猪肉源大肠杆菌对抗生素及消毒剂的耐药性
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摘要
从四川省各市县收集猪肉样品130份,选择性培养基分离大肠杆菌后VITEK进行鉴定,采用K-B法药敏实验测试大肠杆菌对10种药敏纸片的耐药性。琼脂稀释法测定大肠杆菌对3种季铵盐消毒剂的MIC值,PCR扩增10种季铵盐类消毒剂耐药基因。结果表明:130份猪肉样品中分离大肠杆菌96株,分离率为73.85%,其中73株对抗生素产生耐药性,耐药率分别为:TET(64.58%)、AMP(37.50%)、S(32.29%)、K(21.88%)、KF(20.83%)、CIP(18.75%)、CN(9.38%)、SAM(6.25%)、CAZ(2.08%)、CRO(2.08%),共产生了28种耐药谱,TET是最主要的谱型;96株大肠杆菌对季铵盐消毒剂BC、DDAC、CTAB的MIC分别为:16~64μg/mL、8~32μg/mL、64~256μg/mL;QACs耐药基因检出率分别为ydgE/F(81.25%)、mdfA(50%)、sugE(c)(45.83%)、emrE(36.46%)、qacEΔ1(19.79%)、qacF(17.71%)、qacE(14.58%)、sugE(p)(3.13%),qacG-未检出。共检出42种消毒剂耐药基因组合(1.04%~12.50%)。sugE(c)、qacF基因与氨基糖苷类及AMP耐药相关,qacEΔ1基因与AMP耐药相关。四川省肉源大肠杆菌污染情况较严重,菌株对抗生素的耐药率及多重耐药相对较低,对季铵盐类消毒剂MIC较高,消毒剂耐药基因检出率较高,应引起足够重视,加强对其检测。
Escherichia coli strains were isolated from 130 pork samples collected in Sichuan province using selective medium and confi rmed by VITEK. The susceptibility of the isolated E. coli strains to ten antibiotics was tested according to the standard disk diffusion method of the Clinical and Laboratory Standards Institute(CLSI). The minimum inhibitory concentrations(MICs) of three quaternary ammonium compounds(QACs) against these strains were determined by an agar dilution method. QAC resistance genes were amplifi ed using ten different sets of primers. The results showed that 96 E. coli strains were obtained with frequency of 73.85%. The resistance frequency to tetracycline(TET), ampicillin(AMP), streptomycin(S), kanamycin(K), ceftofur(KF), ciprofloxacin(CIP), gentamicin(CN) ampicillin/sulbactam(SAM), c eftazidime(CAZ) and ceftriaxone(CRO) was 64.58%, 37.50%, 32.29%, 21.88%, 20.83%, 18.75%, 9.38%, 6.25%, 2.08% and 2.08%, respectively. There were 28 different antibiotic resistance profiles Among these, TET profile was the dominant one. The MICs of benzalkonium chloride(BC), didecyldimethylammonium chloride(DDAC) and etrimonium bromide(CTAB) against E. coli were 16-64 μg/mL, 8-32 μg/mL, and 64-256 μg/mL, respectively. ydgE/F(81.25%)was the most prevalent QAC resistance gene in E. coli, followed by mdfA(50%), sugE(c)(45.83%), emrE(36.46%), qacEΔ1(19.79%), qacF(17.71%), qacE(14.58%) and sugE(p)(3.13%). And qacG- was not detected in any of the isolates. There were 42 different qac resistance gene groups(1.04%-12.50%). Both sugE(c)and qacF gene were commonly present in aminoglycoside and AMP resistant E. coli, and q acEΔ1 gene was also related with AMP resistance. The resistance frequency and multi-drug resistance were lower than those of other reports. The frequency of QAC resistance genes and the MIC of QACs were high, suggesting that the E. coli from pork may be a reservoir of antibacterial resistance.
