1 •• -■!» J M -Kas.!!;!!)) mm. '^ ^v^. 973 LIBRARY NEW YORK STATE VETERINARY COLLEGE ITHACA, NEW YORK Cornell University Library SF 973.K54i Inoculation experiment with pure culture 3 1924 000 225 411 INOCULATION EXPERIMENT WITH PURE CULTURE OF SPIRO- CHAETA HYOS Studies on Hog-Cholera WALTER E. KING AND RAYMOND H. DRAKE Reprinted from The Journal of Infectious Diseases, Vol. 16, No. 1, Jan., 1915,' pp. 54-57 Cornell University Library The original of tiiis book is in tine Cornell University Library. There are no known copyright restrictions in the United States on the use of the text. http://www.archive.org/details/cu31924000225411 INOCULATION EXPERIMENT WITH PURE CULTURE OF SPIROCHAETA HYOS STUDIES ON HOG-CHOLERA Walter E. King and Raymond H. Drake (^FroMi the I^esearch Laboratory, Parke, Davis and Company, Detroit, Michigan) 111 former publications it has been suggested that the spirochaeta hyos may bear some etiological relationship to hog-cholera. Hereto- fore, it has not been possible to prove the pathogenic significance of this new organism because of the difficulties encountered in attempting to obtain pure cultures. These difficulties have not been completely overcome as yet, but, by painstaking effort, a pure culture of the spirochaeta hyos has been secured and typical hog-cholera of the acute type has been produced with this culture. The protocol of this experi- ment is as follows : On September 23, 1913, a culture was made on Hata medium with rabbit kid- ^viey- from -Berl;e-fcJcHTl+crcd-sti-5pcn5Tan-o-f~ti5aae "from a local lesion on the ear of Hog 653. The local ear lesion, on dark-field examination, showed numerous spirochetes. Hog 653, on autopsy, showed typical lesions of hog-cholera. Culture 653, from the ear, was incubated in a desiccator, under anaerobic conditions, for several weeks at 40 C, and then for several weeks at 37 C. Dark- field examination, on December 13, showed the presence of spirochetes, rela- tively few in number. A portion of the impure culture was macerated in sterile water and filtered twice through the Berkefeld. On January 5, 1914, cultures were made from the filtrate on Hata medium with no kidney tissue. Control cultures from the filtrate gave negative tests, showing that the filtrate was free from bacteria. Culture 653, grown in the same manner as the above, was examined on March 17, on dark-field. It showed growth of the spirochaeta hyos in pure culture. Culture media, inoculated with material from Culture 653 (Transfer 1), gave negative results. A suspension was made of a portion of pure culture of the spirochaeta hyos (Culture 653, Transfer 1) in an equal volume of sterile water. Dark-field examination showed the spirochaeta hyos, uncontaminated, in suspension. On March 17, animal inoculations were made from the suspension as follows : Hog 805, 4.5 CO.; Hog 806, 3.5 c.c, both intramuscularly. A normal hog. Hog 807, was placed with thern in an isolated, disinfected room as a control on Hogs 805 and 806. Hog 805. — On March 17, Hog 805 was injected intramuscularly with 4.5 c.c. of Culture 653 (Transfer 1) ; March, 25, the hog appeared normal, appetite good. 2 Walter E. King and Raymond H. Drake but somewhat inactive ; March 27, normal in every way ; April 6, "oflE feed," appeared sick; April 8, was bright; April IS, very sick; April 20, died. Inguinal, mesenteric, retroperitoneal and other lymphatic glands were enlarged and very hemorrhagic ; lungs, congested and consolidated ; liver, mottled with areas of degeneration ; spleen, much enlarged, dark and soft ; kidneys, ecchymotic ; intestinal mucosa, congested ; no typical ulcers. Spirochaeta hyos present in cecal mucosa. Hog 806. — On March 17, Hog 806 was injected intramuscularly with 3.5 c.c. of Culture 653 (Transfer 1) ; March 26, the hog was inactive, anorexia; March 27, acted better, appetite better ; March 30, anorexia, weak, constipated ; April 2, no appetite, back arched ; April 6, numerous spirochetes in exudate from car ; April 8, very sick ; April 13, moribund, killed. Figure 1.— Hog 806 Lymphatic glands were very enlarged and hemorrhagic ; liver, normal ; lungs, congested and consolidated ; spleen, soft and friable, enlarged ; kidney, ecchy- motic; small button ulcers in mucosa of large intestine. Spirochaeta hyos in mucosa of cecum. Hog 807. — Hog 807 was a control on culture pigs. On March 17 the hog was placed with Hogs 805 and 806; April 1, normal, good appetite; April 2, slight symptoms ; April 6, sick ; April 7, anorexia ; April 8, sick, constipated ; April 15, very sick; April 17, died. Lymphatic glands much enlarged and hemorrhagic; lungs, congested and con- solidated ; spleen, soft and friable, enlarged ; kidneys, ecchymotic ; small ulcers in mucosa of cecum, mucosa of large intestine congested. Spirochaeta hyos present in margins of cecal ulcers. In this experiment Hog 805 showed a mild reaction eight days after inoculation and manifested "secondary" symptoms twenty days after .„-..«. Maroh, 1914. April | " 17 18 19 20 31 zs 12 Si ss 26 27 2829 30 31 1 2 3 4 6 6 7 8 9 10 11 1! 13 14 If) 16 T7 Ifl t i i A <\ / \ fs f ^/ A ^ \ / "V '} 1 1 f / > 1 J J ^ / ^ / <^ J L y J >^ V, M^ r- /^ V ' 1 L^ —t S. J y _J _J L Cliart 1. — Tempetafffre Curve for Hog 805. Vur and Month - Mar, 1914, April OA, IV 18 19 20 ai 22 23 24 25 26 27 28 29 ao 31 1 2 3 4 B 6 1 8 9 10 11 i: U 1- u I z if z < < 0. 06" OS' 04° 03" 02° 00' rN A