Effect of temperature and salinity on virulence of Edwardsiella tarda to Japanese flounder, Paralichthys olivaceus (Temminck et Schlegel)

Experiments were designed to determine the effects of temperature and salinity on the virulence of Edwardsiella tarda to Japanese flounder, Paralichthys olivaceus . In the temperature experiment, a two‐factor design was conducted to evaluate the effects of both pathogen incubation temperature and fish cultivation temperature on pathogen virulence. E. tarda was incubated at 15, 20, 25 and 30±1°C, and the fish (mean weight: 10 g) were reared at 15, 20 and 25±1°C respectively. The fish reared at different temperatures were infected with the E. tarda incubated at different temperatures. The results of a 4‐day LD 50 test showed that temperature significantly affected the virulence of E. tarda ( P <0.01) and the interaction between the two factors was also significant ( P <0.01). For fish reared at 15°C the virulence of E. tarda was the highest at 25°C of pathogen incubation, followed by 20, 15 and 30°C. When the fish rearing temperature was raised to 20 and 25°C, the virulence of E. tarda incubated at all temperatures increased. Isolation testing demonstrated results similar to those of LD 50 . The higher rearing temperature increased the proliferation rate of the pathogen in fish. In the salinity experiment, the incubation salinity of E. tarda was at 0, 10, 20 and 30 g L −1 , respectively, and the fish with mean weight of 50 g were cultured in natural seawater of 30 g L −1 . The results of one‐way anova in 4‐day LD 50 test showed that incubation salinity significantly affected virulence. Virulence was lower when the salinity of the incubation medium was at 0 and 30 g L −1 , higher at 10 and 20 g L −1 . The results of isolation test were in accordance with those of LD 50 . At 20 g L −1 E. tarda had a faster proliferation rate than that at 10 g L −1 .