Characterization and species-specific detection of Vibrio parahaemolyticus-related species associated with mass mortality of ark shell (Scapharca broughtonii)
Ryunkyoung Oh, Min Jeong Lee, Nam-Sil Lee, Young-Ok Kim, Hee Jeong Kong, Ju-Won Kim, Jung Youn Park, Bo-Hye Nam and Dong-Gyun Kim
National Institute of Fisheries Science, Biotechnology Research Division, Busan 46083, Korea National Institute of Fisheries Science, Pathology Division, Busan 46083, Korea
Ark shell (Scapharca broughtonii) is cultured in southern coast of Korea and is a commercially important shell in aquaculture. However its productivity has decreased rapidly in the last decade due to mass mortality. Various microbial investigations were performed to identify the caused bacterial species for economic losses. Using cumulative mortality analysis, 16S rDNA sequence analysis, and API 20 kit assay, Vibrio parahaemolyticus-related species (8M 4-1) was identified as an etiologic bacterium. The LD50 value of the isolated bacterium to ark shell was lower than 106 CFU/mL. After infection, histological diagnosis from hepatopancreatic tissue also provided evidence for pathogenicity. To monitor this ark shell pathogenic bacterium, we designed the PCR detection method with the Vibrio species groEL gene. We also tested various virulence gene-targeted detection methods, but amplified products were not suitable for V. parahaemolyticus recognition. In this study, we identified V. parahaemolyticus-related species as a causative organism of mass mortality, and demonstrated that PCR detection assay could successfully detect V. parahaemolyticus strains from infected tissues with high sensitivity. Using these results, we identified V. parahaemolyticus-related 8M 4-1 species as the causative agent of ark shell mortality and showed that newly developed PCR detection method was useful for pathogen monitoring.
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