Biochemical and immunological characterization of the cell surface of the fish pathogenic bacterium Aeromonas salmonicida.

The identification of lipopolysaccharide as periodic acid-Schiff positive material, present in the membrane fraction of the fish pathogenic Gram-negative bacterium Aeromonas salmonicida, analyzed by SDS-polyacrylamide gel electrophoresis, is shown. Such analysis has revealed several periodic acid-Schiff positive bands and many membrane proteins among which a pathogenicity-related Mr 54000 protein as a constituent of an additional surface layer outside the outer membrane (Evenberg et al., (1982) Biochim. Biophys. Acta 684, 241-248). The latter protein, designated as additional cell envelope protein or ACE protein, has been purified and characterized in our laboratory (Evenberg and Lugtenberg, (1982) Biochim. Biophys. Acta 684, 249-254). Most strains produce both high and low molecular weight lipopolysaccharide species, presumably corresponding with the presence and (virtual) absence, respectively, of an O-antigenic chain. The property to produce high molecular weight lipopolysaccharide can be lost upon subculturing in laboratory growth media and such is greatly enhanced by the prior loss of the ability to produce ACE protein. Lipopolysaccharide and ACE protein were identified as the major antigens. A new polysaccharide-like antigen, designated as PS-antigen, was detected. Moreover, immunological indications for the presence of a lipoprotein in A. salmonicida are described. The surface localization of the antigens was determined by testing whether preadsorption of antisera by intact cells decreased the binding of IgG to these antigens, or decreased the ability of the sera to agglutinate cells. According to these criteria lipopolysaccharide, ACE protein and PS-antigen are the major surface-located antigens. Material cross-reactive with lipopolysaccharide, ACE protein and PS-antigen has been found in a large number of strains. Several lines of evidence indicate the presence of interactions between ACE protein and lipopolysaccharide. Based on these results a molecular model of the cell envelope of virulent A. salmonicida is presented.