272 Experiments on Anthrax at the Brown Institution. 
and whether, by inoculation, the grains might be made the 
means of conveying the poison. For this purpose a small 
quantity (about 1 lb.) of the grains was thoroughly mixed with 
one pint of distilled water and allowed to stand for two hours. 
The liquid thus obtained was filtered and then boiled in a flask, 
the neck of which was plugged with cotton wool while cooling. 
The cultivation was conducted in glass tubes about two 
inches in length, capable of holding about 1 c.c, the ends of 
which were drawn out to capillary points. They were filled by 
breaking off one end under the surface of the liquid. When 
filled, they were at once inoculated by passing in a droplet of 
anthrax blood by means of a fine capillary tube, or by introducing 
a particle of dried anthrax blood on the point of a fine needle. 
The end was then sealed up, and the tubes were kept for periods 
varying from two to five days, at a temperature of 36° C. In 
all these cultivation-experiments a tube containing only the 
cultivating fluid was placed in the incubator* with the inoculated 
tubes. 
In the first experiment, at the end of sixty hours, fluid from 
one of these inoculated tubes was used to inoculate two guinea- 
pigs, both of which died in less than forty-eight hours ; and 
after death the spleen was seen to be large and tense, and the 
blood was full of bacilli. 
As a check-experiment, the grains' infusion to which no anthrax 
blood had been added was injected under the skin of another 
guinea-pig, the result being only local swelling and the forma- 
tion of a small abscess. 
A portion of the active cultivated liquid with which the two 
guinea-pigs were inoculated was added to some freshly-prepared 
grains' infusion, and the same precautions taken as in the former 
cultivation, and at the end of three days this second generation 
of bacilli was found active. 
These cultivations were repeated and carried to the third 
generation from the original inoculation with the anthrax blood, 
and, when injected into the cellular tissue of an animal, they 
caused death in about the same time as if anthrax blood had been 
injected, and after death bacilli were found in the blood of all 
parts of the body, but most abundantly in the spleen. 
Freshly-prepared hay infusion was also used as a cultivating 
medium, and it was found that the anthrax bacillus could be 
cultivated in that fluid to the third generation and retain all its 
virulence, as was proved by animals inoculated with it dying, with 
* The incubator is a chamber in which a teinperaturo a little below blood heat 
is maintained. Aqueous humour is the liquid contained in the anterior chamber 
of the eye. It ia suitable as a soil fur the cultivation of the anthrax virus, because 
it closely resembles in character the liquid part of the blood. 
