VACCINES : DIAGNOSTIC AND CURATIVE 125 



In the light of the results obtained with the vaccines 

 described above, we now prepared a sixth batch. 



Vaccine No. 6. — This was made by growing Johne's 

 bacillus on ordinary glycerine-peptone-beef broth con- 

 taining a glycerine -saline extract of B. phlei (the 

 timothy-grass bacillus). This culture was grown for 

 nine months at 39° C. ; the whole was well shaken to 

 form an emulsion of the bacilli, placed unfiltered into 

 small phials, and heated for one hour at 62° C. The 

 vaccine was first tested on three fully-grown bovines, 

 and on five calves about seven months old, and was 

 inoculated intravenously in doses varying, according to 

 the size of the animal, from 5 to 10 c.c. 



No. I, a Jersey bull, No. 2, a shorthorn cow, and 

 No. 3, a Jersey cow, all showed clinically the typical 

 manifestations of advanced Johne's disease, which they 

 had contracted naturally. Before we obtained these 

 animals, they had been tested several times with 

 diagnostic tuberculin with negative results, though the 

 bull had been in our possession for over eighteen 

 months, and had not been tested for tuberculosis 

 during this period. Each animal received 10 c.c. of 

 the special Vaccine No. 6. No. i gave a maximum 

 temperature of 105° F., which was reached an hour 

 after the inoculation. No. 2 gave a maximum tempera- 

 ture of io6-i° F.five hours after inoculation, and on the 

 following day developed a violent diarrhoea, which 

 persisted after the temperature had fallen. The faeces 

 contained blood and mucus. No. 3 gave a maximum 

 temperature of 104*8° F. This was reached in four 

 hours, and the temperature was not taken again. 



Bovine No. i developed symptoms of tuberculosis, 

 so was killed (October 17, 1912). Post-mortem examina- 

 tion revealed typical Johne's disease in the intestine 

 and mesenteric glands. The disease was in a very 



