断奶前后仔猪和母猪粪便源大肠杆菌耐药性和铜抗性的分析
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摘要
旨在比较断奶前后仔猪和母猪大肠杆菌耐药性和铜抗性的变化,并分析其可能原因,为寻求控制大肠杆菌耐药性形成与传播的营养措施提供依据。试验中,采用CLSI推荐的琼脂稀释法,对分离自仔猪及其哺乳母猪的224株大肠杆菌的7种抗生素耐药性及铜抗性进行检测。结果表明,224株大肠杆菌对7种抗生素均有不同程度的耐药(耐药率范围28.13%~93.30%),70.09%的菌株表现出多重耐药性(MDR)。分离自母猪与哺乳仔猪的大肠杆菌抗生素耐药率相似度较高,仅有对氨苄西林和庆大霉素的耐药率差异显著。而断奶后仔猪源大肠杆菌的耐药率相对于母猪和哺乳仔猪变化明显,耐受头孢噻呋、氟苯尼考和氯霉素的菌株都极显著增加(P<0.01),耐受氨苄西林的菌株则显著增加(P<0.05)。相对于母猪和哺乳仔猪,分离自断奶后仔猪的大肠杆菌对铜的抗性程度明显增加(P<0.01)。大肠杆菌对铜抗性增加时,对抗生素耐药性也有所变化,两者之间有一定的相关性。母猪肠道内的耐药菌及饲料中的抗生素添加剂对仔猪大肠杆菌耐药性产生、维持与加剧发挥了重要作用,仔猪饲料中添加的高水平铜与大肠杆菌耐药水平的变化存在一定的关系。
The experiment was conducted to compare the difference of antibiotic and copper resistance among E.coliisolated from sows and piglets before and after weaning.In the experiment,224 of E.coil were isolated from lactation sows,suckling and weaned piglets,which antibiotic resistance to 7antimicrobial agents and copper resistance was detected using agar dilution method according to the recommendation of the CLSI.The remarkable antibiotic resistance of isolated E.coil to 7selected antibiotics was observed,the percentage of resistance isolates to individual antibiotic was ranged from 28.13% to 93.30%.70.09% of all isolates possessed multi-drug resistance(MDR).The antibiotic resistance of E.coil isolates from sows and piglets was highly similar,except resistance to ampicillin and gentamicin.However,the antibiotic resistance of isolates from weaned pigs changed significantly.The antibiotic resistance to ceftiofur,florfenicol and chloramphenicol increased very significantly(P<0.01),resistance to ampicillin increased significantly(P<0.05).Comparing to sow and suckling piglets,the copper resistance of isolates from weaned pigs increased very significantly(P<0.01).In addition,when the copper resistance of E.coil isolates changed,the antibiotic resistance also changed,there might be a correlation between copper and antibiotic resistance.Drug-resistance bacteria in the gut of sows and in feed antibiotics contribute significantly to the development of antibiotics of E.coilin the gut of piglets.Moreover,it initially reveals the presence of a certain relationship between in feed copper and antibiotic resistance change of E.coil isolates form piglets.
The experiment was conducted to compare the difference of antibiotic and copper resistance among E.coliisolated from sows and piglets before and after weaning.In the experiment,224 of E.coil were isolated from lactation sows,suckling and weaned piglets,which antibiotic resistance to 7antimicrobial agents and copper resistance was detected using agar dilution method according to the recommendation of the CLSI.The remarkable antibiotic resistance of isolated E.coil to 7selected antibiotics was observed,the percentage of resistance isolates to individual antibiotic was ranged from 28.13% to 93.30%.70.09% of all isolates possessed multi-drug resistance(MDR).The antibiotic resistance of E.coil isolates from sows and piglets was highly similar,except resistance to ampicillin and gentamicin.However,the antibiotic resistance of isolates from weaned pigs changed significantly.The antibiotic resistance to ceftiofur,florfenicol and chloramphenicol increased very significantly(P<0.01),resistance to ampicillin increased significantly(P<0.05).Comparing to sow and suckling piglets,the copper resistance of isolates from weaned pigs increased very significantly(P<0.01).In addition,when the copper resistance of E.coil isolates changed,the antibiotic resistance also changed,there might be a correlation between copper and antibiotic resistance.Drug-resistance bacteria in the gut of sows and in feed antibiotics contribute significantly to the development of antibiotics of E.coilin the gut of piglets.Moreover,it initially reveals the presence of a certain relationship between in feed copper and antibiotic resistance change of E.coil isolates form piglets.
