NORMAL HISTOLOGY OF THE KIDNEYS, LIVER, HEART, SPLEEN, LUNG OF THE GUINEA PIG AND HISTO-PATHOLOGICAL CHANGES OF THESE ORGANS IN EXPERIMENTALLY INDUCED BOTULISM BY CHARLES ETHELBURT RANDOLPH THESIS FOR THE DEGREE OF BACHELOR OF SCIENCE IN AGRICULTURE COLLEGE OF AGRICULTURE UNIVERSITY OF ILLINOIS 1922 UNIVERSITY OF ILLINOIS .vhjm ..3 19S2.... THIS IS TO CERTIFY THAT THE THESIS PREPARED UNDER MY SUPERVISION BY CHARLES ETmBURT RANDOLPH ENTITLED.. 9?... 9?.. GUINEA PIG AND HISTO-PATHOLOGICAL CHANGES OF THESE ORGANS IN EXPERIMENTALLY INDUCED .BJPUUSM.^ IS APPROVED BY ME AS FULFILLING THIS PART OF THE REQUIREMENTS FOR THE DEGREE OF. Ba<^3lpr..of .Scie^ HEAD OF DEPARTMENT OF ... . A$Xml . . Husbandry Digitized by the Internet Archive in 2015 https://archive.org/details/normalhistologyoOOrand TABLE OF CONTENTS Int roduct i on — -1 Fixing of Tissues- • ~ — 2 Embedding of Tissues- — • 2 Cutting of Microscopic Sections • 3 Staining of Sections — - — — --3 Preparation of Delafield's Hematoxylin---------- — — — ~ — --—3 Preparation of Eosin Stain--- — 3 Preparation of Xylene-Paraffin Mixture - - --4 Preparation of Paraffin -Beeswax Mixture--- 4 Preparation of Japanese Glycerin-Albumin — — — — 4 Source of Materials----- — — ------4 & 5 Kidney Normal Histology—————— — — - — — — — — ^ — - — ---5 & 6 Histo-Pathological Changes-- — — ----6 Sc 7 Liver Normal Histology------ — — — — •-- — -7 & 8 Histo-Pathological Changes-—— — — >-8 & 9 Heart Normal Histology* — ------- — — - — 9 & 10 Histo-Pathological Changes — ------10 & 11 Spleen Normal Histology — — ------- — — — ---11 & 12 Histo-Pathological Changes---- — — — >12 Lung Normal Hist o logy — — — — — — « — - — 13 Histo-Pathological Changes------ — ————-13 & 14 Summary of Pathological Changes--—- — — — — =-14 Bibliography———— — — - — — « 15 Diagnosis Charts- — — — — — —16 to 20 inclusive NORMAL HISTOLOGY OF THE KIDNEYS, LIVER, HEART, SPLEEN, LUNG OF THE GUINEA PIG AND HISTO-PATHOLOGICAL CHANGES OF THESE ORGANS IN EXPERIMENTALLY INDUCED BOTULISM ************ The sporadic outbreaks of botulism in animals reported in recent years has suggested the desirability of accurate diagnostic methods. In a peracute attack of the atypical type of this disease the symptoms are not pathognomonic, and are probably frequently confused with other diseases. The presence of the spore in the intestinal tract, or in the internal oigans as an agonal in- vasion following death, is not irrefutable diagnostic evidence of botulism. The microscopic changes in the tissues of the guinea pig, after experimentally inducing botulism, have been studied with the view of determining, if possible, the value of histo-pathological changes in the kidneys, liver, heart, spleen and lungs. In the following paper a brief study of the histology of the normal guinea pig and a study of tissues obtained from cases of experimentally induced botulism are recorded. - 2 - METHOD OF FIXING, EMBEDDING, CUTTING AND STAINING OF TISSUES Fixing and Embedding Tissues; It is essential in the sectioning of body tissues to retain the normal relationship of the cells of the organ as present in the individual. Post mor- tem changes take place rapidly so that the tissues should be placed in a fixing fluid (10 Formalin) as soon as possible. After the fixation is complete the tissues are washed and later dehydrated by placing them in a series of solutions of increasing concentrations of alcohol. Prior to the process of embedding, the tissues are cleared in xylene, and after being completely infiltrated with hot paraffin, they are embedded in blocks of the same material. The following technic was employed in fixing, embedding, cutting and staining the preparations. The tissues for sectioning purposes were cut about two centimeters square and after being washed free of excess blood they were placed in a volume of the 1C $ formalin solution about fifty times the volume of the tissue. Fixing and Embedding Technic: 10 % Formalin in physiologic salt solution for 24 hrs. Wash for 24 hrs. 70 $ Alcohol 24 