猪源大肠杆菌耐药性及超广谱β-内酰胺酶流行性分析
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摘要
目的对福建地区分离的猪源大场杆菌进行耐药性测定以及对ESBLs流行性进行分析,以期为临床合理用药科学提供依据。方法采用琼脂稀释法测定373株菌大肠杆菌对12种抗菌药物的敏感性,并用PCR方法检测ESBLs耐药基因的携带情况以及分析CTX-M阳性菌的种系发育关系。结果分离株对甲氧苄啶/磺胺甲噁唑、萘啶酸、氟苯尼考及氨苄西林耐药率较高,分别为93.8%、82.8%、73.7%和70.0%;对恩诺沙星、四环素、庆大霉素、多西环素、环丙沙星、卡那霉素、头孢噻呋和头孢噻肟的耐药率依次为54.4%、53.1%、52.0%、46.6%、44.0%、41.0%、35.9%和30.8%。在373株大肠杆菌中,共有122株菌携带有ESBLs耐药基因,检出率为32.7%,其中CTX-M、TEM及OXA的检出率分别是18.0%(67)、12.1%(45)和7.5%(28),其中CTX-M的阳性菌中以CTX-M-14最为流行,其次是CTX-M-65和CTX-M-55;其种系发育显示分离株主要分布在A群。结论福建地区猪大肠杆菌对抗菌药表现出较强的耐药性,ESBLs在猪源大肠杆菌中广泛流行,CTX-M是其主要流行的基因型,并且多数菌以共生型大肠杆菌为主。
The aim of this research was to study the prevalence of antimicrobial resistance and distribution of extended-spectrum-β-lactamaeses(ESBLs)genes of the Escherichia coli isolated from swine,and provided guidance for the rational use of antibiotics.A total of 373 isolates were tested for antimicrobial susceptibility to 12 antimicrobial agents by agar dilution method,screened for ESBLs genes and analysed phylogenetic group among the CTX-M positive isolates by PCR.Results showed strains were found high resistance rates to trimethoprim/sulfamethoxazole,nalidixic acid,florfenicol and ampicillin,with resistance rates of 93.8%,82.8%,73.7%and 70.0%,respectively.Strains had showed resistance to enrofloxacin,tetracycline,gentamicin,doxycycline,ciprofloxacin,kanamycin,ceftiofur and cefotaxime,and the resistance rates were 54.4%,53.1%,52.0%,46.6%,44.0%,41.0%,35.9% and 30.8% as follows.There were 32.7%(122/373)of the isolates carried ESBLs genes,including CTX-M(18.0%,67),TEM(12.1%,45)and OXA(7.5%,28)genes,and the CTX-M-14 was the most predominant subtype during the encoding CTX-M isolates,then of CTX-M-65 and CTX-M-55.Phylogeny showed that the strains were mainly distributed in the A group.This research revealed that high rates of resistance to antimicrobial agents,and ESBLs was widely prevalent in Escherichia coli isolated from Fujian Province.The CTX-M was the most predominant gene among those strains,and the majority of them belonged to symbiotic Escherichia coli.
The aim of this research was to study the prevalence of antimicrobial resistance and distribution of extended-spectrum-β-lactamaeses(ESBLs)genes of the Escherichia coli isolated from swine,and provided guidance for the rational use of antibiotics.A total of 373 isolates were tested for antimicrobial susceptibility to 12 antimicrobial agents by agar dilution method,screened for ESBLs genes and analysed phylogenetic group among the CTX-M positive isolates by PCR.Results showed strains were found high resistance rates to trimethoprim/sulfamethoxazole,nalidixic acid,florfenicol and ampicillin,with resistance rates of 93.8%,82.8%,73.7%and 70.0%,respectively.Strains had showed resistance to enrofloxacin,tetracycline,gentamicin,doxycycline,ciprofloxacin,kanamycin,ceftiofur and cefotaxime,and the resistance rates were 54.4%,53.1%,52.0%,46.6%,44.0%,41.0%,35.9% and 30.8% as follows.There were 32.7%(122/373)of the isolates carried ESBLs genes,including CTX-M(18.0%,67),TEM(12.1%,45)and OXA(7.5%,28)genes,and the CTX-M-14 was the most predominant subtype during the encoding CTX-M isolates,then of CTX-M-65 and CTX-M-55.Phylogeny showed that the strains were mainly distributed in the A group.This research revealed that high rates of resistance to antimicrobial agents,and ESBLs was widely prevalent in Escherichia coli isolated from Fujian Province.The CTX-M was the most predominant gene among those strains,and the majority of them belonged to symbiotic Escherichia coli.
引文
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[14]蒋鸿超,奎莉越,黄海林,等.儿童产超广谱β-内酰胺酶大肠埃希菌Ⅰ类整合子及其与ESBLs基因关系的研究[J].临床儿科杂志,2015,33(4):345-347.
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[19]左明开.江西猪源产ESBLs ETEC流行及质粒相关性研究[D].南昌:江西农业大学,2016.
[20]靳婵.屠宰前鸡猪源大肠杆菌耐药性和β-内酰胺酶基因调查研究[D].广州:华南农业大学,2016.
[21]高丽丽.猪源产ESBL大肠杆菌的扩散及膨润土促其耐药基因转移机制的研究[D].泰安:山东农业大学,2015.
[22]Picard B,Garcia JS,Gouriou S,et al.The link between phylogeny and virulence in Escherichia coli extraintestinal infection[J].Infect Immun,1999,67(2):546-553.
