CULTURING TREPONEMATA 37 



II. Serum Agar of Muhlens and Hofmann. Fill sterile test-tubes one-third full 

 with horse serum. This is sterilized on three successive days at S5C. Then 

 add an equal amount of a 3% agar containing 0.5% glucose which has been melted 

 down and cooled to 5oC. The mixed serum agar is then kept at S5C. for two hours. 

 Such tubes are inoculated as for ascitic agar rabbit tissue media and incubated under 

 anaerobic conditions, preferably in a flask from which the air has been exhausted and 

 the remaining oxygen absorbed as shown in the anaerobic bottle described and 

 illustrated in Fig. 8. 



