禾花鲤致病性嗜水气单胞菌的分离鉴定、耐药性及毒力基因检测

发布时间：2018-04-24 12:47

本文选题：禾花鲤 + 嗜水气单胞菌　；参考：《西南农业学报》2017年04期

【摘要】：【目的】对广西全州县飞机坪鱼种场患病禾花鲤进行病原菌的分离鉴定,并对其致病性、耐药特性及6种毒力基因进行检测,为细菌引起的禾花鲤疾病及防控技术研究提供理论参考。【方法】从患病禾花鲤的肝脏、心脏和肾脏等部位取样分离细菌,人工感染确定菌株的致病性,API生化鉴定和16S rRNA分子鉴定相结合进行细菌鉴定,细菌的耐药特性试验为K-B纸片扩散法,毒力基因以PCR扩增法检测。【结果】从患病禾花鲤中分离到2株致病力很强的病原菌株TH1和TH3,对健康禾花鲤的致死率均为100.00%;经生化和分子鉴定,2株分离菌株均为嗜水气单胞菌(Aeromonas hydrophila),与标准菌株Aeromonas hydrophila ATCC7966(NR 118944)的亲缘关系最近,同源相似度99.80%;2株菌株对硫酸新霉素等5种药物高度敏感,对青霉素G等4种药物不敏感;TH1携带所检测的hly、Aer、Alt、Act、ahal和ahp 6种毒力基因,TH3只携带hly、Aer、Alt、Act、ahal 5种基因。【结论】引起广西全州县飞机坪鱼种场禾花鲤大批死亡的病原菌是嗜水气单胞菌,2株菌株TH1和TH3的毒力基因型分别为hly~+ Aer~+ Alt~+ Act~+ ahal~+ ahp~+ 和hly~+ Aer~+ Alt~+ Act~+ ahal~+ ahp~-,对健康禾花鲤的致死率均为100.00%,对硫酸新霉素等5种药物高度敏感,对青霉素G等4种药物不敏感。
[Abstract]:[objective] to isolate and identify pathogenic bacteria, pathogenicity, drug resistance and six virulence genes of common carp (Carpio przewalskii) in Jingping Fish seed Farm, Quanzhou County, Guangxi, China. To provide a theoretical reference for the study of disease and prevention and control of common carp caused by bacteria. [methods] sampling and isolating bacteria from liver, heart and kidney of diseased grass carp. Biochemistry and 16s rRNA molecular identification were combined to determine the pathogenicity of the strain. The drug resistance of the bacteria was determined by K-B disk diffusion method. The virulence genes were detected by PCR amplification. [results] two virulent pathogenic strains TH1 and TH3 were isolated from diseased grass carp, the mortality of them were 100.000.The two isolates were identified by biochemical and molecular analysis as follows: The relationship between Aeromonas hydrophila and standard strain Aeromonas hydrophila ATCC7966(NR 118944 was close to that of Aeromonas hydrophila. The homologous similarity of two strains was highly sensitive to neomycin sulfate and other 5 drugs. TH1, which is insensitive to penicillin G and other 4 drugs, carries 6 virulence genes, Hlya Aeroglorius actnahal and ahp, and TH3 only carries 5 genes of hlyapine, Altfus actnahal. [conclusion] the pathogen causing mass death of Cyprinus Carpio in Jingping Fish Farm, Quanzhou County, Guangxi is hydrophilic. [conclusion] the pathogen that caused the death of Cyprinus Carpio in Jingping Fish Farm, Guangxi Province was hydrophilic. The virulence genotypes of two strains of Aeromonas aeruginosa TH1 and TH3 were hlyAerAltActahalahp- and hlyAerAltActActahal-ahp-respectively. The fatality rates of the two strains were 100.000.The virulence genotypes of TH1 and TH3 were highly sensitive to neomycin sulfate and so on. It was insensitive to penicillin G and other 4 drugs.
【作者单位】：广西水生动物病害诊断实验室;全州县水产技术推广站;广西大学动物科学技术学院;广西水产技术推广总站;陆川县水产养殖技术推广站;都安瑶族自治县水产技术推广站;田东县水产技术推广站;
【基金】：广西水产畜牧兽医局专项项目(桂渔牧财[2013]35号,桂渔牧财[2014]52号,桂渔牧财[2015]97号)
【分类号】：S943

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