hrs (or longer). S5 $ Alcohol 24 hrs. Absolute Alcohol 24 hrs. Xylene 2 hrs. Xylene -Paraffin Mixture* **--2 hrs)_ = j n oven at 58°C. Paraffin-Beeswax Mixture ***-4 hrs) Embed in Paraffin-Beeswax Mixture The tissues are embedded in paper boats, which are of common white paper about eight centimeters long and three centimeters wide. Each boat is placed upon a cold surface and filled half full of the warm paraffin-beeswax mixture. Then with warm forceps the square of tissue is placed in the boat, the surface to be cut being embedded down. The filling of the boat is completed with the warm paraffin. This second pouring will drive out bubbles of air which adhere to the bottom. The boat is then floated in cold water to speed the solidification of the paraffin and drive out air. ^**See preparation on page 4. ■ - 3 - Cutting of Microscopic Sections: The paper boats are stripped from the hardened paraffin and the blocks trimmed, taking care that the edges of the cutting surface are parallel to in- sure ribboning of the sections. A horizontal sliding microtome was used in this project. Four microns was the usual thickness of the sections, but with certain blocks it was only possible to get good sections at five or six microns. The cut sections were floated on warm water and mounted on a slide by putting the slide underneath the floating section and raising it out of the water. The sections * * were held to the slide by a Japanese glycerine-albumin mixture previously rubbed on the slide and heated until a steam began to arise from the surface. The mounted sections were allowed to dry in an oven at 58°C for 12 hours. Staining of the Sections: Hematoxylin and eosin ware used for staining all tissues. The hematox- ylin stains the nuclei purple and the cytoplasm takes the eosin or acid stain. Preparation of Delafield's Hematoxylin. Hematoxylin crystals--- A grams Alcohol (absolute) — 25 cc. Saturated solution of ammonium alum-400 cc. The hematoxylin crystals are dissolved in the absolute alcohol. To this solution 400 cc of alum solution was added. The mixture was exposed to the light (sunlight if possible) in an unstoppered bottle for 3 to 4 days. After filtration the following were added: Glycerine----- — - — ---100 cc Alcohol (absolute) — — 100 cc This solution was allowed to stand in the light until the color was sufficiently dark, then filtered and kept in a tightly stoppered bottle. The solution keeps well and is satisfactory as long as it retains a purplish tinge. Preparation of Eosin Stain: Eosin-- — --1 gram Water- — — 100 cc Absolute alcohol 15 cc The eosin was dissolved in the distilled water, the alcohol added, and filtered. ♦♦Preparation on page 4. - 4 - The staining was accomplished by the following technic: Xylene — 3 min. Absolute Alcohol ------ -2 min. 95$ Alcohol-- - 2 min. 60$ Alcohol — ---■ — 2 min. Distilled HgO >• 2 min. Hematoxylin-- — 6to30 min. Tap water — -2 min. Distilled HgO------ 2 min. Eosin ——2 to 3 min. Tap water-— -2 min. Distilled HpO— — 2 min. 70$ Alcohol- — ------- 1 min. 95$ Alcohol ------ -----3 min. Absolute Alcohol-— --4 min. All excess staining matter on the slide around the tissue was removed with a cloth. Xylene — - — --—3 min. The surplus xylene was cleaned off and a drop of Canadian balsam added to the section over which a cover slip was then gently placed taking care to exclude all air bubbles. A light weight was placed on the cover slip to hold it in place until the balsam dried. The slides were sat aside to allow the balsam to dry and seal the cover slip. Preparation of Xylene-Paraff in Mixture: 1 part 56 J C paraffin 1 part 46° C paraffin 2 parts xylene Slowly melt the paraffin and add the xylene to the liquid paraffin. Preparation of Paraffin-Beeswax Mixture Paraffin 850 grams. Stearin 100 grams Beeswax 50 grams This mixture is kept in the oven at 56°C. Preparation of Japanese Glycerin-Albumin, White of Egg) ) — -Equal parts Glycerine ) Beat mixture and filter. Add a few drops of formalin as a preservative. SOURCE OF MATEEIALS