise normal. Microscopical Examinations. (Smear Preparations.) Very numerous tubercle Lung (substance ) bacilli. Spleen ( do. sald (do. ? Moderately numerous Kidney (do. ) tubercle bacilli. Mediastinal Gland (substance) \ Numerous Intestine, Small (scraping of tubercle bacilli mucous membrane). CALF 566. Virus A X. Intravenous inoculation with an emulsion made from the liver of Calf 558. Dose—5°0 cubic centimetres (estimated to contain 150,000,000 tubercle bacilli). Date of Inoculation—October 31, 1907. Killed when ill—December 13, 1907. Clinical History. The calf was killed when moderately ill, emaciating, and breathing hard. Temperature. The temperature was irregular throughout, rising slowly up to the maximum of 40°3° on the 25th day. Ik fell slightly and remained about 39-6° until death. Weights. Kilos. October 31, 1907 60°32 December 13, 1907 49-89 Loss in weight.—10-43 kilos. POST-MORTEM EXAMINATION. General condition poor. Thorax. Pleura.—Normal. Lungs.—The right anterior and raiddle lobes were reddish grey, solid, and airless. On section the lobules were dark red around the periphery and grey in the centre, and many were softened and muco-purulent ; muco-pus could be expressed from the bronchi. The left anterior lobe was partly reddish grey and airless. The posterior lobes did not collapse normally ; they were peppered with minute hemorrhagic foci, and showed scattered pneumonic lobules and small solid areas. The bronchi contained muco-pus. Thoracic Lymphatic Glands.—The bronchial and mediastinal glands were enlarged and firm. On sec- tion the majority showed their cortices replaced by firm yellowish caseated tissue ia a translucent matrix. Heart.—Normal. Abdomen. Omentum and Peritoneum.—Normal. Intestines. —The large intestine appeared normal. In the majority of the Peyer's patches of the small intestine there were from two to six palpable grey elevations which were slightly opaque, yellowish, and .caseous in the centre. Mesenteric Glands.—The mesenteric glands were enlarged and on section the cortices were firm grey [43 days after inoculation.] with a fine yellowish caseous network. The majority showed the whole of their cortices thus affected ; in the rest there were small discrete caseous patches. The Ileo-Colic Glands resembled the mesenteric glands. Spleen.—The spleen was slightly enlarged; sub- stance appeared normal. Liver.—Normal. Portal Glands.—These glands were firm and beset with small yellowish early caseous patches. / Kidneys.—The kidneys were pale, otherwise normal. Suprarenal Bodies.—Normal. Renal, Coeliac, Lumbar, and Iliac Glands.—Normal. Various Lymphatic Glands. Parotid, Submaxillary, Axillary, Prescapular and Prepectoral, Precrural, and Popliteal Glands.— Normal. Cervical Glands.—In the left superior cervical gland there was a nodule 1-7 by 1:0 cm., consisting of firm grey tissne with a caseous network resembling the mesenteric glands. In the right cervical gland there was part of the gland, corresponding to the nodule on the other side, firm and grey but not caseating. Pharyngeal Glands——The right pharyngeal gland was enlarged and firm and a few opaque foci could be seen in the cortex. The left pharyngeal gland was smaller and a few opaque foci could be seen. Microscopical Examinations. (Smear Preparations.) Lung (pneumonic).— A few scattered tubercle bacilli. Lung (muco-pus).—No tubercle bacilli. Spleen (substance).—No tubercle bacilli. Laer (substance).—No tubercle bacilli. Mediastinal Gland (emulsion).—Numerous tubercle bacilli. Mesenteric Gland (emulsion).- -Modcrately numerous tubercle bacilli. Peyer's Patch. — Moderately numerous tubercle bacilli. 34684 3 C 338 CALF 570. Virus A X. Intravenous inoculation with an emulsion made from the liver of Calf 564. Dose—5-0 cubic centimetres. Date of Inoculation—November 8, 1907. [Age about 15 weeks.] Died—December 9, 1907. [31 days after inoculation.] Clinical History. The animal had been slowly emaciating and breath- ing quickly for some time. Death occurred quite ‘suddenly. Temperature. The temperature rose to 39°6° C. on the 10th day, and remained about that level until the 21st day, when it rose to 40°1° C. It dropped again to 39°4° and remained at that level until death. Weights. Kilos. November 8,1907 eee we 64°85 December 9, 1907 awe uaa we 52°60 Total loss in weight.—12:25 kilos. POST-MORTEM EXAMINATION. General condition poor. Thoras. Pleura.—Normal. Lungs.—The right lung was dark red, solid and airless, sinking entirely in water. On section blood and oedema fluid exuded freely, but there was no sign of caseation. The left lung was oedematous and congested, especially in the anterior lobes, but still contained air. Thoracic Lymphatic Glands —The mediastinal and bronchial glands were definitely firmer than normal, but were not caseated ; they were very congested, and on section were dark red in colour. Heart.—Normal. Abdomen Omentum and Peritoneum.—Normal. Intestines.—Normal. Mesenteric Glands.—Normal. Spleen.—The spleen weighed 196 grammes ; it was enlarged and a little firmer than normal ; splenic pulp otherwise normal. Liver.—Normal. Portal Glands.—The portal glands were firmer than normal, and in the cortices scattered small early caseous patches could be seen. Kidneys.—In the cortices of the kidneys there were scattered bluish-black pigmented pits ; the pelves were oedematous and contained some whitish flakes, a smear preparation from which showed a few small clumps and some scattered tubercle bacilli. Suprarenal Bodies.—Normal. Renal, Coeliac, Lumbar, and Iliac Lymphatic Glands. Normal. Tongue, Tonsils, Larynx, and Pharynx.—Normal Various Lymphatic Glands. The prescapular and cervical glands were oedema- tous and a little firmer than normal. The posterior pharyngeal glands were congested and oedematous. Parotid, Submacaillary, Prepectoral, Avillary, Pre- crural, Popliteal, and Giluteal Gilands.—Normal. The tissues about the shoulders and neck were oedematous. Microscopical Examinations. (Smear Preparations.) Lung.—Moderately numerous tubercle bacilli. Liver.—Moderately numerous tubercle bacilli. Spleen.—Numerous tubercle bacilli. CALF 574. Virus A X. Subcutaneous inoculation with an emulsion made from the liver of Calf 566. Dose—10:0 cubic centimetres. Date of Inoculation—December 13, 1907. [Age about 7 months.] Killed when in good health—March 31, 1908. [109 days after inoculation.] Temperature. The temperature was normal throughout. Tuberculin Tests. January 31, 1908. began.] Dose of tuberculin (bovine), 2°0 ce. geaction. March 13, 1908. began.] Dose of tuberculin (bovine), 2°0 cc. of temperature, 1:7° C. [49 days after the experiment No {91 days after the experiment Rise POST-MORTEM EXAMINATION. General condition good. Local Lesion.—There was no local lesion visible. Prescapular Glands.—The right prescapular gland showed at one end just beneath the capsule six soft caseous tubercles about the size of rape seed. The left prescapular gland was normal. Prepectoral G'lands—Normal Cervical Glands.—Normal. Agillary Glands.—Normal. Thorax. Pleura.—Normal. Lungs.—On the pleura of the left lung there was a minute glassy tubercle, which cracked on pressure. Bronchial Glands.—The left bronchial gland was not enlarged but was a little firm at one extremity, and on section showed a nodule composed of a net- work of caseous tissue, not appreciably gritty, ina grey translucent matrix. Immediately around it there were discrete opaque foci and a few grey tubercles with caseous centres up to about a millet seed in size. Mediastinal Glands.—Normal. Heart.—Normal. Abdomen. Omentum and Peritoneum.—Normal. 339 Intestines.—Normal. Gastric, Mesenteric, and Colic Glands.—Normal. Spleen.—Normal. _ Liver.—In the liver there were a few opaque foci im a group surrounded by a zone of congestion, Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies—Normal. Renal, Coeliac, Lumbar, and Iliac Lymphatic Glands, —Normal. Tongue, Tonsils, Larynx, and Pharynz.—Normal, Various Lymphatic Glands. Parotid, Submazillary. Posterior Pharyngeal, Pre- crural, Ischiatec, Gluteal, and Popliteal Glands.— Normal. Microscopical Examinations. (Smear Preparations.) Emulsion of Prescapular Gland (C).—One tubercle bacillus seen. Emulsion of Prescapular Gland (IT).—No tubercle bacilli seen. : Lung (focus).—No tubercle bacilli seen. Bronchial Gland (emulsion).—A moderate number of tubercle bacilli. Liver (foci).—No tuberele bacilli seen. CALF 572. Virus A X. Subcutaneous inoculation with an emulsion made from the liver of Calf 570. Dose—9:0 cubic centimetres. Date of Inoculation—December 9, 1907. [Age about 20 weeks. | Killed when in good health—May 20, 1908. [163 days after inoculation.] Clinical History. The animal showed no sign of ill-health during the experiment. Temperature. Normal throughout. Tuberculin Tests. January 31, 1908. [53 days after the experiment began.] Dose of tuberculin (bovine), 2:0 cc. No reaction. March 13, 1908. [95 days after the experiment began.] Dose of tuberculin (bovine), 2-0 cc. No reaction. May 13, 1908. [156 days after the experiment began.] Dose of tuberculin (avian), 2:0 cc. No reaction. Weights. Kilos. December 12,1907 ... aa ver =: 4°42 69°84 May 20. 1908 ... eos tee ane Total gain in weight.—15°43 kilos. POST-MORTEM EXAMINATION. Careass in good condition. Local Lesion.—In the subcutaneous tissue at the seat of inoculation on the left side of the neck, there was an irregular flattened thickening measuring 6 cm. in length by 3 cm. in greatest breadth and 6 cm. in thickness ; it was firm and on section it was composed of fibrous tissue the central portion of which was beset with discrete caseo-caleareous granules. Prescapular G'lands.—The left prescapular gland measured 8 by 5 by 3 cm. ; it was irregular in outline. On section it consisted of four cystic nodules separated by fibrous tissue. The nodules contained soft caseous gritty substance and the walls were composed of fibrous tissue crossed by trabecule of translucent tissue. The right prescapular gland was normal. Prepectoral Glands.—Normal. Cervical Glands.—Normal. Agillary Glands.—Normal. Thorax. Pleura.—Normal. Lungs.—In the anterior lobe of one lung, there was a nodule composed of soft grey translucent tissue. Bronchial and Mediastinal Glands.—Normal. Heart.—Normal. Abdomen. Peritoneum and Omentum.—Normal. Intestines.—Norma. Mesenteric Glands.—One mesenteric gland showed an opaque yellowish focus, Spleen.—Normal. Liver—In the depth of the liver there was a grey nodule the size of a millet seed with an opaque centre. Portal Glands..—Normal. Kidneys.—Normal. Suprarenal Bodies.--In a suprarenal body there was a singie yellow caseo-calcareous miliary tubercle. Longue, Tonsils, Larynx and Pharynx.—Normal. Renal, Coeliac, Lumbar and Iliac Glands.—Normal. Various Lymphatic Glands. Precrural, Pudic, Gluteal, Popliteal, Submazillary Parotid and Retro-pharyngeal Glands—Normal. Microscopical Examinations. (Smear Preparations.) Lung (nodule).—No tubercle bacilli. - Liver (tubercle).—No tubercle bacilli. Mesenteric Gland (focus).—No tubercle bazilli. Suprarenal Body (tubercle).—Moderately numerous tubercle bacilli. Prescapular tubercle bacilli. Prescapular Gland (Emulsion II.).—Numerous tubercle bacilli. Gland (Emulsion I.),—Numerous 34684 302 340 CALF 576. Virus A X. ‘Subcutaneous inoculation with an emulsion made from the liver of Calf 570. Dose—7‘0 cubic centimetres. Date of Inoculation—December 9, 1907. [Age about 20 weeks.] Killed when in good health—May 20, 1908. [163 days after inoculation.] Clinical History. There was no sign of ill-health during the course of ‘the experiment. Temperature. Normal throughout. Tuberculin Tests. January 31, 1908. began.] Dose of tuberculin (bovine), 2°0 ce. reaction. March 13, 1908. began.] Dose of tuberculin (bovine), 2'0 cc. reaction. May 13, 1908. began.] Dose of tuberculin (avian), 2:0 cc. reaction. [53 days after the experiment No [95 days after the experiment No [156 days after the experiment No Weights. Kilos. December 12,1907 ... we we «= 4444 May 20, 1908 ... us aa we = 76°19 Total gain in weight.—31°75 kilos. POST-MORTEM EXAMINATION. General condition good. Local Lesion.—In the muscle at the seat of inocula- tion there was a slight linear thickening about 1 cm. in breadth which on section was found to be fibrous tissue beset with irregular caseo-calcareous granules. Prescapular Glands.—The left prescapular gland was slightly enlarged, it contained at one extremity just beneath the capsule a collection of caseo-calcareous granules about 6 mm. in diameter. There were also two similar but smaller nodules. The rest of the gland was normal. The right prescapular gland was normal. Prepectoral Glands.—Normal. Cervical Glands.—The left inferior cervical gland was small but the superficial part of the cortex was replaced by a margin of caseo-calcareous tissue. The other cervical glands were normal. Agillary Glands.—Normal. Thoraa. Pleura.—Normal. Lungs.—Normal. Bronchial and Mediastina Glands.—Normal. Heart.—Normal. Abdomen. Peritoneum and Omentum. —Normal. Intestines.—There was a moderate number of minute yellowish foci in the terminal Pever’s patch and a single focus in a second patch. The mucous mem- brane of the small intestine was normal. The large intestine was normal, Gastric Glands.—Normal. Mesenteric Glands—The majority contained either a few discrete caseo-caleareous tubercles about the size of rape seed, or a small ill-defined bright yellow patch just beneath the capsule, which on section was found to be composed of a fine network of yellow caseous tissue. Colic Glands.—Several of the colic glands showed foci similar to those in the mesenteric glands. Spleen.—Normal. Liver—Normal. Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Renal, Coeliac, Lumbar, and Iliac Lymphatic Glands. —Normal. Tongue, Tonsils, Larynx, and Pharynz.—Normal. Various Lymphatic Glands. Precrural, Pudic, Gluteal, Popliteal, Submazillary, Parotid, and Retro-pharyngeal Glands.—Normal. Microscopical Examinations. (Smear Preparations.) Prescapular Gland.—Numerous tubercle bacilli. Left Inferior Cervical Gland—Numerous tubercle bacilli. Mesenteric Gland (1).—A few tubercle bacilli. Mesenteric Gland (1I).—A few tubercle bacilli. Peyer's Patch.—No tubercle bacilli, MONKEY 194. Virus A X. Subcutaneous inoculation of emulsion of mediastinal gland of Calf 566. Date—December 13, 1907. Died—February 2, 1908. Clinical History. Death occurred during very cold weather, which probably had some influence, and after a tuberculin test. Tuberculin Test. The monkey was tested 49 days after the inocu- lation and gave a positive reaction. Rise of temper- ature 1:4° C. _POST-MORTEM EXAMINATION. General condition fair. [51 days after inoculation. ] Local Lesion —At the seat of inoculation in the subcutaneous tissue of the back, posterior to the scapule, there was a prominent tumour measuring 8by 5by2cm. In the skin over it there was an ulcer measuring 25 mm. from which issued a purulent discharge. On section it consisted of a cyst contain- ing a central mass of pinkish grey necrotic tissue beset with purulent foci, together with a little turbid .Serous fluid. Axillary Glands.—Normal. ‘ ete was no sign of tuberculosis elsewhere in the ody. 341 Microscopical Examinations. (Smear Preparations.) “Spleen (substance).—No tubercle bacilli. Liver (substance).—No tubercle bacilli. Local Lesion.— Scattered tubercle bacilli. Cultures.—A culture grown from the local lesion showed numerous typical aviar tubercle colonies PIG 234. Virus A X. Fed once with almost the whole of the spleen of Calf 566. Date of Feeding—December 13, 1907. [Age about 17 weeks.) Killed when in good health—May 25,1908. [164 days after the experiment began.] Tuberculin Tests. March 13, 1908. ‘began.] No reaction. May 13, 1908. [152 days after the experiment began.] No reaction. [91 days after the experiment Weights. Kilos. December 13, 1907 ... eee ac 25°40 May 25, 1908 a5 av rc 36°73 Gain in weight.—10°33 kilos. POST-MORTEM EXAMINATION. General condition good. Tongue, Pharyna, and Larynx.—Normal. Tonsils—In one tonsil there was a soft brownish nodule. Submazillary G'lands.—In a left submaxillary gland there was a soft yellowish caseous miliary nodule. Oesophagus.—In the sub-epithelial tissue of the - oesophagus there was a yellow muco-purulent nodule. Pharyngeal Glands.—Normal. Cervical Glands.—Norwal. Abdomen. Omentum and Peritoneum.—Normal. Intestines. —Normal. Mesenteric and Colic Gilands.—Normal. Spleen.—Normal. Liver and Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Renal, Coeliac, Lumbar, and Iliac Glands—Normal. Thorax. Pleura.—Normal. Lungs.—Normal. Bronchial and Mediastinal Glands.—Normal. Heart —Normal. Various Lymphatic Glands. Prescapular, Precrural, and Inguinal Glands.—- Normal. Microscopical Examinations. (Smear Preparations. ) Tonsil (focus).—No tubercle bacilli seen. Oesophagus (nodule).—No tubercle bacilli. blue stained organisms, Submaxillary Gland (nodule).—A moderate number of tubercle bacilli. A few PIG 236. Virus A X. Intravenous inoculation of an emulsion made from the mesenteric glands of Calf 566. Dose—5°0 cubic centimetres. Date of Inoculation—December 13, 1907. [Age about 17 weeks. ] Died—April 11, 1908. [120 days after inoculation.] Clinical History. The animal was emaciated and in very poor con- ‘dition. The skin was covered with scabs. Temperature. There was no rise of temperature until the 18th of March, when it reached 40°1° C. It remained high until death. Tuberculin Test. March 13, 1908. [91 days after the experiment ‘began.] No reaction. Weaghts. oe Kilos. 1907 ... ass wae 26°58» December 12, 19 oor April 11,1908 ss ee one Loss in eveight.—3'18 kilos. POST-MORTEM EXAMINATION. ; General condition poor. Thorax. Lungs.