Vibrio mimicus strains W-26768 (stool isolate) and N-1301 (environmental isolate) and Vibrio fluvialis strains AA-18239 (stool isolate) and M-940 (environmental isolate) were studied for virulence properties and lipopolysaccharide composition. All four strains were hydrophobic, produced cytotoxin, adhered to HeLa cells and showed mannose-sensitive agglutination of guinea pig erythrocyte. The strains were negative for enterotoxin production and were mostly susceptible to the common antibiotics. The environmental and clinical isolates of both species were antigenically unrelated to each other. Lipopolysaccharide antigen analysis showed that O-antigen polysaccharides of two strains of V. fluvialis and two strains of V. mimicus differed with respect to the sugar components. Only LPS from V. mimicus W-26768 showed the presence of an unusual sugar, 3,6-dideoxy-3-acetamido-hexose. The sugar compositions of these V. fluvialis and V. mimicus strains differed from those of previously reported Japanese isolates. These differences probably reflect differences in the serogroup of strains.