| 鱼源气单胞菌的毒力基因检测、分型及致病力 |
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| 引用本文： | 刘小芳,任燕,张德锋,巩华,石存斌,常藕琴,潘厚军.鱼源气单胞菌的毒力基因检测、分型及致病力[J].水产学报,2021,45(3):462-471. |
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| 作者姓名： | 刘小芳 任燕 张德锋 巩华 石存斌 常藕琴 潘厚军 |
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| 作者单位： | 上海海洋大学水产与生命学院,中国水产科学研究院珠江水产研究所,中国水产科学研究院珠江水产研究所,中国水产科学研究院珠江水产研究所,中国水产科学研究院珠江水产研究所 |
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| 基金项目： | 广州市科技计划项目(201904020004)；国家大宗淡水鱼产业技术体系(CARS-45)；中国水产科学研究院基本科研业务费专项(2020XT0405) |
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| 摘 要： | 为调查鱼源气单胞菌毒力基因与其致病力的相关性,以2009—2018年从不同患病鱼分离的173株气单胞菌为研究对象,通过检测毒力相关基因、测定溶血活性、腹腔注射感染异育银鲫等方法开展评价。通过管家基因gyrB分子鉴定结果显示,173株气单胞菌中维氏气单胞菌(119/173,68.9%)和嗜水气单胞菌(50/173,28.9%)是主要流行的菌株。10个毒力基因aer(162/173,93.64%)、act(131/173,75.72%)、ast(55/173,31.79%)、alt(58/173,33.53%)、lip(152/173,87.86%)、exu(154/173,89.02%)、fla(143/173,82.66%)、gcaT(148/173, 85.55%)、 eprCAI(41/173, 23.70%)和ahyB(51/173, 29.48%)普遍存在于173株气单胞菌中。依据检测到的毒力基因数量从多到少分布情况,这些菌株可分为7大类(Ⅰ～Ⅶ)53个毒力基因型。大部分嗜水气单胞菌检测到8～10个毒力基因,主要分布于Ⅰ、Ⅱ和Ⅲ类基因型;维氏气单胞菌的eprCAI、ahyB、 ast和alt等4个毒力基因检测率低,主要分布于Ⅳ、Ⅴ和Ⅵ类基因型。大部分气单胞菌(94.22%,163/173)具有溶血活性。代表性毒力基因型的38株维氏气单胞菌和20株嗜水气单胞菌腹腔注射异育银鲫攻毒结果显示,3.0×106 CFU/尾的剂量下,3株维氏气单胞菌使鲫死亡率达80%～100%,16株嗜水气单胞菌使鲫死亡率达90%～100%。研究表明,维氏气单胞菌是目前最主要流行的气单胞菌,但其检测到的毒力基因普遍少于嗜水气单胞菌,且对异育银鲫的致病力普遍弱于嗜水气单胞菌。本研究能够为气单胞菌败血症的流行病学调查和疫苗研究提供理论依据。
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| 关 键 词： | 气单胞菌 毒力基因 致病性 患病鱼 |
| 收稿时间： | 2020/3/5 0:00:00 |
| 修稿时间： | 2020/6/20 0:00:00 |
Detection, genotyping and pathogenicity of virulence genes in Aeromonas species isolated from diseased freshwater fish |
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| LIU Xiaofang,REN Yan,ZHANG Defeng,GONG Hu,SHI Cunbin,CHANG Ouqin,PAN Houjun.Detection, genotyping and pathogenicity of virulence genes in Aeromonas species isolated from diseased freshwater fish[J].Journal of Fisheries of China,2021,45(3):462-471. |
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| Authors: | LIU Xiaofang REN Yan ZHANG Defeng GONG Hu SHI Cunbin CHANG Ouqin PAN Houjun |
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| Institution: | College of Fisheries and Life Science,ShangHai Ocean University,ShangHai,Key Lab of Aquatic Animal Immune Technology of Guangdong Province,Key Lab of Fishery Drug Development of Ministry of Agriculture and Rural Affairs,Pearl River Fisheries Research Institute of Chinese Academy of Fishery Sciences,Key Lab of Aquatic Animal Immune Technology of Guangdong Province,Key Lab of Fishery Drug Development of Ministry of Agriculture and Rural Affairs,Pearl River Fisheries Research Institute of Chinese Academy of Fishery Sciences,Key Lab of Aquatic Animal Immune Technology of Guangdong Province,Key Lab of Fishery Drug Development of Ministry of Agriculture and Rural Affairs,Pearl River Fisheries Research Institute of Chinese Academy of Fishery Sciences,Key Lab of Aquatic Animal Immune Technology of Guangdong Province,Key Lab of Fishery Drug Development of Ministry of Agriculture and Rural Affairs,Pearl River Fisheries Research Institute of Chinese Academy of Fishery Sciences |
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| Abstract: | To investigate virulence genes and pathogenicity of Aeromonas species isolated from different diseased freshwater fish in 2009-2018,virulence genes including aer, act, fla, lip, gcaT, exu, ast, alt, eprCAI and ahyB were detected and the bacterial pathogenicity with different virulence genotype was assessed through intraperitoneal (IP) injection challenge Gibel carp. Molecular identification revealed that A.veronii (119/173, 68.8%) and A.hydrophila (50/173, 28.9%) were the most prevalent species in the diseased fish based on sequencing of gyrB. The distribution of 10 virulence factors including aer(162/173, 93.64%), act(131/173, 75.72%), ast(55/173, 31.79%), alt(58/173, 33.53%), lip(152/173, 87.86%), exu(154/173, 89.02%), fla(143/173, 82.66%), gcaT (148/173, 85.55%), eprCAI(41/173, 23.70%) and ahyB(51/173, 29.48%) were determined and 173 strains were sorted into 7 clusters and formed 53 genotypes according to the virulence distribution profile. The challenge tests showed that A.veronii isolates were generally less virulent than A.hydrophila isolates when infecting Gibel carp via IP injection. At the challenge dose of 3.0×106CFU per fish, A.hydrophila strains(16/20) were highly pathogenic and the mortality reached 90%-100%, but most A.veronii isolates (35/38)were less virulent or avirulent and only A.veronii YC170511, XZ180501 and GZ09007 with 5 ~ 7 virulence genes caused 80%-100% experimental fish death. The study suggested the pathogenicity of A.hydrophila isolates had obvious correlation with these virulence genes, but A.veronii isolates not. The research results have important reference significance for epidemiological investigations of motile aeromonad septicemia and vaccine research. |
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| Keywords: | Aeromonas virulence gene pathogenicity diseased freshwater fish |
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