6 
BULLETIN 517, U. S. DEPAETMENT OF AGEICULTUKE. 
as a result, and there remained the alternative conclusion that the 
lest was either nonspecific or that the reacting controls were infected. 
Some of the control birds had been secured in the open market, and 
nothing was known of their previous exposure. Unfortunately con- 
trol fowl 87, which showed a slight reaction on June 23, died frora 
intercurrent causes and in the absence of the authors was not autop- 
sied. As appears in autopsy notes later, control fowls 88 and 95, 
v^dlich also had reacted, were found to be infected birds. 
On August 25 fowls 76, 95, and 96 and a control supposedly nega- 
tive were killed by bleeding and samples held for agglutination test?. 
The ovary of fowl 76 contained angular ova typical of pullorum 
infection, that of fowl 96 dried encapsulated ovum and cysts, not 
certainly due to pullorum infection, and that of fowl 95 one or two 
angular ova and two large cysts. 
Cultures were made from ovaries of fowls 76 and 95 which yielded 
growths characteristic of Bacterium jmllorum^ while that from fowl 
96 yielded a heavy growth not resembling that of Bacterium ful- 
lorum. Fowl 76 gave positive agglutination in 0.01 dilution; fowl 
95 in 0.002 dilution, while fowl 96 was negative at 0.04 dilution. 
An antigen was made from the culture obtained from fowl 96 and 
tested against the serum of fowl 76, but gave a negative agglutina- 
tion at 0.04. The result further strengthens the conclusion that the 
infection in the ovary of fowl 96 was not due to Bacterium fuUorund. 
A check was run on the serum of fowl 76 with Bacillus abortus as 
an antigen, with no agglutination resulting. It is seen that fowl 76, 
[jlthough it did not react well to the intradermal test, gave an agglu- 
tination of 1 : 100. Also, fowl 95 was undoubtedly infected with 
Bacterium^ pullorum. 
With a view of securing a diagnostic agent that would increase 
the size of the swelling in the wattle, the writers next tried a product 
consisting of six strains of B. pullorum. which had been grown in 
plain bouillon at a temperature of 37.5° C. for one month. The 
culture was killed by heating at 60° C. for one hour in a water bath 
and then carbolized to 0.5 per cent. The organisms were not removed 
from the medium, and this product was employed in all subsequent 
tests. 
Table V. — Tests with Jcilled houiUon cultures. 
Fowl No.— 
Edema after 
3 hours, Sept. 2. 
Edema after 24 
hours, Sept. 3. 
Edema 
after 48 
hours, 
Sept. 4. 
Autopsy. 
Culture 
from 
ovary. 
Aggluti- 
nation, 
1 : 100. 
73 
Marked 
Considerable . . 
Slight 
Marked positive... 
Slight 
Positive... 
Trace 
+ 
+ 

+ 
+ 

+ 
77 
+ 
1861 

1 Control. 
