A Bacterial Disease of Alfalfa 17 
ten days growth the cultures were decidedly alkaline. The dex¬ 
trose broth fell from +10 to —3, the saccharose broth from 
+ 8 to —5, the lactose broth from + 8 to —6, the maltose 
broth from + 8 to —5, the glycerine broth from +8 to 
■—4, the mannite broth from + 8 to —7. The results of the 
daily titrations are given in Table No. 1. 
TABLE NO. 1-PRODUCTION OF ACID AND ALKALI. 
Sugar Free Broth 
plus 1$ 
Degree of Reaction After 
No ds 
Id 
2 ds 
3 ds 
4 ds 
5 ds 
6 ds 
7 ds 
8 ds 
9 ds 
10 ds 
Dextrose. 
10 
10 
10 
10 
9 
6 
4 
2 
1 
0 
—3 
Saccharose . 
8 
8 
8 
8 
8 
6 
3 
1 
0 
—4 
-5 
Lactose. 
8 
8 
8 
8 
6 
4 
2 
0 
—1 
—4 
—5 
Maltose.. 
8 
8 
8 
8 
7 
7 
5 
1 
0 
—4 
-5 
Glycerine. 
8 
8 
8 
8 
7 
5 
2 
1 
0 
—3 
—4 
Mannite . 
8 
8 
8 
8 
7 
4 
2 
1 
—1 
—5 
—7 
3 . Production of Ammonia. 
The production of ammonia has been determined in nutrient 
broth, Dunham’s peptone solution, Jordan’s asparagin solution 
and nitrate broth. 100 c.c. portions of nutrient broth and 
Dunham’s solution were placed in 50 0 c.c. Erlenmeyer flasks, 
sterilized, and inoculated with a 4 8 hour agar culture. Simi¬ 
lar portions of each of these media were kept as sterile con¬ 
trols. After ten days, both the cultures and the uninoculated 
checks were analyzed for ammonia by distillation with mag¬ 
nesium oxid. The distillates were collected in N/10 H 2 S0 4 and 
subsequently titrated with N/10 NaOH. Ammonia was produced 
by the culture both in the nutrient broth and in the peptone 
solution, the larger amount being present in the former. The 
results are given in Table No. 2 below. 
TABLE NO. 2—PRODUCTION OF AMMONIA. 
Medium 
Milligrams NH 3 
in 100 c c. of culture 
Milligrams NH 3 
in 100 c. c. uninocu¬ 
lated control 
Milligrams NH 3 
produced by the 
bacteria 
Nutrient Broth.... 
21.63318 
18.39672 
3.23646 
Dunham’s Solution 
.4.00279 
2.81060 
1.19219 . 
Large quantities of ammonia were produced in the asparagin 
solution after ten days. This was easily demonstrated by adding 
one cubic centimeter of Nessler’s solution to inoculated tubes 
of this medium. 
No ammonia was produced in nitrate broth by the reduction 
of the nitrates. 
4. Nitrates in Nitrate Broth. 
Nitrates are not reduced. Tubes of nitrate broth inoculated 
with Ps. medicaginis, n. sp. were tested after five days and 
ten days for nitrites and ammonia, and both were found absent. 
A subsequent test for nitrates showed the original nitrate to 
be present as such. 
5. Indol Production. 
There is no indol produced in Dunham’s peptone solution. 
6 . Relation to Acid and Alkali. 
The organism exhibits much more latitude in the reaction of its 
food stuffs on the acid side of the neutral point than on 
