Monitoring the survival of fish-pathogenic Francisella in water microcosms

In this report, the survival behaviour of fish pathogenic Francisella in water microcosms was investigated under laboratory conditions. Two isolates of Francisella noatunensis (NCIMB14265^T and PQ 1106), from fish held in seawater and freshwater, were inoculated into natural (nonsterile) and sterile sea- and freshwater microcosms, respectively, and monitored under different temperature conditions (4, 8 and 12 °C) over a period of 60 days. The culturability of the strains was inversely related to the water temperature. Strain NCIMB14265^T was found to survive longer in seawater than PQ 1106 held in freshwater at equivalent temperatures. The survival of both strains was higher in sterile than in nonsterile microcosms. These results were confirmed by quantitative PCR analysis targeting the succinate dehydrogenase (sdhA) gene. A cell viability assay coupled with FISH analyses showed that F. noatunensis cells enter a viable but not culturable (VBNC) state after a period in water. However, although metabolically active, the VBNC cells were not pathogenic to cod (Gadhus morhua) following an intraperitoneal challenge, under the conditions tested. The data presented contribute to a better understanding of the behaviour of F. noatunensis in natural seawater and freshwater environments, and show the need for further investigation of the role of VBNC cells in the environmental transmission of this pathogen.