Isolation and Identification of a Aeromonas veronii Strain Causing the Hemorrhagic Disease of Great Blue-spotted Mudskipper (Boleophthalmus pectinirostris)
WANG Wei1,2, LI Chang-Hong1,2,*, ZHAN Ping-Ping2, CHEN Jiong1,2,*
1 State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agroproducts, Ningbo University, Ningbo 315211, China; 2 Laboratory of Biochemistry and Molecular Biology, School of Marine Science, Ningbo University, Ningbo 315832, China
Abstract From August to September 2020, an outbreak of hemorrhagic disease in the great blue-spotted mudskipper (Boleophthalmus pectinirostris) occurred in a farm in Cixi City, Zhejiang Province, causing serious economic losses to farmers. In order to identify the pathogen of the disease, methods including the anatomical observation, bacterial isolation and culture, electron microscope observation, biochemical test, drug sensitivity test, artificial infection test, and phylogenetic analysis of the 16S rRNA and DNA gyrase subunit B gene (gyrB) were used for identification of the pathogen. The results showed that one dominant strain, named AV20211212, was isolated from the diseased fish. Observation by light microscope with gram staining and ultrastructural observation by scanning and transmission electron microscope showed that the isolated strain AV20211212 was Gram-negative with blunt round ends and hollow polar monotrichous flagella. Results from physiological and biochemical characteristics identification showed that AV20211212 had the similar physiological and biochemical characteristics to Aeromonas veronii. Homology analysis demonstrated that the 16S rRNA of the isolated strain shared 93.6%~99.4% homology with those of other A. veronii and the amino acid sequence of the gyrB from the isolated strain shared 99.5%~99.7% homology with those of other A. veronii. Phylogenetic analysis showed that the isolated strain was closely clustered with A. veronii. The isolated strain was highly resistant to 7 antibiotics such as penicillin and ampicillin, moderately sensitive to azithromycin, and sensitive to 12 antibiotics such as norfloxacin and amikacin. The mudskipper infected by the isolated strain AV20211212 showed the same symptoms with naturally diseased mudskipper on the farm, and its medium lethal dose (LD50) was 4.56×103 CFU/g to mudskipper. In the present study, A. veronii was identified as the pathogen of mudskipper for the first time. All the results provided theoretical basis and technical support for the effective prevention and rational medicine use for bacterial fish diseases in Cixi City.
WANG Wei,LI Chang-Hong,ZHAN Ping-Ping, et al. Isolation and Identification of a Aeromonas veronii Strain Causing the Hemorrhagic Disease of Great Blue-spotted Mudskipper (Boleophthalmus pectinirostris)[J]. 农业生物技术学报, 2022, 30(2): 344-355.
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