大菱鲆源杀鱼爱德华氏菌(Edwardsiella piscicida)的分离鉴定
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摘要
从患腹水病大菱鲆Scophthalmus maximus的肝脏中分离到一株优势细菌G1,经人工感染实验表明G1为引发大菱鲆腹水病的致病菌,且半致死浓度为LD50=1.21×105 CFU·g-1。采用常规的生理生化鉴定方法及分子生物学方法对G1进行分析,结果表明,G1的16SrRNA基因序列与杀鱼爱德华氏菌Edwardsiella piscicida的同源性达100%,系统发育分析表明菌株G1与杀鱼爱德华氏菌分支聚为一支,结合生理生化鉴定结果确定G1为杀鱼爱德华氏菌。药敏试验表明菌株G1对头孢曲松、环丙氟哌酸、左氧氟沙星等8种抗菌药物高度敏感。
Strain G1 was isolated from the turbot Scophthalmus maximus with ascites disease.The regression experimental infection showed that G1 was the pathogen causing ascites disease of Scophthalmus maximus,and its median lethal dose(LD50)to Scophthalmus maximus was 1.21×105 CFU·g-1.G1 was identified by conventional physiological and biochemical methods and 16 SrRNA alignment.The results showed that 16 SrRNA sequences of G1 exhibited 100% homology with Edwardsiella piscicida.Phylogenetic trees of 16 SrRNA showed that the strain G1 and E.piscicidaclustered into one branch.Strain G1 was identified as E.piscicidacombined with physiological and biochemical methods.The susceptibility test showed G1 was sensitive to ten antibiotics,including florfenicol,ofloxacin,etc.
Strain G1 was isolated from the turbot Scophthalmus maximus with ascites disease.The regression experimental infection showed that G1 was the pathogen causing ascites disease of Scophthalmus maximus,and its median lethal dose(LD50)to Scophthalmus maximus was 1.21×105 CFU·g-1.G1 was identified by conventional physiological and biochemical methods and 16 SrRNA alignment.The results showed that 16 SrRNA sequences of G1 exhibited 100% homology with Edwardsiella piscicida.Phylogenetic trees of 16 SrRNA showed that the strain G1 and E.piscicidaclustered into one branch.Strain G1 was identified as E.piscicidacombined with physiological and biochemical methods.The susceptibility test showed G1 was sensitive to ten antibiotics,including florfenicol,ofloxacin,etc.
引文
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[2]ˇRehulka J,MarejkováM,Petrás P.Edwardsiellosis in farmed rainbow trout(Oncorhynchus mykiss)[J].Aquaculture Research,2012,43:1628-1634.
[3]Carluhland F,PierreHélie,RobertHiggins.Infections of Edwardsiella tarda among Brook Trout in Quebec[J].Journal of Aquatic Animal Health,2000,12(1):4.
[4]Abayneh T,Colquhoun D J,Srum H.Edwardsiella piscicida sp.nov.,a novel species pathogenic to fish[J].Journal of Applied Microbiology,2013,114(3).
[5]Fogelson S B,Petty B D,Reichley S R,et al.Histologic and molecular characterization of Edwardsiella piscicida infection in largemouth bass(Micropterus salmoides)[J].Journal of Veterinary Diagnostic Investigation,2016:1040638716637639.
[6]Shafiei S,Viljamaa-Dirks S,Sundell K,et al.Recovery of Edwardsiella piscicidafrom farmed whitefish,Coregonus lavaretus(L.),in Finland[J].Aquaculture,2016,454:19-26.
[7]Loch T P,Hawke J P,Reichley S R,et al.Outbreaks of edwardsiellosis caused by Edwardsiella piscicida and Edwardsiella tarda in farmed barramundi(Lates calcarifer)[J].Aquaculture,2017:S0044848617314874.
[8]Reichley S R,Waldbieser G C,Tekedar H C,et al.Complete Genome Sequence of Edwardsiella piscicida Isolate S11-285Recovered from Channel Catfish(Ictalurus punctatus)in Mississippi,USA[J].Genome Announcements,2016,4(6):e01259-16.
[9]蒋魁,徐力文,苏友禄,等.两株珍珠龙趸病原性哈维弧菌(Vibrio harveyi)的分离与鉴定[J].生态科学,2017,36(6):16-24.
[10]胡大胜,黄钧,黄艳华,等.12种药物对罗非鱼无乳链球菌的抑菌试验[J].中国水产,2012(8):60-63.
[11]吴杰,汪开毓,王均,等.斑点叉尾鮰源鮰爱德华氏菌的分离鉴定[C].中国畜牧兽医学会兽医病理学分会学术研讨会暨中国病理生理学会动物病理生理专业委员会学术研讨会.2014.
[12]李筠,颜显辉,陈吉祥,等.养殖大菱鲆腹水病病原的研究[J].中国海洋大学学报(自然科学版),2006,36(4):649-654.
[13]丁正峰,薛晖,边文冀,等.养殖黄颡鱼腹水症病原研究[J].华中农业大学学报,2008,27(5):639-643.
[14]米娜莎,王栋.中国大菱鲆产业现状及发展趋势分析[J].海洋科学,2011,35(6):96-99.