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Identification of Vibrio harveyi, Vibrio ichthyoenteri, and Photobacterium damselae isolated from olive flounder Paralichthys olivaceus in Korea by multiplex PCR developed using the rpoB gene

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  • Aquaculture
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Abstract

Major fish bacterial diseases in Korea are edwardsiellosis, streptococcosis, and vibriosis. Among vibrionaceae, Listonella anguillarum, Vibrio harveyi, V. ichthyoenteri, and Photobacterium damselae were identified as causative organisms of vibriosis in flounder. In this study, we developed a multiplex PCR method using the RNA polymerase β subunit (rpoB) gene, known as a housekeeping gene for identification of Vibrio spp. causing vibriosis in flounder. Three pairs of PCR primers were designed based on the rpoB sequence of three species, V. harveyi, V. ichthyoenteri, and P. damselae. The PCR assay, using a mixture of six primers, yielded amplicons of 601, 434, and 533 bp in V. harveyi, V. ichthyoenteri, and P. damselae. None of the untargeted species yielded an amplicon. The detection limits for pure culture in kidney were 2.5 × 104 cfu/g kidney for V. harveyi, 2.5 × 105 cfu/g kidney for V. ichthyoenteri, and 2.5 × 106 cfu/g kidney for P. damselae. From the colonies on TCBS agar plates of different samples, 632 Vibrio spp. isolated from aquacultured flounder between 2004 and 2010 were identified by the multiplex PCR method. As a result, 265 strains (41.9 %) were V. ichthyoenteri; 115 strains (18.2 %) were V. harveyi and 72 strains (11.4 %) were P. damselae.

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Acknowledgments

This work was supported by the funds of NFRDI in Korea.

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Authors and Affiliations

  1. Pathology Division, National Fisheries Research and Development Institute, Busan, 619-902, Korea

    Myoung Sug Kim & Hye Sung Choi

  2. Department of Marine Biotechnology, Soonchunhyang University, Asan, 336-745, Korea

    Ji Young Cho

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  1. Myoung Sug Kim
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  2. Ji Young Cho
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  3. Hye Sung Choi
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Correspondence to Myoung Sug Kim.

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Kim, M.S., Cho, J.Y. & Choi, H.S. Identification of Vibrio harveyi, Vibrio ichthyoenteri, and Photobacterium damselae isolated from olive flounder Paralichthys olivaceus in Korea by multiplex PCR developed using the rpoB gene. Fish Sci 80, 333–339 (2014). https://doi.org/10.1007/s12562-014-0702-5

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