CULTIVATION OF GLANDERS BACILLUS 319 



place, they are few in number, and it may be impossible to find 

 any in sections. 



Staining. The glanders bacillus differs widely from the 

 tubercle bacillus in its staining reactions. It stains with simple 

 watery solutions of the basic stains, but somewhat faintly (better 

 when an alkali or a mordant, such as carbolic acid, is added), and 

 even when deeply stained it readily loses the colour when a 

 decolorising agent such as alcohol is applied. We have obtained 

 the best results by carbol-thionin-blue (p. 106), and we prefer to 

 dehydrate by the aniline-oil method. In film preparations of 

 fresh glanders nodules 

 the bacilli can be readily 

 found by staining with 

 any of the ordinary com- 

 binations, e.g., carbol- 

 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. 



FIG. 94. Glanders bacilli, from a pure 

 culture on glycerin agar. 



Stained with carbol-fuchsin and partially 

 decolorised to show segmentation of pro- 

 toplasm, x 1000. 



McFadyean recommends 

 that after sections have 

 been stained in Loffler's 

 methylene-blue and slightly 

 decolorised in weak acetic 

 acid, they should be treated 



for fifteen minutes with a saturated solution of tannic acid ; thereafter 

 they are washed thoroughly in water, and as a contrast stain a 1 per 

 cent, solution of acid fuchsin may be applied for half a minute ; they 

 are then dehydrated, cleared, and mounted. 



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 glycerin 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, 



