164 Report oF DEPARTMENT OF BACTERIOLOGY OF THE 
5d. Neutral beef broth; the standard medium used in our 
work and one upon which the organism makes a good growth. 
, 4. The same plus 2% cane sugar; a medium in which the 
growth is more rapid than in plain broth. 
5. Cooked carrot broths. Two kinds have been used: 
(a) Those in which equal weights of pieces of fresh carrot 
roots and of water were cooked and sterilized together by the 
fractional process in the steamer. b 
(b) The same in which, after the first cooking in the steamer, 
the roots were crushed and the liquid. expressed and filtered 
through several thicknesses of paper to remove all of the 
cell-wall substance, then this filtrate returned to the flask 
and sterilized in the steamer by the fractional process. Both 
of these have, except in certain cases discussed below, proved 
to be the best of cooked media for this organism. 
6. Living vegetables. Fresh living roots of carrot and tur- 
nip are both quickly invaded and rotted by this organism, 
furnishing apparently ideal nutritive conditions. The ex- 
pressed juice from such recently decayed vegetables‘ was used 
in comparison with the preceding broth cultures 1-5. 
The results were in general determined by comparing on 
razor sections of carrot and turnip roots the cytolytic action 
of like solutions of the alcoholic precipitates obtained from 
the above culture liquids. ve 
The sixth method, using fresh uncooked vegetable, has given 
the most active enzym product as well as the largest amount 
thereof. In comparison with beef broth, which yields on an 
average about 0.25% of dry precipitate (i. e., 0.5 g. of dry pre- 
cipitate from a 200 c.c. broth culture), this expressed juice 
from decayed turnip after filtration through paper has yielded 
over 0.5% of a precipitate, a 5% aqueous solution of which 
7These were, of course, so handled as to insure pure growths of the 
organism. This was most surely and satisfactorily done by taking several 
roots, washing thoroughly, soaking tw enty minutes in 0. 1% solution of 
corrosive sublimate, rinsing in sterile water, then with precautions against 
contamination removing the surface to a depth of 0.5 to 1 cm., cutting 
thick pteces from the interior tissues and laying in sterile petri dishes. 
Inoculations on the surface of these at temperature 20°—24.° secured their 
decay in two to three days. 
