1911.] Cultivating the Mycobacterium enteritidis, etc. 



523 



bacillus medium. The tubes were capped with gutta-percha tissue and 

 incubated at 39° to 40° C. After two days' incubation all the direct 

 cultures were badly contaminated, yet those inoculated with ericolinised 

 material showed only a few contaminating colonies. Sub-cultures were 

 made from uncontaminated areas of most of the latter tubes on to fresh 

 tubes of the same medium, but, owing to the small amount of the tubercle 

 bacillus medium then prepared, only one of these tubes was sub-cultured 

 — this was one made from a gland. Films from these sub-cultures were 

 prepared at intervals of about four or five days, and examined micro- 

 scopically. After 19 days the sub-culture on the special medium showed 

 quite definite evidence of multiplication, the bacilli had grown larger and 

 thicker, they were well stained, and were present in large close masses. 

 Sub-cultures were made from this tube on fresh tubes of various media, 

 including one tube of the special tubercle bacillus medium. These were 

 examined at intervals as before, and the sub-culture on the special medium 

 showed microscopic evidence of growth in 10 days. Both the first and 

 second sub-cultures showed growth visible to the naked eye after four weeks, 

 which gradually increased, and reached a maximum in about eight weeks. 



These tubes were easily sub-cultured on to fresh tubes of the same medium, 

 but on none of the ordinary laboratory media were we able to get any evidence 

 of growth. 



The third case of pseudo-tuberculous enteritis was obtained from 

 Mr. Hamilton. Specimens of intestine, but no glands, were received on 

 September 23, 1910. They showed the typical lesions of the disease, and a 

 very large number of Johne's bacilli were present in the tissues. "When 

 delivered, the specimens had already commenced to decompose, but from 

 them cultures were made as previously described, both directly, and 

 indirectly after treatment with ericolin solution, on various media, including 

 some tubes of the tubercle bacillus medium. The tubes were capped with 

 gutta-percha tissue and placed in an incubator at 39° to 40° C. The results 

 were the same as in Case No. 2 ; all the direct cultures were badly con- 

 taminated, and those tubes which had been inoculated with material 

 previously treated with ericolin solution grew only a few contaminating 

 colonies. Of the latter, the cultures on the tubes of special medium were 

 sub-cultured from uncontaminated areas on to a number of fresh tubes 

 of various media, including the special medium. The sub-cultures on the 

 ordinary media remained sterile, but those on the tubercle bacillus medium 

 grew Johne's bacillus in pure growth, and were, without difficulty, repeatedly 

 sub-cultured on to fresh tubes of the special medium. Naked eye evidence 

 of growth was present in the first sub-cultures after about six weeks. 



