104 THE ROYAL SOCIETY OF CANADA 



Uschinsky^s Solution. In 24 hours' growth: in 48 hours' growth 

 copious and precipitation of sediment: in four days perceptible clearing 

 of the medium is observed. After a period of three months a heavy- 

 sediment is present, the liquid having a milky looking cloudy appearance. 



Preparation of Sugars. In preparing the sugars especial care was 

 taken in order that any likelihood of chemical change during sterilis- 

 ation should be reduced to a practical minimum. 



The standard broth used was made up of 

 Tap water 

 Witte's peptone, 2% 

 Sodium chloride, .5% 



The reaction to phenolphthalein was not adjusted and on being 

 tested showed .06% of acidity. The Smith fermentation tube was 

 used, and the various sugars — saccharose, glucose and lactose having 

 been added in the ratio of 1%, the sugar broths were filled into the 

 fermentation tubes and sterilised intermittently for three successive days. 



The glycerine broth was prepared similarly. 



In the case of the Dulcite, Mannite and Rafïinose broths a slight 

 variation in the manner of preparation was made. 



The peptone broth, prepared as before, was filled into the fer- 

 mentation tubes and sterilised at 100°C. intermittently for three days. 



Solutions of the three sugars respectively were made in water 

 of such dilution that 1 cc. of the water solution added to 10 c.c. of the 

 prepared broth in the fermentation tube gave a peptone broth sugar 

 solution of 1%. The sugar solutions were then sterilised intermit- 

 tently for three successive days and immediately before inoculation 

 1 c.c. was pipetted into the fermentation tube containing 10 c.c. of 

 the previously prepared peptone broth. 



SUGARS. 



Dulcit. In 24 hours growth in both open and connecting tube: 

 closed arm clear, no gas: alkaline reaction to litmus. 



Mannite. Turbidity throughout in 24 hours. In 72 hours 5% 

 gas: acid reaction to litmus. 



Raffinose. Turbidity throughout in 24 hours. In 72 hours 8% 

 gas: neutral to litmus. 



Glucose. Turbidity throughout in 24 hours with sediment. After 

 72 hours no gas: acid. 



Lactose. Turbidity throughout in 24 hours: after 72 hours 8.5% 

 gas: acid reaction to litmus. 



Saccharose. Turbidity throughout in 24 hours: after 72 hours 

 no gas: acid reaction to litmus. 



