580 TREPONEMATA, SPIRONEMATA, LEPTOSPIRATA 
Brucker and Gelasescoi and Sowade^ injected material from syphilitic 
lesions into the testicles of rabbits and observed considerable prolifera- 
tion of the organisms there. Levaditi and Mcintosh^ placed syphilitic 
material enclosed in celloidin capsules in the abdominal cavities of 
monkeys and observed a proliferation of Treponemata which resembled 
the Treponema pallidum very closely. Cultures were not obtained 
in artificial media. Schereschewsky^ grew the organisms in impure 
culture in anaerobic cultures of gelatinized horse serum, that is, horse 
serum which has been heated to 60° C. for some hours. To Xoguchi,^ 
however, belongs the credit of obtaining Treponema pallidum in 
pure culture, and of demonstrating its etiological relationship to the 
disease syphilis. 
Fig. 79. — Treponema pallidum. 
The medium which gave the best results is prepared in the following 
manner: Two per cent slightly alkaline agar is melted and quickly 
cooled to 45° to 50° C. and sterile ascitic or hydrocele fluid is added 
in the proportion of 2 parts of agar to 1 part of fluid. At the same 
time a small piece of sterile tissue from a rabbit's testis or kidney is 
introduced. The medium is rapidly cooled to room temperature and 
covered with a layer of sterile paraffin oil, 2 to 3 cc. deep, to prevent 
evaporation. The medium is incubated for two days to ensure sterility 
and is then inoculated with appropriate material, after first being 
certain that the material contains the organisms. The syphilitic 
tissue, prior to inoculation, is macerated under sterile conditions 
with 1 per cent sodium citrate solution and then introduced deeply- 
into the agar-ascitic fluid-tissue media. Incubation is maintained at 
37° C. for two to three weeks. The Treponemata in virtue of their 
motility move away from the line of inoculation and cause a more 
• Compt. rend. Soc. de biol., Paris, 1910, 68, 684. 
2 Deutsch. med. Wchnschr., 1911, 37, 682. 
3 .\nn. Inst. Pasteur, 1907, 21, 784. 
' Deutsch. med. Wchnsrhr.. 1909, 35, 835, 1260. 
■■> .Jour. Am. Med. Assn., 1911, 57, 102; Jour. Exp. Med., 1912, 15, 90. 
