| 胡子鲶致病性气单胞菌的分离鉴定及其致病力与毒力基因型相关性 |
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| 引用本文： | 龙苏,韩书煜,牛志伟,梁静真,胡大胜,黄钧,李鸿骥,刘齐,苏江华.胡子鲶致病性气单胞菌的分离鉴定及其致病力与毒力基因型相关性[J].水产学报,2016,40(3):308-317. |
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| 作者姓名： | 龙苏 韩书煜 牛志伟 梁静真 胡大胜 黄钧 李鸿骥 刘齐 苏江华 |
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| 作者单位： | 1. 广西水生动物病害诊断实验室,广西南宁530005;广西大学动物科学技术学院,广西高校水生生物健康养殖与营养调控重点实验室,广西南宁530005;2. 广西水生动物病害诊断实验室,广西南宁530005;广西壮族自治区水产技术推广总站,广西南宁530021 |
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| 基金项目： | 广西水产畜牧兽医局专项项目(桂渔牧财[2012]30号,桂渔牧财[2013]35号,桂渔牧财[2014] 52号) |
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| 摘 要： | 为探讨广西南宁市、浦北县和玉林市暴发性死亡胡子鲶的病原菌及其所携带6种毒力基因对其致病力的影响,用常规方法从病鱼的心脏、肝脏等部位分离细菌,人工感染实验确定病原菌的致病性,以API 20NE生化鉴定和16S r RNA分子鉴定相结合的方法对病原菌进行鉴定,采用PCR扩增法检测病原菌的6种毒力基因携带情况。结果显示,从患病鱼中共分离到5株病原菌,其中嗜水气单胞菌3株,温和气单胞菌2株。3株嗜水气单胞菌与标准菌株Aeromonas hydrophila ATCC 7966(CP000462)的亲缘关系最近,相似性均为99.8%,2株温和气单胞菌与标准菌株Aeromonas sobria NO.106(AB472903.1)的亲缘关系最近,相似性均达99.9%。6种毒力基因在5株病原菌中的阳性检出率分别为Act和Aer基因100%,ahal、hly和Alt基因均为80%,ahp基因仅20%;毒力基因型共3种,在5株气单胞菌中分布情况为Act+ahal+hly+Alt+ahp-Aer+3株,占实验菌株的60%,为主要的毒力基因,Act+ahal+hly+Alt+ahp+Aer+和Act+ahal-hly-Alt-ahp-Aer+各1株,各占20%。携带全部所检6种毒力基因的菌株致病力最强,只携带Act和Aer 2种毒力基因的菌株致病力最弱。ahp基因在菌株的致病力中起重要作用,病原菌的致病力是多种毒力基因协同作用的结果。
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| 关 键 词： | 胡子鲶 暴发性死亡症 嗜水气单胞菌 温和气单胞菌 毒力基因 |
| 收稿时间： | 2015/11/2 0:00:00 |
| 修稿时间： | 2016/1/21 0:00:00 |
Isolation and identification of pathogenic Aeromonas in Clarias fuscus and analysis of the correlation between its pathogenicity and virulence genotypes |
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| LONG Su,HAN Shuyu,NIU Zhiwei,LIANG Jingzhen,HU Dasheng,HUANG Jun,LI Hongji,LIU Qi and SU Jianghua.Isolation and identification of pathogenic Aeromonas in Clarias fuscus and analysis of the correlation between its pathogenicity and virulence genotypes[J].Journal of Fisheries of China,2016,40(3):308-317. |
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| Authors: | LONG Su HAN Shuyu NIU Zhiwei LIANG Jingzhen HU Dasheng HUANG Jun LI Hongji LIU Qi and SU Jianghua |
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| Institution: | Guangxi Aquatic Animal Disease Diagnostic Laboratory,Guangxi Aquatic Animal Disease Diagnostic Laboratory,Guangxi Aquatic Animal Disease Diagnostic Laboratory,Guangxi Aquatic Animal Disease Diagnostic Laboratory,Guangxi Aquatic Animal Disease Diagnostic Laboratory,Guangxi Aquatic Animal Disease Diagnostic Laboratory |
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| Abstract: | This study investigated the pathogenic bacteria of Clarias fuscus with fulminant death in Nanning City, Pubei County and Yulin City of Guangxi, and analysed the effect of the six virulence genes on its pathogenicity. Pathogen was isolated from the heart, liver and some other parts of the diseased fish by using conventional method, then an artificial infection test was measured on the pathogenicity of the isolated strains. Identification of the isolates was confirmed by API 20NE analysis and 16S rRNA gene sequencing, including a PCR assay of detecting six kinds of virulence genes of the isolates. Results showed that five pathogenic strains were isolated from the diseased fish, including three strains of Aeromonas hydrophila and two strains of Aeromonas sobria. The three strains of Aeromonas hydrophila shared the highest similarity (99.8%) with Aeromonas hydrophila standard strain ATCC 7966 (CP000462). Meanwhile the two strains of Aeromonas sobria shared the highest similarity (99.9%) with Aeromonas sobria standard strain NO.106 (AB472903.1). Among the five strains, detection rates of six virulence genes were as follows. Hundred percent of the strains carried gene Act and Aer. Eighty percent carried gene ahal, hly and Alt. Only twenty percent of the strains carried gene ahp. The five Aeromonas strains contained three virulence genotypes in all, including the main genotype Act+ahal+hly+Alt+ahp-Aer+ of three strains (accounting for 60% of all the strains), as well as the genotype Act+ahal+hly+Alt+ahp+Aer+ and Act+ahal-hly-Alt-ahp-Aer+ of one strain (each accounting for 20% of all the strains). Strain which carried all the six detected virulence genes had the highest pathogenicity, whereas the one carrying only Act and Aer had the minimal pathogenicity. Gene ahp played an important role in pathogenicity of strains, and the pathogenicity of pathogenic bacteria resulted from the synergistic reaction of multiple virulence genes. |
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| Keywords: | Clarias fuscas fulminating epidemic disease Aeromonas hydrophila Aeromonas sobria virulence gene |
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