黄沙鳖红底板病病原菌的分离鉴定及其毒力基因检测  

Isolation,Identification and Virulence Genes Detection of Pathogenitic Bacteria from Trionyx sinensis with Red Abdominal Shell Disease

作　　者：黄艳华[1,2] 黄钧[1] 胡大胜 施金谷[1] 彭民毅[1] 彭亚[1] 

机构地区：[1]广西大学动物科学技术学院/广西水生动物病害诊断实验室,广西南宁530005 [2]梧州市水产技术推广站,广西梧州543002 [3]广西水产技术推广总站,广西南宁530022

出　　处：《西南农业学报》2013年第5期2116-2121,F0003,共7页Southwest China Journal of Agricultural Sciences

基　　金：广西自然科学基金项目(2011GXNSFA018065);广西水产畜牧兽医局专项项目(桂渔牧财[2011]52号,桂渔牧财[2012]30号)

摘　　要：以常规方法从患典型红底板病黄沙鳖的心脏、肝脏和腹水等处分离到5株病原菌,API 20NE、16S rRNA基因序列分析进行病原菌鉴定和确定其系统发育地位,PCR检测病原菌的6种毒力基因。结果表明,5株病原菌均为嗜水气单胞菌(Aeromonas hydrophila),与Aeromonas hydrophila QDC01菌株的亲源关系最近。6种毒力基因的阳性率,hly、Aer和Act为100%,ahal、Alt和ahp为80%;毒力基因型共3种,在5株菌株中的分布情况为hly+Aer+Alt+Act+ahal+ahp+3株,hly+Aer+Alt-Act+ahal+ahp-1株,hly+Aer+Alt+Act+ahal-ahp+1株。试验所检测的5株嗜水气单胞菌的毒力基因型主要为hly+Aer+Alt+Act+ahal+ahp+,Alt、ahp和ahal基因的缺失对嗜水气单胞菌的致病力没有影响。Five pathogens from the Trionyx sinensis infected with red abdominal shell disease were isolated from hearts、livers and ascites by routine methods.Their phylogenesises were determined by API 20NE analysis and 16Sr RNA gene PCR amplification followed by PCR assay according to 6 chosen virulence genes.Results showed 5 strains isolates are Aeromonas hydrophila.Phylogenetic tree showed 5 Aeromonas hydrophila have high similarities to Aeromonas hydrophila QDC01 bacteria.The hly Aer and Act gene could be detected in all isolates（100 ％）,also the Ahal Alt and ahp were found in 80 ％ isolates.3 genotypes were found in these isolates,showing 3 hly ＋ Aer ＋ Alt ＋ Act ＋ ahal＋ ahp＋,1 hly＋Aer＋Alt-Act＋ahal＋ahp-,1 hly＋Aer＋Alt＋Alt＋ahal-ahp＋ dominated in 5 strains.The test showed major virulence genotypes was hly＋Aer＋Alt＋Act＋ahal＋ahp＋in 5 hydrophila Aeromonas,Lack of the Alt,ahp and ahal gene had no effect to pathogenicity of Aeromonas hydrophilas.

关 键 词：黄沙鳖 红底板病 嗜水气单胞菌 毒力基因 

分 类 号：S965.1[农业科学—水产养殖]