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Multilocus sequence analysis of Vibrionaceae isolated from farmed amberjack and the development of a multiplex PCR assay for the detection of pathogenic species
Author(s) -
Nishiki Issei,
Minami Takayuki,
Murakami Ayana,
Hoai Truong Dinh,
Fujiwara Atushi
Publication year - 2018
Publication title -
journal of fish diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 85
eISSN - 1365-2761
pISSN - 0140-7775
DOI - 10.1111/jfd.12823
Subject(s) - biology , vibrio harveyi , vibrio anguillarum , microbiology and biotechnology , multiplex polymerase chain reaction , vibrionaceae , 16s ribosomal rna , vibrio , polymerase chain reaction , bacteria , gene , genetics
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.