294 BACTERIOLOGY. 



purpose a pure culture of the anthrax bacillus is injected 

 into the dorsal lymph sac of a frog, and at the of 12 or 18 

 hours the animal is killed with chloroform. 



Cover-glass preparations should be made with the fluid 

 in the dorsal lymph sac and stained, some with simple 

 anilin dyes and others by Gram's method. 



Animal inoculations. — A pure culture of the anthrax 

 bacillus, isolated from the dead animal, can be used to in- 

 oculate a white mouse, a white rat or a rabbit. In the 

 animal thus inoculated the organism can in turn be detected 

 and isolated. The rules of Koch can be easily demonstrated 

 with reference to the anthrax bacillus, thus showing conclu- 

 sively that it is the cause of the disease. 



Summary of Laboratory Work with Anthrax. 



From the guinea-pig' make : 

 Gelatin plates. 



Colonies. — Impression preparations. 

 Stab culture in gelatin. 

 Ag'ar streak culture. 

 Peptonless agar streak. 

 Potato tube culture. 

 Potato tube at 15°. 

 Bouillon tube culture. 

 Agar plates at 37°.— Impression preparations. 

 Hanging-drop culture of heart-blood. — Permanent mounts. 

 Streak preparations of blood, spleen, liver, kidney, lung, stained 

 by simple and by Gram's method. 

 Inoculate a frog with a pure culture of anthrax and examine the 



lymph fluid by the simple and by Gram's method .for phagocytes. 

 Simple and double staining of spores, developed on peptonless agar 



and on potato at 37°. 

 Involution forms on potato at 15°. 

 Sketch characteristic growths and forms. 



Inoculation of other susceptible animals, thus proving that this organ- 

 ism is the cause of a disease. 



Examine each of these 

 >in the hanging-drop, and 

 make permanent mounts. 