The normal tissue specimens which were used for comparative purposes . - 5 - were obtained by destroying a healthy guinea pig and immediately removing the fresh organa. The guinea pigs furnishing the pathological tissue specimens were originally used in botulinua typing tests. A majority of the guinea pigs (13) died in a few hours ( 12-30 hrs) following an acute attack of botulism, while two pigs were destroyed in the agonal stages of the disease in order to avoid the development of post-mortem changes. KIDNEYS Normal Histology: The kidneys are glandular organs located on each side of the dorsomed- ian line. They are invested in a fibrous capsule which blends into the ureter at the hilum. The organ is divided into four regions, the cortex, medulla, in- t . ate zone, and pelvis. Extending into the sinus at the hilum are the re- r .1 papillae, the projections of the renal pyramids. These renal pyramids extend back thru the medulla and have their cu\ .,w..tBwKwi base towards the cortex. Between the pyramids are found -UftW ^ the columns of Bert ini which exter.d to the walls of the ;i 1 nus . On the examination of the cortex under low power magnification it is seen that it is divided into alternating light and dark areas, the medullary rays and * 4 *^ The structure of a renal secretory unit is composed of three general parts namely, the glome: ulus, the convoluted urinfer- ''‘•~-T«xin- v rU.vA t tubules and the collecting tubules or duct system. As U.oU . ' shown in the figure the uriniferous tubules begin in a great ly expanded blind extremity, the capsule. This capsule sur- u«^Wt rounds the tuft of an artery forming the Malpighian body. Th-. • >ule coming from the glomerulus takes a very tortuous and twisted course forming the proximal convoluted tubule. It then extends into the medullary rays and follows a straight course towards the papillae, as the descending limb of Henle. When well into the medullary region it turns upon itself, the loop of fm - 6 - Henle, and returns to the renal pyramid and the medullary rays as the ascending limb of Henle. It again enters the labyrinth and repeats its tortuous convoluted course, forming the distal convoluted tubule. Later it reenters the medullary rays, and joining one of the collecting ducts, it finds its way into the renal pyramid and out through the papillae into the sinus. The structure of the urinary tubules is very simple, consisting of a single layer of cells supported by a basement membrane. The glomerular portion eliminates the water and inorganic constituents, while the organic matter is eliminated thru the tubules, by this arrangement the tubules are continually washed by the liquid which carries the secretory products down the tubule. The urine collects in the sinus which has an outlet thru the ureter. KIDL'EY Histo-Pathological Changes 2 Parenchymatous degeneration of the urinary tubules was the most constant characteristic pathological change found in the kidneys. The degree of degener- ation varied from a very severe one bordering on necrosis to a very mild degen- erative disturbance. In the majority of the cases the parenchymatous degenera- tion was marked. Kidney — Histopathological Changes 1 7 Passive congestion existed in over 80$ of the cases. Over half of the series (.13 cases) presented advanced passive congestion; the congestion backing up to involve the glomeruli. The passive congestion which involved the glomeru- li was classified as severe, while the less severe cases were rated as marked and slight. Ten percent of the kidneys were markedly congested, while 30$ were only slightly congested, A final 10$ showed no passive congestion. There were also other changes present in a few instances. A few hemorr- hages were found in three cases. One kidney showed a beginning necrosis of the urinary tubules around the hemorrhagic area. Another organ also presented a few small areas of necrosis and one showed a slight edema in the cortex region. In general the most constant changes were a marked parenchymatous