中国东海海水中弧菌的分离鉴定及耐药性分析
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摘要
目的:对中国东海海水进行细菌分离,为东海海水中微生物的鉴定及抗生素的合理使用提供理论依据。方法:对取自中国东海海岸线(约北纬30°、东经118°)水域海水样品进行分离纯化,扩增分离菌株,将分离得到的菌株采用Chelex法提取DNA,并进行16S rRNA基因序列测序,邻接法构建16S rRNA序列的系统发育树,确认分离得到弧菌菌株的分类地位,同时分离的弧菌属菌株和弧菌属常见致病菌株进行耐药性比对分析。结果:从中国东海海岸线水域的海水样品中分离得到2株弧菌属菌株DH-1和DH-2,分子学分析鉴定DH-1为Vibrio marisflavi,DH-2为Vibrio hangzhouensis;经抗生素试验比对,海水分离的弧菌属菌株与副溶血性弧菌、霍乱弧菌、创伤弧菌等常见的临床致病菌株对某些抗生素具有相同程度的耐药性。结论:分离的弧菌菌株具有较强的耐药性。
Objective: To provide the theoretical basis for microbial identification and rational use of antibiotics on microorganism from East China Sea. Methods: The bacteria were isolated and identified from East China Sea( 30° N,118° E); amplified and separated the strains,then extracting strains DNA by Chelex; adopting 16 S rRNA gene sequencing,and the phylogenetic tree was based on neighbor-joining assay 16 S rRNA sequences to be constructed. Confirming the categorization isolated strains,comparing the resistance of the isolated vibrio strains with the common pathogenic strains of vibrio. Results: Two vibrio strains isolated from the sample of East China Sea,DH-1 and DH-2 strains were identified as Vibrio marisflavi and Vibrio hangzhouensis by molecular analysis respectively. The antibiotics test showed that vibrio strains isolated from sea water and the common pathogenic strains of vibrio,such as Vibrio parahaemolyticus,Vibrio cholerae,Vibrio vulnificus have same degree of drug resistance to certain type of antibiotics. Conclusions: The isolated vibrio strains have strong antibiotic resistance.
Objective: To provide the theoretical basis for microbial identification and rational use of antibiotics on microorganism from East China Sea. Methods: The bacteria were isolated and identified from East China Sea( 30° N,118° E); amplified and separated the strains,then extracting strains DNA by Chelex; adopting 16 S rRNA gene sequencing,and the phylogenetic tree was based on neighbor-joining assay 16 S rRNA sequences to be constructed. Confirming the categorization isolated strains,comparing the resistance of the isolated vibrio strains with the common pathogenic strains of vibrio. Results: Two vibrio strains isolated from the sample of East China Sea,DH-1 and DH-2 strains were identified as Vibrio marisflavi and Vibrio hangzhouensis by molecular analysis respectively. The antibiotics test showed that vibrio strains isolated from sea water and the common pathogenic strains of vibrio,such as Vibrio parahaemolyticus,Vibrio cholerae,Vibrio vulnificus have same degree of drug resistance to certain type of antibiotics. Conclusions: The isolated vibrio strains have strong antibiotic resistance.
引文
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[2]NQUYEN T V,ALFARO A C,YOUNG T,et al.Metabolomics study of immune responses of New Zealand GreenshellTMmussels(perna canaliculus)infected with pathogenic vibrio sp[J].Mar Biotechnol,2018,3(2):2955-2961.
[3]KONG C,Wang Y,FODJO E K,et al.Correction to:Loop-mediated isothermal amplification for visual detection of Vibrio parahaemolyticus using gold nanoparticles[J].Mikrochim Acta,2017,185(1):81.
[4]WANG H,LIU J W,WANG Y,et al.Vibrio marisflavi sp.nov.,isolated from seawater[J].Int J Syst Bacteriol,2015,3(61):568-573.
