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beriberi, one from a fatal case of acute pernicious beriberi, and eight from 
cases of subacute or chronic beriberi. 
The first sample was shown to be negative by Mr. Bliss, but in order 
absolutely to check his result, the remaining nine were carefully tested for 
arsenic, the method used having the following main features: From samples 1 
to 9, about 1 gram of each was used for the determination; from the remaining 
one, 0.56 gram was employed. Organic nfatter was destroyed in the usual way, 
the solution then diluted, filtered, and evaporated with sulphurie acid, this 
operation being repeated several times after the addition of a small amount of 
nitric acid, until the fumes of sulphuric acid appeared. After all organic 
matter had been destroyed, the test for arsenic was made in a small Marsh 
apparatus. No trace of arsenic could be detected in any of the samples; a 
control by this method was sufficiently delicate to detect 0.01 milligram. 
The ten samples of hair from beriberi patients on analysis, therefore, 
demonstrated the absence of arsenic. 
ANIMAL EXPERIMENTS WITH THE OKATA-KOKUBO COCCUS. 
During the past six months I have extensively studied the Okata- 
Kokubo coccus and the relation it bears.to beriberi. The results of these 
experiments and of the investigations undertaken to find this identical 
organism in beriberi cases occurring in the Philippine Islands may be 
summarized as follows: 
Six strains were selected for cultural and animal experiments. Three 
were isolated by Professor Okata from beriberi autopsies; one was a 
culture grown by Professor Kokubo from the urine of a kakke patient 
and two were isolated by myself from the urine of kakke patients in 
Hiroshima, in which place I was working under the direction of Professor 
Kokubo. One of the last two cultures exclusively showed cocci of the 
large type. The six strains were grown on the following media: agar, 
alkaline to litmus; agar, alkaline to phenolphthalein; glucose-agar ; 
litmus-lactose-agar; litmus-milk; bouillon alkaline to litmus; bouillon, 
alkaline to phenolphthalein; Dunham’s peptone solution; gelatine; 
potatoes. All the cultures were kept in the incubator between 34° to 36° 
except the gelatine tubes which were maintained at a lower temperature. 
A study of the growth of the different strains on the variety of culture 
media employed, in general confirmed the description given by Okata and 
Kokubo in their first paper. However, a few minor differences were 
noticed. ‘These were as follows: 
Examination of gelatine cultures during the first two weeks showed no lique- 
faction. 
After twenty-two days on this medium the following conditions were noticed: 
Stem Kokubo, heavy growth, superficial liquefaction ; 
Stem Herzog (large cocci), scanty growth, no liquefaction. 
Stem Herzog, very abundant growth, liquefaction. 
Stems Okata, I, IT, and III, scanty growth, no liquefaction. 
Non-inoculated control tube. No liquefaction. 
ne 
