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



media by the tubercle bacillus, but not by the bacillus of pseudo-tuberculous 

 enteritis — in other words, that the latter bacillus has lived a pathogenic 

 existence from such remote ages, that it has lost the original power of its 

 wild ancestor — whatever bacillus that may have been — and can no longer 

 build up all its necessary foodstuffs outside the animal body. 



It was thought probable that if these substances could be obtained ready 

 formed and added to some good artificial medium (Dorset's egg medium), the 

 bacillus would grow, and further, that these substances might be elaborated 

 by allied micro-organisms, such as the tubercle bacillus, and even stored up 

 as reserves in their envelopes. On this reasoning, which led to the successful 

 cultivation of the lepra bacillus of man (41), we decided to prepare media 

 containing these allied bacilli which had been killed by heat. 



We had ab the time in our possession about three hundred strains of 

 tubercle bacilli, mostly isolated from human tuberculous material on Dorset's 

 egg medium. A number of these cultures were taken, and after the neces- 

 sary sub-cultures had been made, they were killed by steam. The growth 

 was then scraped off, taking care to avoid any admixture of the medium 

 which might contain the waste products of the bacillary growth and be toxic 

 to the bacillus of pseudo-tuberculous enteritis. More recently we have 

 found this precaution to be unnecessary. The growth of tubercle bacilli 

 thus obtained was ground in a mortar with glycerine and saline, steamed for 

 half an hour, and added to the yolk and white of new laid eggs in the 

 following proportions : Egg 75 parts, 0"8 sodium chloride in re-distilled 

 water, 25 parts; these were thoroughly mixed, and to the mixture were 

 added tubercle bacilli 1 per cent, and glycerine 5 per cent. This 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. for 6 to 12 hours 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 intestine and glands were received on 

 July 28, 1910. Both the intestine and glands showed the typical 

 characters of the condition, and a large number of Johne's bacilli were 

 present in various parts of the tissues. Unfortunately, owing to the hot 

 weather prevailing 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 case, as well as on a number of tubes of the special tubercle 



