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the work in connection with the investigation of the etiology of beri-beri 
had been carried on by Dr. Kokubo, and here the author also worked for 
three weeks in isolating and examining, by Kokubo’s method, the coccus 
which he and Okata maintained to be the specific micro-organism of 
beri-beri. 
Kokubo had been most successful in isolating this coceus from the urine 
of beri-beri cases. His method was as follows: The ordinary, slightly 
alkaline agar medium was first melted in tubes and then poured into 
Petri dishes. After the agar in the plates was thoroughly hardened and 
dried, it was inoculated from the urine which had previously been collected 
in sterile receptacles, by making streaks over the surface of the media 
with a platinum loop. The Petri dishes were then placed in the incubator 
in an inverted position and examined at first after twenty-four hours and 
then successively from day to day. The author examined the urine from 
about thirty to forty cases of beri-beri and in eight was successful in 
obtaining a coccus which apparently was identical with the one which 
Kokubo believed to be the cause of the disease. 
The agglutination test was regarded as the most significant feature in 
the identification of this coccus. Kokubo, by injecting rabbits, had 
prepared an antiserum which in the hanging-drop test, within one to two 
hours or even sooner, promptly agglutinated this particular organism ina 
dilution of 1-100. However, the author was not successful in obtaining 
this coccus from the blood of patients sick with beri-beri. He was never 
personally allowed to draw any blood from the sick soldiers; but Dr, Ko- 
kubo obtained some from several patients in his presence and inoculated a 
number of tubes with it. Dr. Kokubo’s method of taking the blood was 
as follows: 
An area on the back, over the trapezius muscle was cleansed antiseptically and 
then a puncture was made with a sterile, protected blood lancet. A fold of the 
skin on which the small punctured wound was present was then raised between 
the two fingers of the cleansed hand of the operator and firm pressure applied. A 
considerable amount of blood could thus be obtained for the purpose of inoculating 
a number of tubes and procuring cover-glass preparations. 
It appears that this method is not free from objection, because of the 
fact that it may frequently, and in a number of cases will certainly, lead 
to an admixture of pressed-out lymph with the blood and to the danger of 
contamination by means of micro-organisms inhabiting the sweat or 
sebaceous glands. The collection of blood by direct puncture of the 
median cephalic vein was suggested to Professor Kokubo, but the surgeon 
in charge of the kakke hospital refused to employ this method on his sol- 
dier patients. 
The writer spent his mornings in working in the laboratory of the hos- 
pital, whereas the afternoons were devoted to bedside studies which were 
made while accompanying Dr. Kokubo on his daily rounds, and the latter 
was kind enough successively to demonstrate all of the cases at that time 
