7a. ee Ss eel tlie 
ail [Frw Sot Pee abe 
Hospital after an illness of only a few hours and the necropsy (No. 1634) 
was performed two hours after death. Cultures taken from the very 
much enlarged spleen developed colonies of a pale-yellowish diplococcus 
which, in most media, showed considerable similarity to the cocci brought 
from Japan. From these cultures five rabbits were inoculated. 
Tenth series.—Three of the animals (1747, 1748, 1749) received intraperi- 
toneally 1 cubic centimeter of a 2-day-old bouillon culture, and two (1750 and 
1751) received subcutaneously 1 cubic centimeter of the same culture. These 
animals, which were under observation for two months, never showed any signs 
of disease and remained permanently well. 
In order to prepare an anti-serum for the purpose of making agglutina- 
tion tests, other rabbits were inoculated with the Kokubo coccus in suc- 
cessively increasing doses. All of these animals remained well. One of 
the rabbits, when finally bled, after having been injected at intervals on 
numerous occasions during a period of seventy-eight days, furnished a 
serum of only very low agglutinating power. 
On March 7, 1906, a post-mortem examination (No. 1674) was made upon the 
body of a young Japanese who had been ill but a few days. The clinical diagnosis 
of acute, pernicious beriberi was confirmed at autopsy. During this examination 
two agar tubes were inoculated from the cerebro-spinal fluid obtained by puncture 
at the third lumbar vertebra with a sterile syringe, and another was inoculated 
from the heart’s blood. Pieces of the wall of the stomach and of the duodenum, 
together with some of the gastric contents, were placed in a flask containing 
several hundred cubic centimeters of alkaline bouillon the latter was then 
heated to 80° C. and kept at that temperature for about ten minutes. It was 
then cooled in the refrigerator and subsequently incubated. It developed a 
spore-forming bacillus. The tube inoculated from the heart’s blood remained 
permanently sterile, and the tubes inoculated from the cerebro-spinal fluid 
developed colonies of different bacilli, evidently contaminations. 
Monkeys and rabbits were inoculated on March 26, 1906, with the cultures 
obtained from the above necropsy (No, 1674), after the latter had grown in the 
incubator for nineteen days. Of the bouillon culture 1.5 cubie centimeter was 
injected intraperitoneally, of the agar cultures six normal oesen were rubbed 
up with 20 cubic centimeters of salt solution, and each animal received intra- 
peritoneally 1.5 cubic centimeters of this suspension, as follows: 
Kleventh series.— (March 26.) 
Animal number. | Quantity. Culture. « 
monkey: NO; 1916.22) 222 = eee ee ee ates ain aap caren i 5 From stomach. 
Monkey No. 191772 on eek 2. nt ee ee 1.6 Do. 
Rapti Ng. 1018.3 2-2 ek cece eee | 1.5 Do. 
RADDIDNO: 1010 2e- 080) 2c 2 8es eeaeee ee Be | Do. 
Monkey No. 1920 ___--.------------- “ | 1.5. | Cerebro-spinal 1. 
MonkeyoNo. 102) 222-2 ct ee ee aa Do. 
BROMO NO) P0Rte cece) 20 oo - eos tae eee | 1.5 | Do. 
POURING 00282822 2 ac lel oa eee eee 1.5 | Do. | 
MORRO Y ANG. 1924 ooo 2.. 21. See eee | 1.5 Cerebro-spinal 2. | 
POS ULOV ING ROD oo oT 8 le ee ee 1.5 Do. 
HeDDit NO. 1926 cose. os as Se ee Pod. 6 Do 
yi | Do. 
RUC ING, 1007 22 oa ke le oe oo eae ee 
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