Thirty-three strains of Vibrio vulnificus of clinical and environmental origin were examined for production of 12 extracellular enzymes of potential importance to the virulence of this bacterium. Strains of Vibrio vulnificus were consistent in their production of protease, mucinase, lipase, chondroitinase, hyaluronidase, DNase, sulfatase, and hemolysin. No differences between clinical and environmental isolates were noted. Although none of the enzymes appeared to correlate with the ability of these strains to produce lethality in mice, the production of hemolysin and of a protease with activity against native serum albumin may be significant in the pathogenesis of the potentially fatal infections produced by this organism. The production of several of these exoenzymes also appeared to correlate with pathogenicity in the seven other Vibrio species examined. Culture filtrates of all virulent strains of Vibrio vulnificus were cytotoxic for Chinese hamster ovary cells, whereas those of the strains of Vibrio parahaemolyticus and Vibrio alginolyticus examined lacked this activity.