山瑞鳖细菌性败血症病原菌的分离鉴定及其毒力基因检测
发布时间：2019-06-11 01:32
【摘要】：查明广西南宁、贵港和桂平养殖山瑞鳖细菌性败血症的病原菌及其6种毒力基因的携带情况,为有效防控山瑞鳖细菌性败血症提供参考。本研究以常规方法从患病山瑞鳖的心脏和肝脏取样、分离细菌,人工感染方法确定分离菌株的致病性,细菌鉴定采用API 20NE生化鉴定和16S rRNA分子鉴定相结合的方法进行,PCR扩增法对菌株的溶血素基因(hemolysin gene,hly)、气溶素基因(aerolysin gene,Aer)、细胞兴奋性肠毒素基因(cytotonic enterotoxin gene,Alt)、细胞毒性肠毒素基因(cytotoxic enterotoxin gene,Act)、黏附素基因(major adhesin gene,ahal)和丝氨酸蛋白酶基因(serine protease gene,ahp)6种毒力基因进行检测。结果显示,从患病山瑞鳖心脏和肝脏中共分离到4株优势菌SRB125、SRB142、SRB143和SRB345,对健康山瑞鳖的平均致死率为97.50%~100.00%,是引起山瑞鳖细菌性败血症的病原菌;生化和分子鉴定结果显示,4株分离菌均为嗜水气单胞菌(Aeromonas hydrophila),与A.hydrophila L3-5(KP716701)菌株的亲缘关系最近,同源相似性均达到99.9%;6种毒力基因共包含2种毒力基因型,在4株菌株中的分布为hly~+Aer~+Alt~+Act~+ahal~+ahp~+和hly~+Aer~+Alt~+Act~+ahal~+ahp~ 各2株,来源于南宁的SRB143和桂平的SRB345菌株均缺失ahp基因。
[Abstract]:To find out the pathogen of bacterial septicemia and the carrying of six virulence genes in cultured Trionyx sinensis in Nanning, Guigang and Guiping, Guangxi, so as to provide reference for the effective prevention and control of bacterial septicemia of Trionyx sinensis. In this study, bacteria were isolated from the heart and liver of Trionyx sinensis by routine method, and the pathogenicity of the isolated strains was determined by artificial infection. API 20NE biochemical identification and 16s rRNA molecular identification were used to identify the bacteria. The hemolysin gene (hemolysin gene,hly), aerosol gene (aerolysin gene,Aer), cellular excitatory enterotoxin gene (cytotonic enterotoxin gene,Alt), cytotoxicity enterotoxin gene (cytotoxic enterotoxin gene,Act and adhesin gene (major adhesin gene, were amplified by PCR. Ahal) and serine protease gene (serine protease gene,ahp) were detected by six virulence genes. The results showed that four strains of dominant bacteria SRB125,SRB142,SRB143 and SRB345, were isolated from the heart and liver of the diseased soft-shelled turtle (Trionyx sinensis), and the average fatality rate of the four dominant bacteria to the healthy soft-shelled turtle was 97.50% and 100.00%, which was the pathogen of bacterial septicemia in the soft-shelled turtle (Trionyx sinensis). The results of biochemical and molecular identification showed that the four isolates were (Aeromonas hydrophila), and A.hydrophila L3 鈮，
本文编号：2496899
[Abstract]:To find out the pathogen of bacterial septicemia and the carrying of six virulence genes in cultured Trionyx sinensis in Nanning, Guigang and Guiping, Guangxi, so as to provide reference for the effective prevention and control of bacterial septicemia of Trionyx sinensis. In this study, bacteria were isolated from the heart and liver of Trionyx sinensis by routine method, and the pathogenicity of the isolated strains was determined by artificial infection. API 20NE biochemical identification and 16s rRNA molecular identification were used to identify the bacteria. The hemolysin gene (hemolysin gene,hly), aerosol gene (aerolysin gene,Aer), cellular excitatory enterotoxin gene (cytotonic enterotoxin gene,Alt), cytotoxicity enterotoxin gene (cytotoxic enterotoxin gene,Act and adhesin gene (major adhesin gene, were amplified by PCR. Ahal) and serine protease gene (serine protease gene,ahp) were detected by six virulence genes. The results showed that four strains of dominant bacteria SRB125,SRB142,SRB143 and SRB345, were isolated from the heart and liver of the diseased soft-shelled turtle (Trionyx sinensis), and the average fatality rate of the four dominant bacteria to the healthy soft-shelled turtle was 97.50% and 100.00%, which was the pathogen of bacterial septicemia in the soft-shelled turtle (Trionyx sinensis). The results of biochemical and molecular identification showed that the four isolates were (Aeromonas hydrophila), and A.hydrophila L3 鈮，
本文编号：2496899
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