CULTIVATION OF GLANDERS BACILLUS 309 



but in the chronic nodules, especially when softening has taken 

 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 mor- 

 dant, 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. 105), 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 car- 

 bol-fuchsin. By using a 

 stain of suitable strength 



no decolorising agent is necessary, the film being simply washed 

 in water, dried, and mounted. 



McFadyean recommends that after sections have been stained in 

 Lbffler'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. 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 ayar and ylycerin-ayar in stroke cultures growth appears 



FIG. 90. Glanders bacilli, from a pure 

 culture on glycerin agar. Stained with 

 car bol -fuchsin and partially decolorised 

 to show segmentation of protoplasm. 

 xlOOO. 



