1911.] Cultivating the Mycobacterium enteritidis, etc. 



530 



4. The specific bacillus will grow on media containing the dried and 

 powdered growth of certain acid-fast bacilli which have been previously 

 killed, and this is so even when the dead bacilli have been kept for a period 

 of eight years, and subjected to a temperature of 115° C. in the autoclave for 

 1 hour. 



5. The most suitable bacillus to incorporate in the medium is the timothy- 

 grass bacillus, and to a somewhat less degree the smegma bacillus of Moeller 

 and the nasenschleim bacillus of Kaiiinski. The human type of tubercle 

 bacillus is also good, but on media containing the avian type Johne's bacillus 

 grows very slightly, if at all. With the few bovine strains tested in media 

 we were unable to get any definite evidence of growth with Johne's bacillus. 

 Tubercle bacilli isolated from cats also gave negative results. 



6. The essential substance or substances necessary for the growth of 

 Johne's bacillus can be extracted from the various acid-fast bacilli which 

 give positive results by means of hot ethyl alcohol. 



7. We have isolated Johne's bacillus from five consecutive cases of pseudo- 

 tuberculous enteritis, and have proved the morphological and biological 

 characters of the bacilli isolated to be identical in every respect. 



8. The bacilli isolated produce no lesions in mice, rats, guinea-pigs, 

 rabbits, pigeons, or hens, if given by the mouth or inoculated into the 

 peritoneal cavity or into a vein or subcutaneously. 



9. The specific bacillus, when inoculated intravenously or given by the 

 mouth to bovines, reproduces pseudo- tuberculous enteritis in the animal, 

 and this cannot be distinguished from the original disease either clinically 

 during life or post mortem. Further, the bacillus can be recovered from the 

 lesions in the intestine of the inoculated animal, and shows characters in 

 every way identical with the bacilli isolated from the original cases. 



10. Animals suffering from pseudo-tuberculous enteritis, either normally 

 contracted or experimentally produced by the inoculation of pure cultures of 

 Johne's bacillus, give no definite reaction with diagnostic vaccines prepared 

 from cultures of the timothy-grass bacillus or from the avian tubercle 

 bacillus. 



11. Vaccines can be prepared from cultures of Johne's bacillus similar to 

 those prepared from other acid-fast bacilli. 



12. Diagnostic vaccines prepared from cultures of Johne's bacillus grown 

 on tubercle bacillus medium gave positive reactions with tubercular animals, 

 which proved the medium used to be unsuitable for the preparation of a 

 specific diagnostic vaccine for pseudo-tuberculous enteritis. 



13. Vaccines prepared from cultures of Johne's bacillus on a timothy- 

 grass bacillus medium gave negative reactions with normal and with 