degen- eration and a marked passive congestion. LIVER The Normal Histology: The liver, the largest gland in the body, consists of very delicate glan- dular tissue disposed around ramifications of the portal vein. It is covered with a capsule of fibrous tissue giving off trabeculae which extend into the in- terior. The position of the liver in the body cavity is immediately below the diaphragm chiefly on the right side with the smaller lobe extending into the left side of the cavity. Grossly the liver is divided into two large lobes. Each of these lobes is subdivided by the interlobular tissue into many lobules. Centrally located in each lob- ule, the central vein receives the capillary branches of The hepatic cells, the smallest unit of structure of the liver, are poly- gonal in shape and contain a comparatively large nucleus. The cytoplasm is finely granular, and the nucleus contains a reticularly arranged mass of chro- matin material. The liver cells are placed in rows or cords projecting from the the portal vein. ■ . - 6 - central vein-like spokes in a wheel. The biliary sinuses between the rows of cells which collect the secreted bile from the liver cells have no definite vessel walls but their course follows thru trough-like depressions in the walls of the liver cells. The larger bile vessels have true walls and are usually found in close proximity to a hepatic vein and a hepatic artery forming the so-called hepatic trinity. Two blood supplies, namely, the functional and the nutritional, are found in the liver. The functional supply comes from the portal vein. After leaving the portal vein the blood flows into the interlobular branches of the vein which carry it thruout the liver and brings it to the individual lobules. These in- terlobular veins then give off capillary ramifications which extend thruout the lobule and bring the blood in intimate contact with the hepatic cells. Later the capillaries collect at the central vein and conduct the functional blood to the sublcbular branch of the hepatic vein and finally back to the heart. The amount of interstitial tissue varies in different species. In swine the lobules are very distinct due to the large amount of interlobular connective tissue. The nutritional blood is derived from the hepatic artery and follows a course somewhat similar to the functional blood. Later the two unite and both return to the heart by way of the same vessels. LIVES tiisto-Pathological Changes: Passive congestion, tho not present in all the livers was characteristic of them in general. Two livers were severely congested, two others showed mark- ed passive congestion, and ten were only slightly congested while five presented no noticeable congestion. There were no hemorrhages in any of this series (19 cases) . The most characteristic pathological changes in the livers were fatty degeneration and fatty infiltration. Only two cases could be classed as a pure fatty infiltration while fifteen cases showed fatty degeneration with some fatty • ■ _ 9 - infiltration in three cases. It was interesting to note that there were only two cases which were free of pathological fat. L i ver — Hi s t o-Pat h c 1 c gi ca 1 Change s Sixteen of the guinea pigs showed marked parenchymatous degeneration of the liver. Most of the parenchymatous degeneration was present with fatty degen- eration* while there were only two mild cases of parenchymatous degeneration in livers which did not show fatty degeneration. All cases showed either a paren- chymatous or fatty degeneration. A slight cellular hepatitis was found in four cases. Passive congestion, marked fatty degeneration, and parenchymatous degen- eration were the three most characteristic pathological changes in the liver. HE APT Normal Histology: Histologically the heart is composed of three layers of tissue, the epi- cardium, myocardium and endocardium. The endocardium is composed of a single layer of endothelial plate-like cells and underlying connective tissue which is quite rich in elastic fibers with a small quantity of involuntary muscle. This layer is composed chiefly of muscle fibers which are not organized into bundles. 