鲤致病性维氏气单胞菌的分离鉴定及生物学特性分析

Isolation, identification and biological characteristics of Aeromonas veronii from Cyprinus carpio

  • 摘要: 【目的】分离鉴定鲤(Cyprinus carpio)致病性维氏气单胞菌(Aeromonas veronii) ,为探究暴发病害养殖鲤的死亡原因及为防治水产经济鱼类病害提供参考依据。【方法】从暴发病害鲤的内脏组织中分离病原菌,通过形态观察、生理生化鉴定、16S rDNA和gyrB基因测序分析确定其分类地位,通过人工回归感染试验判断其致病性,通过药物敏感性试验判定其对抗生素的耐药性。【结果】细菌形态学观察和生理生化试验结果表明,从暴发病害鲤的内脏中分离的病原菌LY623为维氏气单胞菌。16S rDNA和gyrB基因测序结果显示,LY623菌株与维氏气单胞菌菌株的相似性最高,且在系统发育进化树中与维氏气单胞菌聚为一支。毒力基因和耐药基因检测结果表明,菌株LY623携带Alt、Act、hlyA、lipela 5种毒力基因及tem、ctxM、tetC、qnrAqnrS 5种耐药基因。药敏试验结果表明,LY623菌株对四环素、诺氟沙星和红霉素等14种抗菌药物具有耐药性,对庆大霉素、氟苯尼考和氯霉素等14种抗菌药物高度敏感。人工回归感染试验结果表明,分离获得的维氏气单胞菌LY623对鲤的半数致死量(LD50)为2.64×103 CFU/g,对斑马鱼(Daniorerio)的LD50为4.77×107 CFU/g。【结论】从暴发病害鲤内脏组织中分离到的病原菌LY623鉴定为维氏气单胞菌,其毒力基因包括Alt、Act、hlyA、lipela,耐药基因包括tem、ctxM、tetC、qnrAqnrS。该菌株对鲤具有较高毒力,在其肠道、鳃、肝脏和脾脏中引起明显的组织病理学变化;在鲤实际养殖过程中可选用庆大霉素、氟苯尼考和氯霉素等对该菌感染引起的疾病进行治疗。

     

    Abstract: 【Objective】 Pathogenic Aeromonas veronii for carp was isolated and identified,to study the causes of death in cultured common carp(Cyprinus carpio) subjected to disease outbreaks,with the aim of providing references for the prevention and control of economic fish diseases in aquaculture.【Method】 The pathogenic bacteria were isolated from the visceral tissues of the diseased common carp and the taxonomic status was investigated by using morphological observation,physiological and biochemical identification,and analysis on the gene sequences of 16S rDNA and gyrB. Artificial regression infection experiments were used to assess the pathogenicity and antibiotic susceptibility tests were conducted to investigate the resistances to antibiotics.【Result】 The bacterial pathogen LY623 isolated from the viscera of the diseased carp was A. veronii according to the results of morphological observation and physiological and biochemical experiments. The 16S rDNA and gyrB gene sequences analysis showed that LY623 strain had the highest similarity with A. veronii strains and was clustered together with the strains of A. veronii based on the phylogenetic tree.Analysis results of virulence genesand drug resistance genes showed that the LY623 strain carried five virulence genes including Alt, Act, hlyA, lip and ela and five drug resistance genes including tem, ctxM, tetC, qnrA and qnrS. The results of antibiotic susceptibility test showed that LY623 strain was resistant to fourteen antibiotics such as tetracycline, norfloxacin,and erythromycin,and highly susceptible to fourteen antibiotics such as gentamicin,florfenicol,and chloramphenicol. The results of artificial regression infection experiments showed that the median lethal dosages(LD50) of this strain were 2.64×103 CFU/g and 4.77×107 CFU/g to C. carpio and Danio rerio,respectively.【Conclusion】 The bacterial pathogen LY623 isolated from the visceral tissues of diseased common carp is A. veronii. It carries five virulence genes including Alt, Act, hlyA, lip and ela and five drug resistance genes including tem, ctxM, tetC, qnrA and qnrS. It has high virulence to the common carp,causing obvious histopathological changes in the tissues such as intestine,gill,liver,and spleen. Gentamicin,florfenicol,and chloramphenicolare alternative antibiotics to control the common carp diseases caused by A. veronii infection.

     

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