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SUMMARY: A toxic protein has been isolated from Inaba and Ogawa strains of Vibm'o cholmae and from water vibrios by dissolving the bacteria in 2.5~-urea and subsequent fractional precipitation with ammonium sulphate. This toxic protein accounted for the major portion of the toxicity of whole organism. Immunological and chemical data suggest that the toxic protein is the protein component of the Boivin antigen. The lipopolysaccharide endotoxins of the strains examined were relatively non-toxic as compared with the lipopolysaccharides from other Gram- negative bacteria, though in other biological and chemical properties they resembled the general class. There is a marked tendency on the part of many research workers in the cholera field to assume that the manifestations of the disease are caused by a specific toxin which is liberated from the bacteria during growth. The proof of this hypothesis is difficult for two reasons. First, there is little precise information about the toxins of Vibrio cholma-e, and much confusion exists as to the chemical nature and similarity of those substances which have been described. The second difficulty is that it has not been possible to produce a disease resembling cholera in experimental animals; therefore it is difficult to test toxins for their ability to reproduce the signs of the natural disease. Boivin & Mesrobeanu (.1935) isolated a toxic antigenic complex from Vibrio cholerae by extraction with cold trichloroacetic acid and characterized it as a ' glucolipid '. Similar Boivin antigens have been extracted from many Gram-negative bacteria and have been shown to possess the ' 0 '-antigenic specificity of the parent organisms. It is not clear in the case of V. cholerae how completely this glucolipid complex could account for the toxicity of whole organisms. Bernard & Gallut (1945) confirmed the existence of a toxic Boivin- type antigen and reported a second small-molecular toxin which was later purified to some extent by Gallut & Grabar (1945). Burrows (1944) claimed to have extracted a highly toxic phospholipid from V. cholerae by using hot ethanol. This material contained, on occasion, 500 mouse LD 50 doseslmg. and, moreover, caused an increase in permeability to water of the small intestine of rabbits, when tested in. vitro. This effect on permeability was abolished by using intestines from animals immunized with V. choZerae. Freter (1956) described the isolation of two toxic fractions from V. chohae, one acid-soluble and the other insoluble in HC1 at pH 8.5. The acid-insoluble material had a mouse LD 50 of approximately 500-75Opg. and appeared to be a lipoprotein; the acid-soluble toxic material had a mouse toxicity of

Toxic Proteins from Vibrio cholerae and Water Vibrios which are Lethal for Mice