34 Report OF DEPARTMENT OF BACTERIOLOGY OF THE 
escape by evaporation. Reference has already been made to the 
extensive work of Babcock and Russell. 
The work of Smith (1) emphasizes the necessity of a well-closed 
receptacle since he pointed out that in a cotton stoppered test- 
tube an abundance of liquid chloroform in the bottom of the 
tube would not inhibit the growth of bacteria in the upper layers 
of the supernatant liquid medium. In such cases evaporation 
of the chloroform at the upper surface of the media seemed to 
take place faster than the solution of the same at the lower surface. 
While the earlier work was principally concerned with finding 
substances which would prevent germ action, attention was later 
given to the effect of such substances upon the activity@¢of the 
enzyms. ‘The results of these physiological studies have varied 
widely, depending upon variations in the conditions attending 
the experimental work. Unfortunately, these variations were not 
always recorded, since the worker was naturally often unaware 
of the existence of important factors. Good examples of these 
differences are seen in the conflicting reports regarding the curd- 
ling of milk in the presence of chloroform and in the controversy 
which was waged regarding the coagulating action of chloroform 
upon albuminous substances. 
Green (1, 2) reported the sterilization of vaccine lymph within 
eight days by saturating it with chloroform vapor. He found 
this treatment effective against Bacillus proteus, B. prodigiosus, 
B. pyocyaneous, B. fluorescens, B. coli, B. typhosus, B. diphtheriae, 
B. mallet, B. pestis, B. tuberculosis and Spirillum cholerae. He 
stated that this method of applying the chloroform did not impair 
the activity of the vaccine, while the addition of liquid chloroform 
markedly reduced it. His conclusions have been tested by 
Carini (1) and by Nijland (1). Both found that vegetative bacterial 
forms were promptly destroyed by this treatment but that spores 
survived. Neither considered the treatment with chloroform su- 
perior to the former method of handling lymph with glycerin, 
since in some cases the activity of the vaccine was quickly de- 
stroyed and in general chloroform vaccine lost its virulence more 
auickly on standing than did vaccine treated with glycerin. 
Aare aa Hite: METHOD OF DETERMINING CHLORO- 
FORM. 
The method used in determining the amount of chloroform pres- 
ent in milk or cheese is as follows: 
a a a: 
