Monitoring the Starvation–Survival Response of <i>Edwardsiella piscicida</i> and <i>E. tarda</i> in Freshwater Microcosms, at Various Temperatures

Abstract

Edwardsiella piscicida is an important fish pathogen responsible for economic losses in global aquaculture, and E. tarda is also a human zoonotic pathogen. In this study, the survival of E. piscicida and E. tarda strains kept in filtered and sterilized lake water microcosms was investigated during a 20-week period at 7 °C, 15 °C and 25 °C, as well as its pathogenicity retention during a starvation period. E. tarda V43.2 stayed culturable for 6 weeks at 7 °C, 9 weeks at 25 °C and 12 weeks at 15 °C. Both E. piscicida strains (V12.1 and V57.2) stayed culturable even longer, for at least 12 weeks at 7 °C, 15 °C and 25 °C under the same starvation conditions. After Edwardsiella cells entered into the VBNC state, some became shorter and ”rounded up,” but others aggregated and retained a short rod shape. Aggregates of Edwardsiella cells were common throughout the VBNC period, and a well-formed biofilm was observed for all tested strains at the end of the experiment. The growth capacity of VBNC cells was restored by cultivating microcosm water samples in LB broth at 28 °C. Resuscitated E. piscicida cells were as virulent for the European eel as the controls. Natural waters can be a reservoir for Edwardsiella, and its underestimation in environmental samples poses a risk to public health and aquaculture.

Keywords

• <i>E. piscicida</i>
• <i>E. tarda</i>
• starvation survival
• VBNC state
• biofilm
• retention of virulence