—The lungs did not collapse normally ; they were irregularly congested and emphysematous, and in parts contained reddish grey areas, very small pieces of which sank in water. Just beneath the pleura could be seen numerous discrete irregular translucent tubercles, the majority about the size of rape seed and a few a little larger. They had a yellowish tint, and most of them contained a central yellow calcareous granule. Un section of the lung the tubercles were fairly evenly distributed, and they stood out from the freshly cut surface. Bronchial Glands.—They appeared normal. Spleen.—The substance was a little firm. 342 Liver.—A little pale. Kidneys.—They were pale and bad a greyish mottled appearance. Intestines.—Normal. Mesenteric Glands.—They were darker and firmer than normal ; the cortices were somewhat opaque, and showed a very fine almost invisible yellowish network. Occasionally in between the lobules there were streaks. of yellowish and whitish substance. Other Lymphatic Glands.—A rectal gland was. enlarged and replaced by soft yellow substance. A right iliac gland contained a grey nodule beset with purulent foci. A left submaxillary gland showed a yellowish miliary nodule easily shelling out from the substance. (GOAT 50 (Adult). Virus A X. Subcutaneous inoculation with an emulsion made from the liver »f Calf 570. Dose—6:0 cubic centimetres. Date of Inoculation—December 9, 1907. Killed when in good health—Augusi 5, 1908. Clinical History. The animal showed no sign of ill-health during the experiment. She bore a healthy kid. Temperature. The temperature remained normal throughout. Tuberculin Tests. March 13, 1908. [95 days after the experiment began.] Dose of tuberculin (bovine), 2:2.cc: Rise of temperature 1-0° C. May 13, 1908. [156 days after the experiment began.] Dose of tuberculin (avian), 2-0 cc. Rise of temperatnre 1°9° C. Weights. Kilos. December 9, 1907... eee vee 30°38 August 5, 1908 ase a aoe 22°67 Loss in weight.—7'71 kilos. [240 days after inoculation.] POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion:~At the seat of inoculation in the subcutaneous tissues on the left side of the neck there were a few small fibrous nodules with yellow purulent gritty contents. Prescapular Glands.—The left prescapular gland was enlarged measuring 3°7 by 2:2 by 1:0 cm. On section it was more than half cystic and filled with soft caseous, partly fluid, gritty contents. . The right prescapular gland was normal. Prepectoral Glands.—Normal, Cervical Glands.—Normal. There was no tuberculosis elsewhere. Microscopical examination of the prescapular gland’ showed moderately numerous tubercle bacilli. MONKEY 192. Virus A X. Subcutaneous inoculation of an emulsion of liver of Calf 570. Date—December 9, 1907. Died—February 2, 1908. [55 days after inoculation. ] Tuberculin Test. The animal was tested 53 days after the experiment began and gave a positive reaction. Rise of temperature 1:1° CO. POST-MORTEM EXAMINATION. Local Lesion. At the seat of inoculation in the subcutaneous tissue of the back there was a lenticular tumour measuring 5 by 4 by 15cm. The skin over it was intact. On section the tumour consisted of a mass of caseo-purulent nodules supported by a net- work of soft grey translucent tissue. Axillary Glands—They were a little enlarged, but otherwise were normal, except for one on the right side which showed two opaque foci just beneath the capsule. Thoran. Lungs.—The lungs were normal except for a single translucent tubercle in the left caudal lobe. All other organs and lymphatic glands were normal. Microscopical Examinations. (Smear Preparations.) Lung (tubercle) Spleen (substance) Liver (substance) _ No tubercle bacilli. Local Lesion—Numerous tubercle bacilli. U.—June 14, 1905. riginal material. Total duration of artificial cultivation, 97 days. 5th generation of culture was used, 21 days old. PIG 104.1 196, RABBITS. Intraperitond taneous, Se seen 2.5 Dose : 10-0 m. 5.9 mg. Number. | Method. Result. Killed when hen well : aI ce gd oe tee eased ram July 12, 190 69 lay. By ite 826 | Subcut,| 50 my. | D. 69 days | Chronic days. G.T. P.M. — Small 328 Sabeut.| tOme. | D. zo, Local T. calcareous nodulelealed ulcer at : dominal wall. ulation. The wee Sd peritoneum and omguinal glands were moderately ined a cavity rous caseous nodule soft cheesy kidneys showed One portal grey foci; there tined a single few yellow tuberdo tuberculosis the surface of the In the lungs were grey nodules. Th liac glands were j caseated; many ‘A-PIGS. riseetiy a and ae thoracic glands co _| Buration ot! p asult. yellowish streaks pee | Life. oe a rap. | D. 119 days | G. T. CULTURE : t.| K.45 9 ,, | Loesl T. Derived from Pig | Inoculated on Ma 3 1906, into : GUINEA-PIG 1 Subcut. Died : 21 days : P.M.—Local tuA-PIG 2027. CULTURE. losis only. Subcut. ; Derived from G.P. 1857. M3 of broncbial The 4th generation, 22 days old, was used, on Sans ie Ra: November 17, 1906. 1. 44 days. General tubercu- | Duration of Dose, | GUINEA-PIGS. e of Result. Number. | Method.} Dose. | ss of Result. | HT acres, aelian, 9 Bye eatin, REL gee Mince a 2a nal ar ays | Acute T. 2210 Intrap. | 1:0 mg. | D. 11 days | Acute T. , | Acute T. 2209 | Subcut.| 10mg. | D.35 ,, |G. T. Go Ge aver elliat in type and resembled a bovine tubercle bacillus. Q ae Strain derived from original material. V.—NoveEMBER 30, 1905. Total duration of artificial cultivation, 266 days. The 15th generation of culture was used when 21 days old. — 118. MONKEY 120. MONKEY 122. MONKEY 124. jeous. Subcutaneous. Fed. Fed. _0 mg. Dose : 1:0 mg. Dose : 10:0 mg. Dose: 1:0 mg. very ill: Killed when well : Killed when well : Killed whea well : 1906. March 23, 1906. March 22, 1906. March 22, 1906. ys. 113 days. 112 days. 112 days. ral tuber- P.M. — Small fibrous P.M—One mesenteric P.M.—In the ileum ae lesion at seat of inocula- gland contained a yellow were two hard grey bodies tion. Adjacent glands nodule filled with yellow (? tubercles). Two mes- normal. In the lungs pus. No lesions else- enteric glands contained were two translucent grey where. each a pea-sized collection tubercles. of creamy pus. No lesions elsewhere. RABBITS. “CULTURE. : Derived from the ber. iced Duration | Result. spleen. The 11th genera- | of Life. tion was inoculated on November 16, 1906, when 4 | Subeus. | D.82days| G. T. 21 days old, into : 5 Subeut.|D.89 ,, | G. T. 1 5. GUINEA-PIGS. FOWL 130. : ae Fed. Duration | Result. | | Number. |Method.| Dose. | Duration | Resolt. ees HOS | | Killed : 104 days. D.17 days) a onte 2200 | Intrap. | 1:0 mg. | D. 8days|) 4 ute P.M.—No tuberculosis. D.20 ,, T. 2198 | Intrap.| 01 mg. |D.18_,, a D.30 ,, | G.T. 2199 |Subcut.| 10mg. |D.46 ,, | G. T. p.29 , | G.T. 2197 |Subcut.| 0-1 mg.-|D.46 ,, | G.T. D.63 ,, | GT. 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Virus P II, Intraperitoneal inoculation of culture deri igi i int 2 c D 1 lerived from the original material. (An attempt at an intra- venous inoculation was made, but this failed, a little of the culture suspension ee into the thieanes around the vein (ear)). Dose—10-0 milligrammes. Date—May 4, 1905, Weight a few days after Inoculation—9-97 kilos. [Age 10 weeks. ] Killed when in good health—J uly 12,1905. [69 days after inoculation.] Clinical Notes. _A small nodular swelling developed at the seat of inoculation near the umbilicus. The pig showed no sign of ill health during the experiment. Tuberculin Test. June 30, 1905. [57 days after inoculation.] Doubtful reaction. Rise of temperature, 0:6° C. Weights. Kilos May 10,1905... wg kk 9-97 May 30, 1905... eae ug ws» ~~ :12°70 June 27, 1905 re wits veo «16°78 July 11, 1905... . 19:05 Gain in weight.—9-08 kilos. POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion—In the deep subcutaneous tissues of the abdominal wall, near the umbilicus, situated in the track of the injection needle, there was an irregular nodule the size of a French bean, composed of opaque yellowish white, slightly gritty caseous material. In the subperitoneal fat near the latter there were three or four pinhead-sized yellow tubercles. Inguinal Glands N 1 Precrural Glands i as Abdomen. Omentum.—The omentum was not thickened nor retracted ; it contained moderately numerous spherical opaque yellow nodules, ranging in size from 1 milli- metre or less up to about 2°5 millimetres in diameter ; most of these had a narrow translucent peripheral zone, from which on section a firm cheesy-looking mass could be readily shelled. Peritoneum.—In the peritoneum, between the coils of the large intestine (and to a less extent on that covering the gut), there were numerous similar nodules; these in several places were closely aggregated, forming long continuous rows. On section they were uniformly caseous, and many appeared to be very slightly gritty in the centre. Stretching from coil to coil of the large intestine were many fine strands of fibrinous material ; the serous surface of the large intestine was slightly roughened. On the meso-rectum there were several yellow nodules, mostly small, and tending to occur in little groups. Mesentery.—Normal. Parietal Peritoneum.—The parietal peritoneum on the left side was thickened and velvety, and showed when removed and placed under water long delicate villous processes. On this same side were four loosely attached flattened nodules, two the size of split peas and two that of millet seeds ; the former were dark in colour, and seemed to have had recent hemorrhage into their substance. : On the right side of the abdomen there was a slight roughening of the peritoneum near the liver, but there were no nodules. Bladder.—On the serous surface of the bladder there were three millet seed sized yellow tubercles. Spleen.—Normal. 34684 Liver—The anterior surface of the liver was roughened, and showed about half a dozen yellow tubercles, the largest the size of a millet seed ; these indented, but did not penetrate, the liver substance. Embedded in one of the thin margins of the middle lobe was a series of six similar tubercles (these probably began as surface tubercles). On section the liver substance was normal to the eye. Portal Glands.—Normal. Kidneys.—The left kidney showed on the surface scattered translucent grey foci ; similar foci were seen on the surface of the right kidney. Suprarenal Bodies.—Normal. Coeliac Glands.—The coeliac glands, five in number, were moderately enlarged. On section their tissue was extensively replaced by yellowish white some- what translucent tissue, showing a sprinkling of minute gritty points ; the caseated tissue was brittle, and in places beginning to break down. Lying near the vertebral column there was a gland (lumbar gland ?) the size of a pigeon’s egg, which on section was caseous and softened practically through- out. A gland in the pancreas was enlarged, indurated, and showed irregular yellow foci scattered throughout. Stomach.—Normal. Large Intestine and Smatl Intestine —Mucous mem- brane normal, Mesenteric Glands.—The glands generally appeared a little enlarged, and felt firmer than normal. Those in the anterior part (the first five or six) and the terminal gland were beset with irregular yellow foci and streaks; the rest showed obscure whitish foci, sometimes yellowish streaks. Most of the glands had a peculiar blanched or tallowy appearance, quite unlike that of a normal gland. Gastric Glands,—A gastric gland contained a yellow tubercle. Colic Glands.—The colic glands were not enlarged ; they showed scattered foci, the largest the size of a pin’s head. Rectal Glands.—One of the rectal glands contained a minute yellow focus. Thorax. Heart and Pleura.—Normal. Lungs.—The lung tissue was speckled with minute foci of congestion. In the caudal lobes some of the foci under the pleura had a greyish appearance, re- sembling early tubercles. On the dorsal surface of the left caudal lobe, under the pleura, there were two greyish translucent nodules 1°5 millimetre in diameter ; there was a similar nodule in the right caudal lobe. Bronchial Glands.—One on the right side showed a minute yellowish point in the depth; the rest were normal. Mediastinal Glands.—The long posterior mediastinal gland was enlarged, indurated, and showed irregular yellow tubercles, which at one part of the cortex had become confluent ; in the latter signs of very early calcification were seen. Pharynx, Larynx, Trachea, and Tonsils—Normal. Left Ear.—In the tissues around one of the small veins there was a small irregular yellow patch of necrosis. : A gland near the articulation of the jaw, on the left side, contained two small yellow foci. _ aa A gland near the entrance to the tnorax, lying in the mid line on the under surface of the trachea, 3 D 348 showed discrete yellow pinhead-sized tubercles scattered throughout the cortex. All other lymphatic glands were normal. Microscopical Examination. (Smear preparations.) Omental Nodule esi «+ ) Numerous tuber- Subcutaneous Nodule (abdominal) j cle bacilli. Moderately numer- ous tubercle ba- cilli. Gland at Entrance to Thorax Gland at Base of Left Ear... Bronchial Gland.—No tubercle bacilli. Rectal Gland.—A few tubercle bacilli. Colic Gland ose ia | PIG 106. Virus P II. Subcutaneous inoculation of culture derived from the original material. Dose—5‘0 milligrammes. Date—June 14, 1905. Weight at Inoculation—14'51 kilos. Clinical Notes. A small tumour developed at the seat of inoculation near the umbilicus, ulcerated and finally disappeared. The animal showed no symptom of illness during the experiment. Temperature. The temperature was not recorded for three months following the inoculation. During October there was a slight rise of temperature lasting three weeks and reaching a maximum of 39°6° C. From ‘October 29, 1905, to January 15, 1906, it was normal. Tuberculin Tests. September 20, 1905. [98 days after inoculation. ] Slight reaction. Rise of temperature, 0:7° C. November 17, 1905. [156 days after inoculation. ] Reacted. Rise of temperature, 1-0° C. Weights. Kilos. June 14, 1905 14:51 January 15, 1906 51°69 Total gain in weight.—37'18 kilos. During the first four weeks there was a gain of 3°37 kilos.; then for six weeks the weight was practically stationary (gained 1:36 kilos. only), subse- quently there was a progressive increase. The average gain of weight per week was 1°19 kilos. POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion.—At the seat of inoculation just above the umbilicus there was a shallow healed ulcer about 1 cm. in diameter, over which arched a narrow band of skin the extremities of which were attached to the margins of the ulcer ; the space between this band and the ulcer was filled with a small dry brown mass. On section nothing abnormal was seen in the subcutaneous tissues under the ulcer, but about 2 cm. in front of it there was a yellow slightly gritty tubercle about 15 mm. in diameter; there was a similar smaller tubercle under the skin about 5 cm. posterior to the ulcer. Inguinal Glands. — On the left side the gland nearest the local lesion contained a cavity about 1 cm. in diameter, filled with soft, cheesy, white, slightly gritty material. On the right side the corresponding gland contained a similar, rather larger [Age about 15 weeks.] Killed when in good health—January 15, 1906. [215 days after inoculation. ] nodule and a yellow tubercle the size of a pin’s head. The other inguinal glands were normal. Precrural Glands.—Normal. Abdomen. Iliac, Lumbar, and Coeliac Glands.—Normal. Omentum and Peritoneum.—Normal. Spleen.—Normal. Liver.—There was a doubtful grey speck in the substance of the liver. Portal Glands.—One portal gland contained an opaque whitish tubercle about 1-5 mm. in diameter. Kidneys and Suprarenal Bodies.—Normal. Thoraa. Heart and Pleura.—Normal. Lungs.—Normal. Bronchial and Mediastinal Glands.—Normal. Alimentury Tract. Tongue, Pharynx, Tonsils, Lurynx and Trachea.— Normal. Stomach and Intestines.—Normal. Gastric, Mesenteric, and Colic Glands.—Normal. Various Lymphatic Glands. Cervical Glands——A gland between the thyroid body and the trachea on the left side near the entrance to the thorax, was enlarged and on section yellow, necrotic and partially softened ; a smaller gland on the same side just within the thorax contained a small nodule filled with yellow pus. Other cervical glands were normal. Submazillary, Retro-pharyngeal, Prescapular, and Prepectoral Glands.—Normal. Microscopical Examinations. Smear preparations from the two cervical glands showed numerous streptococcal chains and short plump bacilli stained blue ; no tubercle bacilli were seen. Small Tubercle in Subcutaneous Tissues.—No tubercle bacilli. Liver (small focus).—No tubercle bacilli. Portal Gland.—Four tubercle bacilli. Inguinal Gland.—Numerous tubercle bacilli. 349 PIG 110. Virus P II. Fed with culture derived from the original material. Dose—The growth from seven serum tubes. Date—August 12, 1905. Weight at commencement of experiment—23:12 kilos. Killed when in good health—January 18, 1906. Clinical Notes. There was no sign of illness during the experiment. Temperature. During the first six weeks following the inoculation the temperature was not recorded. During October there was a slight rise of temperature lasting 25 days (maximum 39°9° C.). From October 27, 1905, to January 15, 1906, it remained normal. Tuberculin Tests. September 20, 1905. [39 days after inoculation.] Reacted. Rise of temperature 1°3° C. November 17, 1905. [97 days after inoculation.] No reaction. Weights. Kilos. August 15,1905... ee oon 23°12 January 18,1906... se sae 72°10 The animal was weighed seven times during the course of the experiment and showed a steady increase in weight. The total increase was 44°45 kilos.; the average weekly increase 1°99 kilos. POST-MORTEM. Carcass in good condition. Alimentary Tract. Tongue, Tonsils, and Pharynx.—Normal. Submazillary Glands.— Each submaxillary gland contained two yellow miliary tubercles, slightly gritty from calcification. Pharyngeal and Cervical Glands.