Aims: To evaluate the public health significance of representative strains of two Aeromonas spp., mainly from freshwater fish, on the basis of production of virulence-associated factors and presence of the haemolytic genes aer A and hly A. Methods and Results: Eleven strains of Aer. hydrophila , three strains of Aer . veronii biovar sobria (all from freshwater fish) and one strain of Aer. hydrophila from human diarrhoea were tested for potential virulence traits and for the presence of the haemolytic genes aer A and hly A. Ten Aer. hydrophila isolates were aer A(+) hly A(+) and two aer A(+) hly A(-) . Aeromonas veronii biovar sobria isolates were aer A(-) hly A(-) . Strains from the three genotypes showed enterotoxic activity in the suckling mouse assay. At 28degreesC, four Aer. hydrophila fish strains could be considered as potentially virulent (possessing at least two of these characteristics: haemolytic, cytotoxic and enterotoxic). One Aer . veronii biovar sobria strain and the clinical isolate were cytotoxic on Vero cells. When grown at 4degreesC, these six isolates fulfilled virulence criterion, but at 37degreesC, only one fish strain, an Aer. hydrophila , did. Conclusions: The potential health risk derived from the presence of Aer. hydrophila and Aer. veronii biovar sobria in ice-stored freshwater fish should not be underestimated. Significance and Impact of the Study: Expression of virulence factors is affected by temperature incubation and not always related to the presence of haemolytic genes.