Studies in Forage Poisoning VI. 127 



Agglutination Tests. 



The relation of the anaerobic organism isolated from the 

 ensilage in question to B. botulinus is further suggested by the 

 macroscopic agglutination test. Goat, mule and horse sera 

 highly immune to B. botulinus showed positive agglutinating 

 potency to B. botulinus, as well as to the anaerobic organism 

 resembling B. botulinus isolated from the ensilage, in dilutions 

 varying from 1-100 to 1-2000 (see Charts 6 and 7). Sera drawn 

 from a non-treated horse, sheep, chicken, rabbit and guinea pig 

 failed to agglutinate B. botulinus or the organism isolated from 

 the ensilage, in a dilution of 1-100 (See Charts 6 and 7). Ag- 

 glutination of the organism from the ensilage, in a dilution of 

 1-100, was observed with immune blackleg serum in one in- 

 stance (see Chart 7). 



Horse serum highly immune to the pathogenic bacillus 5 iso- 

 lated from an experimental horse fatally infected from drink- 

 ing an oat hay water, and sheep serum highly immune to the 

 organism isolated from the ensilage in question, showed a posi- 

 tive agglutinating potency to B. botulinus and to the organism 

 isolated from the experimental horse referred to above, as well 

 as to the organism isolated from the ensilage (see Charts 8, 9 

 and 10). 



Serum No. 117 from a horse fatally infected by ingesting 

 2 cc. of an organism resembling B. botulinus, isolated from an oat 

 hay, B. coli immune serum, immune blackleg serum and normal 

 horse serum failed to agglutinate the organism from the ensilage. 

 Serum No. 122 from a mule fatally infected as the result of in- 

 gesting 3 cc. of the sterile nitrate of the organism isolated from 

 the ensilage, completely agglutinated the organism isolated from 

 the ensilage, in a 1-100 dilution (Chart 8). Similar agglutinat- 

 ing potency with the above sera was observed with the organism 

 isolated from the horse that succumbed after drinking the oat 

 hay water (Chart 9), as well as with B. botulinus (Chart 10). 



In conclusion, it is significant that an anaerobic spore-bear- 

 ing bacillus possessing morphological and cultural characters 

 resembling those of B. botulinus was isolated from a silage orig- 



5 Bulletin No. 207. Ky. Exp. Sta., 1917. 



