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



The fourth case was obtained from Mr. de Vine, a specimen of intestine 

 being received at the Institution on January 26, 1911. It showed the 

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

 specific bacilli were present in the lesions. Since the specimen was quite 

 fresh, cultures were made as previously described from the ileum, caecum, 

 and ileo-caecal valve' directly on to nine tubes of the special tubercle 

 bacillus medium ; these were capped and placed at 39° to 40° C. After 

 three weeks' incubation two tubes were found to be contaminated, whilst 

 the remainder were covered with extremely minute colonies of Johne's 

 bacillus without any contaminations; the cultures grew well, and were 

 sub-cultured without any difficulty on to the special medium. Sub-cultures 

 taken on to Dorset's egg medium, glycerine agar, and various other media, 

 gave no growth. 



Case 5 was obtained from Mr. Hamilton, and was received at the 

 Institution on February 8, 1911. The specimen, consisting of ileum and 

 ileo-caecal valve, showed the typical lesions of pseudo-tuberculous enteritis, 

 and a considerable number of acid-fast bacilli were present in the lesions. 

 Cultures were made from several parts of the specimen directly on to 

 12 tubes of Dorset's egg medium. They were taken in the manner already 

 described, but as the specimen was fresh on arrival, previous treatment 

 with ericolin solution was unnecessary. The tubes were capped with gutta- 

 percha tissue, and placed in the incubator at 39° to 40° C. On the 

 following day they were examined and found to be free from contaminating 

 colonies, so the tiny pieces of tissue were removed from three of the tubes 

 and placed on to three tubes of the special tubercle bacillus medium. 

 These were capped and placed with the other tubes in the incubator at 

 39° to 40° C. Six weeks later the three tubes of special medium showed 

 a few tiny colonies of Johne's bacillus. Compared with the previous cases 

 the rapidity of growth was very slow and was slight in amount, due, as 

 was proved later, to the unsuitability of the particular strain of tubercle 

 bacillus incorporated in the medium. Sub-cultures from these tubes on to 

 tubes of a fresh batch of tubercle bacillus medium grew well. All the 

 original cultures on Dorset's egg medium remained sterile, as also did 

 sub-cultures from the special medium on to Dorset's egg medium. 



Four strains of Johne's bacillus having been isolated on media containing 

 dead tubercle bacilli, we next proceeded to test them on slightly modified media. 

 We found that growth was not nearly so good in the absence of glycerine, 

 but the exact percentage most suitable for the growth of Johne's bacillus 

 has not yet been determined, although we have reason to believe that 

 about 4 per cent, by volume gives the best results. A higher per cent. 



