During an investigation on the distribution of fish viruses in wild marine fishes, a rhabdovirus was isolated from wild Japanese flounder (Paralichthys olivaceus) caught in Obama coastal area of the Wakasa Bay, Japan in 1999. The isolated rhabdovirus was neutralized by an antiserum against viral haemorrhagic septicaemia virus (VHSV) but not by antisera against infectious haematopoietic necrosis virus (IHNV) or hirame rhabdovirus (HIRRV). The newly isolated rhabdovirus was composed of five structural proteins with estimated molecular weights of L:>140, G:61.6, N:43.1,M1:28.4 and M2:24.0 kDa. In the Western blot analysis, the anti-VHSV serum strongly reacted with G, N, M1 and M2 proteins of the new viral isolates. In the PCR tests with primers for the G and M1 protein genes of VHSV, amplified products having the same size with each target region were amplified from the rhabdoviral genome. Based on these results, the rhabdovirus isolated from wild Japanese flounder was identified as VHSV. This is the first report on the isolation of VHSV in Asian countries.