8 



L. A. JULIANELLE 



and the cultures were inactivated by chemical or heat, with the enzymes 

 still capable of activity, hemolytic substances were being produced. Each 

 day tests were made for the presence of hemolysins. After the first 

 day, guinea-pig serum and a living (24-hour) culture in 1 c c quantities 

 were added to the 45 C. specimen. This was to supply complement, if 

 it were needed, and any other vital substances necessary for hemolysis 

 that a growing culture might possess. The results are appended in 

 table 2, which shows: 1. No hemolysin was formed in cultures sub- 

 jected to antiseptics or heat. 2. Complement does not appear necessary 

 for hemolysis. 3. A living culture produces hemolysis per se, and is 



TABLE 2 



SHOWING EFFECT OF HEAT AND CHEMICAL AGENTS ON PRODUCTION OF HEMOLYTIC 



SUBSTANCE 



* =: no test conducted. Day 1 is 1st day under treatment, but 6th day of age of culture. 



consequently worthless in such an examination. These results do not 

 show that hemolysin is of an autolytic nature ; neither do they show 

 that it is not of an autolytic nature. The conclusion to be drawn is 

 that in the case of heat, the hemolysin being thermolabile, is possibly 

 dissipated ; while in the case of the antiseptics, the hemolysin is so closely 

 associated with the bacterial cell that destruction of the latter means lack 

 of manifesation from the former. This falls somewhat in line with the 

 work of Gordon on meningocbcci, showing that hemolysins are endocel- 

 lular and are liberated on autolysis of the bacterial cells. 



Effect on Hemolytic Activity of Successive Transplantation in 

 Blood-Free Medium. The object of the next experiment was to 



