猪致病性大肠杆菌耐药性与整合子-基因盒的相关性
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摘要
为了解猪致病性大肠杆菌耐药性与整合子-基因盒的相关性,本研究采用腹腔注射法测定52株大肠杆菌对昆明小鼠的致病性;采用K-B法测定致病性大肠杆菌对15种抗菌药物的敏感性;对致病性大肠杆菌Ⅰ、Ⅱ和Ⅲ型整合子及其基因盒进行PCR检测及测序分析,并分析其整合子-基因盒与耐药性的相关性。结果显示,52株大肠杆菌对小鼠均具有较强致病性,96.15%分离株对15种抗菌药物表现为多重耐药性,61.54%分离株耐药性均在八重耐药以上。Ⅰ、Ⅱ和Ⅲ型整合子阳性率分别为84.62%,15.38%和3.85%。Ⅰ型整合子检测到dfrA17-aadA5(18.18%)、dfrA12-orfF-aadA2(4.55%)和dfrA1-catB3-aacA4(4.55%)基因盒,Ⅱ型整合子检测到dfrA1-sat2-aadA1(50.00%)基因盒,Ⅲ型整合子未检测到基因盒,基因盒阳性率与细菌耐药性不存在相关性。
To understand the correlations between integron gene cassettes and antimicrobial resistance of pathogenic E.coli isolated from pigs.The pathogenicity of 52 E.coli isolates to Kunming mice was tested by intraperitoneal injection.Antibiotic susceptibility of pathogenic Ec.oli was tested with the K-B diffusion assay.Integron gene cassettes of pathogenic E.coli were amplified by PCR and sequenced,and the correlation between integron gene cassettes and antimicrobial resistance was analyzed.The results showed that all the isolates were pathogenic to Kunming mice,and96.15%isolates showed multi resistance to 15 antibiotics.61.54%isolates were resistant to 8 or more antibiotics.The positive rate of classⅠ,ⅡandⅢintegron were 84.62%,15.38%and 3.85%,respectively.DfrA17-aadA5(18.18%),dfrA12-orfF-aadA2(4.55%)and dfrA1-catB3-aacA4(4.55%)cassette arrays were detected in classⅠintegrons,dfrA1-sat2-aadA1(50.00%)cassette array was detected in classⅡintegrons and none of the cassette array was detected in classⅢintegrons.The positive rate of cassettes did not relate to antimicrobial resistance of pathogenic Ec.oli isolated from pigs.
To understand the correlations between integron gene cassettes and antimicrobial resistance of pathogenic E.coli isolated from pigs.The pathogenicity of 52 E.coli isolates to Kunming mice was tested by intraperitoneal injection.Antibiotic susceptibility of pathogenic Ec.oli was tested with the K-B diffusion assay.Integron gene cassettes of pathogenic E.coli were amplified by PCR and sequenced,and the correlation between integron gene cassettes and antimicrobial resistance was analyzed.The results showed that all the isolates were pathogenic to Kunming mice,and96.15%isolates showed multi resistance to 15 antibiotics.61.54%isolates were resistant to 8 or more antibiotics.The positive rate of classⅠ,ⅡandⅢintegron were 84.62%,15.38%and 3.85%,respectively.DfrA17-aadA5(18.18%),dfrA12-orfF-aadA2(4.55%)and dfrA1-catB3-aacA4(4.55%)cassette arrays were detected in classⅠintegrons,dfrA1-sat2-aadA1(50.00%)cassette array was detected in classⅡintegrons and none of the cassette array was detected in classⅢintegrons.The positive rate of cassettes did not relate to antimicrobial resistance of pathogenic Ec.oli isolated from pigs.
引文
[1]BULMAN Z P,CHEN L,WALSH T J,et al.Polymyxin combinations combat Escherichia coli harboring Mcr-1and blaNDM-5:preparation for a postantibiotic era[J].MBio,2017,8(4):17.
[2]HUYNH B T,PADGET M,GARIN B,et al.Burden of bacterial resistance among neonatal infections in low income countries:how convincing is the epidemiological evidence?[J].BMC Infect Dis,2015,127(15):843.
[3]OBRIEN T F.Emergence,spread,and environmental effect of antimicrobial resistance:how use of an antimicrobial anywhere can increase resistance to any antimicrobial anywhere else[J].Clin Infect Dis,2002,34(3):S78-S84.
[4]张济培,韦庆兰,谭华龙,等.广东省水禽源大肠杆菌Ⅰ型整合子的检测分析[J].中国兽医学报,2016,36(7):1119-1139.