Escherichia coli strains were isolated from 130 pork samples collected in Sichuan province using selective medium and confi rmed by VITEK. The susceptibility of the isolated E. coli strains to ten antibiotics was tested according to the standard disk diffusion method of the Clinical and Laboratory Standards Institute(CLSI). The minimum inhibitory concentrations(MICs) of three quaternary ammonium compounds(QACs) against these strains were determined by an agar dilution method. QAC resistance genes were amplifi ed using ten different sets of primers. The results showed that 96 E. coli strains were obtained with frequency of 73.85%. The resistance frequency to tetracycline(TET), ampicillin(AMP), streptomycin(S), kanamycin(K), ceftofur(KF), ciprofloxacin(CIP), gentamicin(CN) ampicillin/sulbactam(SAM), c eftazidime(CAZ) and ceftriaxone(CRO) was 64.58%, 37.50%, 32.29%, 21.88%, 20.83%, 18.75%, 9.38%, 6.25%, 2.08% and 2.08%, respectively. There were 28 different antibiotic resistance profiles Among these, TET profile was the dominant one. The MICs of benzalkonium chloride(BC), didecyldimethylammonium chloride(DDAC) and etrimonium bromide(CTAB) against E. coli were 16-64 μg/mL, 8-32 μg/mL, and 64-256 μg/mL, respectively. ydgE/F(81.25%)was the most prevalent QAC resistance gene in E. coli, followed by mdfA(50%), sugE(c)(45.83%), emrE(36.46%), qacEΔ1(19.79%), qacF(17.71%), qacE(14.58%) and sugE(p)(3.13%). And qacG- was not detected in any of the isolates. There were 42 different qac resistance gene groups(1.04%-12.50%). Both sugE(c)and qacF gene were commonly present in aminoglycoside and AMP resistant E. coli, and q acEΔ1 gene was also related with AMP resistance. The resistance frequency and multi-drug resistance were lower than those of other reports. The frequency of QAC resistance genes and the MIC of QACs were high, suggesting that the E. coli from pork may be a reservoir of antibacterial resistance.
引文
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[19]邹立扣,蒲妍君,杨莉,等.四川省猪肉源大肠杆菌和沙门氏菌的分离与耐药性分析[J].食品科学,2012,33(13):202-206.
[20]SIDHU M S,S?RUM H,HOLCK A.Resistance to quaternary ammonium compounds in food-related bacteria[J].Microbial D rug Resistance,2002,8(4):393-399.
[21]BUFFET-BATAILLON S,BRANGER B,CORMIER M,et al.Effect of higher minimum inhibitory concentrations of quaternary ammonium compounds in clinical E.coli isolates on antibiotic susceptibilities and clinical outcomes[J].Journal of Hospital Infection,2011,79(2):141-146.
[22]WANG C,ZHAN Q,MI Z,et al.Distribution of the ant iseptic-resistance gene qacEΔ1 in 283 clinical isolates of Gram-negative bacteria in China[J].Journal of Hospital Infection,2008,69(4):394-396.
[23]WELCH T J,FRICKE W F,McDERMOTT P F,et al.Multiple an timicrobial resistance in plague:an emerging public health risk[J].Public Library of Science One,2007,2(3):e309.
[24]ZHAO Weihua,CHEN Gelin,ITO R,et al.Identifi cation of a plasmidborne blaIMP-11 gene in clinical isolates of Escherichia coli and Klebsiella pneumoniae[J].Journal of Medical Microbiology,2012,61(2):246-251.
[25]CARSON R T,LARSON E,LEVY S B,et al.Use of antibacterial consumer products containing quaternary ammonium compounds and drug resis tance in the community[J].Journal of Antimicrobial Chemotherapy,2008,62(5):1160-1162.
[26]WELCH T J,EVENHUIS J,WHITE D G,et al.IncA/C plasmid-mediated florfenicol resistance in the catfish pathogen Edwardsiella ictaluri[J].A ntimicrobial Agents and Chemotherapy,2009,53(2):845-846.
[2]毛雪丹,胡俊峰,刘秀梅.2003—2007年中国1060起细菌性食源性疾病流行病学特征分析[J].中国食品卫生杂志,2010,22(3):224-228.
[3]庞璐,张哲,徐进.2006—2010年我国食源性疾病暴发简介[J].中国食品卫生杂志,2011,23(6):560-563.
[4]唐国策.中国肉类食品安全报告[J].中国牧业通讯,2005(16):13-17.
[5]孙艳,汤雪梅,丛柏林,等.沈阳市零售肉类大肠埃希菌与沙门菌污染调查[J].中国公共卫生,2005(10):1233-1234.
[6]席美丽.食源性革兰氏阴性肠道病原菌PFGE分型和大肠杆菌耐药性研究[D].杨凌:西北农林科技大学,2009.
[7]K?RKK?INEN U,IK?HEIMO R,KATILA M L,et al.Pfi mbriation of Escherichia coli strains from patients with urosepsis demonstrated by a commercial agglutination test(PF TEST)[J].Journal of Clinical Microbiology,1991,29(1):221-224.
