Multilocus sequence analysis of Vibrionaceae isolated from farmed amberjack and the development of a multiplex PCR assay for the detection of pathogenic species

Issei Nishiki; Takayuki Minami; Ayana Murakami; Truong Dinh Hoai; Atushi Fujiwara

doi:10.1111/jfd.12823

Multilocus sequence analysis of Vibrionaceae isolated from farmed amberjack and the development of a multiplex PCR assay for the detection of pathogenic species

J Fish Dis. 2018 Jun 7. doi: 10.1111/jfd.12823. Online ahead of print.

Authors

Issei Nishiki¹, Takayuki Minami², Ayana Murakami³, Truong Dinh Hoai^{4

5}, Atushi Fujiwara¹

Affiliations

¹ Research Center for Bioinformatics and Biosciences, National Research Institute of Fisheries Science, Japan Fisheries Research and Education Agency, Yokohama, Japan.
² Miyazaki Prefectural Fisheries Research Institute, Miyazaki, Japan.
³ Advanced Technology Development Center, Kyoritsu Seiyaku Corporation, Tsukuba, Japan.
⁴ Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan.
⁵ Faculty of Fisheries, Vietnam National University of Agriculture, Hanoi, Vietnam.

PMID: 29882274
DOI: 10.1111/jfd.12823

Abstract

Since 2011, high mortality rates and symptoms consistent with vibriosis have been observed in farmed amberjack (Seriola dumerili) in Japan. To identify 41 strains isolated from diseased amberjack, a multilocus sequence analysis using nine concatenated genes (ftsZ, gapA, gyrB, mreB, pyrH, recA, rpoA, topA and 16S rRNA) was conducted. Twenty-seven strains were identified as Vibrio harveyi, suggesting an epidemic of V. harveyi infection in amberjack farms. Other strains were identified as Vibrio anguillarum, Vibrio owensii and Photobacterium damselae subsp. damselae. To develop an efficient diagnostic method for vibriosis in amberjack, a multiplex PCR system was developed to identify V. anguillarum, V. harveyi and P. damselae subsp. damselae. The method successfully discriminated between these three bacterial species, with amplification products of 350 bp for V. anguillarum, 545 bp for V. harveyi and 887 bp for P. damselae subsp. damselae and can be used for diagnosis in aquaculture farms.

Keywords: Vibrio anguillarum; Vibrio harveyi; Photobacterium damselae subsp. damselae; amberjack; multilocus sequence analysis; multiplex PCR.