—Normal. Stomach.—Normal. Small Intestine—The mucous membrane of the long Peyer’s patch in the ileum had a finely fissured or cracked appearance (suggesting previous ulceration and almost complete regeneration of the mucous membrane) ; the substance and the serous surface of the patch were normal. Nothing abnormal was seen in any other part of the intestine. Large Intestine.—Normal. Gastric Glands.—Normal. [Age about 194 weeks.] [159 days after commencement of experiment.] Mesenteric Glands.—The mesentric glands were slightly enlarged and felt indurated ; on section the greater number showed the substance almost com- pletely replaced by dry coarsely granular caseo-cal- careous tissue ; the glands at the extremities of the mesentery were least affected, several of the terminal glands showing only a few discrete yellow gritty tubercles. Lleo-colic Glands.—The ileo-colic glands were not enlarged ; they contained each a few discrete irregular yellow gritty tubercles, about the size of pins’ heads. Colic Glands.—One colic gland on the first part of the colon contained three tubercles similar to those in the ileo-colic glands ; in the rest of the colic glands three somewhat smaller tubercles were seen. Rectal Gland.—Normal. Abdomen. Omentum and Peritoneum.—Normal. Spleen.—Normal. Liver and Portal Glands.—Normal Kidneys.—Normal. Suprarenal Bodies.—Normal. Thoras. Heart and Pleura.—Normal. Lungs.—The lungs were crepitant and collapsed normally ; in the ventral flap of the left cephalic lobe there was a grey translucent nodule 2°5 mm. in diameter ; there was a similar smaller nodule in the right caudal lobe ; lungs otherwise normal. Bronchial and Mediastinal Glands.—Normal. Various Lympnatic Glands. Coeliac, Prescapular, Prepectoral, Lumbar, Iliac, Superficial Inguinal, and Precrural Glands.—Normal. Microscopical Examination. Mesenteric Gland.—Numerous T.B. Guinea-pigs Inoculated. No. 1859 (s) 2 with emulsion of mesenteric gland No. 1860 (i) developed slight tuberculosis only. ‘PIG 136. Virus P IL. Subcutaneous inoculation of culture derived from the original material through Pig 110 (mesenteric glands). Dose—100°0 milligrammes. Date—March 24, 1906. Weight at Inoculation—18'14 kilos. [Age about 124 weeks.] Killed when in good health—August 13, 1906. [142 days after inoculation. | Clinical Notes. A small slightly raised tumour developed at the seat of inoculation near the umbilicus, and the adjacent inguinal gland became slightly enlarged. ; The Be remained in good health during the period of the experiment. 34683 Témperature. Normal throughout. Tuberculin Test. The pig was not tested subsequent to inoculation. 3D2 Weights Kilos. March 24 18°14 April 18 21°31 May 15 24:03 August 13 on 42°18 Total gain of weight during experiment.—24-04 kilos. POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion.—In the middle line of the abdomen, a few inches in front of the umbilicus, there was a smooth somewhat flattened elastic swelling the size of a pigeon’s egg, composed of soft cheesy caseous sub- stance surrounded by a thin capsule of fibrous tissue, smooth and vascular on the internal surface. The tumour was situated in the subcutaneous fat, and was not adherent to the skin. In the fat surrounding the tumour there were a few discrete hard tubercles, the largest the size of a millet seed, composed of dry yellowish white friable substance. Inguinal Glands.—On the left side the gland nearest the local lesion contained four or five whitish caseous nodules, somewhat irregular in outline, the largest rather larger than a hemp seed; they could be readily shelled out from delicate fibrous capsules and were gritty from calcification chiefly around the margins. The corresponding gland on the right side contained two similar nodules. The resi were normal. Abdomen Omentum and Peritoneum.—Normal. Spleen.—Normal. Liver.—The liver contained in the substance a grey 3 a7 0 0 translucent nodule the size and shape of a barley geain, with a soft whitish centre, slightly gritty from calcification. The liver was otherwise normal. Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal Renal, Lumbar, and Iliac Glands.—Normal. Intestines. —Normal. Mesenteric Glands.—Normal. Thorax. Pleura.—Normal. Lungs.—The ventral parts of the small anterior lobes showed several lobules collapsed and carnified. Bronchial and Mediastinal Glands.—Normal. A gland just above the manubrium sterni, not enlarged, contained a number of minute discrete yellow calcareous particles. Heart.—Normal. Tongue, Pharyna, Palate, Tonsils.—Normal. Eyes.—Normal. Various Lymphatic Glands. Submazillary, Prescapular, Cervical, Prepectoral, Azillary, Popliteal, Precrural.—Normal. Microscopical Examination, Nodule from Liver.—No tubercle bacilli seen, Emulsion of Left Inguinal Gland.—Tubercle bacilli numerous. CAT 62. Virus P II. Intraperitoneal inoculation of culture derived from the original material. Dose—1°0 milligramme. Date of Inoculation—September 25, 1905. Died—December 7, 1905. [73 days after inoculation. | POST-MORTEM EXAMINATION. Local Lesion—In the subcutaneous tissues at the seat of inoculation there was a broad-bean sized mass of firm tissue, greyish and somewhat resembling glandular tissue. In the muscle wall near it there was a similar smaller mass. Abdomen. Omentum.—The omentum was enormously enlarged and composed of greyish tissue resembling glandular tissue, the whole mass having the appearance of a lipoma ; on section the mass was homogeneous throughout, and there were no areas of necrosis or softening. Pyloric Gland—The pyloric gland was enlarged and firm, but otherwise unaltered. In the Small Omentum and in the gastro-splenic omentum there were masses of tissue similar to the omental tumour. The surface of the Stomach, particularly near the pylorus, was closely covered with semi-transparent, slightly raised flattened granules, the largest about i mm. or a little over in diameter. Onthe surfaces of the Large and Small Intestines there were numerous granulations similar to those on the stomach, but larger and elongated in the length of the intestine. On the lower end of the intestine there were larger grey nodules up to a split pea in size made up of substance resembling the thickened omentum. The Mesentery and Mesocolon showed numerous greyish translucent filmy foci varying in diameter up to about 2 mm., just perceptible as slight thickenings between the finger and thumb. Mesenteric and Colic Glands.—The mesenteric and colic glands were enlarged and firm, but not necrosed. Spleen.—The spleen was not enlarged and on section appeared normal, Liver.—The liver showed a few doubtful minute yellow points, but was otherwise normal in appearance. Portal Glands.—The portal gland was much en- larged ; there was no necrosis. Kidneys.—Normal. Coelic, Lumbar and Iliac Glands.—Slightly enlarged ; no necrosis. Inguinal G'lands.—The inguinal glands were larger than normal, but were not opaque. Thoran. Lungs.—The lungs were congested and crepitant and slightly emphysematous in the anterior lobes, but showed no tubercles. Bronchial Glands.—Normal. Sternal Glands.—The sternal glands were greatly enlarged, on section congested and firm but not opaque. 351 Microscopical Examination. (Smear preparations.) _ Sternal Gland.—Extremely numerous short tubercle bacilli. Subcutaneous Nodule.—Extremely numerous tubercle bacilli, rather larger than in the sternal gland; the ‘smear resembled in places a pure culture. _ Inguinal Gland.—Numerous tubercle bacilli, many in clumps, Spleen—Very numerous tubercle bacilli; many large loose clumps and also small spherical clumps of bacilli closely gathered together. Marrow.—Numerous short tubercle bacilli; a few large loose clumps. CAT 64. Virus P II. Subcutaneous inoculation of culture derived from the original material. Dose—1°0 milligramme. Date of Inoculation—September 25, 1905. Killed—February 6, 1906. [134 days after inoculation. | POST-MORTEM EXAMINATION. Local Lesion.—In the subcutaneous tissues of the abdominal wall there was an elongated flat thickening, composed of brownish translucent tissue externally and tough opaque white tissue internally. Inguinal Glands.—The inguinal glands were normal in size and general appearance. Thoraz. Lungs.—In the right lung under the pleura there was a slightly projecting spherical white tubercle about 1:0 mm. in diameter; this when crushed was found to be softened throughout. Bronchial and Mediastinal Glands.—Normal. Abdomen. Peritoneum, Spleen and Kidneys.—Normail. Liver.—The liver showed in the substance under the capsule an opaque white tubercle the size of a pin’s head. Portal Glands.—Normal. Intestines and Mesenteric Glands.—Normal. All other organs and glands were examined and found to be normal. Microscopical Examinations. (Smears from) Local Tumour.—Numerous tubercle bacilli, Liver—A moderate number of tubercle bacilli. Lung.—No tubercle bacilli. Left Inguinal Gland.—No tubercle bacilli. RHESUS MONKEY 118. Virus P II. Subcutaneous inoculation of culture derived from the original material. Dose—10°0 milligrammes. Date—November 30, 1905 Killed when very ill—March 7, 1906. Clinical Notes. Fifteen days after the inoculation between the ‘shoulders, there was a softened ill-defined swelling ‘slightly raised above the surface, 3:7 cm. in greatest diameter. : On the 22nd day there was a tense fluctuating local tumour measuring 5 by 3°5 cm., and about 2 cm. in thickness. The axillary glands were slightly enlarged. Up to this time there had been no change in the general condition of the animal. ; ; On the 25th day the tumour burst, discharging a _yellowish-brown purulent matter. [Age—Young adult.] [97 days after inoculation. | On the 36th day the tumour was the size of a hen’s egg and there were two small openings in the skin, one of which was discharging; there was a group of slightly enlarged glands in each axilla. On the 87th day the monkey was first noticed to be really ill; its appetite was poor and the respiration increased. For several weeks previously the monkey had been observed to be gradually becoming smaller and thinner. Six days later its condition was much worse; the respirations were quick and jerky, and the appetite very poor. On the 97th day it was killed ; it would probably not have lived more than two or three days longer. 302 Temperature. JAneub. Jest JANJARY FE RY From the commencement of the experiment to January 23, 1906, a period of eight weeks, the temperature- remained approximately normal. Tuberculin Test. February 2. [64 days after inoculation.] Reacted. Rise of temperature, 0°8° C. POST-MORTEM EXAMINATION. Local Lesion.—In the subcutaneous tissues over the upper lumbar vertebrae there was a small flattened extremely thin-walled cavity containing two tough yellowish caseous masses and a small quantity of caseo-pus. The inner surface of the cyst was smooth and shiny Axillary Glands.—An axillary giand on each side contained one soft greyish white tubercle about 1:5 mm. in diameter. Prepectoral Glanis.—Normal. Thorax. Lungs.—On opening the thorax the right middle and lower lobes were found to be enlarged and to fill the lower part of the right pleural cavity ; the right lung was adherent to the chest wall chiefly on its lower dorsal and diaphragmatic surfaces ; the parietal pleura in these regions was thickeaed, and showed a honeycombed surface. On section the greater part of the right lung was firm, airless, and in a condition of caseous pneumonia ; the posterior portion of the right caudal lobe was uniformly caseous, but in the anterior part and in the right middle lobe, chiefly around the margins, there was a small amount of dark red consolidated lung tissue surrounding the caseous patches ; at the apex of the caudal lobe there was a small amount of air- containing tissue and the anterior lobe was crepitant throughout ; the latter was intimately adherent to the middle lobe. The crepitant portions of the lung contained firm discrete greyish yellow tubercles ranging from 1 to 2 mm. in diameter. The left lung collapsed normally and contained moderately numerous firm discrete caseous nodules ranging in size from 1 to nearly 3 mm. in diameter ; the ventral flap of the anterior lobe was consolidated, and showed on section firm yellow caseous nodules closely aggregated together and surrounded by dark red collapsed lung tissue. Thoracic Glands.—The bronchial glands on the right side were greatly enlarged and caseous throughout. The subsequent temperatures are given in the chart. The irtertracheo-bronchial glands showed the greatest degree of enlargement and were fused together. On section the caseous substance was beginning to break down, forming greenish yellow pus, chiefly around the periphery of the gland. On the left side a praetracheo-bronchial gland, the size of a small bean, was more than half caseous and softened ; one small intertracheo bronchial gland was normal. The sternal glands showed soft caseous tubercles. Pleura.—tIn the intercostal spaces on the right side near the sternum there was a series of firm caseous nodules, the largest the size of a kidney bean; near the diaphragm were several prominent yellow caseous tubercles. Vertebral Glands—Two vertebral glands on the right side, in the posterior interspaces, contained each one or two soft pinhead-sized caseous foci. Marrow.—The marrow of the femur and tibia con- tained moderately numerous soft yellow tubercles, mostly about 1:0 or 15 mm, in diameter, but some were larger. Abdomen. There was no fluid in the peritoneal cavity. Omentum.—The omentum contained a few scattered greyish-white opaque tubercles 1 to 1°5 mm. in, diameter and a number of minute transparent tubercles, Spleen.—The -spleen was slightly enlarged, and crowded with soft yellowish white nodules with thin capsules, the largest 5 mm. in diameter. Splenic Glands.—Two splenic glands contained each one or two small opaque whitish tubercles, Liver.—The liver was displaced by the enlarged right lung and was closely beset with firm discrete Opaque white tubercles ranging in size from a mere speck up to 2 mm. in diameter; the majority were small and about 0°5 mm. in diameter Portal Glands.—In the hilum cf the liver there were two slightly enlarged glands; one contained a number of discrete soft yellowish white tubercles up- te a little over 1 mm. in diameter; the other con- tained only two or tbree. 353 Coeliac Glands. — Two slightly enlarged coeliac ‘glands showed arranged around the cortices a number of soft yellow purulent nodules, 1 to 2 mm, in diameter, in one gland becoming confluent. A precardial gland contained two yellowish soft tubercles. Kidneys.—Each kidney showed in the cortex, on the surface and in the depth, numerous opaque yellowish-white tubercles ranging in size from 1 to 15 mm, in diameter ; many of the tubercles projected from ee surface. The medullary zone and pelvis were normal. Suprarenal Bodies—The right suprarenal body contained two opaque miliary tubercles. The left was normal, Iliac Glands.—A gland on the left side, on the ramus of the pubis, contained three soft yellow nodules, the largest 15 mm. in diameter. Other glands in this region were normal. Lumbar Glands.—Normal. Alimentary Tract. Tonsils, Pharynx, and Larynz.—Normal. The Subdmasaillary and Glands were normal. Submental Retro-pharyngeal Glands.—The right retro-pharyn- geal gland contained a nodule about 2°5 mm. in diameter filled with yellow pus. The left was normal. Parotid Salivary Gland.—In the parotid salivary gland lying over the insertion of the masseter muscle there was a firm whitish nodule, 2:0 mm. in diameter, beginning to soften. Cervical Glands.—Normal. Intestines—Normal. Mesenteric Glands.—Two mesenteric glands con- tained each a soft yellow nodule about 3 mm. in diameter. Other mesenteric glands were normal. Colic Glands —Two colic glands on the ascending colon were slightly enlarged and partly caseous; the rest of the cclic glands were normal. Testes.—Normal. In the subcutaneous tissues of the abdominal wall there were scattered firm spherical tubercles ranging up to nearly 2 mm. On section they were opaque, white, and some were softened. There was a soft greyish white tubercle (1°5 mm.) in the right latissimus dorsi muscle. Brain.—Normal. MONKEY 120. Virus P II. Subcutaneous inoculation of culture derived from the original material. Dose—1-0 milligramme. Date—November 30, 1905. Killed when in good health—March 23, 1906. Clinical Notes. Fifteen days after the inoculation between the shoulders there was a small local tumour not so large ag a broad bean. On the 36th day there was a soft fluctuating tumour half the size of a pigeon’s egg, the axillary glands were ‘slightly enlarged. The tumour subsequently burst and discharged, and the. ulcer afterwards healed, the tumour being represented by a patch of thickening. The monkey was killed on the 113th day. It had shown no definite symptoms of ill health, but it appeared to have become a little thinner and smaller during the experiment. : Temperature. The temperature remained normal throughout, the maximum range of variation during a period of three months being only 37°7° C.-39°3° C. (1°6° C.). Tuberculin Test. February 2. [64 days after inoculation.] Reaction. ‘Rise of temperature 0°9° C. POST-MORTEM EXAMINATION. Local Lesion.—At the seat of inoculation there was [Age—Young adult. ] [113 days after inoculation. ] a thin patch of fibrous thickening, the size of a sixpenny piece ; the fibrous tissue was congested. Azillary and Inguinal Glands.—Normal. Liver, Kidneys, and Suprarenal Bodies.—Normal. Spleen.—The spleen was normal in size ; the mal- phigian bodies were very conspicuous but showed no sign of necrosis (no tubercle bacilli found). Thorax. Lungs.