[8]宋立,宁宜宝,沈建忠,等.中国不同年代食品动物大肠杆菌耐药性调查研究[J].中国科学C辑:生命科学,2009,397:692-698.
[9]HENRIQUES I S,FONSECA F,ALVES A,et al.Occurrence and diversity of integrons and beta-lactamase genes among ampicillinresistant isolates from estuarine waters[J].Research in Microbiology,2006,157(10):938-947.
[10]CHAPLIN C E.Bacterial resistance to quaternary ammonium disinfectants[J].Journal of Bacteriology,1952,63(4):453-458.
[11]PASTRANA-CARRASCO J,GARZA-RAMOS J,BARRIOS H,et al.qacEΔ1 gene frequency and biocide resistance in exten ded-spectrum beta-lactamase producing enterobacteriaceae clinical isolates[J].Revista de Investigaci on Clinica,2012,64(6):535-540.
[12]张本,刘衡川,张朝武,等.四川省大肠杆菌O157:H7分子流行病学、耐药性及对消毒剂抗性研究[J].卫生研究,2005(5):606-610.
[13]SOUM ET C,FOURREAU E,LEGRANDOIS P,et al.Resistance to phenicol compounds following adaptation to quate rnary ammonium compounds in Escherichia coli[J].Veterinary Microbio logy,2012,158(1/2):147-152.
[14]Clinical and Laboratory Standards Institute(CLSI).Performance standards for antimicrobial susceptibility testing:19th informational supplement M100-S19[S].Wayne,PA:Clin ical and Laboratory Standards Institute,2009.
[15]JORGENSEN J H,CRAWFORD S A,FIEBELKORN K R.Susceptibility of Neisseria meningitidis to 16 antimicrobial agents and characterization of resistance mechanisms affecting some agents[J].Jou rnal of Clinical Microbiology,2005,43(7):3162-3171.
[16]只帅,席美丽,刘攻关,等.陕西部分地区不同食源性大肠杆菌耐药性检测[J].中国食品学报,2011,11(1):196-201.
[17]史秋梅,张艳英,高桂生,等.河北省部分地区肉、蛋食品大肠杆菌污染状况的检测[J].河北科技师范学院学报,2012(4):7-11.
[18]何嘉仪,卢沛炯,赖红青.肉类大肠杆菌耐药性调查[J].中国畜牧兽医,2009(3):201-202.
[19]邹立扣,蒲妍君,杨莉,等.四川省猪肉源大肠杆菌和沙门氏菌的分离与耐药性分析[J].食品科学,2012,33(13):202-206.
[20]SIDHU M S,S?RUM H,HOLCK A.Resistance to quaternary ammonium compounds in food-related bacteria[J].Microbial D rug Resistance,2002,8(4):393-399.
[21]BUFFET-BATAILLON S,BRANGER B,CORMIER M,et al.Effect of higher minimum inhibitory concentrations of quaternary ammonium compounds in clinical E.coli isolates on antibiotic susceptibilities and clinical outcomes[J].Journal of Hospital Infection,2011,79(2):141-146.
[22]WANG C,ZHAN Q,MI Z,et al.Distribution of the ant iseptic-resistance gene qacEΔ1 in 283 clinical isolates of Gram-negative bacteria in China[J].Journal of Hospital Infection,2008,69(4):394-396.
[23]WELCH T J,FRICKE W F,McDERMOTT P F,et al.Multiple an timicrobial resistance in plague:an emerging public health risk[J].Public Library of Science One,2007,2(3):e309.
[24]ZHAO Weihua,CHEN Gelin,ITO R,et al.Identifi cation of a plasmidborne blaIMP-11 gene in clinical isolates of Escherichia coli and Klebsiella pneumoniae[J].Journal of Medical Microbiology,2012,61(2):246-251.
[25]CARSON R T,LARSON E,LEVY S B,et al.Use of antibacterial consumer products containing quaternary ammonium compounds and drug resis tance in the community[J].Journal of Antimicrobial Chemotherapy,2008,62(5):1160-1162.
[26]WELCH T J,EVENHUIS J,WHITE D G,et al.IncA/C plasmid-mediated florfenicol resistance in the catfish pathogen Edwardsiella ictaluri[J].A ntimicrobial Agents and Chemotherapy,2009,53(2):845-846.