22 



PRACTICAL BACTERIOLOGY. 



Examine with the dry and oil-immersion lenses. 

 Anthrax bacilli and nuclei are stained blue, the red 

 corpuscles and eosinophile granules of leucocytes, pink. 



Spores are not seen in the animal body. 



3. Staining by Gram's method and eosin. 



(a) If films have been fixed by heat place in alcohol 

 first for 1 minute; if by formalin place at once in anilin 

 gentian violet for 2 to 3 minutes. 



(6) Rinse in water and transfer to Gram's iodine for 

 1 minute. 



(c) Rinse again in water and then in alcohol till 

 stain ceases to come away. 



(d) Place in eosin, | to 1 minute. 



(e) Wash in water, dry and mount in Canada balsam. 



Plate Cultures. 



4. Make plate cultivations from blood of heart of 

 guinea pig, dead of Anthrax septicaemia. 



(a) Liquefy two or three tubes of gelatine in water 

 bath, or incubator at 37°C. 



(6) Observing all aseptic precautions, with a steril- 

 ized forceps break oflF the fused end of pipette contain- 

 ing blood from the heart, and by carefully applying 

 heat to bulb of pipette, allow one drop of blood to fall 

 into one of the liquefied gelatine tubes. 



Label this tube No. 1. 



(c) From tube No. 1, after careful admixture of the 

 blood drop, transfer three loops to a second liquefied 

 tube (No. 2). If microscopic examination has shown a 