—The lungs were crepitant and collapsed normally ; they showed under the pleura two trans- lucent grey tubercles, 1:5 mm. in diameter, with minute opaque yellow centres. In the left caudal lobe there was a loculated bronchiectatic cavity, with smooth shining walls, containing a small quantity of yellow pus (this showed no tubercle bacilli but several elliptical bodies resembling ova) ; the pleura over the cavity showed a depressed scar. Rest of lung normal. Bronchial Glands——Normal. Neck Glands, Pharynx, and Tonsils—Normal. Portal, Coeliac, Lumbar, and Iliac Glands— Normal. Wicroscopical Examination. No tubercle bacilli were found in the spleen nor in the pus from the lung. MONKRY 122. Virus P II. Fed with culture derived from the original material. Dose—About 10:0 milligrammes. Date—November 30, 1905. [Age—Young adult. ] Killed when in good health—March 22, 1906. [112 days after feeding. ] Clinical Notes. The monkey remained well during the experiment. Temperature. The temperature remained approximately normal throughout, with the exception of one week (January 28 to February 3), during which it was slightly raised, the maximum being 39°8° C. Tuberculin Test. February 2, 1906. [64 daysafter feeding.] No re- action Rise of temperature 0°3° C. 3 POST-MORTEM EXAMINATION, Body in very good condition. Alimentary Tract. Tongue, Tonsils, Larynx, and Pharynx.—Normal. Submawillary, Pharyngeal and Cervical Glands.— Normal. Stomach, Small and Large Intestines —Normal. Mesenteric Glands.—One mesenteric gland contained in the centre a yellow nodule, filled with soft pus, about 2 mm. in diameter ; other mesenteric glands were normal in size and appearance. Lleo-colic and Colic Glands.—Normal. Abdomen. Omentum and Peritoneum.—Normal. 54 Spleen, Liver, Kidneys, and Suprarenal Bodies— Normal. Portal, Lumbar, and Iliac Glands.—Normal. Testes.—Normal. Thoraz. Heart and Pleura.—Normal, Lungs.—The lungs were normal except for two or three cysts with fibrous walls containing a few: parasites. Bronchial Glands.—Normal. Microscopical Examination, Pus from Mesenteric Gland.—No tubercle bacilli. MONKEY 124. Virus P IT. Fed with culture derived from the original material. Dose—1°0 milligramme. Date—November 30, 1905. [Age—Young adult.] Killed when in good health—March 22, 1906. [112 days after feeding. ] Clinical Notes. The monkey remained well during the experiment. Temperature. The temperature remained approximately normal throughout. Tuberculin Test. February 2,1906,. [64 days after feeding.] Reacted. Rise of temperature, 0°8° C. POST-MORTEM EXAMINATION. Body in very good condition. Alimentary Tract. Tongue, Tonsils, Larynx, and Pharyna.—Normal. Submazaillary, Pharyngeal, and Cervical Glands.— Normal, Stomach and Large Intestine —Normal. Small Intestine.—The small intestine showed in the ileum under the mucous membrane two small hard bodies which on section were grey and translucent. Mesenteric Glands—Two mesenteric glands con- tained each a collection of creamy white pus, the size of a small pea ; between the peritoneum and another gland there was a small quantity of pus which could be moved about under the membrane by pressure. Other glands were normal in size and appearance. Ileo-colic and Colic Glands.—Normal. Abdomen. Omentum and Peritoneum.—Normal. Spleen, Liver, Portal Glands, Kidneys, andj Supra- renal Bodies.—Normal. Lumbar, Iliac, and Bronchial Glands.—Normal. Thorax. Lungs—The lungs were normal except for one parasitic cyst. Heart and Pleura.—Normal, Testes.—Normal. Microscopical Examination. Pus from Mesenteric Gland.—No tubercle bacilli. GOAT 34. Virus P II. Fed once with culture derived from the original material. Dose—15°0 milligrammes. Date—February 8, 1906. Killed—June 20, 1906. [Age about 1 year.] Clinical Notes. The animal remained in good health during the experiment. Temperature. On March 5, 25 days after inoculation, the tempera- ture rose to 39°6° C but quickly fell to normal. With this exception the temperature was normal throughout. [182 days after inoculation. ] Tuberculin Test. May 31,1906. [112 days after inoculation.] Re-- acted. Rise of temperature, 1-4° C. Weights. Kilos. February 21,1906 ... é ee = 22.22. March 20, 1906 ues aes we =: 19°95 April 18, 1906 ie oo we «=: 16°78 May 15, 1906... ans we ws: 16°32 June 20, 1906 ans ee wwe =—- 1587 Total loss of weight during experiment.—6'35 kilos. 355 POST-MORTEM EXAMINATION. ‘Condition fair. ‘Tongue, Tonsils, and Pharyna.—Normal. Submazillary, Retro-pharyngeal,and Cervical Glands. -—Normal. ‘Thyroid Body.—Normal. Abdomen. ‘Omentum.—-Normal. ‘Peritoneum.—The peritoneum covering the left side of the diaphragm and the serous surface of the stomach was roughened with numerous small connec- tive tissue tags. Intestines. — The mucous membrane of the ileum ‘showed some scattered greyish yellow foci, which ‘resembled solitary follicles ; otherwise the small intes- tine was normal. Large intestine appeared normal. ‘Gastric Glands.—Normal. _ Agland on the first part of the duodenum showed in the cortex a yellowish white calcareous tubercle the size of a millet seed and two or three calcareous grains. Mesenteric Glands.— Several small glands in the anterior part of the mesentery were normal ; all the others were normal in size, but showed in the cortices just under the capsule numerous irregular yellow calcareous tubercles ranging in size from a mere point to that of a hemp-seed ; they were most numerous in the large terminal gland. Leo - colic Glands. — The ileo - colic glands, not enlarged, contained a few irregular yellow calcareous tubercles, the largest the size of a millet seed. Colic Glands.—Normal. Spleen. - Normal. Liver.—Normal. Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Thorax. Lungs.—The right lobes showed large patches of recent hemorrhage ; in the left lobes there were some small hemorrhagic patches. In the thin posterior borders of the caudal lobes there were a few small parasitic nodules. No tubercles were seen anywhere. Bronchial and Mediastinal Glands.—Normal. Heart and Pleura.—Normal. Precrural, Pudic, Lumbar, Iliac, Prescapular, Pre- pectoral, and Axillary Glands.—Normal. Mammary Gland and Uterus.—Normal. Microscopical Examinations. (Smear Preparations.) Duodenal Gland Ileo-Colic Gland Small Intestine (yellow foci).—A few tubercle bacilli. i A few tubercle bacilli. GOAT 36. Virus P II. Subcutaneous inoculation of culture derived from the original material. Dose—10°0 milligrammes. Date—February 8, 1906. [Age—Old adult.] Killed when in good health—June 19, 1906. Clinical Notes. For two or three days after inoculation the goat was unwell, lay with its head under the manger and ate very little food. On February 19 she aborted. On February 22 the tumour at the seat of inocu- lation measured 7°5 by 4°5 cm. by about 2°5 cm. thick. The right prescapular gland was slightly enlarged, 5cm.in length. The goat was then quite well. Subsequently the tumour became soft and fluctuat- ing; on April 10 it was egg shaped, measuring 9 cm. in long diameter. The prescapular gland was 5 cm. in length. The goat was quite well. No further changes were noted in the local con- ditions. : The goat remained well during the remaining period of the experiment. Temperature. On the 5th day after inoculation the temperature rose to 39°6° C. and reached 40°1° C. on the 8th day ; on the 12th day it was again normal, and remained so until the animal was killed. Tuberculin Test. May 31, 1906. [112 days after inoculation.] Reacted. Rise of temperature, 1°4° C. Weights. Kilos. February 21, 1906 ... 40°36 March 20, 1906 ste bois 40°36 April 18, 1906 ams ae ae 15, 1906 we ale : Tare I! 29:93 June 19, 1906 ans ais ive Total loss of weight during eaperiment.—10°43 kilos. 34684 [131 days after inoculation. ] POST-MORTEM EXAMINATION. Condition fair. Local Lesion.—Iin the subcutaneous tissues on the right side of the neck there was a fluctuating tumour, the size and shape of a hen’s egg ; on section it was found to be a cyst with a thin fibrous wall and yellowish white softened pultaceous contents; the internal surface of the cyst wall was smooth and shiny, except at one part, where there was a patch of granulation tissue ; to the latter there was attached a small irregular firm caseous mass; the skin was lightly adherent to the outer surface of the cyst wall. Right Prescapular Gland.—The right prescapular gland measured 3°5 by 2 cm. and a little over 1 cm. in greatest thickness; on section it contained a firm pinkish yellow caseous mass, slightly gritty from calcification, replacing about three quarters of the gland substance ; the mass had begun to soften around the margins and was almost completely separated from the thin fibrous capsule which surrounded it. Left Prescapular Gland.—Measured 2:7 by 1 by about 0°5 cm. and was normal on section. Prepectoral and Cervical Glands.—Normal. Thorax. Lungs.—In the left caudai lobe just under the pleura there was a nodule, the size of a hemp seed, with a fibrous wall and soft caseous gritty contents ; near the root of this lobe, projecting slightly from the ventral surface, there was an irregular nodular mass, 1 cm. in greatest diameter, composed apparently of several small nodules joined together ; on section it showed a pea-sized cavity filled with yellow caseo- pus and several soft yellow foci surrounded by grey 3E 356 translucent fibrous tissue. In the right middle lobe there was a pea-sized nodule with a fibrous capsule and caseo-purulent contents. In the thin posterior parts of the caudal lobes and under the pleura on the dorsal surface were several firm grey patches, composed apparently of aggregated grey foci (verminous pseudo- tuberculous pneumonia) ; sparsely scattered over the surface of both lungs just under the pleura were tubercles, varying in size from a mere point to about 1 mm. in diameter ; the smaller ones were grey and semitransparent ; the larger ones had yellow centres, in some calcareous, in others softened ; several of the latter were examined microscopically but no tubercle bacilli were found ; these were probably of a parasitic nature. Zhoracie Glands.—The bronchial glands appeared normal, The long dorsal mediastinal glands contained four firm yellow nodules, composed of a friable caseous substance, one the size of a pea, the other that ofa millet seed; all were slightly gritty. The gland immediately anterior to the latter was enlarged and contained two similar nodules, one 1 cm. in diameter, which almost completely replaced the gland substance. An intra-tracheal gland, the size of a kidney bean, ‘contained two pea-sized caseous nodules fibrous around the margins, Pleura and Heart—Normal. Abdomen. Omentum and Peritoneum.—Normal. Spleen.—Normal. Liver.—Normal. Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies—Normal. Alimentary Tract. Tongue, Tonsils, and Pharynx.—Normal. Submaslilary, Pharyngeal, and Cervical Glands.— Normal. Intestines.—Normal. Mesenteric Glands.—One showed in the cortex a small irregular calcareous tubercle ; in the medulla of almost every one of the glands were scattered spherical grey foci, the largest about 1 mm. in diameter, with minute opaque centres (no T.B. found, probably parasitic). Ileo-Colic and Colic Glands.—Normal. Various Lymphatic Glands. Lumbar, Iliac, Precrural, Popliteal, Gluteal, and Pudic Glands.—Normal. Thyroid Body.—Normal. Microscopical Examinations. (Smear Preparations.) Lung (two large nodules).—Tubercle bacilli easily found in both. Several small tubercles. No tubercle bacilli. Mesenteric Glands (calcareous tubercle).—? 1 tubercle bacillus. (medullary foci)—No tubercle bacilli. ” ” CALF 424. Virus P II. Subcutaneous inoculation of culture derived from the original material. Dose—50-0 willigrammes. Date of Inoculation—May 22, 1906. Weight at Inoculation—26°75 kilos. [Age about 5 weeks. ] Killed when in good health—August 30, 1906. [100 days after inoculation.] Clinical Notes. Eighteen days after inoculation on the left side of the neck, there was a firm tumour 7:5 by 6°5 cm. by about 2 cm. in greatest thickness ; the adjacent pres- capular gland was large and hard, 9 cm. in length. On the 25th day the tumour was firm and prominent, but smaller, measuring 6°5 by 5 cm. ; the prescapular gland had also diminished in size, and was now about 7°5 cm. in length. No further changes of importance took place in the jlocal conditions. The calf remained in good health during the experiment. Temperature. A period of slight pyrexia commenced on the 11th day and lasted 9 days, the maximum temperature being 39°7° OC. Subsequently the temperature remained normal. Tuberculin Test—The calf was not}tested subse- quent to inoculation. Weights. Kilos. May 22 ite aes ane 26°75 July 17 as ee is a 45°80 August 30 4... aes a 58°49 Total gain of weight.—31°74 kilos. Average rate of gain per week.—2°26 kilos POST-MORTEM EXAMINATION. General condition good. Local Lesion—At the seat of inoculation on the left side of the neck there was an oval tumour measuring 5 by 4:5 by 25cm. On section it was a fibrous walled cyst adherent to, but not infiltrating, skin and muscle. The cavity was lined by pinkish granulation tissue, and contained two caseous, slightly gritty, necrotic masses, surrounded by a glairy purulent fluid. Left Prescapular Gland.—The left prescapular gland measured 7°5 by 5:0 by 3°8cm. On section the gland substance had disappeared, and it consisted of a cyst containing caseo-necrotic masses bathed in yellow caseo-pus. There were several fibrous bands stretch- ing across from side to side. Right Prescapular, Prepectoral and Axillary Glands. —Normal. Thorax. Pleura and Heart.—Normal. Lungs and Thoracic Glands.—Normal. Abdomen. Peritoneum and Omentum.—Normal. Spleen and Liver.—Normal. Gall Bladder and Portal Glands—Normal. 357 Kidneys and Suprarenals.—Normal. Renal, Lumbar, and Iliac Glands.—Normal. Alimentary Tract. Tongue, Pharynz, Palate, Tonsils—Normal. Pharyngeal, Submawillary, Hyoid, and Parotid Glands.—Normal. Intestines.—In the first part of the small intestine, beneath the mucous membrane of the Peyer's patches, there were four slightly irregular yellowish nodules up to a millet seed in size; on section they contained a creamy purulent substance. The large intestine was normal. Mesenteric, Ileo-Colic, and Colic Glands.—Normal. Eyes.—Normal. Testes.—Normal. Various Lymphatic Glands. Precrural, Popliteal, Gluteal, Ischiatic, Pudic.— Normal. Microscopical Examinations. Nodule from Small Intestine—Two tubercle bacilli seen. Left Prescapular Gland.—Tubercle bacilli fairly numerous. CALF 556. Virus P II. Fed once with culture derived from the original material. Dose—1,000-0 milligrammes. Date of feeding—July 16,1907. [Age 8 weeks.] Killed when in good health—February 6, 1908. Clinical History. The animal showed no sign of ill-health during the course of the experiment. Temperature. The temperature was normal throughout. Tuberculin Tests. September 18, 1907, [64 days after the experiment began]. Dose of tuberculin (Bovine), 2°0 cc. No reaction. : October 29, 1907, [105 days after the experiment 7 beganj. Dose of tuberculin (Avian), 40 cc. Rise of temperature 0°9° C. Weights. Kilos. July 16,1907... se vee ee AT February 6, 1908 ... ate we = 8117 Total gain in weight.—39°45 kilos. [205 days after inoculation. ] POST-MORTEM EXAMINATION. General condition good. Liver.—In the substance and just beneath the capsule there were about twelve soft irregular yellowish foci, a few of which were surrounded by slight zones of congestion. Rectal Glands.—In each of three there were from one to three minute opaque soft yellowish foci. There was no sign of disease in any other organ or gland in the body. Microscopical Examinations. (Smear Preparations.) Rectal Gland.—No tubercle bacilli. Liver (focus 1).—No tubercle bacilli. Liver (focus 2).—No tubercle bacilli. Culture made from rectal gland was] avian Jin character. 34684 3 Ez 358 “*sUIMeHOIg, SNOdseO -O1QY [[@Us & SVM 9107} YOU oy} JO opis ‘an}uem0 eq} UL se_npou pues syveTys snosied -[eo pue ‘1aATT 94} UL sepoteqn} 490 -nysued} Jo tequinu eyeiepom ve ‘ssun{ eq} UL sefnpou Mores omy put soeTo -1oqNn} qusoON[SUeI, MOT wv OOM OIOT J, snoeseo peure}u0o Spue]s O1[0d OY} PUe OIIo}WISOU OUINY ‘pejooge §=ATQqSI[s e10M si0q}O aNoF ‘suorse] 449119 SnOeseo PeuleyU0d spurls ‘1003 £49113 snoesto jo dnois v sea pisuo} euo U[—''d ‘shep eT "8067 ‘GT eung : port "BU 0.009 : pealeder JUNOMe [eYOT, ‘shup 9 IoF poy ‘?T ANOd “LON | * OOTH | dexjur | Toze TAT] JO" ‘LS | “ or ‘a | denay | ooze ‘LON | “ oor | desquy | coze | Suny jo -g ‘ “purrs “Leo | skep COTM | “Feyyuy | goze | ooo Jo -q “4jnsey yo mee “‘poqpe fsoqunny| ‘[erreyepy “L [007 “68a | Sur o.or | ‘gnoqng| 0468 3 ‘SDId-VANING “L Teoory | sfep gg -q | ur g.or | moqng| 696% | = | “4[nsoy jou Saentigy ‘es0q =| pouyey | ‘soquinyy 4JO] OY} UO OPOBNUL oy} UT “SVId-VAENING ‘sofnpou =. 444119 ‘Ly at ‘sIsOTNOIEqny “sIso[noTeqny amorgg}] “ gota }Sar 100-0] sear | 9T9OT ‘LD! “ 26°a [8m 100.0] aeyuy | OPE eyn0V— Wd e4noy—'hd ee “sue810 “skep OF ‘skep G¢ ara y | skep es “MH | 8m 10.0 | aetar | ATOT “LD | shep Fo -q | Sur 10.0 | ‘aeaquy | BPE “8061 ‘8061 ‘6 TON: pd == ‘F qoaeyy : porq ‘ay 2 : : : : : ; . : : : , MSH | oy g cer esoq, |:poyyepg | zeny wmsey | 4, area asog |poyyeyy | zequny | “8M 0.0T : e80q Sut 0.J : esq “Snouoavlzyuy “SNOTOACIIUT Tees ce ‘9G aes “Bq fee “plo sfep 1g ‘uoneseued 44)z 949 Se posn ONy[NO OT, ‘skep 990'T Jo potsed [2}0} B WOTPBAT4[NO UI Useq peYy pUL ‘[elIE}eUT [VUTSIIO 9) WOAT poAliop SV TIB14S OTL 7, "8061 ‘6% AUVONVE—TA “‘panugquoo—SNOTLVTIOONI TAALTNO ‘ponurquoo—(Btq Jo puxyy Arey[IxeMaNg) “TIT d SNAIA FEEDING EXPERIMENT WITH CULTURE, COMMENOING ON JULY 25, 1905. hin derived from original material. Total duration of artificial cultivation, 138 days. The 8th generation of culture was used. PIG 112. Feeding. Number. Meth}, Fed for seven days with culture, the owth from one tube being used each by. 334 331 GUINE Sub Inoculat r,derived f Killed P.M.