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III. THE DROPSICAL ACCUMULATIONS.
Fluid from the pleural and abdominal cavities and subcutaneous tissues has been examined bacteriologically (culturally and experimentally). In a certain proportion of the tests the presence of the inhibiting alexins in such fluids was destroyed by heat. Subsequently all the flasks containing the fluid and nutrient media were incubated. All tests made in this way were negative.
IV.—THE CEREBRO-SPINAL FLUID (lumbar puncture and post-mortem).—This fluid was also examined microscopically and bacteriologically, the examinations. being made after thorough centrifugalisation in order to obtain a suitable deposit. Bacteriologically the tests were similar to those given under the foregoing headings. The results were negative as regards organisms in every instance.
Post-mortem Results.--Microscopic examinations of smears of almost every organ and tissue of fresh Beri-beri cadavers have been made in a large number of cases.
Indeed in every case of Beri-beri examined post-mortem smears of the blood spleen and liver are always prepared stained by LEISHMAN's method and examined microscopically. All our microscopic examinations in these cases have given a negative result.
Bacteriologically.-The tests applied were those usually adopted in order to isolate micro-organisms, namely, plate cultivations. tube inoculations and the em- ployment of a variety of nutrient media. The experimental tests were also applied (vide Experimental part of the research).
In fresh cadavers inoculations were made into nutrient media from every organ. In many,
of the cases cocci and bacteria of various species were obtained in pure culture, thus agreeing with the results of PEKELHARING and WINKLER. On applying further tests, inclusive of experimentation, with these micro-organisms, the results obtained shewed that these cocci and bacteria, bore no causal relation- ship to Beri-beri.
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Repeatedly cultures were made from the gastro-duodenal mucous membrane especially from the areas of necrosis and hæmorrhagic extravasations. Bacteria were isolated according to the aerobic and anaerobic methods, but these on further examination proved to be organisms having nothing in common with the disease and whose natural hatitat is the intestine.
From these examinations which have extended over several years, and from the general negative results as regards a specific biological excitant of the disease, we feel assured that the organisms hitherto isolated by many observers are of extraneous origin, and have no causal relationship to Beri-beri. Again, the bacteria noted by HAMILTON WRIGHT in his recent memoirs on this subject can be demonstrated again and again in the pathological areas of the gastro intestinal mucosa, but their presence must be regarded as the result of secondary bacterial invasion of the pathological mucosa.
Supplementary evidence of these all-ronnd negative results is supplied in the experimental part of our research. There, it will be observed, experiments of the most varied character were undertaken, and notwithstanding the application of all the methods known to induce the incidence of the disease amongst animals, the results were uniformly negative throughout.
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In conclusion, therefore, it may be said that the bacteriological methods so far applied to cases of Beri-beri have failed to isolate for us a specific pathogener of the disease. Naturally we cannot definitely say that the disease is of a non- bacteriological nature, yet we are strongly of opinion that in the absence of posi- tive results after many searching biological investigations by ourselves and others, a specific infectivity as regards Beri-beri must lose much of its significance, and the disease, in the future, relegated to the confines of the pathological chemist.