634 
Journal of Agricultural Research 
Vol. XI, No. 12 
Agar slants. —On peptone-beef agar slants, stroke cultures make a moderate, 
filiform, wax-yellow, opaque, smooth, shining, convex growth. 
Gelatin plates. —Surface colonies show a characteristic margin. This may be 
due to liquefaction, which takes place slowly at i8° C. In about io days liquefaction 
is complete and the colonies float. 
Gelatin stabs. —At 20° C. in +10 peptone gelatin the surface growth is abundant 
after several days with a medium pit of liquefaction. At the end of four days the 
growth becomes stratiform and slowly liquefies its way toward the bottom of the tube. 
It progresses somewhat more than halfway in 40 days. 
Beep bouillon. —In +10 peptone-beef bouillon uniform clouding occurs within 
48 hours. This clouding subsequently becomes rather strong; a membranous pellicle 
forms at about the fourth day and then slow clearing begins, becoming first evident 
just under the pellicle. Sediment is flocculent. Crystals have been noted. 
Potato cylinders. —Growth on steamed potato cylinders in two days is abundant, 
spreading, slimy, flat, glistening, smooth, without odor, and in color wax-yellow with 
a tinge of old-gold. Later the growth becomes very abundant and the elevation 
convex, but no change in medium is noticed at the end of two weeks. 
Milk. —Inoculated milk coagulates slowly and then clears, beginning at the top. 
In a month there is partial digestion, some soft coagulum, and a heavy, watery, yellow 
growth at the top. Reaction showed increase in acidity. 
Litmus milk. —No marked color change is noticeable during the first two weeks, 
although a heavy yellow growth develops at the top. Then reduction begins, and 
in a month the medium is amber-colored. 
Fermentation tubes. —The tests were made in 2 per cent Witte’s peptone water 
and 2 per cent, respectively, of each of the following carbon compounds: Dextrose, 
lactose, saccharose, maltose, glycerin, and mannit. Clouding was first noted at 
end of two days. At end of six days nearly all were clouded in the open end. 
Mannit and glycerin were the slowest to cloud. In all cases there was a definite line 
of demarcation across the inner part of the U and no visible growth in the closed 
arm. No gas was formed in the closed arm in any case. 
When 20 days old, they were titrated with phenolphthalein as indicator, boiling 
to drive off carbon dioxid. In each case there was slight acid production. They 
were similarly tested again when 30 days old, and found to have produced more acid, 
When the test was made without boiling, they were found to be almost neutral to 
phenolphthalein. Both ammonia and acids are produced, the ammonia apparently 
slightly in excess. 
A strong test for ammonia was found by Nessler’s reagent and also by testing the 
gas evolved with red litmus paper when a portion of the culture was heated with 
potassium hydroxid. 
No gas was formed in fermentation tubes containing sterile milk. The milk in the 
open end cleared gradually, followed later by the slow clearing in the closed end. 
The reaction was acid to litmus. 
Nitrate bouillon in fermentation tubes gave a good clouding in open end; none 
in closed end. No gas was produced. There was a strong test for ammonia and 
nitrates. Since the organism produces ammonia from peptone, the test does not 
show whether nitrates were reduced or not. There was a slight production of acid 
when tested after boiling. No nitrites found. 
Toleration of sodium chlorid.— Neutral peptone-beef bouillons containing, 
respectively, 2,5,6, and 7 per cent of chemically pure sodium chlorid were inoculated 
from young bouillon cultures. There was uniform clouding after four days in the 
2 per cent strength. All other strengths inhibited growth. The experiment was 
repeated, using 2, 3, and 4 per cent of sodium chlorid. Growth occurred in the 2 per 
cent strength only, and in this the organism developed long chains and soon died. 
