present in the hospital, the patients who were more seriously and more 
acutely affected being repeatedly seen and examined. It was impossible 
to see more than one post-mortem examination; this oceurred in the 
service of Surgeon Lieutenant-Colonel Shimada.” Surgeon-General Okata 
was also present at this autopsy and he subsequently informed the writer 
that from the kidneys and the cerebro-spinal fluid he had succeeded in 
isolating from this case the coccus which Kokubo and he had previously 
obtained inter vitam from the urine and from the blood of kakke cases. 
In September, 1905, Okata and Kokubo published their first prelimi- 
nary report on their beri-beri investigation, it appearing in the Journal 
of the Military Surgical Association, printed in Japanese. Surgeon- 
General Okata, who is professor of bacteriology in the Military Medical 
School at Tokyo, was kind enough to furnish the writer with a copy." The 
following extract includes all of its main features : 
REPORT ON THE OKATA-KOKUBO BERI-BERI COCCUS. 
On examining the blood of beri-beri patients we sometimes find cocci in micro- 
scopical preparations; these are oceasionally met with intracorpuscularly and 
sometimes outside the corpuscles. They generally appear as diplocoeci, but are 
also seen as individuals. Occasionally they are observed in the form of staphy- 
lococci. These cocci do not stain uniformly, but show an uncolored slit in the 
center. They are not very numerous in general, there being only one or two 
observed in the field. These cocci have no capsule and are not motile. 
For the purpose of obtaining cultures from beri-beri cases, the blood is collected 
as follows: (1) The region over the trapezius is cleaned with soap and water, 
(2) washed with bichloride solution, (3) with physiological salt solution, (4) with 
distilled water, (5) with sterile alcohol, and (6) finally with sterile water. The 
area is then punctured with a sterile lancet and the flowing blood is utilized to 
inoculate a number of tubes and to make some cover-glass preparations. 
The number of patients examined by this method was 129. We had 65 cases in 
which both microscopical cover-glass examinations and cultures gave positive 
results. In 34 cases both were negative, in 11 eases the microscopical examina- 
tion was positive and the cultures negative, and in 19 eases the microscopical 
examination was negative and the cultures were positive. 
Staining —All the aniline stains, color the coccus deeply. It is well stained 
with Loeffler’s alkaline methylene blue, and still better: by the method of Semeno- 
wiez and Marzinowsky* (a combination of the staining solutions of Ziehl and 
Loeffler ) . 
Culture.—All ordinary agar media may be used. The most suitable temperature 
is 30° to 37° C. When the cultures are kept at this temperature a growth, visible 
to the naked eye, develops after eighteen hours. At room temperature (10° C.) 
a very slight growth is observed after three weeks. When bouillon tubes are 
inoculated and placed in the incubator, a slight turbidity shows itself after fifteen 
hours. After twenty-four hours a grayish-white sediment forms at the bottom 
?In the Japanese military service, autopsies can be held only after special per- 
mission from the family of the dead person is received. This permission is only 
very rarely given, 
* For a translation of this report the author is indebted to Mr, ©. J. Arnell, of 
this Bureau. . 
‘Centralblatt fiir Bakteriologie (1897), 21, 874. 
