| 史磊,罗伟杰,黄瑜,等.斜带石斑鱼源致病溶藻弧菌的分离、鉴定及病理性分析[J].广东农业科学,2024,(7-8):- |
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斜带石斑鱼源致病溶藻弧菌的分离、鉴定及病理性分析 |
| The isolation, identification, and pathological analysis of Vibrio alginolyticus from cultured Epinephelus coioides |
| 投稿时间：2024-06-12 修订日期：2024-08-01 |
| DOI： |
| 中文关键词: 斜带石斑鱼，溶藻弧菌，16S rRNA，药敏试验，基因检测 |
| 英文关键词: Epinephelus coioides Vibrio alginolyticu 16S rRNA Drug susceptibility testing Virulence genes |
| 基金项目:National Natural Science Foundation of China (No. U20A2065) |
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| 中文摘要: |
| 【背景】溶藻弧菌(Vibrio alginolyticus)广泛分布于海水环境与动物体内,是海水养殖鱼类频发细菌性鱼病的主要致病菌之一【目的】为确定深圳市某石斑鱼养殖基地斜带石斑鱼(Epinephelus coioides)突发性死亡病因,本研究特从患病濒死斜带石斑鱼病灶组织中分离得到一株优势菌株SZGP226,【方法】对分离菌株形态特征、生理生化特性、16S rRNA基因进化毒力基因和耐药基因及药物敏感性进行研究,并对其进行回归感染。【结果】该菌对蔗糖、甘露醇、葡萄糖、尿素、氧化酶活性和以及VP等生理生化指标鉴定为阳性；SZGP226的16 S rRNA扩增序列与数据库上溶藻弧菌(Vibrio alginolyticus) 2014V-1072株具有同源性,同时携带tlh、toxR、ctsA、sto和vpi 5种毒力基因；综上结果,鉴定该菌为致病性溶藻弧菌。回归感染实验表明菌株SZGP226对斜带石斑鱼具有强致病性,LD50为1.63×107 CFU/mL,而且可引起石斑鱼的肝脏、肾脏和脾脏发生明显组织病理性损伤；药敏结果显示,该分离株耐受氨苄西林、苯唑西林等9种抗生素耐药,并携带qnrB、floR、tetC、tetM、qnrD、gyrA等9种耐药基因,最小抑菌实验结果发现杀菌效果最好的消毒剂为氢氧化钠,其最小杀菌(MIC)浓度8 mg/mL。【结论】综合以上研究结果,本次从病鱼分离到致病性溶藻弧菌,可用作对比以前研究已发表本地溶藻弧菌相关文献,为当地斜带石斑鱼养殖弧菌耐药性和致病力进化进行补充,同时以上实验结果可用于本次临床治疗,以期为控制斜带石斑鱼致病性溶藻弧菌感染防控和致病机理研究提供参考依据。 |
| 英文摘要: |
| [Background] Vibrio alginolyticus is widely distributed in the seawater environment and animals, and is one of the main causative organisms of frequent bacterial fish diseases in marine aquaculture fish [Objective] In order to determine the cause of sudden death of Epinephelus coioides in a grouper aquaculture base in Shenzhen City, a dominant strain, SZGP226, was isolated from the lesion tissue of a diseased grouper, and its physiological and biochemical characteristics, 16S rRNA genes, evolutionary virulence genes and drug-resistance genes were investigated. A dominant strain SZGP226 was isolated from the diseased tissues of grouper foci. [Methods] Morphological characteristics, physiological and biochemical properties, 16S rRNA gene evolution virulence genes and drug resistance genes, and drug susceptibility of the isolate were investigated, and regression infection was performed. [Results] The bacterium was identified as positive for sucrose, mannitol, glucose, urea, oxidase activity and and VP, and other physiological and biochemical indicators; the 16S rRNA amplified sequence of SZGP226 was homologous to Vibrio alginolyticus strain 2014V-1072 on the database, and it also carried the tlh, toxR, ctsA, sto and vpi; In summary, the results identified the bacterium as a pathogenic Vibrio alginolyticus. Regression infection experiments showed that strain SZGP226 was strongly pathogenic to slash-and-burn grouper, with an LD50 of 1.63×107 CFU/mL, and that it could cause obvious histopathological damage to the liver, kidney, and spleen of grouper; drug sensitivity results showed that the isolate was resistant to nine antibiotics such as ampicillin and benzoxycillin, and carried qnrB, floR, tetC, tetM, qnrD, gyrA and other 9 resistance genes, and the results of the minimal inhibition experiment found that the disinfectant with the best bactericidal effect was sodium hydroxide, with a minimal bactericidal (MIC) concentration of 8 mg/mL.[Conclusion] Taking the above findings together, this isolation of pathogenic Vibrio alginolyticus from diseased fish can be used as a comparison of previous studies of published local Vibrio alginolyticus related literature to supplement the evolution of Vibrio resistance and pathogenicity for local culture of Vibrio alginolyticus in slant grouper, meanwhile, the results of the above experiments can be used in the present clinical treatment with a view to providing a reference for the control of the prevention and control of the infection and the research on the mechanism of pathogenicity of pathogenic Vibrio alginolyticus in slant grouper. |
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