引文
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[10]RUAN X,ZHANG W.Oral immunization of a live attenuated Escherichia coli strain expressing a holotoxin-structured adhesin-toxoid fusion(1FaeG-FedFLTA2:5LTB)protected young pigs against enterotoxigenic E.coli(ETEC)infection[J].Vaccine,2013,31(11):1458-1463.
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[16]汤景元,王红宁,张鹏举,等.95个猪场大肠杆菌耐药表型及氨基糖苷类药物耐药基因型调查[J].畜牧兽医学报,2008,39(4):472-477.TANG J Y,WANG H N,ZHANG P J,et al.Phenotypic and aminoglycosides genotypic antimicrobial resistance characterization of Escherichia coli isolated from 95pig farms[J].Acta Veterinaria et Zootechnica Sinica,2008,39(4):472-477.(in Chinese)
[17]曾博,王红宁,邹立扣,等.2008-2009年西南地区规模化猪场病原菌流行情况及耐药性调查[C].第四届中国畜牧科技论坛论文集,2009:491-494.ZENG B,WANG H N,ZOU L K,et al.Prevalence and antimicrobial susceptibility of bacterial pathogens isolates from diseased swine in southwest China during 2008-2009[C].Proceedings of the Fourth China Livestock Technology Forum,2009:491-494.(in Chinese)
[18]于卓腾,姚文,朱伟云.体外培养发现二花脸猪粪样菌群具有降解大豆黄酮产生雌马酚的能力[J].南京农业大学学报,2009,32(1):164-167.YU Z T,YAO W,ZHU W Y.In vitro culture reveals the conversion of daidzein to equol by fecal microflorof erhualian pigs[J].Journal of Nanjing Agricultural University,2009,32(1):164-167.(in Chinese)
[19]尹会方,池圣清,黄伟斌.硫酸庆大霉素与百球清预防哺乳仔猪腹泻效果观察[J].湖北畜牧兽医,2013,34(4):15-17.YIN H F,CHI S Q,HUANG W B.The effect of gentamicin sulfate and toltrazuril prevent piglets diarrhea[J].Hubei Journal of Animal and Veterinary Sciences,2013,34(4):15-17.(in Chinese)
[20]吕惠序.庆大霉素在猪病上的合理使用[J].养猪,2009(5):55-56.LH X.The rational use of gentamicin in the swine disease[J].Swine Production,2009(5):55-56.(in Chinese)
[21]MEDARDUS J J,MOLLA B Z,NICOL M,et al.Infeed use of heavy metal micronutrients in US swine production systems and its role in persistence of multidrug-resistant Salmonellae[J].Appl Environ Microbiol,2014,80(7):2317-2325.
[22]HLZEL C S,MLLER C,HARMS K S,et al.Heavy metals in liquid pig manure in light of bacterial antimicrobial resistance[J/OL].Environ Res,2012,113:21-27.[2015-1-26].http://www.sciencedirect.com/science/article/pii/S0013935112000059.
[2]National Research Council.Nutrient Requirements of Swine[M].Washington DC:National Academic Press,2012.
[3]YANG T,ZENG Z,RAO L,et al.The association between occurrence of plasmid-mediated quinolone resistance and ciprofloxacin resistance in Escherichia coli isolates of different origins[J].Vet Microbiol,2014,170(1-2):89-96.
[4]SZMOLKA A,NAGY B.Multidrug resistant commensal Escherichia coli in animals and its impact for public health[J/OL].Front Microbiol,2013,4:258.[2015-1-26].http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3759790/.
[5]朱力军.动物大肠杆菌耐药性的变化趋势[J].中国兽药杂志,2001,35(2):16-18.ZHU L J.Changes of antimicrobial resistance of E.coli[J].Chinese Journal of Veterinary Drug,2001,35(2):16-18.(in Chinese)
[6]JACELA J Y,DEROUCHEY J M,GOODBAND R D,et al.Feed additives for swine:Fact sheets-high dietary levels of copper and zinc for young pigs,and phytase[J].Swine Health Prod,2010,18:87-91.
[7]BAKER-AUSTIN C,WRIGHT M S,STEPANAUSKAS R,et al.Co-selection of antibiotic and metal resistance[J].Trends Microbiol,2006,14(4):176-182.
[8]HASMAN H,KEMPF I,CHIDAINE B,et al.Copper resistance in Enterococcus faecium,mediated by the tcrBgene,is selected by supplementation of pig feed with copper sulfate[J].Appl Environ Microbiol,2006,72(9):5784-5789.
