94 



J. T. CONNELL AND L. E. HOLLY 



working with because of the action (digestive and otherwise) 

 upon it of the microorganisms. What probably is more impor- 

 tant is that it contained emulsifying substances which were 



TABLE 2 



A. pH of broth 7.1 with 60 mgm. 



B. pH of broth 7.1 with 60 mgm. 



C. pH of broth 7.4 with 60 mgm. 



D. pH of broth 7.4 with 60 mgm. 



E. pH of broth 7.9 with 60 mgm. 



F. pH of broth 7.9 with 60 mgm. 



K salt antigen per liter 

 acid antigen per liter 

 K salt antigen per liter 

 acid antigen per liter 

 K salt antigen per liter 

 acid antigen per liter 



1 CO. of A plus 1 cc. rabbit cell emulsion. 

 1 cc. of B plus 1 cc. rabbit cell emulsion. 



1 cc. of C plus 1 cc. 

 1 cc. of D plus 1 cc. 

 1 cc. of E plus 1 cc, 

 1 cc. of F plus 1 cc. 



rabbit cell emulsion, 

 rabbit cell emulsion, 

 rabbit cell emulsion, 

 rabbit cell emulsion. 



APT EH 

 eS^C FOR 

 THIRTY 



MINUTES 



+ +23' 

 + +28' 

 -1 hr. 

 -1 hr. 



+ +40' 



+ +45' 



From the above experiment it will be seen that the artificial hemolysin is in- 

 activated at 65°C. for thirty minutes at a pH of 7.4, but at concentrations of 7.1 

 and 7.9 the inactivation is only partial. 



delivered into it with the disintegration of the bacteria. With 

 this idea in mind we tried the effects of some emulsifying sub- 

 stances to see if we could imitate the natural process more 

 closely, and to determine whether the broth would not hold 

 more of the fats without clouding. The substances chosen 

 were such proteins as hemoglobin, casein, and typhoid protein, 

 with which the fat antigen was emulsified, — imitating the con- 

 ditions we believe to exist in the germ bodies^prior to adding 

 to the broth. The following table is an example of the results. 



TABLE 3 



A. 35 CC. of broth to which was added slowly in fractions 5 cc. of a solution con- 



taining 10 mgm. of typhoid protein and 2.4 mgm. of K salt antigen. 



B. 35 cc. of broth to which was added in the same manner 5 cc. of a solution of 



10 mgm. of casein with 2.4 mgm. of K salt antigen. 



C. 35 cc. of broth to which was added in the same manner 5 cc. of a solution of 



hemoglobin with 2.4 mgm. of K salt antigen. 



