CULTIVATION OF THE BACILLUS 71 



obtained was ground in a mortar with glycerine and 

 saline, steamed for half an hour, and added to the white 

 and yolk of new-laid eggs in the following proportions : 

 ^SSt 75 parts; oS per cent, sodium chloride in re- 

 distilled water, 25 parts. These were thoroughly 

 mixed, and to the mixture were added tubercle bacilli 

 I per cent, and glycerine, 5 per cent. The medium was 

 placed in sterile test-tubes; these were plugged with 

 cotton-wool, and heated in a hot-water bath at 60° C. 

 for one hour on three successive days, the tubes being 

 incubated at 37° C. in the intervals between steaming. 

 Finally the tubes were inspissated in slopes at 85° 

 to 90° C. 



A second case of pseudo-tuberculous enteritis was 

 now obtained from Mr. De Vine. Specimens of in- 

 testine and glands were received on July 28, 1910. 

 Both the intestine and glands showed the typical 

 characters of the condition, and large numbers of 

 Johne's bacilli were present in various parts of the 

 tissues. Unfortunately, owing to the hot weather pre- 

 vailing at the time, the specimens on delivery had 

 commenced to decompose; but, in spite of this, we 

 prepared some cultures in the manner previously 

 described, both directly, and indirectly after treating 

 with ericolin solution. The cultures were made on 

 several of the media tested with the first cases, as well 

 as on a number of tubes of the special tubercle bacillus 

 medium. The tubes were capped with gutta-percha 

 tissue, and incubated at 39° to 40° C. After two days' 

 incubation all the direct tubes were badly contami- 

 nated, 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 



