6 



BULLETIN 517, U. S. DEPARTMENT OF AGRICULTURE. 



as a result, and there reniained 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 from 

 intercurrent causes and in the absence of the authors was not autop- 

 sied. As appears in autopsy notes later, control fowls 88 and 95, 

 vvhich 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 tests. 

 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 jmlloruTn, while that from fowl 

 96 yielded a heavy growth not resembling that of Bacterium pul- 

 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 fuUor-um. 

 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, 

 although 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. — T(sst8 with Icillcd bouillon 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 1 Marked positive . . . 



Positive... 

 Trace 



+ 

 4- 

 



+ 

 + 

 



-1- 



77 



Considerable . . 

 Slight 



Slight 



+ 



186' 

















» Control. 