— in inguil tuberculo| 34684 2T Intr, Killed when “ : September 14, 905 Intre 51 days. P.M.—The left submaxillary glands rere enlarged and hard; in one a 1inute focus was seen. The small atestine contained numerous shallow Icers showing neither congestion nor aseation (smear, T.B.). The mes- nteric and ileo-colic glands were nlarged and mottled with whitish treaks and foci. GUINEA-PIG 1691. Intraperitoneal. E. of mesenteric gland. Died : 17 days. P.M.—Acute tuberculosis (avian type). FOWL 30. 2 Fed: 25°0 mg. Killed: 65 days. P.M.—No tuberculosis. RABBIT 379. Intravenous. Inoculated with small dose on July 27, 1905. Died ; 22 days. P.M.—Acute tuberculosis 3 EF TII.—Aprit 12, 1906. The strain was derived from the original material and had been in cultivation a total period of 399 days. The culture used was the 18th generation, 15 days old. \B MONKEY 134. DOG 122 (Puppy). BITS. Subcutaneous. Subcutaneous. Duration Dose : 10:0 mg. Dose : 25:0 mg. Number. ates of Life, | Result. Killed ; August 21, 1906. Killed : 74 days. : 131 days. P.M.—Flattened cavity 79 Intramg-'| D. 21 days P.M.—At the seat of containing fluid and lined f sone T. inoculation was a small by granulation tissue in mg-| D.41 ,, cicatrix. Adjacent glands Se a soe ‘ 80 =| Suber normal. On the wall of ine ee TD LUNGS: (an the caceum were two cysts Evers containing caseo-purulent 78 Suber matter ; on the mucous surface opposite one was a minute ulcer, and the wall was infiltrated. No tuberculosis elsewhere. IV.—Auveusr 22, 1906. The strain was derived from the original material and had been in cultivation a total period of 561 days. The culture used was the 21st generation, 23 days old. FOWLS. | | Durati Number. | Method. Dese. of Life. Result. | 102 Intrav. | 10:0 mg. | D. 29 days | G.T. 7 — 100 Intrav. 10mg. |] D. 33, G.T. 98 Intrav.| O-1mg.| D. 41, G.T. | E. ee é V.—NoOVEMBER 28, 1906. E. The strain was derived from the original material, and had E been in cultivation a total period of 659 days. te The culture used was the 20th generation, 21 days old. | oe GUINEA-PIGS. RABBIT. | Duration papas . Method.; Dose. of Life. Result. | Number. | Method.| Dose. | ee | Result. Fed | 500 mg.| K.96 days | NoT. | 1133 | Fed | 10-0 mg. | D. 42 days | Local T. Fed | 100 mg.| K.96_,, No T. : a 361 CALF 282. Virus P III. Tnoculation into the sheath of the Dose—10-0 milligrammes. Date—June 17, 1905. jugular vein of culture derived from the original material. Weight at Inoculation—37-64 kilos. [Age about 7 weeks.] Killed when apparently in good health—July 4, 1905. [17 days after inoculation.] Clinical Notes. _A hard swelling developed in the sheath of the jugular vein and completely surrounded it, Temperature. The temperature rose to 39:8°C. on the 3rd day after inoculation and reached 40:5° C. on the 10th day. The following day the temperature was normal. Tuberculin was then inoculated in a dose of 1°5 cc. and the temperature rose to 40°1° C., a rise of 0°6° C Weights. Kilos. June 27 sce wate cats 37°64 July 4 se’ eg ciate 35:37 Loss in weight.—2°27 kilos. POST-MORTEM EXAMINATION. Local Lesion.—In the deep tissues of the neck, on the right side, there was an elongated tumour 10 by 5 by 4cm., almost cartilaginous in hardness. On re- moval and section it was found to surround the right jugular vein, which ran through its centre, and to consist of very dense, somewhat translucent, fibroid tissue, showing here and there yellow necrotic streaks. The jugular vein in the middle of the tumour was constricted, and it was filled above and toa certain extent below with red blood clot. Cervical Glands.—A long narrow gland (one and a half inches in length), situated near the tumour, showed about half the substance composed of firm grey translucent tissue, penetrated with a firm yellow network. The rest of the gland seemed firmer than normal, but no opaque foci were visible. The right superior cervical, the size of a broad bean, was similarly affected. A gland, the size of a sparrow’s egg, situated between the latter two, was indurated, and composed throughout of grey translucent tissue showing signs of early necrosis. On the left side a mid cervical gland was a little enlarged and firmer than normal; no necrotic foci were seen. Other left cervical glands were normal. Right Prescapular Gland.—The right prescapular gland was a little larger than the left. Near the superior extremity there was an indurated patch in the cortex, which showed a few extremely minute yellowish points. Left Prescapular Gland.—Normal. Prepectoral Glands.—The right spherical prepectoral gland was enlarged (1 centimetre in diameter), and indurated. Other prepectoral glands were normal. Axillary Glands—Normal. Thorax. Heart.—Normal. Pleura.—Normal. Lungs.—The lungs were speckled with very nume- rous minute dark red foci of congestion. In the caudal lobes, under the pleura, there were diffuse pinkish areas of congestion, and the cephalic lobes contained several dark red collapsed lobules, the caudal lobes a few; with these exceptions the lungs were crepitant throughout. Bronchial and Mediastinal Glands.—The bronchial and mediastinal glands were not obviously enlarged. On section their tissue was oedematous and congested. Abdomen. Omentum.—Normal. Peritoneum.—Normal. Spleen (140 grammes).—Normal. Liver.—Normal. Portal Glands.—Normal. Kidneys——Each kidney showed one or two grey points on the surface ; these on section were prolonged into the cortex as fine radial streaks. Suprarenal Bodies.—Normal. Intestines.—Normal. Other lymphatic glands (not mentioned above) were normal. Microscopical Examinations. (Smear preparations.) Local Tumour.—Numerous slender tubercle bacilli. Left Mid-cervical Gland.—Moderate number of tubercle bacilli. Small Right Mid-cervical Gland.—Very numerous short tubercle bacilli, mostly single forms, but many large loose clumps seen. Spleen.—A few small groups of tubercle bacilli, and a few scattered single forms (short diplobacilli stained blue ?). Lungs.—Numerous short fat bacilli stained blue, also some longer slender forms ; no tubercle bacilli. Kidney.—Short blue stained bacilli in small number. Liver.—No organisms. Left Bronchial Giland.—No tubercle bacilli ; a few organisms stained blue. CALF 392. Virus P III. Subcutaneous inoculation of culture derived from Calf 282 (inoculated with culture of original material) through G.P. 1612. Dose—50°0 milligrammes. Date--March 20, 1906. Weight at Inoculation—52'16 kilos. Killed when in good health—June 22, 1906. Clinical Notes. Four days after the inoculation on the left side of [Age about 11 weeks.] [94 days after inoculation.] the neck a slight local thickening could be felt the size of a two shilling piece. On the 21st day a firm tumour had developed, 362 measuring 7°5 by 9 cm. and not quite 2°5 cm. thick ; the prescapular gland was enlarged, about 7°5 cm. in length. Very little change took place subsequently in the local conditions ; both tumour and gland diminished a little in size. The calf remained well during the experiment. Temperature. There was a slight pyrexia aud irregularity of tem- perature during the third and fourth weeks after inoculation, the maximum range of variation being between 38:2° and 39°4°C. Otherwise the temperature remained normal during the experiment. Tuberculin Test. May 31, 1906. [72 days after inoculation.] Re- acted. Rise of temperature, 1°5° C. Weights. The animal gained weight rapidly during the ex- periment. Kilos. March 20 52°16 April 24 87:98 May 15 ace soe ea 96°14 June 21 Rs iis eee 111°65 Total gain of weight.—59°49 kilos. Average rate of gain per week.—4'48 kilos. POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion.—In the subcutaneous tissues on the left side of the neck there was a fluctuating tumour, measuring 7°5 by 5 by 4 cm.; on section this was found to be a cyst with thin fibrous walls and thick tenacious caseo-purulent contents; the cavity was lined internally by a thin layer of organised granula- tion tissue and was crossed by a number of slender fibrous trabeculae ; the skin was lightly adherent to the capsule ; in the muscles under the cyst there was a caseous mass the size of a broad bean with a capsule of fibrous tissue. Left Prescapular Gland.— The left prescapular gland measured 5°3 by 2°8 by 1°2 cm. and showed in the cortex four calcareous nodules, one about the size of a kidney bean, the others much smaller ; one of the nodules contained a small focus of caseo-pus. Right Prescapular Gland.—The right prescapular gland measured 5:3 by 2°5 by 1°3 cm. and was normal on section. Prepectoral and Cervical Glands on both sides were normal. Thorax. Lungs.—The lungs were perfectly normal in general appearance and no tubercles could be seen on the surface or on section. Thoracic Glands.—The dorsal mediastinal glands and the left bronchial gland showed in the cortices a few minute yellow points one or two of which were just perceptibly gritty. The right bronchial gland was normal. Heart, Pericardium, and Pleura.—Normal. Abdomen. Omentum and Peritoneum.—Normal Spleen (252 grammes).—Normal Liver.—Normal. Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Alimentary Tract. Tongue, Pharynx, and Tonsils.—Normal. Intestines, Large and Small—Normal. Submazillary, Pharyngeal, Parotid, Gastric, Mes- enteric, Ileo-colic, and Colic Glands.—Normal. Various Lymphatic Glands. Renal, Lumbar, Iliac, Axillary, Precrural, Popliteal, Pudic, Gluteal, and Ischiatic Glands.—Normal. Testes.—Normal PIG 112. Virus P III. Fed for seven days (from July 25 to July 29, 1905, and on July 31 and August 1) with culture derived from the original material ; one serum tube used on each day. Weight at commencement of experiment—14°06 kilos. [Age about 17 weeks.] Killed when in good health—September 14,1905. [51 days after the experiment began.] Clinical Notes. The pig remained in good health during the experiment. Weights. Kilos. July 25, 1905... en 14:06 August 8, 1905 ee as w=: 16°32 September 12, 1905 ... outs ww. =: 24°04 Total gain in weight during experiment.—9'98 kilos. Average rate of gain per weck.—1°35 kilos. POST-MORTEM EXAMINATION. Carcass in very good condition. Alimentary Tract. Tonsils—Normal. Tongue.—Normal. Pharynz.—Normal. Submaaillary Glands.—The left submaxillary glands were slightly enlarged and harder than those on the right side. Nothing abnormal was seen on section, but on crushing a small piece of one of the glands between ground glass plates a minute whitish point was observed. Pharyngeal Glands.—Normal., Cervical Glands.—Normal. Stomach.—Normal. Small Intestine—The mucous membrane of the small intestine showed numerous shallow ulcers the margins of which in some cases had a whitish appear- ance ; there was no appreciable thickening, however, of the margins or of the bases, and the peritoneal surface was normal. The ulcers were very numerous in the upper and middle parts of the intestine, and were punched out, circular or irregular in outline, ranging up to 2 or 3 millimetres in diameter ; where they were most numerous they had become united. 363 In the lower part of the intestine the ulcers were not 80 numerous nor so definite in outline, and appeared as superficial erosions. They usually involved the mucous membrane only, but in one or two cases the peritoneum was seen to be exposed in the centre of the ulcer. The Peyer’s patches, except in the lower part, were “severely affected, and in many the whole of the central part was eroded; in others there were irregular serpiginous ulcers, often surrounding isolated islands of mucous membrane. The ulcers extended to the muscle wall, the substance of the patch appearing -as if eaten away in these places. There was no thickening of the margins of the ulcers nor of the bases, and there was no surrounding congestion. In the patches in the lower part of the intestine they became less pronounced, and in the long terminal et one saw merely a number of small irregular pits. Large Intestine. — The solitary lymphoid follicles were conspicuous, but not apparently enlarged ; they had an opaque white appearance, and their central openings showed a little larger than usual ; no ulcers were seen. Mesenteric Glands—All the mesenteric glands were firm, and were two or three times their normal in size. On the surface they showed a faint whitish mottling. On section they showed creamy white streaks or whitish foci throughout their substance ; in some a kind of network was formed. Lleo-colic Glands. — The ileo-colic glands were slightly enlarged, and contained a smaller number of - streaks and foci. Colic Glands.—The colic glands were normal. Abdomen Peritoneum.—Normal. Liver——In the substance of the liver, under the capsule, a pinhead-sized white spherical body was seen ; otherwise normal. Portal Glands.—Normal, Spleen.—Normal. Kidneys.—One kidney showed on the surface a grey point of doubtful nature. The other was normal. Suprarenal Bodies.—Normal. Thorax. Lungs.—Normal. Bronchial Glands.—Normal. Mediastinal Glands.—Normal. Heart.—Normal. Pleura.—Normal. Peripheral Lymphatic Glands.—Normal. Testes.—Normal. Microscopical Examinations. (Smear preparations. ) Scraping from Mucous Membrane of Small Intestine. —Moderate number of tubercle bacilli. Left Submasillary Gland.—A few tubercle bacilli, Mesenteric Gland.—Numerous tubercle bacilli. Spleen ... see aes eee Liver ... eee ose ove Kidney ai ous «» >} No tubercle bacilli. Portal Gland... aes Lymphoid Follicle from Colon J CALF 318. Virus P III. Subcutaneous inoculation of culture derived from the original material. Dose—50-0 milligrammes. Date—September 7, 1905. Weight at Inoculation—97°90 kilos. [Age about 53 months.] Killed when in good health—December 11, 1905. [95 days after inoculation.] Clinical Notes. Five days after inoculation on the left side of the neck a flattened thickening could be felt, and the -adjacent prescapular gland was slightly enlarged. On the 15th day there was a slightly raised hard tumour measuring 14 by 10 cm.; the prescapular -gland was very large, 12°5 cm. in length. On the 23rd day both the tumour and the prescapular gland had shrunk a little. The calf which “up to this time had been well, seem a little unwell and ‘was quieter than usual. On the 55th day the tumour measured 9 by 7:5 cm., Temperature. SEPTEMBER it was adherent to skin and subjacent muscle and appeared to be softening. The prescapular gland was 11°5 cm. in length; the prepectoral gland was slightly enlarged. The general condition of the animal was now good. Subsequently one extremity of the left prescapular gland become soft and fluctuating. It increased in size and became adherent to the skin and finally (88th day) burst, discharging yellow pus ; the opening then became covered with a scab. The calf was quite well. No further change of importance took place in the local conditions, (Chart to November 2.) OCTOBER NOVEMBER, ... From November 2 to December 11, when the calf was killed, the temperature remained nermal. 364 Tuberculin Test. November 17, 1905. [71 days after inoculation.] Slight reaction. Rise of temperature 0:7° C. Weights. Kilos. September 12 eee ass 97:90 September 26 aoe ees 99°35 October 10 ... ui dea 98°35 November 21 vee ies we =—:119°26 December 11... aaa iow .. —:130°60 During the first four weeks the weight remained stationary ; afterwards the calf grew rapidly. Total gain of weight during experiment—32°70 kilos. Average rate of gain per week—2°26 kilos. POST-MORTEM EXAMINATION. Carcass in good condition. Local Lesion At the seat of inoculation, on the left side of the neck, there was a tense fluctuating tumour, the shape of an egg, measuring 8°3 by 5 by 5 cm., and weighing with attached skin a little over 6 ozs. On section the tumour was a cyst containing yellowish pus (resembling muco-pus), and an irregular partially detached mass of dense yellowish white caseated tissue ; gritty particles were just perceptible in the firm cageous tissue as well as in the pus. The walls of the cyst were fibrous, moderately. thick, and were lined internally with pale granulation tissue. Left Prescapular Gland.—The left prescapular gland was much enlarged, and weighed 127 grammes. On section the greater part of the gland substance was composed of yellowish caseo-necrotic tissue, contain- ing scattered minute gritty particles; the necrotic tissue was for the most part firm, but was beginning to soften, and was in places converted into thick creamy white pus; the whole mass was surrounded by a thick capsule of fibrous tissue. At the lower ex- tremity of the gland there was a small portion of normal looking gland tissue containing two or three yellow tubercles. The skin just in front of the left shoulder showed a raised scab, which, when removed, revealed an ulcer a little more than 1 cm. in diameter ; the floor of the ulcer presented exuberant granulation tissue, and showed in the centre the orifice of a sinus ; the latter extended into the prescapular gland, the capsule of which was pierced by a small opening towards the lower extremity. The sinus contained a little yellow pus, and was lined by a thick layer of brownish, some- what translucent granulation tissue, which was sur- rounded by dense white fibrous tissue. Right Prescapular Gland.—Normal. Prepectoral Glands.—The rounded prepectoral gland on the left side measured 1 cm. in diameter, and was composed of dense necrosed tissue, softened around the periphery, and converted into thick yellow pus. Right prepectoral gland normal. Cervical Glands.—Normal. Thoran. Pleura.—Costal pleura normal. On the pleural surface of the diaphragm there was a loosely attached hemorrhagic lenticular body about 8 millimetres in diameter. Lungs.