[5]LIU W J,LIN H J,LI C Y.ATP-Binding Cassette Transporter vca M from vibrio cholerae is dependent on the outer membrane factor family for its function[J].Int J Mol Sci,2018,19(4):171-176.
[6]黄淑华,刁保卫,陈磊,等.开封市13株创伤弧菌分离株耐药性及PFGE分型研究[J].中国热带医学,2016,12(16):1163-1165.
[7]GEMMELL B J,BACOSA H P,DICKEY B O,et al.Rapid alterations to marine microbiota communities following an oil spill[J].Ecotoxicology,2018,3(19):1-4.
[8]PAUL C K,MARYANN T,DANA E H,et al.Vibrio metoecus sp.nov.,a close relative of Vibrio cholerae isolated from coastal brackish ponds and clinical specimens[J].Int J Syst Evol Microbiol,2017,1(64):3208-3214.
[9]HIDETAKA D,AKITO C,HIROO F,et al.Vibrio algivorus sp.nov.,an alginate-and agarose-assimilating bacterium isolated from the gut flora of a turban shell marine snail[J].Int J Syst Evol Microbiol,2016,01(66):3164-3169.
[10]李忠东.超级细菌和抗生素[J].生命与灾害,2017,1(4):24-27.
[11]MANAFI M,WALDHERR K.Evaluation of CP Chromo Select agar for the enumeration of clostridium perfringens from water[J].Int J Food Microbiol,2016,167(1):92-95.
[12]KIM O S,CHO Y J,et al.Introducing Ez Taxon-e:A prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species[J].Int J Syst Evol Microbiol,2012,62(3):716-721.
[13]THOMPSON J D,HIGGINS D G,GIBSON,et al.Gleaning structural and functional information from correlations in protein multiple sequence alignment[J].Curr Opin Struct Biol,2016,12(38):1-8.
[14]LI X J,MA T B,WANG P.Molecular epidemiology of Cryptosporidium infection in infants with diarrhea in Wuhan City[J].Zhongguo Xue Xi Chong Bing Za Zhi,2017,29(2):188-191.
[15]吴继纲.海洋环境污染损害赔偿法律机制研究[D].上海:上海海洋大学,2017.
[16]崔晓波,曲文彦,高秀文.水体抗生素污染现状及藻类生态风险评价[J].山西农业科学,2017,45(12):2056-2062.
[17]TIMOTHY R W,JANIS W,DAVID M L,et al.Dissemination of NDM-1 positive bacteria in the new development and its implications for human health:an environment point prevalence study[J].The Lancet Infectious Disease,2015(11):355-362.
[18]MOLLER T S,OVERQAARD M.Relation between tetR and tet A expression in tetracycline resistant Escherichia coli[J].Applied and Environmental Microbiology,2016,3(12):36-39.
[19]娄阳.上海市不同来源副溶血性弧菌耐药性研究及潜在风险分析[D].上海:上海海洋大学,2016.
[20]王跃萍,王桠男,邓慧雅等.副溶血弧菌引发食源性疾病得病原学检测及同源性分析[J].中国热代医学,2015,15(4):402-404.
[21]NQ T H,LU C W,LIN S S.The Rho signaling pathway mediates the pathogenicity of AHPND-causing V.parahaemolyticus in shrimp[J].Cell Microbiol,2018,3(6):65-66.
[22]陈若恒,张秋丽,付丽.Application of a phage in decontaminating Vibrio parahaemolyticus in oysters[J].Int J Food Microbiol,2018,3(275):24-31.
[23]LEIGHT A K,CRUMP B C.Assessment of Fecal Indicator bacteria and potential pathogen Co-occurrence at a shellfish growing area[J].Front Microbiol,2018,14(9):384-389.
[24]陈荣凤.广东省O1/O139群霍乱弧菌耐药分析研究[D].广州:南方医科大学,2016.
[25]陈盛峰,陈佳,龙寒,等.北海近岸养殖海域3种弧菌的分离鉴定及耐药性分析[J].动物医学进展,2016,37(6):29-34.