150 REporT oF DEPARTMENT OF BACTERIOLOGY OF THE 
The details of a single trial will suffice to illustrate the 
method and results. Chloroform was added as follows to 
each of six c. c. broth tube cultures, six days old, shaking 
very thoroughly; to tubes 1 & 1’ added 1 c. c. of chloroform ; 
to tubes 2 & 2’ added 0.5 c. c. of chloroform; to tubes 3 & 3” 
added 0.8 c. c. of chloroform. On the third day thereafter 
transfers from these showed 1 & 1’ to be sterile, whereas the 
others contained living organisms. On the fifth day transfers 
from these latter again showed living organisms. Cubical 
blocks of fresh carrot were placed in tubes thus sterilized with 
10% additions of chloroform and in other similar series steril- 
ized by 25% and 50% additions respectively, and in all cases 
alike they were quickly and fully softened. 
There was no appreciable retardation in the rate of soften- 
ing in any such case as compared with tubes sterilized with 
thymo] or phenol, or even with cultures containing the living 
organisms. 
This matter was of so much importance that further com- 
parative trials were made by using razor sections of turnip. 
There was no appreciable difference in the rate of softening 
as between living cultures and those sterilized by chloroform, 
Thus in one trial, broth cultures four days old were used; 
10% of chloroform was added to each of two of these; after 
thorough shaking immediate trials were made comparing the 
activity with that of similar cultures; no difference was found. 
Again, at the end of the sixth day when the chloroform tubes 
were sterile, comparative trials showed the sterile broth to 
equal in enzymic activity the living control cultures, now ten 
days old. | 
Comparisons and final conclusions.— These results showed 
chloroform to have no inhibiting effect upon the enzym when 
used even in great excess and proved the efficiency of chloro- 
form as a germicidal agent in such work as we were doing. 
At the same time they emphasize the need of painstaking and 
caution if chloroform is relied upon, either to sterilize cul- 
tures or to preserve sterility of enzym solutions as is so fre- 
quently done. 
A comparison of these results with those obtained by others 
will again prove helpful. 
