100 J. T. CONNELL AND L. E. HOLLY 



vated at 56°C. for thirty minutes following which they were 

 tested for antilytic power. Table 6 shows a typical protocol of 

 several experiments. 



TABLE 6 

 HS. Serum of rabbit immune to natural hemolysin. 

 AS. Serum of rabbit immune to artificial lysin. 

 NS. Serum of normal rabbit. 

 A. Supernatant fluid from a centrifugated eighteen-hour 

 culture of B. megatherium. 



0.5 cc. A plus 1 CO. cell suspension ++ in 8 minutes 



0'5 cc. A plus 1 cc. cell suspension ++ in 8 minutes 



0.5 cc. A plus 0.2 cc. HS plus 1 cc. cell suspension — in IJ hours 



0.5 cc. A plus 0.2 cc. HS plus 1 cc. cell suspension — in H hours 



0.5 cc. A plus 0.2 cc. AS plus 1 cc. cell suspension — in 1§ hours 



0.5 cc. A plus 0.2 cc. AS plus 1 cc. cell suspension — in \\ hours 



0.5 cc. A plus 0.2 cc. NS plus 1 cc. cell suspension ++ in 60 minutes 



0.5 cc. A plus 0.2 cc. NS plus 1 cc. cell suspension ++ in 60 minutes 



Further experiments on the immune bodies resulting from 

 immunization of animals with the artificial hemolysin of B. 

 megatherium are given in the paper on the Nature of Toxin by 

 Warden and ourselves. 



DISCUSSION 



We have dealt merely with the hemolysins of two organisms 

 streptolysin and megatheriolysin, both of which are very power- 

 ful and occur early in the growth of the organisms. The former 

 is only produced under special conditions and disappears m 

 twenty-four hours. The latter is elaborated readily upon veiy 

 simple media and remains active for weeks. The difference in 

 the ease with which these hemolysins are produced appears to 

 bear a direct relation to the rapid and abundant growth of the 

 bacteria, the hemolysin not being present in quantities sufficient 

 to detect until the growth is quite abundant. We are aware 

 that with the artificial antigens we have not obtained hemolysins 

 as active as those formed by the germs, but a discrepancy of the 

 sort was to be expected. The microorganism delivers its antigen 

 into the broth in a state of emulsification difficult to imitate with 

 only the colloid properties of the broth itself, and those of the 

 few proteins used as adjuvants to aid us in our attempts to 