[5]李晓娜,杨慧君,余婷,等.牛源大肠杆菌耐药表型与Ⅰ型整合子-基因盒的关系[J].中国兽医学报,2017,37(3):438-442.
[6]林居纯,舒刚,张辉建,等.健康畜禽肠道大肠杆菌耐药性及整合子-基因盒检测[J].中国兽医学报,2014,34(1):56-60.
[7]ODETOYIN B W,LABAR A S,LAMIKANRA A,et al.Classes 1and 2integrons in faecal Escherichia coli strains isolated from mother-child pairs in Nigeria[J].PLoS One,2017,12(8):e0183383.
[8]SIQUEIRA A K,MICHAEL G B,DOMINGOS D F,et al.Diversity of class 1and 2integrons detected in Escherichia coli isolates from diseased and apparently healthy dogs[J].Vet Microbiol,2016,194:79-83.
[9]CAVICCHIO L,DOTTO G,GIACOMELLI M,et al.Class 1 and class 2integrons in avian pathogenic Escherichia coli from poultry in Italy[J].Poult Sci,2015,94(6):1202-1208.
[10]VENTURINI C,HASSAN K A,ROY CHOWDHU-RY P,et al.Sequences of two related multiple antibiotic resistance virulence plasmids sharing a unique IS26-related molecular signature isolated from different Escherichia coli pathotypes from different hosts[J].PLoS One,2013,8(11):e78862.
[11]REN C,ZHAO Y,SHEN Y.Analysis of the effect of integrons on drug-resistant Staphylococcus aureus by multiplex PCR detection[J].Mol Med Rep,2013,7(3):719-724.
[12]LEVESQUE C,PICHE L,LAROSE C,et al.PCRmapping of integrons reveals several novel combinations of resistance genes[J].Antimicrob Agents Chemother,1995,39(1):185-191.
[13]WHITE P A,MCLVER C J,RAWLINSON W D.Integrons and gene cassettes in the Enterobacteriaceae[J].Antimicrob Agents Chemother,2001,45(9):2658-2661.
[14]CORREIA M,BOAVIDA F,GROSSO F,et al.Molecular characterization of a new class 3integron in Klebsiella pneumoniae[J].Antimicrob Agents Chemother,2003,47(9):2838-2843.
[15]TANG X,TAN C,ZHANG X,et al.Antimicrobial resistances of extraintestinal pathogenic Escherichia coli isolates from swine in China[J].Microb Pathog,2011,50(5):207-212.
[16]毕保良,李富祥,胡骑,等.云南多重耐药猪源大肠杆菌的分离鉴定及其耐药机制初步研究[J].上海畜牧兽医通讯,2012(5):6-7.
[17]李琳,代兴杨,汪莹,等.安徽鸡源大肠杆菌整合子-基因盒分布及同源性分析[J].中国兽医科学,2016,46(10):1277-1282.
[18]林居纯,曹三杰,张辉建,等.动物源大肠杆菌整合子携带氨基糖苷类耐药基因盒的研究[J].中国兽医学报,2011,31(1):1587-1590.
[19]赵凤菊,李清竹,关淼,等.辽宁地区猪源大肠埃希菌整合子携带情况调查与分析[J].动物医学进展,2017,38(1):115-118.
[20]POIREL L,CARATTOLI A,BERNABEU S,et al.Anovel IncQ plasmid type harbouring a class 3integron from Escherichia coli[J].J Antimicrob Chemother,2010,65(8):1594-1598.
[21]李景云,崔生辉,张力,等.鸡来源产超广谱β-内酰胺酶大肠埃希菌携带整合子基因的研究[J].中华流行病学杂志,2008,29(10):1020-1023.
[22]MACHADO E,CANTON R,BAQUERO F,et al.Integron content of extended-spectrum-beta-lactamaseproducing Escherichia coli strains over 12years in a single hospital in Madrid,Spain[J].Antimicrob A-gents Chemother,2005,49(5):1823-1829.
[23]MAY T,ITO A,OKABE S.Induction of multidrug resistance mechanism in Escherichia coli biofilms by interplay between tetracycline and ampicillin resistance genes[J].Antimicrob Agents Chemother,2009,53(11):4628-4639.
[2]HUYNH B T,PADGET M,GARIN B,et al.Burden of bacterial resistance among neonatal infections in low income countries:how convincing is the epidemiological evidence?[J].BMC Infect Dis,2015,127(15):843.
