520 



Messrs. Twort and Ingram. Isolating and [Nov. 7, 



a decoction of grass (Phleum pratense) and glycerine agar a pure culture 

 of an acid-fast bacillus which Koch and Eabinowitsch (17) declared to be 

 identical with the bacillus of avian tubercle. Mettam also, in a private 

 communication, states that he has sub-cultured for 12 generations an acid- 

 fast micro-organism obtained from a cow suffering from Johne's disease, 

 and that it agrees in every respect with the avian tubercle bacillus. These 

 must be regarded as cases of accidental contamination with the avian 

 tubercle bacillus. 



Most authors agree that animals suffering from this disease give no 

 reaction with ordinary diagnostic tuberculin, but 0. Bang (38) has obtained 

 in some cases a more or less decided reaction with avian tuberculin. In 

 this country Male (39), of Eeading, has since used avian tuberculin prepared 

 by Stockman. He tested 19 cattle, giving 5 c.c. doses of tuberculin, and in 

 four cases obtained a reaction of 3*6°, 3°, 4° and 2*8° F. respectively. He 

 does not state, however, whether the pre-inoculation temperature given is 

 that of the morning or evening, and it must be remembered that both 

 Meissner and Mettam have obtained avian tubercle bacilli from the intestine 

 of cows infected with Johne's bacillus. 



In June, 1910, we started some experiments with the object of cultivating 

 Johne's bacillus and of preparing a diagnostic vaccine from the culture 

 obtained. A preliminary note on the results of this work was included in 

 a paper by one of us (41) on the cultivation of the lepra bacillus of man, 

 published in 1910. 



We have to thank Mr. Brennan de Vine, of Birmingham, and Mr. D. 

 Hamilton, of Hamilton, for the pathological specimens used in these 

 experiments. 



Portions of infected intestine and glands were obtained in as fresh a 

 condition as possible, but owing to the time taken in transit, some of the 

 specimens were found to be contaminated in the deeper tissues, and it was 

 found necessary to kill off these contaminations with a solution of ericolin — 

 a method which was originally devised for isolating tubercle bacilli (42). 

 The gut and glands were thoroughly washed in water, and the surface of 

 an infected area seared with a hot spatula ; microscopic films were made 

 from the tissues beneath to prove the presence of the specific bacillus. 

 Small pieces of tissue were then removed with sterile scissors and rubbed 

 over the culture medium to be tested, either directly, or indirectly after 

 being incubated in a 1 per cent, watery solution of ericolin for one to two 

 hours at 37° 0. 



The first case received from Mr. de Vine, on June 15, 1910, showed the 

 typical lesions of pseudo-tuberculous enteritis, and contained a large number 



