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ON THE PRESENCE OF MICRO-ORGANISMS IN THE BLOOD STREAM OF BERI-BERICS.
Previous workers on this subject have described micro-organisms which were formed in cultures made direct from the blood. The organisms described by these investigators were usually of the nature of cocci and rod-shaped bacteria. The cocci were often of the chromogenic variety, and so far as one can gather from the descriptions given of such organisms, they would appear to differ but slightly, indeed if at all, from the micro-organisms found ubiquitous in nature, and in the deeper folds and sebaceous ducts of the normal human skin.
The bacteria described up to the present would also appear to come under the same category, and so far as one can gather have but little to do with the disease under consideration, and much less with its etiology.
During this research experiments were undertaken on an extensive scale, and under very favourable conditions for the final determination of the presence or absence of any forms of micro-organisms in the blood stream of Beri-beri patients. In our examinations acute cases of Beri-beri were chosen to begin with, and later on the experiments were supplemented by the examination of the blood streams of the dropsical and atrophic varieties of the disease.
The technique employed was the following:-Peptone bouillon was prepared according to EYRE's method, and transferred to tubes. Each tube contained approximately 20 e.c. of bouillon. These bouillon tubes were kept under incuba- tion for at least a week previous to being used for purpose of cultivation. Great care was always exercised in maintaining absolute sterility of the medium used. The tubes of bouillon were prepared in the Bacteriological Laboratory and transferred to the Tung Wah Hospital for inoculation. The patient was prepared for a venesection of the arm. The arm having been rendered aseptic by the use of thorough washing, alcohol, sublimate, etc., one of the superficial veins was com- pletely severed at the bend of the elbow. The blood was allowed to flow into a porcelain dish and escape slowly for at least 30 seconds. Thereupon the bouillon tubes were brought to the bedside, the cotton wool plugs carefully removed and from 10-20 c.c. of blood were allowed to pass into each tube. Several tubes were inoculated with the blood of each patient. After the operation, pressure was applied over the wound and in no instance did adverse conditions shew them- selves subsequently. The tubes were removed to the Laboratory as soon as possible and incubated for definite periods. In other cases tubes of agar- agar and other media were used and filled with blood extracted from the patient in the same way. All these were subsequently incubated for long but indefinite periods. Further flasks of a capacity of 1,000°c.c. were filled with bouillon to the extent of 250 c.c. These were tested before use as to their sterility by incubation at 37° C. for over a week. They were inoculated with blood from typical cases of the disease—one flask being used for each patient, and each receiving at least 50 c. c. of blood.
By these methods it is obvious that if micro-organisms were present in the blood in cases of Beri-beri these means would undoubtedly settle the question. A large number of experiments were made because of the difficulty of rendering the skin absolutely aseptic and the prevention of extraneous organisms entering the flask when exposed in the airy wards of a tropical and Chinese Hospital.
The following is a description of the results obtained :--
Experiment No. I.
Nature of case.
Media..
No. of Tube.
Quantity of Blood.
Period of Incubation.
Result.
Acute Beri-beri, 7 days ill.
Bouillon.
1
10 c.c.
7 days.
2
15 c.c.
Mould. Sterile.
3
10 c.c.
14 days.
11
4
10 c.c.
19
5
10 c.c.
14
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19