54 HISTORICAL NOTES ON BEE DISEASES. 



spores of Bacillus alvei. When lie inoculated gelatin, agar, or bouillon 

 with some of this ropy larval mass, no growtli took place and when he 

 used larvae which were more recently attacked, the same failure to 

 obtain a growth was observed. He took these facts to mean that 

 there is something present in the foul-brood larvae which prevents, by 

 its antiseptic property, the growth of the bacteria. He proceeded, 

 therefore, to inoculate a considerable quantity of sterile bouillon, in 

 order that the supposed antiseptic present might be diluted, thus per- 

 mitting the germination of the spores. By following tliis technique 

 a growth was obtained and he interpreted it to be the growth of the 

 spores wliich he had observed microscopically in such large numbers 

 in the diseased brood. 



The bacillus wliich he thus obtained he isolated in a similar manner 

 from a large number of samples of "foul brood," and by a study of 

 its morphology and cultural characters Lambotte identified it as 

 being the one described by Cheyne as Bacillus alvei. Furthermore, 

 he studied the morphology and cultural characters of a number of 

 cultures of Bacillus mesentericus, and by a comparison of these with 

 those of Bacillus alvei he reached the conclusion that the two are 

 very similar. 



To show more conclusively that Bacillus alvei and Bacillus mesen- 

 tericus are very similar, Lambotte made use of the phenomenon of 

 agglutination. Guinea pigs were used in his experiments. In 

 immunizing the animals he used a suspension of an agar culture of 

 Bacillus alvei and Bacillus mesentericus, respectively, in physiological 

 salt solution. In each case the animal received four inoculations at 

 weekl}^ intervals. He reports that the serum of an untreated guinea 

 pig did not exhibit, upon examination, the phenomenon of agglutina- 

 tion. The serum of a guinea pig, on the other hand, which had been 

 treated by inoculations with Bacillus alvei, agglutinated a culture of 

 this species at a dilution of 1 to 350, and the serum from the same ani- 

 mal agglutinated cultures of Bacillus mesentericus at a dilution of 1 to 

 250. Furthermore, the serum of a guinea pig that received the inocu- 

 lation of Bacillus mesentericus agglutinated Bacillus mesentericus as 

 well as Bacillus alvei at a dilution of 1 to 250. Neither of these two 

 sera agglutinated any other bacilli at these dilutions. This caused 

 Lambotte to conclude that Bacillus alvei and Bacillus mesentericus 

 vulgaris are the same species. 



Having convinced himself that tliis very intimate relation exists 

 between Bacillus alvei and Bacillus mesentericus vulgaris, he at- 

 tempted to prove by the inoculation of healthy colonies that foul 

 brood could be produced with cultures of Bacillus mesentericus. He 

 killed some of the larvae by pricking them and placing a suspension 

 of Bacillus mesentericus in the cell with the dead larvae. He hoped 