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[24]Boyd EF,Hartl DL.Chromosomal regions specific to pathogenic isolates of Escherichia coli have a phylogenetically clustered distribution[J].J Bacteriol,1998,180(5):1159-1165.
[25]甄盼盼,汤电,任艳娜,等.食品动物源产CTX-M-14大肠杆菌传播分子机制的演变[J].中国畜牧兽医,2012,39(9):195-202.
[2]Ma J,Zeng Z,Chen Z,et al.High prevalence of plasmid-mediated quinolone resistance determinants qnr,aac(6′)-Ib-cr,and qepAamong ceftiofur-resistant Enterobacteriaceae isolates from companion and food-producing animals[J].Antimicrob Agents Chemother,2009,53(2):519-524.DOI:10.1128/AAC.00886-08
[3]何涛.动物源大肠杆菌对乙酰甲喹和氟苯尼考的敏感性折点及分子耐药机制研究[D].北京:中国农业大学,2015.
[4]Clermont O,Bonacorsi S,Bingen E.Rapid and simple determination of the Escherichia coli phylogenetic group[J].Appl Environ Microbiol,2000,66(10):4555-4558.DOI:10.1128/AEM.66.10.4555-4558.2000
[5]Weill FX,Demartin M,Tande D,et al.SHV-12-like extendedspectrum-beta-lactamase-producing strains of Salmonella enterica serotypes Babelsberg and Enteritidis isolated in France among infants adopted from Mali[J].J Clin Microbiol,2004,42(6):2432-2437.DOI:10.1128/JCM.42.6.2432-2437.2004
[6]Brinas L,Moreno MA,Zarazaga M,et al.Detection of CMY-2,CTX-M-14,and SHV-12beta-lactamases in Escherichia coli fecal-sample isolates from healthy chickens[J].Antimicrob A-gents Chemother,2003,47(6):2056-2058.DOI:10.1128/AAC.47.6.2056-2058.2003
[7]曲志娜,刘红玉,王娟,等.青岛地区产ESBLs鸡源大肠杆菌耐药性调查与优势基因型分析[J].中国农业科学,2015,48(10):2058-2066.
[8]Liu JH,Wei SY,Ma JY,et al.Detection and characterisation of CTX-M and CMY-2beta-lactamases among Escherichia coli isolates from farm animals in Guangdong province of China[J].Int J Antimicrob Agents,2007,29(5):576-581.DOI:10.1016/j.ijantimicag.2006.12.015
[9]岳秀英,葛荣,吴晓岚,等.四川省猪、鸡源大肠杆菌对抗生素耐药性研究[J].中国兽医杂志,2017,53(1):93-95.
[10]曹敏,谭艾娟,吕世明,等.贵州部分地区猪源大肠杆菌耐药性分析及ESBLs基因型检测[J].中国畜牧兽医,2016,43(4):1098-1104.
[11]孙坚.肠外致病性大肠杆菌ESBLs耐药与毒力关系研究[D].广州:华南农业大学,2012.
[12]Ho PL,Chow KH,Lai EL,et al.Extensive dissemination of CTX-M-producing Escherichia coli with multidrug resistance to'critically important'antibiotics among food animals in Hong Kong,2008-10[J].J Antimicrob Chemother,2011,66(4):765-768.DOI:10.1093/jac/dkq539
[13]曾莉.不同来源大肠杆菌ESBLs基因分子流行病学研究[D].广州:华南农业大学,2016.
[14]蒋鸿超,奎莉越,黄海林,等.儿童产超广谱β-内酰胺酶大肠埃希菌Ⅰ类整合子及其与ESBLs基因关系的研究[J].临床儿科杂志,2015,33(4):345-347.
[15]Canton R,Gonzalez-Alba JM,Galan JC.CTX-M enzymes:origin and diffusion[J].Front Microbiol,2012,3:110.DOI:10.3389/fmicb.2012.00110
[16]郑红青.食品动物源大肠杆菌ESBLs传播机制研究[D].广州:华南农业大学,2011.
[17]郑新添,张孝辉,吴玲,等.猪源产ESBLs大肠杆菌三种耐药基因检测及分析[J].龙岩学院学报,2015,33(2):100-103.
[18]王基伟.猪源大肠杆菌分离鉴定及耐药性研究[D].长春:吉林农业大学,2015.
[19]左明开.江西猪源产ESBLs ETEC流行及质粒相关性研究[D].南昌:江西农业大学,2016.
[20]靳婵.屠宰前鸡猪源大肠杆菌耐药性和β-内酰胺酶基因调查研究[D].广州:华南农业大学,2016.
[21]高丽丽.猪源产ESBL大肠杆菌的扩散及膨润土促其耐药基因转移机制的研究[D].泰安:山东农业大学,2015.
[22]Picard B,Garcia JS,Gouriou S,et al.The link between phylogeny and virulence in Escherichia coli extraintestinal infection[J].Infect Immun,1999,67(2):546-553.
[23]Johnson JR,Stell AL.Extended virulence genotypes of Escherichia coli strains from patients with urosepsis in relation to phylogeny and host compromise[J].J Infect Dis,2000,181(1):261-272.DOI:10.1086/315217
[24]Boyd EF,Hartl DL.Chromosomal regions specific to pathogenic isolates of Escherichia coli have a phylogenetically clustered distribution[J].J Bacteriol,1998,180(5):1159-1165.
[25]甄盼盼,汤电,任艳娜,等.食品动物源产CTX-M-14大肠杆菌传播分子机制的演变[J].中国畜牧兽医,2012,39(9):195-202.