患病蝶尾金鱼维氏气单胞菌温和生物变种的分离鉴定  

Isolation,Identification and Antibiotic Sensitivity Analysis of Aeromonasveronii Biovar Sobria from Diseased Butterfly Tail Goldfish

作　　者：胡秀彩 孙金辉 孙敬锋 宋亚娇 吕爱军 陈成勋 

机构地区：[1]天津农学院水产学院、天津市水产生态及养殖重点实验室,天津300384 [2]河南师范大学水产学院,河南新乡453007

出　　处：《水生态学杂志》2018年第1期105-110,共6页Journal of Hydroecology

基　　金：天津市自然科学基金重点项目(16JCZDJC33500,15JCZDJC34000);国家自然科学基金(31272692);天津市水产产业技术体系创新团队(ITTFRS2017009);河南省高等学校重点科研项目(17A240001)

摘　　要：研究从患病蝶尾金鱼（Butterfly）肝脏中分离获得的1株病菌,为金鱼疾病防治提供参考。分离纯化获得1株细菌,编号为DW-1,通过细菌形态观察、理化特性、16S rDNA序列分析等方法进行鉴定。结果表明DW-1菌株为革兰氏阴性短杆菌,可发酵葡萄糖产气,赖氨酸脱羧酶、硝酸盐还原等为阳性;进一步通过PCR方法扩增16S rDNA序列,测得长度为1 385 bp,与维氏气单胞菌（Aeromonas veronii）相似性为99%~100%;构建系统发育树与维氏气单胞菌温和生物变种（A.veronii biovar sobria）模式菌株ATCC9071聚为一支。最终鉴定该菌株为维氏气单胞菌温和生物变种。人工感染试验结果表明,对斑马鱼半数致死量（LD_（50））为1.17×10~6CFU/m L。药敏试验结果显示该菌株对头孢克肟、头孢哌酮、链霉素、恩诺沙星、诺氟沙星、左氟沙星、复方新诺明等23种药物敏感。Aeromonas veronii has recently become a serious threat to aquaculture and healthy fish farming.Fish infected by Aeromonas spp.often display hemorrhagic septicemia,resulting in high mortality and severe economic loss.However,there are few reports about the isolation and identification of pathogens isolated from butterfly tail goldfish infected with Aeromonas veronii.In this study,the morphological,physiological and biochemical characteristics of an Aeromonas veronii strain isolated from the goldfish was analyzed.Amplification of the bacterial small subunit ribosomal DNA（16 S rDNA） gene sequence was carried out using the polymerase chain reaction（PCR）technique using universal primers for bacteria.A bacterial strain was isolated from the liver of the diseased goldfish and named DW-1.The strain DW-1 is a short,gram-negative bacillus that produces gas from glucose and tested positive for oxidase,lysine decarboxylase,indole,nitrate reduction,gluconate and methyl red（MR）,while tests were negative for ornithine decarboxylase,urease,phenylalaninase,citrate,hydrogen sulfide and Vogues Proskauer（VP）.A 1 385 bp DNA fragment was obtained upon PCR amplification of the 16 S r DNA gene.Phylogenetic tree analysis shows that the strain is most closely related to Aeromonas veronii biovar sobria,strain ATCC9071,and homology of the 16 S rDNA sequences was 99%-100%.Thus,the DW-1 strain was positively identified as Aeromonas veronii biovar sobria.Animal infection experiments indicated that the mortality was 100% to zebrafish and the median lethal dose（LD_（50）） was 1.17 × 10~6 CFU/m L.Antibiotic sensitivity results show that the isolated strain was sensitive to 23 antibiotics including cefixime,cefoperazone,cefotaxime,cephalexin,streptomycin,kanamycin,norfloxacin,enrofloxacin,levofloxacin,vancomycin,azithromycin,gentamicin,neomycin,amikacin,chloramphenicol,florfenicol and meropenem,but was resistant to imipenem,nalidixic acid,enoxacin,amoxicillin,ampicillin,rifampicin and tetracycline.The study provides a scient

关 键 词：蝶尾金鱼 维氏气单胞菌 分离鉴定 药敏试验 

分 类 号：S941.42[农业科学—水产养殖]