MORPHOLOGY 483 



parts of the protoplasm remaining unstained : these unstained parts do not 

 however represent spores. 



Sections. For staining the bacilli in sections either Nicolle's tannin method 

 or one of the following may be employed : 



Kuhne's method. 1. On taking the sections 

 out of alcohol, wash them in water and stain 

 for a few minutes with carbol-blue. 



2. Pass the sections rapidly through a 1 per 

 cent, aqueous solution of hydrochloric acid and 

 wash in water. 



3. Dehydrate very quickly in alcohol and 

 aniline oil : wash carefully in xylol and mount 

 in balsam. 



Loeffler's method. 1. Stain for a few minutes 

 in aniline-fuchsin (prepared in a similar manner 

 to aniline-violet) to which 1 part in 10,000 of 



, . -, i i 111 FIG. 235. Film preparation from 



Caustic potash has been added. an infected testicle showing glanders 



2. Wash rapidly in a 1 per cent, solution of ^f 1 - 1 ' Carb i-\ hionin - (Reich ;obj. 



. . -. 4 1T -, . T aiu m. , oc. iv.; 



acetic acid. Wash in water. 



3. Dehydrate rapidly in alcohol and aniline oil, wash carefully in xylol 

 and mount in balsam. 



2. Cultural characteristics. 



Conditions of growth. The glanders bacillus is an aerobic organism ; it 

 hardly grows at all below 25 C., but on glycerin-agar it yields a scanty 

 growth when incubated at 23-24 C. Growth is arrested at 42 C. The 

 optimum temperature for cultivation is 35-38 C. 



Characters of growth on ordinary media. Broth. When sown in broth 

 and incubated at 37 C. for 24 hours the bacillus produces first a cloudiness of 

 the medium and later a white, mucous deposit. A culture on this medium 

 has no characteristic feature. 



Agar. Glycerin-agar. After incubating for about 24 hours a narrow 

 whitish streak is seen along the line of sowing. The culture is at first semi- 

 transparent but as the layer thickens it becomes opaque. On glycerin- 

 agar growth takes place more freely and may spread over the whole surface 

 of the medium. 



Coagulated serum. Horse serum is the best for the growth of the glanders 



bacillus. Semi-transparent colonies appear in 

 about a couple of days which become white 

 and opaque as growth progresses. 



Gelatin. On a 12 or 15 per cent, gelatin 

 which will remain solid at 25 C. a very 

 scanty almost invisible growth is formed after 

 incubating at 25 C. for several days. 



Potato. Under suitable conditions the 

 rowth of the g landers bacillus on potato is 

 characteristic. Potatoes which are either 

 naturally rich in starch or which have been made alkaline should be used if the 

 characteristic appearances are to be developed to the best advantage. After 

 incubating for 48 hours at 37 C. a thick yellowish viscous film appears 

 along the line of sowing, which as the growth extends during the next few 

 days becomes brown and then acquires a chocolate colour, while the potato 

 in the neighbourhood of the growth turns black. 

 Milk. Milk is coagulated in about 10-12 days. 