—In the left caudal lobe, under the pleura, there was a spherical yellow tubercle about 1 milli- metre in diameter, and a greyish red nodule the size of a pea. Scattered about over the surface were minute dark grey points of a doubtful nature. On section the lungs were normal. Bronchial Glands.—The right bronchial gland con- tained a soft yellow tubercle 1:5 millimetre in diameter, in which minute gritty particles could be detected. Mediastinal Glands.—The caudal mediastinal gland contained two pinhead sized yellowish tubercles. The gland above the bifurcation of the trachea. contained a soft yellow tubercle 1°5 millimetre in diameter. 2 An infratracheal gland contained a similar tubercle, not perceptibly gritty. Heart.—In the muscle substance of the left ventricle there was an irregular yellow gritty nodule about the- size of a wheat grain, which easily shelled out ; other- wise the heart was normal. Trachea.—Normal. Abdomen. Omentum and Peritoneum.—Normal. Small Intestine.—There were a few scattered soft yellow tubercles, some slightly gritty, in the Peyer's patches, and several firm projecting nodules up to the size of a pea. The latter were most numerous in the- ileum, and the majority showed in the centre a small ulcer ; the nodules were firm, grey or greyish red, and homogeneous, except one, in which a yellow point was seen. Large Intestine —Normal. Mesenteric Glands.—The mesenteric glands were not enlarged. They contained scattered opaque yellow softened tubercles, the largest about 1°5 millimetre in diameter ; they were rather numerous in the terminal gland. Lleo-colic Glands.—The ileo-colic glands contained similar tubercles, grouped together in places. to form nodules as large as a pea, of soft yellow, slightly gritty caseo-pus. Spleen.—The spleen weighed 280 grammes, and was normal in size and general appearance. Liver.—The liver was normal on the surface; it showed on section a few scattered minute opaque yellow foci (fatty ?). Gall Bladder. —Normal. Portal Glands.—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Renal Gland.—The renal gland contained two yellow pinhead-sized tubercles, one of which seemed gritty. Coeliac Glands—One contained two firm opaque yellow spherical tubercles, each about 1°5 millimetre in diameter ; another contained one similar tubercle, and a second the size of a pin’s head. Testes.—Normal. Lymphatic Glands. Right Gluteal Gland.—In the right gluteal gland there was a soft yellow tubercle, about 1°5 millimetre in diameter. Submaxillary and Retro-pharyngeal. — The right submaxillary and the left retro-pharyngeal glands. contained each a soft yellow miliary tubercle. All other lymphatic glands were normal. Tonsils.—Normal. Pharynx.—Normal. Larynz.—Normal. Microscopical Examinations. Prescapular Gland.—Moderately numerous tubercle bacilli. Heart Nodule (two smears).—One tubercle bacillus. seen in one. Liver ... ode oa ath Intestinal Nodule ... }No mbprele baci Dorsal Mediastinal Gland Renal Gland en's eile Coeliae Gland ais Submuaillary Gland \ A fow tubercle bacilli. ei } No tubercle bacilli. 365 CALF 530. Virus P III. Subcutaneous inoculation with culture derived from Calf 318. Dose—5,000:0 milligrammes. Date of Inoculation—April 25, 1907. Killed when in good health—October 10, 1907. Clinical History. The animal showed no sign of ill-health. Temperature. The temperature remained normal throughout. Tuberculin Test. September 18, 1907. [146 days after the experi- ment began.| Dose of tuberculin (Bovine), 2°0 cc. No reaction. Weights. Kilos. April 25, 1907 asi ack 28°12 October 10, 1907 sa a 79°81 Total gain in weight.—51-69 kilos. [Age 12: weeks. ] [168 days after inoculation. ] POST-MORTEM EXAMINATION. Condition good. Local Lesion.—At the seat of inoculation in the subcutaneous tissue on the left side of the neck there were six nodules varying in size from a pea up to a sparrow’s egg. On section they were filled with a creamy yellow caseo-purulent substance. Left Prescapular Gland.—Normal. There was no sign of disease elsewhere in the body. Cultures from liver avian T -and mediastinal gland were 2626 | D. 49, , Spleen and | Avian. avian in character. omentum. 2627 | K.169 ,, | Local T. None. — GUINEA-PIG 2905. Intraperitoneal. Emulsion of spleen from Guinea-pig 2626. Died : 143 days. P M.—Nodules in omentum containing pus. Emulsion of liver con- taining moderately numerous T.B. wie 570. Intravenous. Dose : 5-0 ce. Died: December 9, 1907. 31 days. P.M.—Acute tubercu- losis. (Table continued on oppo- site page.) | GUINEA-PIGS. Animal otherwise healthy. Material. | Number. i ei OF Result. eee Source. Characters, Bmolsion f 2800 | D. 55 days} Acute avian ee ak Avian, nae | eerie a [ ST ad No growth. Emulsion of kate ea — | is _ eae \ SEE | ele None. a 393 ‘ “pars = “GUO NT ‘Lreoy] “ corm | 92Tg d119qU98eUL puvpb (Z) pars a oo ‘LON | “ COTM] O8Tg | Fo Torspnmg ‘uDlay qournbuy “Lreooy | “ Gorm | Gare aze[ndeoseid “puvd} jo worspnmy “uDlapy pounbuz “LreooyT | “ Gora] rete “pues “UDmy wunjuaUc “Lieooy | “ Gorm | 2zte [J [Rotated “pune = au ‘LteootT | “ corm | zere Jo uorsjnug ‘unay qoumbuy “Lreooy | * GOL | e2ig 1) uaeie “pun? Avpndeosew' “UDA qounbuy ‘Lieot | “ corm | gzt¢e ‘peels A “UDIAy | “unpuowc “Lieoy | “ cola | F21E yO wotsTawy ‘aynpou zepndeoser “ZT UBiar “WOIS9] [BOOT ‘upp | ppauojiwag *L Teooy | skepcor ay | ezte jo WoIs;nMy _ ‘quugy | aqnoe-qug |skep te “qd ] ZLzI¢e jo WOIsjnUy \ “adh I, *a0unog 01] “SL920DLDYD "a0.Lnog ; -aytr] ; ; meT | 50 coreg [AON [erleyeyy q[nsey jo uoyaing (ION [else ey sainjyng Saunqngy “‘SDId-VONIND ‘SYId-VENIND “Lteooy | =“ Tea | Or0e ‘LON| * 09H | alee ‘ZL were eynoy | shepgtd | 6008 “LAJIDLDYD Ut UDIAD asan “Lieoog] “ o9 a | greg spunj6 orwaquasau pun 6 ‘yoo90 = ‘Logndposaud : VL (eoory | sep 09 TIS “q[nsey Jo aoa nd “IeqMnyy mous saanyng “qoyed ‘MapoDLDYyo up uDIAD ; | sNOssevd [[BUIS & 10 se[o ata, ae ee ‘oy . = -Iaqny snod1evd[Bd-08sBd pud_ punjb = sojnanos “qusey : “TEqQuIN yy SdId qe Moz z poureytucs qove 7 - ed 7 eee Jo woreing SpUL[S o1[oo pue 1104 : : = “MOISO[ [BOO] FO WOIS[NU -uesom Auepl “snosrvo pue ‘ L061 ‘F taqurasacy ‘Lp , sep oF pod : ‘Bar 10.0 | | osstT | ‘Lp | sep 22 pea ‘ ‘Sut 10.0 GLGT ‘Lp | skep 1¢ pod | : “But 1.0 | GLC ‘1,9 | skep 0g pata ‘ ‘Bur 1.0 9LST ‘L°D | stp 96 peta snomaavaquy ‘sa 1.0 [J U SAGT ‘L'p | sdep 1 pera snousava} Uy ‘Sato {J GAcT “pro skep 0% ‘Wndsg UlAs04TH UO 919M ZZ YOIYA JO UOTeIOUENH ACE ‘oany[uQ wIMNIIg uyaeo AT ‘pto skep 0g ‘woryeIeusy WIOF ‘AINA[ND wWnIeg ‘shBp FFZ'I—UOYVAT[NO [eIOYTAe JO UOWeInp [eI], “XI ‘d@ SNUIA ‘L‘) | sep og pera : “SUL 0.01 |) (| P9GT LD | shop oF pag : "Bur 0.01 |] ( OAGT ‘LD | sep 1g ped snosuemnoqng “Bur 0.01 | 69ST ‘L ‘9 | skep oot por | snoeueynoqug | -Sur 0.0T 89ST ‘Lp | sep zg paid : "SUI 10.0 TAGT L ‘9 | sfep og pad : ‘BUI 10.0 69ST ‘Lp | shep 1% pad : “301 10-0 | ¢ L061 ‘grequte.eq = { 9ST + LOGT ‘g toqutaseq 4 ‘L ‘9 | shep re para : "BUI 0.0 LOST ‘LO | sep ig por “Su 10-0 G9GT LD | skep gz por ‘ “Bu 1.0 99CT ‘LD | s&ep 32 pod : “SUL 1.0 | | TOT ‘L ‘9 | skep ez pera SnNOmsAvIqUL ‘su T.0 |J L gOGT ‘L ‘9 | skep og pot *‘SNOWOABIYUT ‘su 1.0 | J l 09ST k, “OFT ‘HOTFB[NOOUT - : - Es “OFTT “TOT}e[NOOUT 7 s A fe qrnsey jo wore Jo poureny asoq uolMepnoouy fo a3eq IOC UN NT q[nsoy yo uomerng Jo pomeyy asog WOTZe[NOOUT Jo 99eq TOC|UAN Ny ‘pjo skep 0g ‘twnseg urTII0ATH UO OIOM FZ YOIYA JO uOMeI9TEH pigp ‘erngynH wnseg upIe0ATH ‘po skep Og ‘WoNeIeUey YI9gE ‘9INA[ND wWNnAGG ‘shep LSP [—UOeA[NS TeOyNIe Jo UoMVINp [IO], P L8F I “AI “@ SNUIA ATHALLOGGSAU NOUS NIUMOATO CNV WNugs NO NMOUSN ITTIOVA WIOMMANL ANIAOM AO SHUNLTINO HLIM GHLVINOONI SLIGavu 435 “spunom : ordes 02 onp yyeeq | sep ZI parq | ME OL || (| 6ST LO | sfep $9 potty : “BU O.01 |) ‘L “9 | skep F9 porq snosaurqnoqng ‘SUL 0.01 | | G6ST ‘LS skep GE polq snosurqnogng ‘SU O OT | “qyeap Jo esnvp 7 | skep 2 perq : ‘BUI 10.0 6ST ‘ED | séep 0g porg "SUL 10.0 ; LOGT ‘OT oqmaseq 4 t LOGT ‘OT/Iequreoaq “qavep Jo esnep 7 | sep » porg F “Sur 10.0 2 T6ST ‘L‘9 | shep 4% porq ‘ “Su 10.0 ‘L ‘9 | shep og por “ ‘SUI T.0 PEST LD | sep zz por . “BU 1.0 'L ‘O | skep $g porq SNOUIABIIUT ‘su to |) L O6ST ‘'L ‘YD sep QT psig SNOUSARIFUT suo [J Sects ‘OFTT “dor4e[noouy ‘ ‘ ‘ ‘ ‘OTT “WOTyE[NOOUT ; : 4[Ns0 yy jo worerng jo pouseyy esoqd WOTFeNOOUT Jo ae 1eq wun Ny q[asey yo wore yo pomoyy esoq UWOTYe[NooUyT Jo ayeq ‘plo skep 0g ‘mnzeg UlIe0ATH Uo o10M Cg YOTYA Jo Tone1euey YIOF ‘“e1nj[NO wnasg ure04[H ‘plo skep 0% ‘uonvioueg W463 «“eANyIND wnIeg | L ‘ponurquoo—KTGFATLOWASAU WOYAST NIUHOATO CNV Was NOGNMOYD TITIOVA ATIOUMAOL ANIAOG JO SHYNALINO H1IM GHLVINOONI SLICGvy ‘skep 68T‘I—toreanno [elogigze Jo woWemMp ye,OJ, TX ‘€ SNUIA A8cT 98GT 68ST 88ST G8GT PBST ‘aquin yy 437 INVESTIGATION OF ARTIFICIALLY MIXED CULTURES. BY F. GRIFFITH, M.B. TABLE OF CONTENTS. PAGE Experiments with artificially mixed Cultures on Calves wn ees w. = 438 ig i 3 4 on Rabbits... ei «xo 488 Summary... seis ies a3 ee ve ae it io ww. 488 Chart of the Experiment on Calves with mixed Cultures of Virus H. 56. “ F.T.” and Virus B. XXVI. 439 ‘5 » on Rabbits with mixed Cultures of Virus H. 56. “ F.T.” and Virus B. XXVI. 440 Post-Mortem Notes of :— Calf 460 ... ies as see se a i ee is 441 Calf 462 ... po ses ‘on ee ae ses id ne 442 Calf 468 ... se as aoe ee ote oy te ee 442 Calf 470 ... aes ao es a oom aa ia wa 443 Calf 472 ... as ise tee eee fae on ise esis 444 438 INVESTIGATION OF ARTIFICIALLY MIXED CULTURES. On CALVES. In view of the possibility of meeting with mixed infections in the investigation of tuberculosis in man it was considered desirable to determine what takes place in the body of a calf after the inoculation of a mixture of cultures possessing different degrees of virulence. Two cultures were chosen whose properties and characters had been fully worked out. Of these, the one (H. 56 F.T.) derived from a case of phthisis in man was easy growing and slightly virulent for calves and rabbits; the other (B. XX VI.), isolated from an ox, grew poorly and was fully virulent. With a mixture of these cultures two calves were inoculated subcutaneously, each receiving ‘02 mg. of the virulent virus and 50 mg. of the slightly virulent virus. This mixed culture grew readily on artificial media, exhibiting the cultural characters of the slightly virulent element, while the results of the inoculations showed it possessed the virulence of the bovine element. In fact, the presence of the slightly virulent bacillus in no way diminished the effective- ness of the bovine tubercle bacillus, since one of the two calves (Calf 460) died of general tuberculosis in 113 days, an exceptionally severe result from the subcutaneous inoculation of so small a dose of culture as ‘02 mg. The second calf (Calf 462) was killed after a period of 59 days, and showed in addition to the local lesion and tuberculosis of the nearest glands only slight dis- seminated tuberculosis. From the prescapular gland of this animal an emulsion was made and inoculated subcutaneously into another calf (Calf 468, the second in the series) which died of general tuberculosis in 79 days. Two more calves (Calves 472 and 470, the third in the series) were inoculated, one from the spleen, the other from the prescapular gland of the preceding calf. They were killed 152 and 150 days later and showed chronic general tuberculosis. From the lesions in all these calves cultures were isolated both directly and through guinea-pigs and rabbits, the main purpose of the experiment being to ascertain whether these cultures resembled the mixed culture or exhibited the characters of either one or other of the component parts of the mixture. The characters of the cultures were determined in the usual way from the comparison of the growths obtained on the various differential media :—Serum, serum with the addition of glycerin, glycerin broth, glycerin agar and glycerinated potato. The result of the cultural investigation was to establish that every culture isolated, even those from the prescapular glands, exhibited the distinctive characters of the bovine element of the mixture and in none was there any evidence of the presence of the easy growing human tubercle bacillus. On Rapseits. At the same time as the calves, a rabbit (No. 925) was inoculated subcutaneously with an equivalent dose of the mixture. It was killed after 58 days and showed general progressive tuberculosis. From the tuberculous lesions in the lung, spleen and kidney cultures were isolated; the culture from the lung grew luxuriantly, resembling the slightly virulent bacillus, and when tested in rabbits was found to be virulent, that is to say, it possessed the characters and virulence of the original mixture ; the cultures from the spleen and kidney grew poorly and were identical with the bovine element of the mixture. The mixed culture grown from the lung was, as stated above, tested by inoculation into rabbits and from the lung of the rabbit subcutaneously inoculated (No. 1162), a culture was isolated which was identical with the bovine tubercle bacillus. As will be seen from reference to the chart, two series of passage experiments in rabbits with emul- sions of tuberculous tissue were made, beginning with the lung of Rabbit 925, from which the mixed culture was obtained. All the rabbits in these series deve- loped general tuberculosis, and the cultures isolated from different sites were typically bovine in character. A rabbit (No. 926) inoculated subcutaneously with the original mixture, the dose being :02 mg. of Virus B. XXVI. and 20 mg. of Virus H. 56 E.T., died of an intercurrent disorder within 24 hours. Cultures were sown direct from the spleen and several discrete colonies developed. Two of these were sub- cultured separately and tested on the differential media ; one grew poorly like the bovine bacillus, the other luxuriantly. A guinea-pig inoculated with an emulsion of the spleen of the rabbit became tuberculous and the culture isolated grew luxuriantly. This experiment shows that both the virulent and the slightly virulent bacillus were disseminated within 24 hours. SUMMARY. The result of the experiment on the calf shows that if a calf is inoculated subcutaneously with a mixture of a slightly virulent culture and a virulent culture, it is possible to recover from the lesions produced cultures in which the virulent element can alone be demonstrated, even though in the original mixture the slightly virulent bacilli were in preponderating numbers. This elimina- tion is demonstrable when, as in this case, there is a wide difference between the cultural characters of the two cultures. A similar result can be obtained though less quickly by means of the subcutaneous inoculation of rabbits. In considering this result it is important to point out that in the test inoculations of calves with Virus H_ 56, the subcutaneous inoculation of 50 mg. of culture gave rise to only a few discrete nodules in the prescapular glands and no disseminated lesions. It is well known that the slightly virulent bacillus (GroupII.) is disseminated over the body of the calf after subcutaneous inoculation and can retain its vitality in the tissues for more than 59 days. Therefore the fact that the presence of slightly virulent bacilli could not be demonstrated in any of the cultures isolated even in those from the prescapular glands cannot be taken as proof that the slightly virulent bacilli were completely eliminated. Their apparent disappearance may be explained on the assumption that the cultures were isolated from tuber- culous tissue which owed its origin entirely to the action of the virulent bacilli and in which the latter would be present if not alone at any rate in superior numbers. On the other hand since tubercles are often found in the lung of a rabbit after the subcutaneous inoculation of a large dose of slightly virulent tubercle bacilli, it is easy to understand the isolation of a mixed culture from that organ, as occurred in the rabbit experiment. The details of the experiments are represented graphically in the appended tables, in which all refer- ences to cultures are printed in italics. The terms dysgonic and eugonic have been used to denote briefly the characters of the virulent and the slightly virulent culture respectively. F. GRIFFITH. ‘8 B XXVI. CALF 4 Subcutan Dese : 50 mgs. +°02 mg. Viru Died:: October (113 da P.M.—General Material used. Prescapular q gland. | Lee Cultures. : Material ‘ Duration of asad. Animal. Life. Result. = Source. Character. : ‘ Axillary gland| Dysgonie. Rabbit 1033 | Died 52 days G.T. | Kidney ... | Dysgonie. Se Po 1034|Died 61 ,, | GT. || No culture ... ves : . Inguinal gland _ || G.-Pig 2158 | Killea77_,, c.f oan Ged aan Caves Rabbit 1035 | Died 80 ,, | G.T. || Kidney ... | Dysgonic. gland. )| @ pig 2160| Died 82, '| G.T. || No culture ... we Woediastinal | Rabbit 1036 | Died 87 __,, G.T. || No culture ... — gland. : Local lesion | Dysgonic. » 1037] Died 81, ar. { fae ia Spleen ... | G.-Pig 2159 | Killed 90 ,, | Healthy —_ — ' Local lesion. Do K Cultures. F Duration of — PJ Animal. Life. Result. Source. Character. G.-Pig 2187 | Killed 18 days...} S. T. Omentum | Dysgonic. a Fe 2188 » 64, ..) 8 T. Omentum Dysgonic. Material | used. Geviea {) 2 gland. renin $4684 3 P ND VIRUS B. XXVI. CuLTuRE From LuNG or Raspit 925. Number of Method of | Duration of | 7 Rabbit. Dose. Inoculation. Life. Result. 1162 1:0 mg.| Subcut. | Died 53 days} G.T. 1163 1:0 mg. Tntrap. Died 19 ,, | G.T. Cultures from Rabbit 1162. : Source. Character. Kidney ... | Dysgonic. Lung... Dysgonic. 032: ee Direct cultures fi irect cultures from "th, 1906. Rabbit 1032. ) berculosis. | Source. Character. Lung ssevee Dysgonic. Agillary Dysgonic. gland. — 1042. )43. Subcutaneous. vus. . Killed : October 25th, 1906. 18th, 1906. Bea ah le (17 days.) ) P.M.—Flattened translucent losis. tumour, partly caseous ; opaque ee Source. Character. foci in ‘one axillary lanl, A few translucent foci in lungs. Lung sss Dysgonic. Kidney ... | Dysgonic. Direct cultures from Rabbit 1042. | Source. Character. Spleen ...... | Dysgonic. 