DIAGNOSIS OF INFECTION WITH B. PULLORUM. 17 



hens were cystic, exhibiting cysts of varying sizes, and the color was 

 from normal yellow to grayish green. Hen No. 5 showed the least jxitho- 

 logical condition of the ovaries. Many ova here were quite normal, and 

 exhibited the usual high-colored picture so characteristic of a healthy 

 ovary. Material from hens Nos. 1, 5, 6, 792, 464, 13 and 452 (previously 

 examined) yielded cultures of Bacterium pullorum. Hens Nos. 34, 315, 

 49, 312, GO, 22, 77, 4, 35, 618 and 46, whose sera had never agglutinated, and 

 in whose eggs Bacterium pullorum had not been found, were autopsied. 

 All ova were normal except that of 77, which showed grossly congestion. 

 More than 100 agar tubes were streaked from these ovaries and from 

 materials crushed in sterile bouillon and incubated, and all gave nega- 

 tive results for B. pullorum. Hen No. 53, a positive reactor, and hen No. 

 46, a non-reactor, were killed accidentally, so data on ovaries from these 

 hens were not obtained. The pathological findings of the birds that ag- 

 glutinated correspond well with those previously described, and especially 

 with previous observations by the author in 1910,^ and also substantiate 

 the work of Jones. ^ All ovaries from these birds exhibited one or more 

 retention cysts and several irregular lobulated cysts, and the color varied 

 from shades of yellow to green. 



Specificity of Bacterium Pullorum Agglutinin. 



The first recognition of the agglutination reaction as a separate function 

 of immune sera was by Gruber and Durham.^ From the first these in- 

 vestigators had claimed specificity for the agglutination reaction, and 

 for this reason it was utilized by Widal for the diagnosis of typhoid fever. 

 Even by early workers it was observed that serum of animals immunized 

 against one micro-organism would often agglutinate to a much less marked 

 degree other closely related species. The serum of a typhoid-immune 

 animal maj^ agglutinate the typhoid bacillus in dilutions of 1-1,000 and 

 higher, and B. coli in dilutions as high as 1-200. The normal agglutinative 

 power of B. coli does not exceed 1-20. Therefore the specificity of the 

 reaction for practical purposes is not destroyed if the proper dilutions 

 are carried out, the degree or amount of agglutinin formation being always 

 far higher for the specific organism causing the formation of the aggluti- 

 nin than for closely related species. 



After carrying out 300 tests with normal sera from birds known to have 

 no infection, we feel justified in stating that in some instances we were 

 able to obtain slight agglutinative reactions in dilutions of 1-25, but in 

 no instance was there ever exhibited the slightest sign of the agglutination 

 of Bacterium pullortnn when dilutions 1-100 of normal serum from non- 



1 Gage, Geo. Edward: " Notes on ovarian infection with Bacterium pullorum (Rettger) in 

 the domestic fowl." Journal Medical Research, Vol. XXIV., No. 3; N. S., Vol. XIX., No. 3; 

 June, 1911, pp. 491-496. 



2 Jones, F. S.: " The value of the macroscopic agglutination test in detecting fowls that are 

 harboring Bad. pullorum." Journal Medical Research, Vol. XXVII., No. 4; N. S., Vol. XXII., 

 No. 4, pp. 485-495. 



' Gruber and Durham: Miinch med. Woch, 1906. 