[3]OBRIEN T F.Emergence,spread,and environmental effect of antimicrobial resistance:how use of an antimicrobial anywhere can increase resistance to any antimicrobial anywhere else[J].Clin Infect Dis,2002,34(3):S78-S84.
[4]张济培,韦庆兰,谭华龙,等.广东省水禽源大肠杆菌Ⅰ型整合子的检测分析[J].中国兽医学报,2016,36(7):1119-1139.
[5]李晓娜,杨慧君,余婷,等.牛源大肠杆菌耐药表型与Ⅰ型整合子-基因盒的关系[J].中国兽医学报,2017,37(3):438-442.
[6]林居纯,舒刚,张辉建,等.健康畜禽肠道大肠杆菌耐药性及整合子-基因盒检测[J].中国兽医学报,2014,34(1):56-60.
[7]ODETOYIN B W,LABAR A S,LAMIKANRA A,et al.Classes 1and 2integrons in faecal Escherichia coli strains isolated from mother-child pairs in Nigeria[J].PLoS One,2017,12(8):e0183383.
[8]SIQUEIRA A K,MICHAEL G B,DOMINGOS D F,et al.Diversity of class 1and 2integrons detected in Escherichia coli isolates from diseased and apparently healthy dogs[J].Vet Microbiol,2016,194:79-83.
[9]CAVICCHIO L,DOTTO G,GIACOMELLI M,et al.Class 1 and class 2integrons in avian pathogenic Escherichia coli from poultry in Italy[J].Poult Sci,2015,94(6):1202-1208.
[10]VENTURINI C,HASSAN K A,ROY CHOWDHU-RY P,et al.Sequences of two related multiple antibiotic resistance virulence plasmids sharing a unique IS26-related molecular signature isolated from different Escherichia coli pathotypes from different hosts[J].PLoS One,2013,8(11):e78862.
[11]REN C,ZHAO Y,SHEN Y.Analysis of the effect of integrons on drug-resistant Staphylococcus aureus by multiplex PCR detection[J].Mol Med Rep,2013,7(3):719-724.
[12]LEVESQUE C,PICHE L,LAROSE C,et al.PCRmapping of integrons reveals several novel combinations of resistance genes[J].Antimicrob Agents Chemother,1995,39(1):185-191.
[13]WHITE P A,MCLVER C J,RAWLINSON W D.Integrons and gene cassettes in the Enterobacteriaceae[J].Antimicrob Agents Chemother,2001,45(9):2658-2661.
[14]CORREIA M,BOAVIDA F,GROSSO F,et al.Molecular characterization of a new class 3integron in Klebsiella pneumoniae[J].Antimicrob Agents Chemother,2003,47(9):2838-2843.
[15]TANG X,TAN C,ZHANG X,et al.Antimicrobial resistances of extraintestinal pathogenic Escherichia coli isolates from swine in China[J].Microb Pathog,2011,50(5):207-212.
[16]毕保良,李富祥,胡骑,等.云南多重耐药猪源大肠杆菌的分离鉴定及其耐药机制初步研究[J].上海畜牧兽医通讯,2012(5):6-7.
[17]李琳,代兴杨,汪莹,等.安徽鸡源大肠杆菌整合子-基因盒分布及同源性分析[J].中国兽医科学,2016,46(10):1277-1282.
[18]林居纯,曹三杰,张辉建,等.动物源大肠杆菌整合子携带氨基糖苷类耐药基因盒的研究[J].中国兽医学报,2011,31(1):1587-1590.
[19]赵凤菊,李清竹,关淼,等.辽宁地区猪源大肠埃希菌整合子携带情况调查与分析[J].动物医学进展,2017,38(1):115-118.
[20]POIREL L,CARATTOLI A,BERNABEU S,et al.Anovel IncQ plasmid type harbouring a class 3integron from Escherichia coli[J].J Antimicrob Chemother,2010,65(8):1594-1598.
[21]李景云,崔生辉,张力,等.鸡来源产超广谱β-内酰胺酶大肠埃希菌携带整合子基因的研究[J].中华流行病学杂志,2008,29(10):1020-1023.
[22]MACHADO E,CANTON R,BAQUERO F,et al.Integron content of extended-spectrum-beta-lactamaseproducing Escherichia coli strains over 12years in a single hospital in Madrid,Spain[J].Antimicrob A-gents Chemother,2005,49(5):1823-1829.
[23]MAY T,ITO A,OKABE S.Induction of multidrug resistance mechanism in Escherichia coli biofilms by interplay between tetracycline and ampicillin resistance genes[J].Antimicrob Agents Chemother,2009,53(11):4628-4639.