10 but form a very complex system of branching and intermingling fibers or intercal- ated discs. Between the branching cardiac muscle structure the connective tissue aids in binding the fibers together. There is no well defined endomysium as in voluntary muscle. The outer layer of the heart, the epicardium, consists of a single layer of endothelial cells having a substratum of f ibro-elast ic tissue. The epicardi- um completely covers the heart and blends into the adventitia of the great veins. HEART Histo-Pathclogical Changes: The only constant pathological change in the heart was a parenchymatous degeneration. Most of the heart series (16 cases) were only slightly degenerated, while two showed marked degeneration, and one severe degeneration. Heart--Histo-Pathological Changes The striations of the muscle fibers were not visible due tc the parenchymatous degeneration. Congested coronary vessels were found in six specimens of this organ and one showed a passive congestion of the capillaries of the cardiac mus- cle. Another change present in three of the hearts, which might have been due 11 to the method of fixation, was a separation of the muscle fibers at the cement lines. The fibers of one of the hearts appeared as tho they were shrunken. Summarizing the series of the heart it was found that parenchymatous de- generation was characteristic thruout and a congestion of the coronary vessels in a few cases. THE SPLEEN Normal Histology: The spleen may be considered as a huge hamolymph node, possessing the functions of producing lymphocytes and of destroying erythrocytes. It is dark red in color due to the fact that it contains a large quantity of blood. This organ is located on the left side of the abdominal wall between the stomach and the diaphragm. The spleen is invested in a capsule composed of dense elastic tissue with a few scattered fibers of involuntary muscle. Extending from the capsule into the spleen are the trabeculae which branch into a delicate frame- s^...X^ S o\e. work giving support and a degree of rigidity to the organ. syW&lL Histologically the spleen is composed of splenic lobules. These are imperfectly defined by the interlobular trabecu- -'Kay ecu la if lae. Branching from these interlobular trabeculae the in- ■ T'-- ' ' • tralcbular trabeculae divide the 3plenic lobule into about ten primary compartments. The space between this frame-work of trabeculae is filled by splenic pulp and lymph follicles. The splenic pulp is composed of essentially four elements namely, lymphocytes, leucocytes, erythrocytes and the large phagocytic cells. The blood enters at the hilum and after traversing in the hilum connec- tive tissue for a short distance the arteries breakup into branches which supply individual splenic lobules. Prolongations of the connective tissue of the trab- eculae follow the arterial branches into the pulp, thus surrounding them with an extra fibrous envelope. Between this envelope and the artery wall local accumu- lations of lymphocytes occur giving rise to a spherical or fusiform mass of lymphoid tissue, the Malpighian bodies. The functional blood supply after con- . 12 tinuing in vessels for a time, after entering at the hilurn, a part leaves the vessels and flows freely thru the splenic pulp, while another part continues thru thin walled venous sinuses. There is a slow movement of the blood thru the spleer after which it is again collected in vessels and carried away. The nutritional blood supply on the other hand is conducted thruout its course in closed vessels. SPLEEN Histo-Pathological Changes: Follicular hyperplasia was the outstanding feature of over 85$ of the spleens. Most of the cases, however, were mild and only two were marked. Con- gestion was present in 65$ of the cases. In one spleen there was an increased number of leucocytes while three others had eosinophilic infiltrations. A mild splenitis was found in one specimen. Spleen--Histo -Pathological Changes The increase in the size of the splenic follicles and the congestion were the most outstanding changes. - . 