[9]AMACHAWADI R G,SHELTON N W,SHI X,et al.Selection of fecal enterococci exhibiting tcrB-mediated copper resistance in pigs fed diets supplemented with copper[J].Appl Environ Microbiol,2011,77(16):5597-5603.
[10]RUAN X,ZHANG W.Oral immunization of a live attenuated Escherichia coli strain expressing a holotoxin-structured adhesin-toxoid fusion(1FaeG-FedFLTA2:5LTB)protected young pigs against enterotoxigenic E.coli(ETEC)infection[J].Vaccine,2013,31(11):1458-1463.
[11]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.DONG X Z,CAI M Y.Identification mannual of common systematic bacteriology[M].Beijing:Science Press,2001.(in Chinese)
[12]CLSI.Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals[M]//Approved Standard.CLSI Document M31-A3.Third Edition.Clinica Laboratory Standards Institute,Wayne,PA,2008.
[13]AGGA G E,SCOTT H M,AMACHAWADI R G,et al.Effects of chlortetracycline and copper supplementation on antimicrobial resistance of fecal Escherichia coli from weaned pigs[J].Prev Vet Med,2014,114(3-4):231-246.
[14]FOOD AND DRUG ADMINISTRATION.National Antimicrobial resistance monitoring system-enteric bacteria(NARMS):2007 Executive Report[M].Rockville,MD,US:Department of Health and Human Services,US FDA,2010.
[15]薛原,张秀英,陈建飞,等.猪源大肠杆菌多重耐药性的检测与分析[J].中国兽医杂志,2010,46(3):41-43.XUE Y,ZHANG X Y,CHEN J F,et al.Detection and analysis of multidrug-resistant Escherichia coli isolated from pigs[J].Chinese Journal of Veterinary Medicine,2010,46(3):41-43.(in Chinese)
[16]汤景元,王红宁,张鹏举,等.95个猪场大肠杆菌耐药表型及氨基糖苷类药物耐药基因型调查[J].畜牧兽医学报,2008,39(4):472-477.TANG J Y,WANG H N,ZHANG P J,et al.Phenotypic and aminoglycosides genotypic antimicrobial resistance characterization of Escherichia coli isolated from 95pig farms[J].Acta Veterinaria et Zootechnica Sinica,2008,39(4):472-477.(in Chinese)
[17]曾博,王红宁,邹立扣,等.2008-2009年西南地区规模化猪场病原菌流行情况及耐药性调查[C].第四届中国畜牧科技论坛论文集,2009:491-494.ZENG B,WANG H N,ZOU L K,et al.Prevalence and antimicrobial susceptibility of bacterial pathogens isolates from diseased swine in southwest China during 2008-2009[C].Proceedings of the Fourth China Livestock Technology Forum,2009:491-494.(in Chinese)
[18]于卓腾,姚文,朱伟云.体外培养发现二花脸猪粪样菌群具有降解大豆黄酮产生雌马酚的能力[J].南京农业大学学报,2009,32(1):164-167.YU Z T,YAO W,ZHU W Y.In vitro culture reveals the conversion of daidzein to equol by fecal microflorof erhualian pigs[J].Journal of Nanjing Agricultural University,2009,32(1):164-167.(in Chinese)
[19]尹会方,池圣清,黄伟斌.硫酸庆大霉素与百球清预防哺乳仔猪腹泻效果观察[J].湖北畜牧兽医,2013,34(4):15-17.YIN H F,CHI S Q,HUANG W B.The effect of gentamicin sulfate and toltrazuril prevent piglets diarrhea[J].Hubei Journal of Animal and Veterinary Sciences,2013,34(4):15-17.(in Chinese)
[20]吕惠序.庆大霉素在猪病上的合理使用[J].养猪,2009(5):55-56.LH X.The rational use of gentamicin in the swine disease[J].Swine Production,2009(5):55-56.(in Chinese)
[21]MEDARDUS J J,MOLLA B Z,NICOL M,et al.Infeed use of heavy metal micronutrients in US swine production systems and its role in persistence of multidrug-resistant Salmonellae[J].Appl Environ Microbiol,2014,80(7):2317-2325.
[22]HLZEL C S,MLLER C,HARMS K S,et al.Heavy metals in liquid pig manure in light of bacterial antimicrobial resistance[J/OL].Environ Res,2012,113:21-27.[2015-1-26].http://www.sciencedirect.com/science/article/pii/S0013935112000059.