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| 大黄鱼(Pseudosciaena crocea)致病性维氏气单胞菌的分离鉴定与药敏特性研究 |
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李忠琴1,2,3, 张坤1,2,3, 林茂1,3, 赖晓健1,2,3, 江兴龙1,2
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1.集美大学水产学院 厦门 361021;2.鳗鲡现代产业技术教育部工程研究中心 厦门 361021;3.厦门市渔用药物工程技术研究中心 厦门 361021
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| 摘要: |
| 本研究从自然发病的养殖大黄鱼（Pseudosciaena crocea）肝脏处分离得到1株优势菌9L2，经形态学观察和传统生理生化鉴定，显示该菌为发酵型革兰氏阴性短杆菌，有动力，氧化酶阳性，水解赖氨酸及色氨酸等。利用Biolog细菌鉴定方法进行种属鉴定，确定9L2为维氏气单胞菌（Aeromonas veronii），相似性达100%。同时对其16S rDNA基因进行PCR扩增、测序和系统发育分析，结果表明该菌与气单胞菌属的菌株亲缘关系最近，与维氏气单胞菌（登录号为LMG13695）的16S rRNA基因序列相似性达99.79%。经人工腹腔注射感染吉富罗非鱼（Oreochromis niloticus）的试验结果表明，9L2对罗非鱼的半数致死剂量LD50为1.08×103CFU/g。24种抗生素药敏试验结果表明，该菌株对诺氟沙星、新生霉素、呋喃妥因、头孢曲松等13种药物高度敏感，对洛美沙星、卡那霉素等5种药物中度敏感，对利福平、青霉素等6种药物产生耐药性。20种中药体外抑菌试验结果表明，其中11种对致病菌体外抑菌效果较好，平均抑菌浓度范围为0.11-7.20mg/mL。 |
| 关键词: 大黄鱼 维氏气单胞菌 药敏实验 病原菌 |
| DOI：10.11693/hyhz20160400094 |
| 分类号: |
| 基金项目:福建省教育厅项目，JA15260号；公益性行业（农业）科研专项，201203085号；海洋经济发展区域示范项目，14PYY050SF03号；国家海洋局海洋经济创新发展区域示范专项，2012FJ03号；厦门市重点项目，3502Z20110014号；鳗鲡现代产业技术教育部工程研究中心开放基金，RE201606号，RE201607号；大学生创新创业训练计划项目，201510390053号。 |
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| ISOLATION AND IDENTIFICATION OF PATHOGENIC AEROMONAS VERONII FROM PSEUDOSCIAENA CROCEA |
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LI Zhong-Qin1,2,3, ZHANG Kun1,2,3, LIN Mao1,3, LAI Xiao-Jian1,2,3, JIANG Xing-Long1,2
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1.Fisheries College, Jimei University, Xiamen 361021, China;2.Engineering Research Centre of the Modern Industry Technology for Ministry of Education, Xiamen 361021, China;3.Engineering Technology Research Centre of Fishery Medicine in Xiamen, Xiamen 361021, China
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| Abstract: |
| A dominant bacterial strain, named 9L2, was isolated from the liver of diseased Pseudosciaena crocea and identified by biochemical characterization, Biolog identification, and 16S rRNA gene sequence analysis. The bacteria was fermented, Gram-negative, motile, and oxidase-positive, could degrade lysine and tryptophan based on morphological, physiological, biochemical tests. The analysis of 16S rRNA gene sequence show that the organism and the type strain of Aeromonas veronii (LMG13695) formed consistently a monophyletic clade with a distant sequence similarity of 99.79%. According to the results, the bacterium was identified as A. veronii. Moreover, lethal dose and 50% trials were carried out to demonstrate the virulence of 9L2 in Oreochromis niloticus when administered at 1.08×103CFU/g. Drug sensitivity test on 24 antimicrobial agents and 20 Chinese herbs showed that the strain were very sensitive to 13 agents (norfloxacin, novobiocin, nitrofurantoin, ceftriaxone, etc.), while mild or not sensitive to the rest; of 20 Chinese herbs, 11 herbs showed better antibacterial effect in vitro in average minimum inhibitory concentration ranging from 0.11 to 7.20mg/mL. |
| Key words: Pseudosciaena crocea Aeromonas veronii antibiotic sensitivity pathogenic bacterium |