24 



lein, 2 per cent, of the following sugars, saccharose, lactose, glucose 

 was added to the above medium, and the tubes sterilized at 100'^ C. 

 on three successive days. 



Saccharose bouillon. Both arms of the tube became cloudy, 

 considerable sediment formed but no pellicle. Reaction after 10 

 day's growth + 1.8 per cent. 



Lactose bouillon. After 24 hours both arms of the tube became 

 cloudy, the closed one with less turbidity, there was some sediment 

 but no pellicle or gas. After 48 hours, the amount of sediment 

 increased and 1 per cent, of gas formed, subsequently the closed arm 

 became clear, but there was no increase of gas. Reaction after 10 

 day's growth, + 1.43 per cent. 



Glucose bouillon. There was more growth in this medium than 

 in the others, 0.5 per cent, of gas collected on the 2nd day, with no 

 subsequent increase. Sediment very copious. Reaction after 10 

 day's growth, + 1. 8 per cent. 



Enzymes. Proteolytic enzymes, cytase, and diastase are pro- 

 duced by the organism. Evidence as to the formation of these 

 enzymes is afforded by the following experiments. 



Proteolytic Enzymes. These enzymes are produced in small 

 quantities. Gelatin is slowly liquelied, blood serum even more 

 slowly, milk is partially peptonized. 



Fresh milk serum sterilized by filtration was inoculated with a 

 culture of the bacillus, and the medium held at 25 degrees C, for 10 

 days. At the end of this time a portion tested for peptones gave the 

 biuret reaction. The proteid bodies except peptones in the larger 

 portions were precipitated with ammonium sulphate and the filtrate 

 treated with caustic potash solution and copper sulphate gave a 

 violet color indicating the presence of peptones. 



Diastase. Diastase is produced in small quantities in ordinary 

 bouillon. Equal parts of sugar free starch paste and thymol were 

 mixed with a 10-days old bouillon culture and left at 25 degree C, for 

 12- -24 hours. A test of the filtrate of this mixture with Fehling's 

 solution showed small traces of sugar to be present. 



The organism when grown on potato also destroyed starch. 

 Slices of raw potato inoculated with the organism did not give any 

 coloration when treated with iodine, which indicated the destruction 

 of the starch. 



