244 GLANDERS. 



ising for a few seconds in water, 10 c.c., to which were 

 added ten drops of a concentrated solution of sulphurous acid 

 and one drop of a 5 per cent solution of oxalic acid. We 

 have, however, obtained the best results by carbol-thionin- 

 blue (p. 98), 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 combinations, 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. 



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., and though a 

 certain amount of growth occurs down to 2 1 C, a tempera- 

 ture 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 sub-cultures 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 which 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. 