50: 2 ig: Peeled Kidney Dysgonic. irect cultures from Oth, 1907, Rabbit 1150. ) yerculosis. Source. Character. Lung weve Dysgonie. Spleen ....++ Dysgonic. Kidney ... | Dysgonic. 44] CALF 460. Subcutaneous inoculation of mixed cultures derived from the original materials of Virus B. XXVI. and Virus H. 56 F.T Dose—50 milligrammes of Virus H. 56 and -02 milligramme of Virus B. XXVI. Date of Inoculation—June 26, 1906, [Age 8°5 weeks. ] Died—October 17, 1906. [113 days after inoculation. ] Weights. kilos. June 26, 1906 dis 59°54 October 17, 1906 ... 52:27 Total loss in weight—7°27 kilos. POST-MORTEM EXAMINATION, _ Local Lesion.—In the subcutaneous tissues on the left side of the neck there was a pear-shaped prominent swelling 12-7 by 7 by 5:1 cm., weighing 227 grammes with skin and muscle to which it was adherent, On section the tumour was almost wholly cystic, contain- ing curdy caseous masses in aclear serous fiuid : the walls were formed by fibro-caseous gritty tissue lined by a softened caseous substance internally and varying from ‘8 to 3 cm. in thickness. Left Prescapular Gland.—The left prescapular gland measured 10:2 by 5 by 1 by 3 cm. The gland was firm, and on section the cortex, except for a small part at one extremity, was replaced by translucent tissue with a close caseous network studded with calcareous foci. Right Prescapular Gland.—The right prescapular gland was congested; the cortex was firm without any sign of caseation. Prepectoral Glands.—The rounded left prepectoral gland was almost entirely replaced by necrotic sub- stance resembling the left prescapular gland. The left reniform prepectoral gland contained a caseo-calcareous nodule replacing three-quarters of the gland. The right prepectoral glands were dark and a little firm, Cervical Glands.—The left inferior cervical gland was large, firm, and almost entirely caseous with numerous calcareous foci. A middle and a left upper cervical gland contained discrete fibro-caseous nodules. The right inferior cervical gland resembled the left prepectoral glands. Other cervical glands on this side appeared normal. The Axillary Glands appeared normal, Thorax. In the pleural cavities there was some clear yellowish fluid. Pleura.—The lymphatic fringes of the pleura con- tained flattened fibroid tubercles up to 1:5 mm, in diameter ; they were more numerous and smaller on the diaphragm. Lungs.—The lungs did not collapse normally. The left lung weighed 1:7 kilo. and the right lung 2-4 kilos. They just floated in water. The anterior and middle lobes and the anterior portions of the caudal lobes were firm and quite solid ; they contained numerous closely placed caseous nodules with cal- careous centres and narrow grey margins measuring up to nearly 2m. in diameter. The lung substance in between was dark and airless, containing numerous grey foci and smaller caseous tubercles. In the posterior portions of the caudal lobes the nodules were more discrete and showed a great uniformity in size ; the lung substance in between was oedematous, but air-containing, and in it could be seen numerous grey foci. Thoracic Lymphatic Glands. — The mediastinal glands were large and firm; the long gland measured 12:7 by 5:1 by 38 cm. On section the gland 34684 substance was toa great extent replaced by translu- cant tissue with a close yellowish caseous network with more opaque yellow calcareous streaks. The other glands were similarly and even more extensively affected. Two Ventral Mediastinal Glands were enlarged, con- gested, and contained numerous minute foci. _ Bronchial Glands——The bronchial glands on both sides were enlarged and very firm; on section the gland substance was entirely replaced, closely re- sembling the anterior mediastinal glands. The Pericardium and the ligament attaching it to the diaphragm were congested and showed moderately numerous minute grey tubercles ; on the inner surface of the pericardium there were patches containing numerous translucent foci surrounded by an area of congestion ; other patches were composed of groups of prominent tubercles. On the aorta there were two loosely attached fibro- caseous growths. Endocardium.—Normal. Abdomen. The Omentum and Peritoneum were normal. Intestines—The small intestine and the large in- testine were slightly and irregularly congested. Mesenteric Glands.—In an anterior mesenteric gland there was a pea-sized translucent nodule with a caseo- calcareous network. Spleen.—The spleen weighed 212 grammes and con- tained two tubercles, which could be seen beneath the surface, the size of a millet seed and a hemp seed respectively ; they were yellowish and calcareous in the centre. Liver.—On the surface of the liver and throughout the substance were sparsely scattered minute grey foci without any visible central caseation. Gall Bladder.—Normal. Portal Glands.—The portal glands were five in number and also showed discrete nodules up to 1°3 cm. in diameter ; they consisted of a margin of grey tissue, becoming more opaque towards the centre, where there were yellowish calcareous streaks. Kidneys.—On the surface of the left kidney there were three minute grey tubercles, and on section there were visible in the cortex sparsely scattered grey tubercles up to°5 mm. On the surface of the right kidney and scattered throughout the substance were more numerous similar but slightly larger tubercles, some with minute opacities in the centre. Suprarenal Bodies.—In the cortex of each suprarenal body there were three or four minute grey tubercles. Renal Glands.—In each renal gland there were in the cortex firm translucent nodules with more opaque yellowish grey network in the centre. Coeliac Glands.—Of the coeliac glands, four in number, three were small and normal. The fourth was enlarged and contained five separate nodules, re- placing the greater part of the gland substance. Lumbar Glands.—In one lumbar gland there was a minute yellow gritty focus. A second gland contained in the cortex two early caseous nodules with yellow foci. On the left side of Pharyna near to the tonsil was a small ulcer with thin undermined edges covered with a yellowish scab. 3Q 442 The Tonsils were normal. 7 The Epiglottis and Vocal Chords were congested. The Iliac, Precrural, Pudic, Popliteal, Gluieal, and Ischiatic Glands were normal, Parotid, Submazillary, and Posterior Pharyngeal Glands were congested and the cortices were somewhat firm. Microscopical Examination. (Smear preparations.) Left Suprarenal tubercle bacilli. Pharynx (ulcer).—No tubercle bacilli seen. Lung, Mediastinal Gland, Prescapular Gland. — Numerous tubercle bacilli in each. Body. — Moderately numerous CALF 462. Subcutaneous inoculation of mixed cultures derived from the original materials of Virus B. XXVI. and Virus H, 56 F.T. Dose—50 milligrammes of Virus H. 56 and -02 milligramme of Virus B. XXVI. Date of Inoculation—June 26, 1906. Killed—August 24, 1906. Weights. kilos. June 26, 1906 91°81 August 24, 1906 110°91 Total gain in weight—19-10 kilos. POST-MORTEM EXAMINATION. Local Lesion.—On the left side of the neck there was a swelling the size of a goose’s egg which on sec- tion was found to be a cyst with thin fibrous walls, lined with reddish granulation tissue, and filled with caseo-pus. Left Prescapular Gland. — The left prescapular gland measured 5°8 by 3°8 by 1:2 cm., and on section showed in the cortex a few scattered yellow foci. Right Prescapular Gland.—Normal. Prepectoral Glands.—Normal. Cervical Glands.—A mid-cervical gland on the left side, the size of a French bean, was firm and contained numerous small yellow tubercles (up to a pinhead) forming a small group at one extremity. Other cervical glands were normal. Axillary Glands.—Normal. Thorax. Pleura.—Normal. Lungs.—The lungs were crepitant throughout and contained sparsely scattered and evenly distributed irregular reddish nodules ranging in size up to about a hemp seed ; on section all these showed in the centre yellow foci, in some quite large. The Bronchial and Mediastinal Glands were not [Age 8°5 weeks.] [59 days after inoculation:] enlarged and showed in the cortices scattered irregular yellow foci up to 1 mm.,a few of which were per- ceptibly gritty. The Pericardium and Heart were normal. Abdomen. Peritoneum and Omentum.—Normal. Intestines —Normal. Gastric, Mesenteric, and Colic Glands.—Normal. Spleen.—The spleen showed a doubtful point. Liver.—Normal. Portal Glands—Normal. Kidneys.—Normal. Suprarenal Bodies.—Normal. Renal, Lumbar, and Iliac Glands.—Normal. Coeliae Glands.—One coeliac gland showed a group of irregular yellow foci scattered over an area about 1 cm. in diameter. The Tongue, Lonstls, Larynx, Pharynz, and Trachea were normal. The Precrural, Pudic, Gluteal, Popliteal, Submazil- lary; Retropharyngeal, and Parotid Glands were normal. Microscopical Evaminations. (Smear preparations.) Cervical Gland.— A few tubercle bacilli. Jediastinal Gland.—No tubercle bacilli. Lung.—A few tubercle bacilli. CALF 468. Subcutaneous inoculation with an emulsion made from the Prescapular Gland of Calf 462. Dose—9'0 cubic centimetres. Date of Inoculation—August 24, 1906. Died—November 11, 1906. Weights. kilos. August 24,1906... 57:27 November 11, 1906... 53°63 Total loss in weight—3'64 kilos, POST-MORTEM EXAMINATION. Local Lesion.—In the subcutaneous tissue on the [Age 15 weeks. ] [79 days after inoculation. ] left side of neck there was an oval somewhat promi- nent tumour weighing 12 ozs. and measuring 12°8 by 9-5 by 3:8 cm. It was adherent to and infiltrated both skin and muscle, and a caseous cord passed to the left prescapular gland. On section it consisted throughout of eee caseated tissue wich scattered calcareous streaks, Left Prescapulur Gland.—The left prescapular gland weighed 7 ozs. and measured 10 by 5 by 6°3 cm. ; 443 it consisted throughout, except for a small part of the pelvis, of dense caseous tissue with calcareous streaks and foci. The Right Preseapular Gland measured 6°3 by 25cm. It was congested and firm at one extremity where there was an early caseous network; in the centre there were two pea-sized caseo-calcareous nodules. Prepectoral Glands.—The left reniform prepectoral gland was firm at either extremity, where there were discrete caseous tubercles. The right prepectoral glands were normal. The Tissues in the neighbourhood of glands and tumour were oedematous. The Awillary Glands were normal. Cervical Glands.—The left inferior, middle and Superior cervical glands were enlarged and caseo- calcareous throughout. In addition, there were numerous small glands on the left side varying from a pea up toa small marble in size, containing caseous tubercles and nodules. _In one right superior cervical gland there was a single caseous nodule. Thorax. There was some excess of fluid in the pleural cavities. On the pleural surface of the Diaphragm there were scattered loosely attached fibro-caseous nodules up to 3 and 4 mm., situated in congested lymphatic fringes. The outer surface of the Pericardium was similar ; the internal surface was normal. The Tissues of the Posterior Mediastinum were oedematous. Lungs.—The lungs were dark and congested, and the left was quite airless, sinking in water. On section the whole lobe consisted of irregular caseo-calcareous nodules closely placed and separated by pneumonic lung tissue. The right lung was similar but just floated on account of some air in the emphysematous margins. Thoracic Glands.—The bronchial glands were en- jarged and consisted practically throughout of firm waxy caseous tissue with opaque yellowish calcareous streaks and foci. Mediastinal Glands—The long mediastinal gland measured 11°5 by 5:2 by 38cm. It was firm and on section contained irregular caseo-calcareous masses replacing the greater part of the gland; other media- stinal glands were similar. Heart—Normal. Abdomen. On the Peritoneum there were several flat trans- lucent tubercles larger than a wheat grain. On the Omentum there were sparsely scattered small grey tubercles, the largest the size of a millet seed. Small Intestine—In each of two Peyer’s patches there was a single caseous nodule the size of a wheat grain. Lhe Large Intestine was normal. Gastric Glands.—The majority of the gastric glands contained caseous nodules. Mesenteric Glands.—The majority of the mesenteric glands contained from 1 to 5 or 6 waxy caseo-cal- careous nodules varying from a pea up to 1 cm. in diameter. Colic Glands.—Several of the colic glands were normal; two contained caseo-caleareous nodules re- placing half the substance. The Rectal Glands were similar. Spleen.—The spleen weighed 7 ozs. On section it contained in the pulp discrete caseo-caleareous nodules up to 5 mm. in diameter. Liver.—On the surface of the liver there were visible sparsely scattered tubercles varying from ‘5 up to 3 mm. in diameter, most numerous on the convex surface ; some projected slightly and had become flattened. On section of the liver they were more sparsely distributed than on the surface, but in the depth there was a caseous nodule with calcareous foci, 8 mm. in diameter. Portal Glands—The portal glands were greatly enlarged, one measuring 7-7 by 3:8 cm. They were firm and on section were closely filled with nodules formed of a caseous network with calcareous streaks. embedded in translucent tissue. Gall Bladder was normal. Kidneys.—The left kidney showed four nodules in the cortex, the largest measuring 6 mm. in diameter, composed of translucent tissue caseating in the centre. In the right kidney there were sparsely scattered nodules some slightly prominent on the surface and passing radially through the cortex. Suprarenal Bodies.— The left suprarenal body showed one, the right several opaque grey tubercles the size of rape seeds. The Renal Lymphatic Glands were large and almosé entirely composed of caseated tissue with calcareous foci. The Coeliac, Lumbar and Iliac Lymphatic Glands were to a great extent replaced by caseo-calcareous nodules. The Tongue, Tonsils, Larynx, and Pharynx were normal. The Right Precrural Gland contained a caseo-cal- careous nodule in the centre, the size of a° wheat grain. The Left Precrural Gland was normal. The Left Submaaillary and a Haemo-lymph Gland were caseous throughout. The Right Parotid, Left Posterior Pharyngeal, Left Popliteal, and both Gluteal Glands bad parts of their cortices replaced by caseo-calcareous nodules. The Left Parotid, Right Posterior Pharyngeal, Right Popliteal, and Ischiatic Glands were normal. CALF 470. Subcutaneous inoculation of an emulsion from the Prescapular Gland of Calf 468. Dose—9:0 cubic centimetres. Date of Inoculation—November 11, 1906. Killed—April 10, 1907. Tuberculin Test —February 19, 1907. 100 days after experiment began. Rise of temperature, 1:2°C. Weights. Kilos. November 11,1906 .., Pe ady 101°82 “April 10,1907... 0. 0a 129°54 -- Lotal gain in weight—2772. 34684 [Age 26 weeks. ] [150 days after inoculation.] POST-MORTEM EXAMINATION, Local Lesion.—The local tumour on the right side of the neck measured 12°7 by 10°2 by 55cm. In the skin at the bottom of a funnel-shaped depression there was a small ulcer, from which the skin around was covered with a dried discharge. On section the tumour was adherent to and slightly infiltrated the skin and muscle; it ccnsisted of caseo-necrotic sub- 3Q 2 44 stance which was softening around the periphery, leaving shaggy walls of pinkish granulation tissue. Right Prescapular Gland.—The right prescapular gland measured 9 by 5:1 by 5:1cm. On section about two-thirds of the cortex was replaced by pinkish caseo-necrotic substance ; the rest contained discrete caseous nodules. There was great oedema of the .subcutaneous tissues on the left side of the neck, about the site of a recent inoculation of tuberculin. The Left Prescapular Gland was enlarged and firm, and the greater part of the cortex was filled with a fine caseous network. Prepectoral Glands.—The right round prepectoral gland contained a moderate number of discrete caseo- calcareous tubercles with grey margins. The Right Reniform Prepectoral Gland contained a few smaller tubercles. The Left Prepectoral Glands were normal. The Right Azillary Gland showed two caseo- calcareous nodules. The left axillary gland was normal. Cervical Glands.—The right inferior cervical and a middle cervical gland were almost replaced by caseo- calcareous tissue. The right superior cervical gland showed at one extremity discrete fibro-caseous nodules. The left cervical glands and the gland within the thorax were normal, Thoran. ‘Pleura. — The pleural lymphatic fringes were congested and on the parietal pleura there were two flattened fibro-caseous nodules, 6 mm. in diameter. Lungs.—The lungs showed on the surface and throughout the substance numerous discrete nodules varying from a small pea up to a mass 3 cm. in diameter ; they had grey translucent margins with yellow caseous centres, which in the larger nodules were quite soft and caseo-purulent. Thoracie Lymphatic Glands—The long mediastinal gland measured 15:2 by 5:1, and on section the cortex was firm and closely filled with fibro-caseous gritty nodules with confluent grey margins. The other mediastinal glands contained similar nodules with marked calcification. The Bronchial Glands contained similar but more discrete nodules. Abdomen. Peritoneum.—On the peritoneum there were five loosely attached translucent tubercles. Omentum.—The omentum contained one fibro-caseous nodule. 4 = Diaphragm. — On the peritoneal surface of the diaphragm there was a firm fibrous nodule diffusely caseous in the centre, 2 cm. in diameter, and a second nodule the size of a pea. Intestines.—Normal. The Mesenteric Glands appeared normal. Spleen.