26 
Colorado Experiment Station 
nesia. The asparagin solution yielded considerably more ammonia 
than the nutrient bouillon. The results follow: 
TABLE II 
Ammonia Production 
Nutrient solution 
Milligrams N as 
NH, in 100 C.C. 
culture after 5 
days 
Milligrams N as 
NH, in 100 C.C. 
sterile culture 
solution 
Milligrams N as 
NHj produced by 
the micro¬ 
organisms 
Nutrient bouillon... 
15.1308 
15.2709 
9.03645 
6.09435 
6.23445 
Asparagin solution.. 
27.4596 
29.0707 
0.9807 • 
26.47890 
28.09000 
3. NITRATES IN NITRATE BROTH.— Nitrates are not reduced 
after five days either at 20° C. or 28° C. 
4. INDOL PRODUCTION.— Neither indol nor the cholera red 
reaction is produced in Dunham’s peptone solution after 10 days 
at 20° C. 
5. ACID production FROM SUGARS AND GLYCERINE. -One 
hundred c.c. portions of +10 sugar free broth to which 1% of 
the different fermentable substances was added were inoculated 
with two loops of 24-hour broth culture and kept at 20° C. Five 
c.c. were taken from each flask with a sterile pipette daily and the 
reaction to phenolphthaiein determiined with n/io NaOH or n/io 
HCl. The titrations were made by adding the 5 c.c. of the sample 
to 45 c.c. of hot, freshly boiled distilled water, rather than by boil¬ 
ing the sample with the water. This was done in order to avoid 
the loss of any volatile acid that might be present. At the end 
of ten days there was a slight increase in the acidity of the broths 
containing saccharose, galactose and dextrose, from +10 to +13, 
altho in all probability this does not begin to represent the actual 
amount of acid produced, since considerable quantities must have 
been neutralized by the ammonia which we know is formed. The 
remaining broths were more alkaline at the end of the ten days than 
at the beginning. The detailed titrations are given in Table III. 
TABLE III 
Production of Acid from Sugars and Glycerine 
250 c.c. Erlenmeyer flasks 
containing 100 c.c. sugar- 
free broth and 1 percent 
Check . . . 
Dextrose 
Galactose 
Saccharose 
Mannite . 
Laevulose 
Maltose . 
Inulin . .. 
Lactose . . 
Glycerine 
Reaction in degrees Fuller’s scale after 
given number of days 
0 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
10 
9 
7 
6 
5 
5 
5 
4 
1 
10 
10 
10 
10 
9 
9 
9 
9 
11 
13 
10 
10 
10 
9 
9 
10 
11 
11 
12 
13 
10 
10 
10 
8 
8 
9 
9 
11 
13 
12 
10 
8 
8 
8 
6 
5 
4 
4 
4 
2 
10 
8 
8 
8 
6 
6 
5 
4 
4 
4 
10 
8 
8 
6 
6 
5 
5 
5 
3 
_2 
10 
8 
6 
6 
6 
6 
5 
5 
3 
3 
10 
10 
10 
7 
6 
6 
5 
5 
5 
1 
10 
8 
8 
8 
5 
5 
4 
4 
4 
4 
10 
—2 
13 
13 
13 
—5 
O C<J CC 
