| 克氏原螯虾副溶血弧菌的分离鉴定及药敏特性 |
| Isolation, Identification and Antibiotic Sensitivity of Vibrio parahaemolyticus Isolated from Procambarus clarkii |
| 投稿时间：2019-11-20 修订日期：2022-01-20 |
| DOI：10.15928/j.1674-3075.201911200289 |
| 中文关键词:克氏原螯虾 副溶血弧菌 生化特性 药敏特性 |
| 英文关键词:Procambarus clarkii Vibrio parahaemolyticus biochemical characteristics antibiotic sensitivity |
| 基金项目:湖北省技术创新专项（2018ABA103）；中央高校基本科研业务费专项资金（2662018YJ022） |
| 作者 | 单位 | | 霍诗天 | 华中农业大学水产学院水生动物医学系, 湖北 武汉430070；湖北省水生动物病害防控工程技术研究中心, 湖北 武汉 430070；水产养殖国家级实验教学示范中心, 湖北 武汉 430070 | | 焦厚琪 | 华中农业大学水产学院水生动物医学系, 湖北 武汉430070；湖北省水生动物病害防控工程技术研究中心, 湖北 武汉 430070；水产养殖国家级实验教学示范中心, 湖北 武汉 430070 | | 李 清 | 水产养殖国家级实验教学示范中心, 湖北 武汉 430070 | | 兰江风 | 华中农业大学水产学院水生动物医学系, 湖北 武汉430070；湖北省水生动物病害防控工程技术研究中心, 湖北 武汉 430070；水产养殖国家级实验教学示范中心, 湖北 武汉 430070 | | 顾泽茂 | 华中农业大学水产学院水生动物医学系, 湖北 武汉430070；湖北省水生动物病害防控工程技术研究中心, 湖北 武汉 430070；水产养殖国家级实验教学示范中心, 湖北 武汉 430070 | | 刘学芹 | 华中农业大学水产学院水生动物医学系, 湖北 武汉430070；湖北省水生动物病害防控工程技术研究中心, 湖北 武汉 430070；水产养殖国家级实验教学示范中心, 湖北 武汉 430070 |
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| 中文摘要: |
| 克氏原螯虾（Procambarus clarkii）感染副溶血弧菌（Vibrio parahaemolyticus）会导致大量死亡，给养殖业造成了巨大的经济损失，亟待寻求有效的防治措施。从湖北省公安县某养殖场采集到一批患病的克氏原螯虾，解剖后发现病虾的肝胰腺肿大发黄，肠道内无食物，有类似肠炎症状，从肝胰腺中分离出的致病菌经TCBS培养，得到光滑蔓延生长的菌落，呈蓝绿色斗笠状，菌落大小不一。显微观察发现细菌形态为棒状或弧状，长短不一，在革兰氏染色呈红色，属于革兰氏阴性菌，细菌形态单一；通过分析其16S rDNA序列和生理生化特征，鉴定为副溶血弧菌并命名为Vp-2，其与弧菌属种类同源性最高，并与菌株Vibrio parahaemolyticus DQ068942.1的亲缘关系较近。通过纸片扩散法进行药敏试验，该菌株对氟苯尼考、卡那霉素、复方新诺明等7种抗生素敏感；对利福平、磺胺异恶唑、庆大霉素3种药物耐药。生理生化特性测定结果显示，菌株Vp-2的理化特性基本与标准菌株一致。人工回感实验表明，发病克氏原螯虾的症状与自然发病症状类似，解剖发现死亡的克氏原螯虾肝胰腺肿大发黄，肠道内有炎症现象，并从中分离到该菌。试验表明，分离到的副溶血弧菌菌株对克氏原螯虾有致病性，生产中可用氟苯尼考、四环素等药物进行防治。 |
| 英文摘要: |
| Infection of Procambarus clarkii (red swamp crayfish) by Vibrio parahaemolyticus results in high mortality and large economic losses to the P. clarkii breeding industry. Thus, there is an urgent need for effective measures to prevent and control this pathogenic bacteria. In this study, diseased P. clarkii were collected from a farm in Gong'an County, Hubei Province, showing clinical symptom of enteritis: an enlarged, yellow hepatopancreas and no food in the intestines. To identify the pathogen infecting P. clarkii and support efforts to find effective inspection and prevention strategies for the disease, the dominant pathogenic strain was isolated from the hepatopancreas of diseased P. clarkii and labeled the Vp-2 strain. The Vp-2 strain was identified through morphological examination, physiological and biochemical analysis, and phylogenetic tree analysis based on 16S rDNA sequencing. An artificial infection study was carried out on healthy P. clarkii by injecting a suspension of the Vp-2 strain, and an antibiotic susceptibility test was carried out using 11 antibiotics. The bacterial strain Vp-2 was characterized as gram-negative, rod or arc-shaped, of variable length, red with gram stain, and the16S rDNA sequence is 1 449 bp. The Vp-2 strain, grown on TCBS media, developed smooth, spreading, blue-green hat-shaped colonies of varying size. Its phylogenic tree, constructed from the 16S rDNA sequence, displayed the highest homology with other species of the Vibrio genus and is closely related to the strain Vibrio parahaemolyticus (DQ068942.1). Combined with the morphological characteristics and physiological and biochemical characteristics of the colony, the Vp-2 strain was conclusively identified as Vibrio parahaemolyticus. The susceptibility of the Vp-2 strain was tested by disc diffusion and the results show that the strain was sensitive to seven of the antibiotics (florfenicol, kanamycin and trimethoprims) and showed resistance to three (rifampicin, sulfisoxazole and gentamicin) . The artificial infection study was carried out on healthy P. clarkii by injecting a Vp-2 strain suspension (1×108 CFU/mL) and symptoms appeared that were similar to those of the naturally infected P. clarkii. Further, the morphological, physiological and biochemical characteristics of the pathogenic strain isolated from artificially infected P. clarkii were identical with strain Vp-2. The V. parahaemolyticus strain isolated in this experiment is pathogenic to P. clarkii, and drugs such as florfenicol and tetracycline can be used to prevent and control the disease during production. |
| 霍诗天,焦厚琪,李 清,兰江风,顾泽茂,刘学芹.2022.克氏原螯虾副溶血弧菌的分离鉴定及药敏特性[J].水生态学杂志,43(1):124-129. |
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