—The spleen weighed 11 ozs. On section it showed sparsely scattered caseo-caleareous tubercles with grey margins, varying from a millet seed up toa pea in size. : Liver—In the liver just beneath the surface there were scattered nodules up to 1°3 cm. in diameter ; three projected slightly and had fibrous fringes attached to them. On section these nodules consisted of a thick translucent capsule enclosing a caseous centre which was occasionally completely softened. In addition there were numerous very irregular yellowish grey foci (not tuberculous). Portal Glands.—The portal glands were enlarged and showed moderately numerous fibro - caseous nodules. Gall Bladder.—Normal. The Kidneys appeared normal. Suprarenal Bodies Normal. The Renal Glands showed a few discrete caseo- calcareous tubercles up to a hemp seed in size. Iliac Glands each contained two fibro - caseous nodules gritty in the centre, up to a pea in size. Tongue, Tonsils, Larynx, Pharynz and Trachea were normal. The Right Preerural, Left Submasillary, Left Posterior Pharyngeal, Right Parotid, and Right Gluteal Glands each contained one or two caseous nodules varying up to 8 mm. in diameter, some calcareous, others softening in the centre. Coeliac Glands. — One contained discrete caseous nodules with fibroid margins and gritty centres, up to a pea in size. Two others were normal. The Left Popliteal Gland showed a part of its cortex firm translucent and congested around the margins, The Right Popliteal, Left Parotid, Right Posterior Pharyngeal, Left Precrural, Pudic and Ischiatic Glands were normal. Testicles.— Normal. Microscopical Examinations. _(Smear preparations.) Lung.—One tubercle bacillus seen. Portal Gland.—No tubercle bacilli seen. Bronchial Gland.—T wo tubercle bacilli seen. Liver.—No tubercle bacilli seen. CALF 472. Subcutaneous inoculation of an emulsion made from the Spleen of Calf 468. Dose—9'0 cubic centimetres. Date of inoculation—November 11, 1906. Tuberculin Test.—February 19, 1907. Dose, 4:0 cc. of tuberculin, 100 days after experiment began. Re- acted. Rise of temperature, 1°9° C. Weights. Kilos. November 11, 1906... Bes ns 69°09 April 12, 1907 as we oe 68°18 Loss on initial weight—0-91 kilo, [Age—18°5 weeks. ] Killed—April 12, 1907. [152 days after inoculation.] POST-MORTEM EXAMINATION. Local Lesion.—On the right side there was a pear- shaped swelling measuring 12°7 by 71 by 51cm. On section it was partly cystic, containing turbid fluid and a pinkish necrotic sequestrum partly adherent to the walls; the walls were roughened by projecting calcareous granules. There was slight infiltration of the muscle. Right Prescapular Gland.—The right prescapular 44 gland measured 9 by 5:1 by 51cm. On section it con- sisted throughout of dense pinkish caseo-necrotic substance with calcareous foci. Left Prescapular Gland.—The left prescapular gland ‘contained a single caseo-calcareous nodule. Prepectoral Glands.—The right reniform prepectoral gland had about half its substance filled with tubercles. The right round prepectoral gland was firm and resembled the preceding. The left prepectoral glands were normal. The Right Awillary Gland showed a few gritty tubercles. Cervical Glands.—-The right inferior cervical gland was enlarged and firm and was replaced by caseo- calcareous tissue. The right middle cervical gland contained collections of caseo-calcareous tukercles. The right superior cervical gland showed a large easeous nodule. The left inferior and middle cervical glands showed a few discrete submiliary tubercles. The left superior cervical gland contained one caseo- calcareous nodule. Thorax, Lungs.—The lungs did not collapse normally ; they _ were oedematous and contained numerous caseo- calcareous nodules surrounded, in a majority of cases, by areas of consolidation ; they were most closely placed in the posterior lobes, which felt firm to the touch. Many of the larger nodules contained in the centre mucopurulent substance with gritty foci. Thoracic Lymphatic Glands.—The long mediastinal gland was enlarged, and the greater part of the cortex was replaced by caseo-calcareous tissue consisting of aggregations of tubercles. The other mediastinal and bronchial glands were also extensively replaced. Abdomen. Intestines.—Small intestine :—Situated in the long Peyer’s patch were seventeen submucous nodules rang- ing in size from 3 mm. up to8 mm. On section these contained soft yellow caseous substance. There were only three others in the rest of the small intestine, of which two were in the mucous membrane. The large intestine appeared normal. The Mesenteric and Gastric Glands contained hard gritty nodules and tubercles. The Colic Glands appeared normal. v oO Spleen—The spleen weighed 5 ozs. There were several projecting nodules beneath the surface. On section the substance contained scattered caseo- calcareous nodules, up to a hemp seed in size. Liver.—On the surface of the liver there were several flattened translucent nodules with caseous foci ; there were also a few caseo-calcareous tubercles slightly embedded in the substance. On section no tubercles were seen in the deep parts of the substance. Portal Glands—The portal glands contained moderately numerous irregular caseo-calcareous tubercles. Kidneys.—In the cortices of the kidneys there were sparsely scattered calcareous tubercles the size of millet seeds. Both Renal and one Lumbar Gland were closely filled with caseo-calcareous tubercles ; other lumbar glands appeared normal. The Iliac Glands showed a few discrete submiliary calcareous tubercles. The Coeliac Glands showed collections of caseo-cal- careous tubercles, one gland being almost replaced. Diaphragm.—On the peritoneal surface around the tendon of the diaphragm, on the right side, there were moderately numerous somewhat flattened translucent nodules with caseous centres. The Tongue, Tonsils, Larynx, Pharynz, and Trachea were normal, Various Lymphatic Glands. The Left Precrural, Right Popliteal, Right Gluteal, and Left Submawillary Glands each showed one or two caseo-calcareous nodules. The Right Precrural, Left Popliteal, Left Gluteal, and Right Submazillary Glands were normal. The Posterior Pharyngeal Glands contained moderately numerous caseo-calcareous tubercles, up to a hemp seed in size. Microscopical Examinations. (Smear preparations.) Lung.—Moderately numerous tubercle bacilli, Mediastinal Gland—A moderate number of tubercle bacilli. Prepectoral Gland, Cervical Gland.—A few tubercle bacilli in each, 446 INVESTIGATION OF TUBERCLE BACILLI DERIVED FROM TUBERCULOSIS OCCURRING NATURALLY IN THE HORSE. INDEX TO THE ANIMALS. Species and Number of Animal. Page. Species and Number of Animal, Page. Calves :— CALVES—cont, :— Consideration of Results :-— 12-14 No. 626 ks iy dist 7 ie 117 Labular Summaries of Buperiments :— » 628 dies ei es va wi 113 (a) With Cultures from Original 17 Material :— » 630 ne es a at ad 130. (6) With Passage Cultures :-— 94-95 » 632 oe Lee pee set pis 111 No. 404 tae 53 gp OBBE. tee i ee oe, one HBR: B08! Gy MA, Make. See. i 58 » 6860 eee ee | BE » 408 ii ae ee ana a 79 », 638 ep i re a sas 120 » 410 aes sis ss wes ee 79 . 640 ine oe ie wate si 118. SURO), agin mG etn se ge 42 TGS die Jo Meee eee’, ae 133 Gt oot, ae oe ee, 69 oR ee SS a a A » 498 ia nis si te ‘a 82 » 646 5G oa ee ii Ae 112 32309 ee ke. ake Ge Se 81 yp OAR ae SE wp0l) ah 25. 4h ah de Bd 9 BBO eee |B 506 ee ais a pis ies 55 » 652 id ie as a ats 142° » 508 a ds cis sh 70 BII0™ see. GS. Fee ce 45 Pigs :— .» =528 or ah ae ae ag 42 Consideration of Results :—- 12-14 » 578 sn ste — es — 116 Labular Summary of Experiments :—- 19 rg BOW eel aie ome ce et Nocdeh, 25: - 2 ge oA 8 59 Sy SBBD) gin RB cies | 185 a NBO) chek: eG. AGE. di Ah 60: » 584 oe a5 aa a 2 134 5, 188 a us ay a hey 84 » 586 sale ity ahs Pe ee 133 » 190 we uid a te es 85 Basa coe, coe Aa fl oy 132 pr OO) acs Ce ae | ey 43 590 ss ee ai dat sin 110 » 194 aa sii aa A ae 71 » 594 ie ae ice is ig 114 4 B96? es a Re Ne. © eee 8 Ponies :— 598 ise ce ee si i 119 Consideration of Results :-— 12-14 » 600 tl ihe sc a4 ea 121 Tabular Summary of Experiments :-— 19 » 602 i si a et Sa 129 No. 6 2 ant i ea aes 145. » 604 i oe 238 = es 127 » 8 oo she sh es ase 145. 606 sae hi set aa 56 yo 12 aes vas sas as du 141 oe] », 608 ee re ses hs aati 56 » 16 ate eas wid an a 44 , 610... ca sas A oe 57 ee TSE. eh, Se kesh ad es 57 gp BER eh Bn cele; ty: shew th AB? OO weal ae das. Gas ie 58. Git kk a cae we tS mp 281 8g. SBS hh Bef: 88 Blor fe. ake. Se Ua es 88 ero. Gee. CRA, ba 89 GIS. cee owe ee GE cot] 4 pO ay. ae ee ee veel) TED » 620 as ae i Sad ne 87 » BU ae es ea site sis 143 447 INDEX TO THE ANIMALS—continued, Species and Number of Animal. | Page, Species and Number of Animal, Page. Rhesus Monkeys :— Be : Lemurs :— Consideration of Results :— 12-14 Consideration of Results :— 12 Labular Summaries of Baperiments :— \ Tabular Summary of Haperiments :-— 18 (a) With Cultures from Original Material :-— 18 No, 14 eae ais 2a ae ae 72 (0) With Passage Cultures:— 95 » 16 dee us se oe ae 65 No. 166 oe les i ae ai 61 » 18 i i ae sas sets 65 » 168 oe : ae st Aig 61 » 170 463 ales ree ss 85 4 17S Sy = ok ai sa 86 » 10 2. i me a a 62 Rabbits :— » 182 sip iis ise a Se 86 Consideration of Results :— 12-14 » 184 das ioe ‘ii 2a ie 62 Labular Summaries of Ha periments :— ee te 63 (a) With Cultures from Original | 20-38 Material :— » 202 ass whe es ais a 63 ‘ (1) With Passage Cultures -— 96-103 » 206 sf ~ 5 6 si 110 » 208 er ne se ah or 64 + 210 te vd = a si 91 +» 212 whe is ah sie she 110 gy BT Sh ee SS » 216 she ay eh a re 64 : : Guinea-pigs :— ene a oor a ie a0 Consideration of Results :— 12-14 saa a re ai - 3 nee Tabular Summaries of Experiments :— » 222 ae an ee a a re (a) With Cultures from Original 21, 26, 28, 4 Ds a se a = 144 aaterial :— 37, 38. (h) With Passage Cultures :— 99, 104 Bahboons :— Consideration of Results :— 13 Labular Summary of Euperi- ments :— 18 No. 72 80 ; » 7 a, 8% unt ooh, av 80 INVESTIGATION OF VIRUSES OBTAINED FROM CASUAL TUBERCULOSIS OF VARIOUS MAMMALS. INDEX TO THE ANIMALS. Species and Number of Animal, Page. Species and Number of Animal. Page. a fees : alves : Pigs = Summaries of Results :— a No. 386 aa wee es: se ‘ibe 153 Summaries of Results :— 151, 156 ‘i aia 154 moter. She Ses a No. 170 0 nate tees ee | 160 ne sab 159 » 510 wis ita sis oat ie _ . _ | ~ 516 sie ee oa Feo a 159 ” 448 INDEX TO THE ANIMALS—-continued. Species and Number of Animal. Page. Species and Number of Animal. Page. Rabbits :— Fowls :— Summaries of Results :— 151, 153, Summaries of Results :— 151, 159, 156, 162, 164, 165. 164, 165. | Guinea-pigs :— Summaries of Results :— 151, 153, 159, 162, : 164, 165. ' INVESTIGATION OF AVIAN TUBERCLE BACILLI OBTAINED FROM BIRDS AND SWINE. INDEX TO THE ANIMALS. Species and Number of Animal. ' | Page. Species and Number of Animal. Page. | Calves — | CALVES—cont, :— Consideration of Results :-— 186 No. 556 357 Labular Summaries of Experiments :— 242-245, » 558 336 252, 256- 257, 4 B60 335 No. 282 361 » 562 334 y 318 363 » 564 336. » 390 376 » ©6566 337 , Boe | 361 » 568 300- » 402 | 286 570 338. , 412 373 » 572 339 424 | B56 | » 574 338 » 482 ee 29) » 576 340 yy £56 ie 299 » 458 ae 302 | Pigs :— » 490 oa 380 Consideration of Results :— 186 » 492 .. | 378 Labular Summaries of Experiments :— 245-247, » 494 | 299 253-254, 257-258, », 520 bist 294 No. 104 347 4 524 a |. BT » 106 ia 348 » 3530 B65 » lo 349 » od at 313 » 12 362 » «=546 329 4 1386 349 » 548 ie 331 oy =<140 377 4 BBO | 382 » 176 306 BOE Y 333 » 178 306 t 449 INDEX TO THE ANIMALS—continued Species and Number of Animal. Page. Species and Number of Animal. Page. Si aca aas Rhesus Monkeys :— Consideration of Results :— 188 No. 180 0 sae wie se - 307 Tabular Summaries of Experiments :— 250-251 9 =-182 ies sis a ste 308 sl li TO) ce ee ee ee ee a mea cher » 198 = eS Z aay » 120 ude wie iss ave ese 353 go oe oe ee, Ses Coe a a a) « Be wm “ . . : Bi » «124 aes see Sas vee wae 354 Ne ge Oe ee 341 Cae ao a RE 246 2 ir 7 A any » 136 oie ive aie Se es 372 ne ak. ee. 2 cow aie) SB i al ie » 260 sor 5, 154 Bes ie ae8 wwe aT 374 262 508 » 156 sas Bre xs see wi 375 264 sis » 158 wwe vee san oie 286. » 160... ae age eta ns 287 » 162 wes ae eee aoe a 288. » 164 ae oe as as see 288. Goats :— yp ABB a eae | BRE Consideration of Results :— 189 » 190 see ee aa as 326 Tabular Summaries of Experiments :— rae », 192 ee ees 368 2s5 ais 342° ’ ‘i os » 194 sia or ae wae see 340 Ice Gp, Sess ck 8 as 5 RG es we. a ae BE oe me SAGE ees ee. RE GS 327 a ee 295 Chimpanzee :— a ee ee ee am Consideration of Result :— 191 _ : ov oe a a Tabular Summary of Experiment :— 250: » 50 wie 342 No, 20 ee aes Sse ae 293. Bk ae 309 '| Dogs :— n 58 se os i ae Consideration of Results :-— 191 ee = Jabular Summary of Eaperiments :-— 282: RR on ag ees z : » 64 a ae sis 331 No. 122... 88 fe ass 366 » 124 owe ose a see 7 372 » 152 ae ete sss ose exe 314 Horses (Ponies) :-— 5 EE Gh ee me ee] BUS Consideration of Results :— 187 yp 162 renee nee tte nee 325 Jabular Summaries of Experiments :— 252, 255 » 164 ace sue Seis as id 325 No 2 « , i as 314 Cats :— » 4 r ove 3 315 Consideration of Results :— 188 » 10 er ied be ot Ag 309 Tabular Summary of Experiments :— 259-260 ee “EPS | an Nob? he G6. “sie ke 4 350 i a ; a » 64 ue. ass ; - oe | 351 3R 34684 450 INDEX TO THE ANIMALS—continued, Species and Number of Animal. Page. Species and Number of Animal, Page. ‘CATS—oont. :— Pigeons :— Consideration of Results :— 181 No. 92 297 Labular Summaries of Experi- » «94 297 ments :— 215-217 » 96 309 Rabbits :— Parrots :— ey ara sd Consideration of Results :— 182 sues Semmens of Baperi« 990-237 Tabular Summaries of Euperi- : ments -— 218-219 Guinea-pigs :— Rats:— Consideration of Results :— 189 Consideration of Results :-— “190 : & Tabular Summaries of teehee emia of Experi weecone Realiecm 277-281 Mice :— Fowls :— Consideration of Resuits :— 185 i i — 179 Consideration of Results Labular en ae Tabular Summaries of Experi- Results :-— 237-242 ments :— 194-215 MODIFICATION EXPERIMENTS WITH TUBERCLE BACILLI DERIVED FROM ANIMALS OTHER THAN MAN. INDEX TO THE ANIMALS. Species and Number of Animal. Page. Species and Number of Animal, Page. Calves :— ’ CALVES—cont, :— Charts of Passage HEaperi- ments :— ; No. 548 331 With Virus A,X... 392-395 350 332 With Virus P. IV. 432 y BBE 333 No. 282 361 , 558 336 » B12 7 » 560 335 » 314 Included in 5» 562 334° Volume III. 350 ‘ of the 5 564 336 Appendix, » 356 which see. » 566 337 + 358 J » 570 ‘ 338 ys 390 402 and 376 872 339 » 426 Included in Vol. III, » «574 402 and 338 of the Appendix, 4 456 402 and 299 » 576 340 45] INDEX TO THE ANIMALS—continued. Species and Number of Animal, Page, Pigs :— No. 234 os 341 » 236 iv ote isa 34) ‘Goats :— No. 36 402 and 355 » 50 see ets 342 Rhesus Monkeys :— No. 118 ans wae os 402 and 351 » 192 aes oes ene 342 yy 194 fs 340 Chimpanzees :— Chart of Experiments :— 424 No. 24 425 Baboons :— Chart of Experiments :— 424 No. 70 427 » 76 426 Dog's :— Charts of the Passage Eaperi- ments :— With Virus B. VIII. 416 With Virus B. IX, 417 With Virus B. IV. 418 With Virus B. XXIT. 419 With Virus B. 1V. (throug Chimpanzee 8) ... $e 420-421 Tabular Summaries of the eae Experiments :— Species and Number of Animal. Page. Rabbits :— Charts of Passage Experi- ments :— With Virus Z, ITT. With Virus P. IV. Labular Summaries of Haperi- ments with Cultures of Bovine T.B. grown on Serum and Glycerin Serum re- spectively :— No. 777 Guinea-pigss :— Charts of Passage Experi- ments :— With Virus A. Vo. With Virus P. XXI, Tabular Summary of Eaperi- ments with Collodion Cap- sles :— No. 1612 » 1857 Fowls :— Charts of Passage Experiments with Virus B, XXVI,:— List of Fowls, inoculated with Mammalian 1.B., from which Cultures were recovered Tabular Summary of Experi- ments with Collodion Cap- sules :— Pigeons :— Tabular Summary of Experi- ments with Collodion Cap- sles s— 428-430 431 434-435 402 and 232 396 397 406-408 402 and 359 402 and 343 409-410 411-414 404-406 404-406 INVESTIGATION OF ARTIFICIALLY MIXED CULTURES, INDEX TO THE ANIMALS. Species and Number of Animal. Page. Species and Number of Animal. Page. eee os a Rabbits :— Celta retion es HORNA N= Consideration of Results :— 438 No. 460 441 5, 462 a wee eos 442 » 468 442 » 470 4143 444 472 LONDON: PRINTED FOR HIS MAJESTY’S STATIONERY OFFICE,. By DARLING & SON, Lrp., 34-40, Bacon Street, E. 1911. Pek Ad, Brann 1909; Breus ae Paopnerion, ‘VAL, &e, 1907-1910. 1.0 i . 5 ds ‘Boann, 1910411. ° First Report, H.0,.292. 7d, oi f Me ie ; Anuy Muvinw. Vol EF No. 2. Oct, 1911, 1s, RiMLURY StoRM ACCOUNTS AND THE SUPERVISION LOB A R.G,A, S0B-bistRw?. Nores on, . Amend- = ments. 1d. Cinchiding See 8 > OtotHing: Ann Nucussartns ‘Priced: Vocabulary of. 1911. ; ** Do, Amendments, &;, Oct. 1911. Id, » > Croptine Ruautarions, Paris 1, 2;-and 3, “Amend- 42 mente. Sept. 1911. id Oxmwa oF © | Av Home Srarions, Regulations for the. Appoint- > ao meut, Pay, ind Promotion of: 1911, 2d. =) Dover Garrison. Standing Orders. Gd, >. -PQuiewess ReGunartions —. Part 2 Bec. xf. Signal Company (Air-line). (Regu- lar Army). 2d. 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In 100: vols. : 103. ‘each FINAL REPORT OF THE ROYAL COMM APPOINTED TO IN aoe INTO THE RELATIONS OF HUMAN AND ANIMAL TUBERCULO: PART II. APPENDIX. VOLUME IV. “naturally in Certain Animals other than Man: and Modification Experiments, oe. F. GRIFFITH, M.B. Preseinted to both Houses of Parliament by Command of His Majesty. . / LONDON: PUBLISHED BY HIS MAJESTY’S STATIONERY OFFIOE. To bé purchased, either directly or through any Bookseller, from WYMAN Anp SONS, Lrp., Ferrer Lang, E.C., and 32, ABINGDON STRUBT, 8.W. ; or . OLIVER anpD BOYD, TWEEDDALE. CouRT, EDINBURGH; or E. oe LD.; 116, GRAFTON STREET, DUBLIN. : PRINTED BY ithe DARLING AND “SON, ‘Lrp., BACON STRERT, E. 1911. [Cd. 5894.] Price 4s. 5d. a 4 \ ; me