Survey of viral haemorrhagic septicaemia virus in wild fishes in the southeastern Black Sea

Species diversity in the Black Sea ecosystem has been declining rapidly over the last 2 decades. To assess the occurrence and distribution of viral haemorrhagic septicaemia virus (VHSV) in various wild fish species, a wild marine fish survey was carried out in 2009, 2010, and 2011. The pooled or individual samples of kidney, liver, and spleen of 5025 specimens, belonging to 17 fish species, were examined virologically using cell culture. The cells showing cytopathic effects (CPE) were subjected to ELISA and multiplex reverse transcriptase polymerase chain reaction (RT-mPCR), for VHSV and infectious pancreatic necrosis virus (IPNV), after blind passaging to determine the virus species causing CPE. The virus species and possibility of co-infection with IPNV were verified by the RT-mPCR developed in this study. Twelve species of fish (pontic shad Alosa immaculata, red mullet Mullus barbatus, three-bearded rockling Gaidropsarus vulgaris, black scorpionfish Scorpaena porcus, Mediterranean horse mackerel Trachurus mediterraneus, whiting Merlangius merlangus euxinus, stargazer Uranoscopus scaber, pilchard Sardina pilchardus, garfish Belone belone, round goby Neogobius melanostomus, thornback ray Raja clavata, and anchovy Engraulis encrasicolus) tested positive for VHSV Genotype Ie (VHSV-Ie). Except whiting, pilchard, and round goby, the rest are new host records for VHSV. The extent and spread of VHSV-Ie was significantly higher among bottom fish than among pelagic fish. Sensitivity and specificity of the RT-mPCR developed was sufficiently high, suggesting that this assay may be used for both diagnostic and surveillance testing. According to the RT-mPCR results, IPNV was not present in wild fish. These results support the hypothesis that the VHSV-Ie genotype, highly prevalent among fish species in the Black Sea, may have a serious impact on the population dynamics of wild fish stocks.