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8. Post-mortem upon the guinea pig inoculated at the previous period. 
Carefully observe the method of procedure in the post-mortem examination 
of animals and the location of the various internal organs. 
9. With the sterile platinum loop inoculate an agar slope from the heart 
blood. Place it in the incubator for 48 hours. Make the culture at the post¬ 
mortem table, not at the desk. Make a similar culture from heart blood of a 
healthy pig killed by chloroform. 
10 Clean four slides very clean. Label two heart blood, one spleen, and one 
liver. 
11. At the post-mortem table make as thin smears as possible with the loop 
from the heart blood, spleen, and liver of the animal. Be sure the loop is 
sterilized after each operation and handle the slides with care. Make a slide 
from the heart blood of a healthy animal. 
12. Permit the smears to air-dry. When perfectly dry, fix the smears of 
liver and spleen by heating in the usual way. Stain the liver and spleen 
smears by Gram’s method. 
13. Fix the heart blood smear by standing it in methyl alcohol from 10 
to 15 minutes. 
14. Transfer without washing to eosin for 5 or 10 minutes. Wash and blot 
dry. 
15. Stain in aqueous methylene blue until the smear has a lavender or rose 
tint, usually about 3 to 5 minutes. Wash and dry. 
16. Examine first with the 16 m. m. objective until a good field is found. 
17. Examine with the oil-immersion objective. The red blood cells will be 
red. The nuclei of the white blood cells will be blue and their cyptoplasm will 
be pink with red granules. The anthrax bacilli will be blue and the capsules 
will appear clear as if unstained. 
18. Draw and describe a good field in the above preparation. 
19. Examine the agar slope culture made from the heart blood. Draw and 
describe it. Is there any growth on the slopes inoculated from the blood of the 
healthy animal? 
20. Prepare a slide from the above culture and stain it with carbol fuchsin. 
Draw and describe the organisms as seen under the oil-immersion objective. 
21. Examine, draw and describe the organisms as seen in stained sections of 
tissue. 
22. Leave all of the cultures in a tumbler on the desk in order that 
they may be sterilized. 
23. 
(a) On what kind of media, liquid or solid, do spores form most 
rapidly and why? 
(b ) There are many organisms in soil and in the dust of the air 
that grow like anthrax and appear very much like anthrax in 
cultures? How could you prove such a culture was not 
anthrax? 
(c) Why does anthrax produce a growth like a tuft of cotton in 
bouillon? 
( d ) Why is it so difficult to eliminate anthrax from an infected 
farm? 
( e ) What is the best method to prove positively that anthrax is 
the cause of the death of an animal? 
