| 大黄鱼致病嗜水气单胞菌的分离鉴定及其多克隆抗体制备 |
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| 引用本文： | 何亮银,周逢芳,史晓丽,黄伟卿,阮俊峰,田文贞,李进寿.大黄鱼致病嗜水气单胞菌的分离鉴定及其多克隆抗体制备[J].水产科学,2022,41(1):69-75. |
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| 作者姓名： | 何亮银 周逢芳 史晓丽 黄伟卿 阮俊峰 田文贞 李进寿 |
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| 作者单位： | 宁德师范学院生命科学学院,福建宁德352100 |
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| 基金项目： | 福建省科技厅自然科学基金项目(2018J05062);宁德师范学院引进人才科研经费资助项目(2017Y03);宁德师范学院个人发展基金项目(2018FZ07);宁德师范学院闽东优势农产品高值化利用协同创新中心项目(2017Z02). |
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| 摘 要： | 为探究引起网箱养殖大黄鱼肠炎病的病原及其特性,自患病大黄鱼肝脏组织分离到1株优势菌NDLc-P,经回归感染试验确认该菌为大黄鱼致病菌。人工感染7 d后,该菌对体质量(46.4±3.5)g的健康大黄鱼的半数致死密度为3.98×107 cfu/mL,表明该菌株对大黄鱼有一定的致病性。利用梅里埃微量多项鉴定系统对病原菌NDLc-P的理化特性进行分析,初步判定其为嗜水气单胞菌;病原菌的16S rDNA序列经在线同源比对,与数据库中嗜水气单胞菌的同源性达99.87%,基于16S rDNA的病原菌进化树也显示,病原菌与嗜水气单胞菌聚为一支。综合病原菌的生理生化特性及16S rDNA序列分析结果,判定菌株NDLc-P为嗜水气单胞菌。制备嗜水气单胞菌的鼠源多克隆抗体,酶联免疫吸附试验测定其效价为40000,显示抗血清与病原的良好结合特性。试验结果将为大黄鱼嗜水气单胞菌病的病原特征研究、诊断检测及免疫防治提供参考。
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| 关 键 词： | 大黄鱼 嗜水气单胞菌 分离鉴定 多克隆抗体 |
Isolation,Identification and Polyclonal Antibody Preparation of Aeromonas hydrophila in Large Yellow Croaker Larimichthys crocea |
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| HE Liangyin,ZHOU Fengfang,SHI Xiaoli,HUANG Weiqing,RUAN Junfeng,TIAN Wenzhen,LI Jinshou.Isolation,Identification and Polyclonal Antibody Preparation of Aeromonas hydrophila in Large Yellow Croaker Larimichthys crocea[J].Fisheries Science,2022,41(1):69-75. |
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| Authors: | HE Liangyin ZHOU Fengfang SHI Xiaoli HUANG Weiqing RUAN Junfeng TIAN Wenzhen LI Jinshou |
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| Institution: | (College of Life Science, Ningde Normal University, Ningde 352100, China) |
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| Abstract: | A dominant bacterial strain NDLc-P was isolated from the liver of diseased large yellow croaker Larimichthys crocea in Ningde,Fujian Province.An artificial infection with the isolates caused the similar symptom in the fish,with 50%lethal concentration of 3.98×107 cfu/mL within 7 d trial.Physiological and biochemical tests by commercially produced kit systems(bioMerieux)revealed that strain NDLc-P was identified as Aeromonas hydrophila.Then,the 16S rDNA sequence analysis showed that the bacterium had a 99.87%identify with A.hydrophila.The phylogenetic analysis also showed that the NDLc-P isolate was clustered most clearly with A.hydrophila.Furthermore,healthy mice were immunized with formalin inactivated strain NDLc-P,and the ELISA showed that the titer of antisera was determined to be 40000.These findings provided a reference for research on pathogenic characteristics,diagnosis and prevention and treatment of large yellow croaker disease caused by A.hydrophila. |
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| Keywords: | Larimichthys crocea Aeromonas hydrophila isolation and identification polyclonal antibody |
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