CULTIVATION OF GLANDERS BACILLUS. 273 



thionin-blue or weak carbol-fuchsin. By using a stain of 

 suitable strength no decolorising agent is necessary, the film 

 being simply washed in water, dried, and mounted. Gram's 

 method is quite inapplicable, the glanders bacilli rapidly 

 losing the stain in the process. 



Cultivation. (For the methods of separation vide infra.) 

 The glanders bacillus grows readily on most of the ordinary 

 media, but a somewhat high temperature is necessary, 

 growth taking place most rapidly at 35 to 37 C. Though 

 a certain amount of growth occurs down to 21 C., a 

 temperature above 25 C. is always desirable. 



On agar and glycerine agar in stroke cultures growth 

 appears along the line as a uniform streak of greyish-white 

 colour and somewhat transparent appearance, with moist- 

 looking surface, and when touched with a needle is found 

 to be of rather slimy consistence. Later it spreads laterally 

 for some distance, and the layer becomes of slightly brownish 

 tint. On serum the growth is somewhat similar but more 

 transparent, the separate colonies being in the form of 

 round and almost clear drops. In subcultures on these 

 media at the body temperature growth is visible within 

 twenty-four hours, but when fresh cultures are made from 

 the tissues it is not visible till the second day. Serum, 

 however, is much more suitable for cultivating from the 

 tissues than the agar media ; on the latter it is sometimes 

 difficult to obtain growth. 



In broth, growth forms at first a uniform turbidity, but 

 soon settles to the bottom, and after a few days forms a 

 pretty thick flocculent deposit of slimy and somewhat 

 tenacious consistence. 



On potato the glanders bacillus flourishes well and 

 produces a characteristic appearance, incubation at a high 

 temperature, however, being necessary. If inoculation is 

 made to potato from another medium, growth proceeds 

 rapidly, and on the third day has usually formed a trans- 

 parent layer of slightly yellowish tint, like clear honey in 

 appearance. On subsequent days, the growth still extends 

 and becomes darker in colour and more opaque, till about 

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