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A novel research on isolation and characterization of Photobacterium damselae subsp. damselae from Pacific white shrimp,Penaeus vannamei,displaying black gill disease cultured in China
Authors:Ziyan Wang  Chengyin Shi  Hailiang Wang  Xiaoyuan Wan  Qingli Zhang  Xiaoling Song  Ge Li  Miao Gong  Shigen Ye  Guosi Xie  Jie Huang
Institution:1. College of Fisheries and Life Science, Dalian Ocean University, Dalian, China

Laboratory for Marine Fisheries Science and Food Production Processes, National Laboratory for Marine Science and Technology (Qingdao), Key Laboratory of Maricultural Organism Disease Control, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, China;2. Laboratory for Marine Fisheries Science and Food Production Processes, National Laboratory for Marine Science and Technology (Qingdao), Key Laboratory of Maricultural Organism Disease Control, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, China;3. College of Fisheries and Life Science, Dalian Ocean University, Dalian, China

Abstract:In June 2019, massive mortalities of cultured Penaeus vannamei occurred in a local farm in Hainan Province, China. The diseased shrimp displayed evident black gills. Three bacterial strains 20190611001, 20190611007 and 20190611022 were isolated from hepatopancreas and gills of the diseased shrimp and identified as Photobacterium damselae subsp. damselae based on the sequence analysis of 16S rRNA and toxR genes. These three isolates showed haemolytic activities. Of them, strain 20190611022 isolated from hepatopancreas was selected and processed for pathogenic analysis. The calculated median lethal dose (LD50) was 9.75 ± 4.29 × 105CFU/g (body weight) by challenging P. vannameivia reverse gavage. The diseased shrimp displayed enlarged hepatopancreatic tubules and sloughing of epithelial cells in tubular lumens. The strain 20190611022 was also characterized by the testing of API 20NE systems and antibiotic susceptibility. The results of disc diffusion test showed that strain 20190611022 was sensitive to chloramphenicol, compound sulfamethoxazole, cefoperazone, ceftriaxone, ceftazidime and cefuroxime. To our knowledge, this is the first report of isolation and characterization of Photobacterium damselae subsp. damselae from natural diseased P. vannamei. Our findings can serve as a basis for further studies of its pathogenicity and provide technological support for disease controlling in shrimp aquaculture.
Keywords:black gill disease  pathogenicity  Penaeus vannamei  Photobacterium damselae subsp  damselae
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