13 THE LUNGS Normal Histology: The lungs, the respiratory organs of the body, are composed of thin walled air sacs surrounded by thin walled capillaries which allow the air to come in close contact with the blood and permit a gaseous exchange. On examining the outer surface of the lung it is seen that it is divided into polygonal areas, or lobules which are the unit of structure of the lung, beginning at the trachea the air passages divide as seen in the figure below. The trachea extends well down into the thorax and then divides into two bronchi, one going to each lung. The bronchus on entering the lung begins to divide and subdivide. The branches are called bronchioles. The larger bronchioles have cartilaginous rings as in the walls of the main bronchi. The medium sized bronchioles contain only scattered plates of cartilage. These smaller bronchi- oles divide and finally end in terminal bronchioles. The terminal bronchioles are connected with the alveoli or air sacs by means of the atria. On cross section the lung appears as shown in figure below. The walls of the air sacs are composed of flattened spherical cells. The blood supply follows the divisions of the bronchioles and extends into the interstitial tissue between the air sacs. The walls of the bronchioles are composed of three parts, .namely, the fibrous tunic which contains the cartilaginous ' fllveal ‘' rings, the central layer of smooth muscular and connective TW —' ' I tissues, and the mucous membrane lining the inner surface of the lumen. THE LUNGS Histo-Pathological Changes: Passive congestion of the lunsrs was present in all of the cases but one. Three of the lungs apparently showed hemorrhages, which might be the result of agonal struggling in those few cases destroyed by concussion, Seren of the lungs were severely congested and six only slightly congested. One lung was normal. ' ■ 14 Lung- -His to -Pathological Changes A slight catarrhal exudate was present on the edges of the air sacs in one lung and another had a catarrhal bronchial exudate. Five lungs presenting advanced passive congestion also showed sub-pleural edema. SUMMARY The most consistent histo-pathological change in the kidney, liver, heart, spleen and lung of guinea* pigs suffering from an acute form of botulinu3 poisoning was a passive congestion, with parenchymatous degeneration in varying degrees. The later in a few instances bordered on necrosis. Occasionally be- ginning inflammatory changes were encountered, but in the series as a whole this was not characteristic. Similar changes may be observed in other toxemic dis- eases, and are of little or no value in rendering a differential diagnosis. The acute character of the disease in the guinea pigs that furnished the tissues for. these studies may not be typical of changes in a chronic form of botuiinus in- t oxicat ion, ***************** • : . - • - lb BIBLIOGRAPHY' Textbooks in Histology Normal Histology' — Pier sol Essentials of Histology----- — ---LeRoy A Compend of Histology — • — Eadasch Textbooks in Pathology Special Pathologic Studies(Two volumes) Durck General Pathology-- -Durck Principles of Pathologic Histology -Mallory Manual of Pathology — McConnel References in Botulinus Poisoning McCaskey, G.W. , 19 IS, Bacillus Botulinus Poisoning. Am. Jour. Mel. Sc. July 1919 No. l Vol.clviii p 5? Dick son, Ernest C. . 19 13, Botulism, Archives of Internal Medicine Oct . 1913, Vo 1. XXII, pp 483-495. Dickson, Ernest C. , 19 13, Botulism, Monographs of the Rockefeller Institute for Internal Medicine, No. 8 July 31,1913. . 16 DIAGH0S1S CHART OF KiDHiL'YS Ho. Blood. Cloudy Swelling Remarks 8-1 -A Congestion-Localized hem* Slight hem. in glomeruli. C.5**in varying degrees. Slight edema Some border on necrosis 9-1 -A Passive Congestion. Slight cong.in glom# Marked c.s.of T.