BACILLUS ANTHEACIS, 17 



DEMONSTRATION VII. 



Bacillus Anthracis in pure cultures — Tissues of animal dead of 



Anthrax Septicaemia — Inoculation of animal with 



Bacillus Anthracis. 



1. Make cover glass preparations from the agar, and 

 the broth cultures of Bacillus anthracis. 



Stain with LoefHer's methylene blue, diluted gentian 

 violet or by Gram's method. 



2. Make cover class preparations from the carbolized 

 broth culture of Bacillus anthracis. 



Stain with Loeffler's methylene blue. No spores 

 are seen. 



3. Inoculate two litmus milk tubes, one from the 

 ordinary broth culture and the second from the car- 

 bolized broth culture of Bacillus anthracis. 



Incubate at 37°C and examine day by day. The 

 latter produces less acid. (By animal experiments it 

 could also be shown to be less virulent). 



Tissues of Animal dead of Anthrax Septicaemia. 

 Sections from lung, liver and kidney are supplied. 



4. Staining with Loeffler's methylene blue. 



(a) Place sections in the stain for 5 to 10 minutes. 

 (6) Remove excess of stain by washing in water, 

 (c) Place in ^ to 1% acetic acid for 10 to 20 seconds, 

 till the sections become a light blue, 



