90 ^^^ Philippine Journal of Science im 



serum of low potency; also certain strains were found to agglu- 

 tinate with immune rabbit serum, but not with immune horse 

 serum. Zlatogoroff (45) was able to produce agglutination in an 

 imperfectly agglutinable strain of cholera by repeated passage 

 through animals. On the other hand, McLaughlin and Whit- 

 more (31) were unable to confirm Zlatogoroff 's work. 



Our own work, based on the study of cases of cholera and 

 cholera carriers after death, will now be presented. 



From July, 1915, until the present time the intestinal contents 

 and gall bladder from all autopsies have been examined for 

 cholera vibrios as a routine procedure. 



Technic. — A loop of small intestine about 0.5 meter from the 

 caecum was ligatured, severed from its connections, placed in a 

 new, clean, self-sealing jar, and transported to the laboratory 

 for bacteriological examination. In the same way the bile ducts 

 were ligatured and severed, and the gall bladder was removed 

 without opening, placed in another new, clean jar, and trans- 

 ported to the laboratory. The specimen as received at the lab- 

 oratory was handled with sterile instruments. The outer surface 

 of the intestine was seared with a hot spatula, and with a 

 sterile scalpel an incision was made into the lumen of the gut. If 

 the contents were fluid, 1 cubic centimeter was removed with 

 a sterile pipette and inoculated into a cholera peptone tube. 

 The last drop in the pipette was allowed to fall on a Dieudonne 

 plate and was then spread with a sterile platinum loop. If the 

 fpeces were solid, 1 standard loop (4 millimeters) was inoculated 

 into peptone and also smeared on a Dieudonne plate. The same 

 procedure was followed with the gall bladder. These direct 

 plates and peptone tubes were incubated overnight at 37.5° C. 

 In the event of the plate remaining sterile the next morning, the 

 peptone tubes were examined that morning for organisms of 

 suspicious motility and new Dieudonne plates were seeded from 

 those plates with suspicious organisms. Suspicious colonies on 

 Dieudonne plates were picked, emulsified in physiologic salt solu- 

 tion, and tested for agglutination with a cholera-immune goat 

 serum of titer 1 : 6000, in dilution of 1 : 500. 



There have been 269 cases in which there has been reason 

 clinically, anatomically, or bacteriologically to suspect the pre- 

 sence of Asiatic cholera. In 22 of these, vibrios were not found, 

 and 5 cases were known to have been carriers of the vibrios 

 at varying periods before death. These latter 27 cases will be 

 discussed later. Of the remaining 242 cases, 32 were interpreted 

 as cholera carriers and 210 as true cholera cases. 



The classification of all of these cases is presented in Table II. 



