A Leukocytolytic Factor Isolated from Cultures of Aeromonas salmonicida

Abstract

A substance characterized as a glycoprotein, isolated from the supernatant fluids of broth cultures of Aeromonas salmonicida by a combination of ammonium sulfate and ethanol precipitations followed by chromatography on DEAE-cellulose, was cytolytic for rainbow trout (Salmo gairdneri) leukocytes, and antigenic when injected into rabbits. The ratio of protein to hexose determined by analysis of the purified fraction was between 0.35 and 0.45, and small amounts of amino sugars were detected. A virulent strain of A. salmonicida produced much more of this factor than an avirulent strain. This factor was cytolytic for leukocytes in vitro and also produced a pronounced leukopenia when injected intravenously in adult rainbow trout. When injected in small coho salmon (Oncorhyncus kisutch) 8–13 cm long together with about one LD50 of live A. salmonicida 36 of 40 fish succumbed to the combination, whereas only 14 of 40 died from an injection of the bacterium alone. Thus, the pathogenicity of the organism was enhanced, presumably by increasing the susceptibility of the host. Hence, this glycoprotein apparently is one of the virulence factors of this bacterium. Key words: leukocytolytic factor, Aeromonas salmonicida, glycoprotein, virulence factor