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龙梦, 李彤彤, 江遥, 张倩倩, 张德峰, 张富铁, 王剑伟, 李爱华. 长江上游网箱养殖圆口铜鱼杀鲑气单胞菌杀鲑亚种的分离与鉴定[J]. 水生生物学报, 2020, 44(1): 153-161. DOI: 10.7541/2020.018
引用本文: 龙梦, 李彤彤, 江遥, 张倩倩, 张德峰, 张富铁, 王剑伟, 李爱华. 长江上游网箱养殖圆口铜鱼杀鲑气单胞菌杀鲑亚种的分离与鉴定[J]. 水生生物学报, 2020, 44(1): 153-161. DOI: 10.7541/2020.018
LONG Meng, LI Tong-Tong, JIANG Yao, ZHANG Qian-Qian, ZHANG De-Feng, ZHANG Fu-Tie, WANG Jian-Wei, LI Ai-Hua. ISOLATION AND CHARACTERIZATION OF AEROMONAS SALMONICIDA SUBSPECIES SALMONICIDA FROM LARGEMOUTH BRONZE GUDGEON (COREIUS GUICHENOTI) CAGE-CULTURED IN THE UPPER REACHES OF YANGTZE RIVER[J]. ACTA HYDROBIOLOGICA SINICA, 2020, 44(1): 153-161. DOI: 10.7541/2020.018
Citation: LONG Meng, LI Tong-Tong, JIANG Yao, ZHANG Qian-Qian, ZHANG De-Feng, ZHANG Fu-Tie, WANG Jian-Wei, LI Ai-Hua. ISOLATION AND CHARACTERIZATION OF AEROMONAS SALMONICIDA SUBSPECIES SALMONICIDA FROM LARGEMOUTH BRONZE GUDGEON (COREIUS GUICHENOTI) CAGE-CULTURED IN THE UPPER REACHES OF YANGTZE RIVER[J]. ACTA HYDROBIOLOGICA SINICA, 2020, 44(1): 153-161. DOI: 10.7541/2020.018

长江上游网箱养殖圆口铜鱼杀鲑气单胞菌杀鲑亚种的分离与鉴定

ISOLATION AND CHARACTERIZATION OF AEROMONAS SALMONICIDA SUBSPECIES SALMONICIDA FROM LARGEMOUTH BRONZE GUDGEON (COREIUS GUICHENOTI) CAGE-CULTURED IN THE UPPER REACHES OF YANGTZE RIVER

  • 摘要: 研究首次报道了圆口铜鱼(Coreius guichenoti)疥疮病, 从患病圆口铜鱼的肝脏中分离到优势菌株YTL1, 并运用形态学观察、生理生化检测、16S rRNA和6个管家基因的系统发育分析等对分离菌株进行鉴定。基于以上实验结果, YTL1被最终鉴定为杀鲑气单胞菌杀鲑亚种(Aeromonas salmonicida subsp. salmonicida)。通过标准Kirby-Bauer纸片扩散法进行抗菌药物敏感性试验, 筛选治疗该暴发病的有效药物, 结果显示YTL1对氟苯尼考, 诺氟沙星和氨苄青霉素等13种抗生素敏感, 对6种抗生素如杆菌肽, 链霉素和卡那霉素有耐药性, 对红霉素具有中等敏感性。因此, 氟苯尼考被建议用来伴饵投喂, 并取得了较好的疾病控制效果。草鱼幼鱼(Ctenopharyngodon idella)和斑马鱼(Danio rerio)的人工感染试验结果显示, 经腹腔注射7.6×106—7.6×108 CFU/mL的YTL1菌液后, 感染鱼的症状与患病圆口铜鱼症状相似。研究证明基于6个管家基因的多序列位点分型是鉴定杀鲑气单胞菌至亚种水平的一种有效方法, 杀鲑气单胞菌是圆口铜鱼人工养殖的最大威胁之一, 并发现鲤科鱼类, 如草鱼和斑马鱼均是杀鲑气单胞菌杀鲑亚种的易感宿主。

     

    Abstract: Largemouth bronze gudgeon (Coreius guichenoti) is a potamodromous and endemic fish in the upper reaches of Yangtze River. An epidemic was found in largemouth bronze gudgeon at a farm in Luzhou, Sichuan province, southwest China, at the end of March 2012. In this study, we reported the first observation of furunculosis found in largemouth bronze gudgeon. One dominant bacteria strain, YTL1, was isolated from the liver of diseased largemouth bronze gudgeon, and a series of methods including morphological observation, biochemical tests, and phylogenetic analysis of 16S rRNA and six housekeeping genes were used to identify the pathogen. The strain was finally identified as A. salmonicida subsp. salmonicida based on the results. Antimicrobial susceptibility tests were carried out by the standard Kirby-Bauer disc diffusion method to screen effective drugs for the therapy of the disease, with results showing that YTL1 was sensitive to thirteen antibiotics such as florfenicol, norfloxacin, and ampicillin, resistant to 6 antibiotics such as bacitracin, streptomycin, and kanamycin, and mid-sensitive to erythromycin. Accordingly, florfenicol was added into diets to control furunculosis in largemouth bronze gudgeon with a good result. Artificial infection experiments in grass carp fingerlings and zebrafish resulted in the similar symptoms as diseased largemouth bronze gudgeon. In conclusion, our study demonstrated that multilocus sequence typing based on six housekeeping genes is an effective method to identify A. salmonicida strains to the subspecies level, confirmed that A. salmonicida infection is one of the greatest threats to artificial breeding and aquaculture of largemouth bronze gudgeon, and has expanded the susceptible hosts of A. salmonicida subsp. salmonicida to more cyprinid fishes.

     

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