*** Collection of red cells under cap- suled! ight nec- rosis at hem. 13-b Severe passive cong. Marked c.s.of T. 14-c Passive Congestion Marked c.s.of T. 15-C Very severe cong. Marked c.s.of T . lo-c Slight pass.cong. Mild c .s.of T. 1 7-2-A Severe pass.cong. Mild c.s.of T. 18-1- A Severe pass.cong. Marked c.s.of T. Areas of necrosis 19-A Severe pass.cong. C.s.of T. 21-1-ABeginning pass.cong. Slight c.s.of T. 23- 2- A Severe pass.cong. Severe c ,s .of T. 24-A Marked c.s.of T. 25-A Severe pass.cong. Slight c.s.of T. 2b-C Slight c.s.of T. 27-s Slight c.s.oi x. 2Z-a Marked pass.cong. Marked c.s.of T. 29 -A Slight pass.cong. Slight c.s.of T 30 -D Slight pas. cong. Slight c . s .of T . 31 -A Marked pas. cong. Slight c.s.of T. *Hemorrhage C. s.**uloudy Swelling of Urinary Tubules fGlomeruli . 17 No . Blood DIAGNOSIS CHART Cloudy Swelling OF THE LIVERS Degenerations Remarks . 8 Severe pass Slight c.s.* Fatty infilt .& Slight cell- 13 -A cong. Pass. cong. Severe c.s. degene rat ion Fatty infilt & ular hepatitis. 14-D Slight p.c"* Severe c.s. deg. (slight ) Slight fatty deg. 15-B Marked p.c. Slight c.s. Slight fatty deg. 16 -B Slight p.c. C.s. Severe fatty deg. Slight cellular 17-2-C Mild p.c. C.s. Fatty deg.& Infilt. hepatitis. 18-l-C Slight p.c. Marked c.s. Fatty deg. Deg. near necrosis 19-C Slight P.c. Marked fatty deg. Slight cellular 21 -C 23 -l-C Severe p.c. Marked c.s. Marked fatty deg. Marked fatty deg. hepatitis 24-C Slight c.s. Severe fatty deg. Slight cellular 25 -D Marked p.c. Marked c.s. Marked fatty deg. hepatitis 26 - B 27- C Mild p.c. Mild c.s. Mild fatty deg. Mo fatty deg. 28- E 29 - A 30- B Slight p.c. Slight p.c. Marked c.s. Slight c.s. Slight c.s. Fatty infilt. on piripheral areas Marked fatty deg. 31 -D Slight p.c. Marked c.s. Slight fatty deg. c.s*cloudy swelling ♦♦Passive Congestion . . . . . . No. 13- 0- 14- B 15- A 17- 1- 18- 1- 21 -B 23- 1- 24- B 2 i>-B 26- E 27- A 28- D 29- D 30- C 31- C DIAGNOSIS CHART OF THE HEARTS Blood Cloudy Swelling Remarks Cong. of coronciry vessels i Pass.cong.of cap- illaries of car- diac muscle . { Coronary vessels congested Cong. of coronary vessels . No congestion Cong. of coronary vessels Striations not visible due to c.s.* Marked c.s. All striations gone Severe c.s. Marked c.s. Striations gone Slight c.s. Striations gone Mild c.s. Very slight c.s. Striations visible in part of the fibers Slight c.s. Very mild c.s. Slight c.s. Si ight c.s. Veyy little c.s. Slight c.s. Slight c.s. Slight separation at cement lines°Fibers shrunken . Slight separation at cement lines. Separation at cement I ines . *Cloudy Swelling ' ■ DIAGNOSIS CHART OF THE SPLEENS No. Blood Follicular Hyperplasia Remarks 9-i-c General cong. Increase in leuk* Mild f.h.** 13-F Congestion 14-F f .h. 15-2-A Slight f.h. 16-E No congestion Slight f.h. 17-l-B Blight f.h. 18-2-B Mild congestion 19-E Slight cong. Slight f.h. 21-3-B Mild interstitial splinitis 23-2-C Slight cong. f.h. 24- F Congestion f.h. 26-F Focal collections of eosinphiles . 27-F f.h. Eosinphilic infiltration 28-F Very slight cong. Slight f.h. Increase of eosinphiles 29-F Very slight cong. Marked f.h. 31-E Mild f.h. ♦Leucocytes ♦♦Follicular Hyperplasia , - 20 - DIAGNOSIS CHARTS OF THE LUNGS No. Blood Exudate Remarks 8-1 -D Passive cong* Slight catarrhal the edges of air exudate on sac 3 . 9-2-d Very severe passive congestion Mild compensatory emphysema 13-E Slight congestion 14-A Severe cong. in most areas°Hem .over a large area. Sub-Pleura edema A farw eosinphiles. 15-D Slight passive cong. Suggestive of emph ysema. 16-A Slight passive cong. 17-1-E Severe passive cong. Slight sub-pleura edema . 18-1-D Severe passive cong. 19-D Slight passive cong. 21 -2- A Hypostatic cong. Desquamation of epth. cells of air sacs. 23-1 -B Very severe cong. Ed ema- D e s quarna t i 0 n of alveloir cells. 24-1-E Marked congestion Slight edema 25-C Very severe cong. 26 -A Slight congestion 27- E Normal 28-A Hemorrhage 29-E Very marked cong. Slight exudate in bronchi 30-A Slight passive cong. 31-B Hemorrhage Edema- Blood in bronchi . ♦Congestion