152 REporRT OF DEPARTMENT OF BACTERIOLOGY OF THD 
of phenol (about 0.5 g., i. e., 5%). Two other tubes were in- 
cluded in the series; one of these, No. 5, contained sterile 
broth in which the organism had grown for seven days and 
which had then been rendered sterile by passage through the 
Pasteur-Chamberland filter. The other, No. 6, contained a 
living culture of the organism, five days old. Three days later 
transfers made from each tube except the last proved sterility. 
On the same day a sterile cube cut from living carrot root was 
inserted into each of these tubes. Forty-eight hours there- 
after the carrot tissue in tubes 1, 1’, 38, 3’, 4, 4 and 6 were 
alike well softened, there being no evidence of inhibition by 
any of these chemicals and no greater softening in the presence 
of the living organisms than in these sterile tubes. The carrot 
in the filtrate (5) was considerably less acted upon and for- 
malin (2 & 2’) showed still less softening. Further examina- 
tion showed full softening in these latter tubes (2, 2’ & 5) 
at the end of nine days. Transfers at the close of the ex- 
periment proved continued sterility in all the tubes except 
one of those containing chloroform. 
These confirmed the evidence from previous trials and led 
us to conclude that neither chloroform, thymol nor phenol 
had any inhibiting effect on the enzym; that well developed 
broth cultures sterilized by the addition of any of these pos- 
sessed as active cytolytic properties as that in which the 
organisms continued alive; that formalin inhibited the en- 
zymic activity; that filtration through porcelain reduced the 
enzym content. 
To preclude the possibility of error because of the softening 
action upon the vegetable tissues of the broth itself, or of any 
of these chemicals, a series of control tubes was held in which 
carrot blocks were immersed in sterile broth without any 
added chemical and in similar ones in which the various chem 
icals were added in the amounts indicated in the above experi- 
ments. These carrot tissues in all cases remained unsoftened. 
SECURING THE ENZYM BY DIFFUSION. | 
Our observations upon decaying vegetables have shown that 
the cell walls are affected some distance in advance of the 
invasion of the organisms. This would indicate the diffusion 
