﻿REPORT ON CHOLERA IN MANILA. 437 



rather be regarded as an endotoxin, for convincing evidence to the con- 

 trary at least has not yet been brought forward. 



During the year, opportunity was afforded to study the antitoxic serum 

 of Denier and to witness its practical application in the treatment of 

 Asiatic cholera. A request having been made by Dr. Denier to carry on 

 the experimental serum treatment of cholera in the Government cholera 

 hospital in Manila, I was called upon to examine the sera and report upon 

 them before this method of treatment was undertaken. The two sera 

 which Dr. Denier employed were of a different nature. One serum 

 designated as "A" was prepared by injecting the horse with the cholera 

 toxin entirely free from the bacteria, and the second one, "B," was 

 produced ...by injecting the horse with the living organisms. These sera 

 were, upon examination, found to possess specific agglutinative and bac- 

 tericidal properties, serum "B" showing a much higher value in this 

 respect. No study was made of the neutralizing power of the sera for 

 lethal amounts of the filtered cholera toxin. Guinea pigs inoculated with 

 1 cubic centimeter of serum "B" and at the same time with 1 or even 2 

 oesen of a cholera vibrio, of which the lethal dose was 1/10 oese, survived 

 the inoculation ; however, when they were inoculated with 5 oesen and 2 

 cubic centimeters of the serum, they invariably succumbed. Pfeiffer's 

 phenomenon seemed to have been complete, as was shown by the post- 

 mortem examination of a number of these animals, since microscopic 

 preparations from the exudate in the abdominal cavity showed no motile 

 vibrios and the animals had apparently died rather from an intoxication 

 than from an infection. However, these experiments obviously do not 

 demonstrate whether death had occurred from the effect of the endotoxin 

 contained in such a large amount of the spirillum (5 oesen) or from 

 the effects of another soluble toxin. 



Serum "B" was found to protect against larger doses of the living- 

 organism than serum "A" as was proved by testing the bactericidal 

 power of the two sera. The bactericidal value of the sera was ap- 

 parently, at all events so far as the living organisms were concerned, 

 the most important factor in protecting the animals, at least up to a 

 certain dose. In many of the animals which died and which had not 

 received excessively large doses of the cholera spirillum, Pfeiffer's pheno- 

 menon was also found to be complete or almost so. 



In all, 52 human cases of cholera were treated by Dr. Denier with the 

 sera. In each instance a bacteriologic diagnosis of cholera was made 

 by Dr. Denier and also by this laboratory, as was customary with all 

 cases in the Government hospital. The injections of the sera were given 

 intravenously and in large quantities, as much as 250 cubic centimeters in 

 a liter of Hayem's solution being inoculated at a single dose. Following 

 this primary inoculation, 100 cubic centimeters of serum were injected 

 in an equal amount of saline solution every three hours until a reaction 



