养禽场周围水环境中磺胺类抗性菌、抗性基因分布
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摘要
为进行养禽场周围水环境中磺胺类抗性菌、抗性基因分布的研究,从3家养禽场多个区域水环境中分离的56株大肠杆菌(Escherichia coli,E.coli),应用肉汤微量稀释法分析磺胺类药物敏感性,应用PCR方法检测磺胺类药物抗性基因。结果有18株(32.14%)磺胺类抗性菌,对利福平的耐受性最弱,对氯霉素的耐受能性最强;共检测到64个抗性基因,三个养禽场分别为:19(29.69%)、29(45.31%)、16(25.00%);分别为sul1 18(28.12%)、sul2 13(20.32%)、sul3 7(10.94%)、Int1 18(28.12%)、Int2 8(12.50%)。含一种抗性基因的菌株为1株、含2种抗性基因的菌株为1株、含三种抗性基因的菌株为6株、含四种抗性基因的菌株为5株、含五种抗性基因的菌株为4株,而1株无抗生素抗性的菌株检出抗性基因。结果表明在养禽场场内检测到抗性基因和抗性菌株数量较高、养禽场周围水环境中存在磺胺类抗性菌株,且具有多重抗性,可能对生态环境的产生一定的不良影响。
In order to study the distribution of sulfanilamide resistant bacteria and resistance genes in the water around the poultry farms,56 strains of Escherichia coli( E. coli) were isolated from water in three poultry farms.Broth microdilution and polymerase chain reaction( PCR) were used for the sulfonamides susceptibility testing and detection of sulphonamides resistance genes respectively. Between all the isolated E. coli,18( 32. 14%) isolates were susceptible to resistant bacteria,which had the weakest resistance to rifampicin and the strongest resistance to chloramphenicol. 64 resistant genes were detected in the three poultry farms,including 19( 29. 69%),29( 45. 31%) and 16( 25. 00%),respectively. Among them,sul1,sul2,sul3,Int1 and Int2 were 18( 28. 12%),13( 20. 32%),7( 10. 94%),18( 28. 12%),and 8( 12. 50%),respectively. 1 strain was contained one resistance gene,1 strain was contained two resistance genes,6 strains were contained three resistance genes,5 strains were contained four resistance genes,5 strains were contained five resistance genes,while 1 strain of without resistance gene was detected in resistance gene. The results showed that the sulfonamide resistance of bacteria had multiple antibiotic resistances in the poultry farm. In conclusion,antibiotic resistance bacteria and resistance genes related with a lot of factors. Obviously,the sulfonamides resistance did not coincide with the presence of such genes completely.
In order to study the distribution of sulfanilamide resistant bacteria and resistance genes in the water around the poultry farms,56 strains of Escherichia coli( E. coli) were isolated from water in three poultry farms.Broth microdilution and polymerase chain reaction( PCR) were used for the sulfonamides susceptibility testing and detection of sulphonamides resistance genes respectively. Between all the isolated E. coli,18( 32. 14%) isolates were susceptible to resistant bacteria,which had the weakest resistance to rifampicin and the strongest resistance to chloramphenicol. 64 resistant genes were detected in the three poultry farms,including 19( 29. 69%),29( 45. 31%) and 16( 25. 00%),respectively. Among them,sul1,sul2,sul3,Int1 and Int2 were 18( 28. 12%),13( 20. 32%),7( 10. 94%),18( 28. 12%),and 8( 12. 50%),respectively. 1 strain was contained one resistance gene,1 strain was contained two resistance genes,6 strains were contained three resistance genes,5 strains were contained four resistance genes,5 strains were contained five resistance genes,while 1 strain of without resistance gene was detected in resistance gene. The results showed that the sulfonamide resistance of bacteria had multiple antibiotic resistances in the poultry farm. In conclusion,antibiotic resistance bacteria and resistance genes related with a lot of factors. Obviously,the sulfonamides resistance did not coincide with the presence of such genes completely.
引文
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11 Clinical and Laboratory Standards Institute.Method for broth dilution susceptibility testing of bacteria isolated:M7-A8[S].Wayne,Pennsylvania,USA:Clinical and Laboratory Standards Institute,2006
12 National Committee for Clinical Laboratory Standards.Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals:M31-A2[S].Wayne,Pennsylvania USA:National Committee for Clinical Laboratory Standards,2001
13 Biro2ováL,Mackul’ak T,Bodík I,et al.Pilot study of seasonal occurrence and distribution of antibiotics and drug resistant bacteria in wastewater treatment plants in Slovakia[J].Science of the Total Environment,2014,490:440-444
14 Lopes G V,Pissetti C,Pellegrini D da Cruz Pay珓ao,et al.Resistance phenotypes and genotypes of salmonella enterica subsp.enterica isolates from feed,pigs,and carcasses in Brazil[J].J Food Prot,2015,78(2):407-413
15 Gao P,Mao D Q,Luo Y,et al.Occurrence of sulfonamide and tetracycline resistant bacteria and resistance genes in aquaculture environment[J].Water Research,2012,46:2355-2364
16 Radhouani H,Poeta P,Igrejas G,et al.Antimicrobial resistance and phylogenetic groups in isolates of Escherichia coli from seagulls at the berlengas nature reserve[J].Veterinary Record,2009,165:138-142
17 Xu B J,Luo Y,Zhou Q X,et al.Sources,dissemination,and ecological risks of antibiotic resistance genes(ARGs)in the environment[J].Environmental Chemistry,2010,29(2):169-178
18 Chen H,Shu W,Chang X,et al.The profile of antibiotics resistance and integrons of extended-spectrum beta-lactamase producing thermotolerant coliforms isolated from the Yangtze River basin in Chongqing[J].Environmental Pollution,2010,158:2459-2464
19 Looft T,Johnson T A,Allen H K,et al.In-feed antibiotic effects on the swine intestinal microbiome[J].Proc Natl Acad Sci USA,2012,109(5):1691-1696
20 张崇淼,庄凯,巨欣,等.紫外线消毒对3种大肠杆菌的灭活效果和耐药性影响[J].环境工程学报,2015,9(9):4097-4101Zhang Chongmiao,Zhuang Kai,Ju Xin,et al.Inactivation and antibiotic resistance variation of three Escherichia coli under ultraviolet disinfection[J].Chinese Journal of Environmental Engineering,2015,9(9):4097-4101.
21 李静.紫外消毒致耐药基因水平转移规律和影响因素的研究[D].天津:天津大学,2016Li Jing.Effect of UV disinfection on horizontal gene transfer of antibiotic resistance genes[D].Tianjin:Tianjin University,2016
22 Wen L H,Xu Y B,Zhou Y,et al.Effects of heavy metal stress on antibiotics resistance of Pseudomonas alcaligenes LH7[J].Acta Ecologica Sinica,2015,35(10):1-12
2 Pruden A1,Larsson D G,Amézquita A,et al.Management options for reducing the release of antibiotics and antibiotic resistance genes to the environment[J].Environ Health Perspect,2013,121(8):878-885
3曹慧,詹健,刘振中.磁性碳纳米管对水中低浓度罗红霉素的吸附特性研究[J].科学技术与工程,2015,15(16):232-237Chao Hui,Zhan Jian,Liu Zhenzhong.Adsorption properties of magnetic multi-wall carbon nanotubes for low concentration erythromycin in water[J].Science Technology and Engineering,2015,15(16):232-237
4 韩瑞,阚旺晨,宋倩,等.内蒙古额吉淖尔盐碱湖嗜盐古菌的抗生素抗性及分泌的嗜盐菌素的抑菌性[J].科学技术与工程,2013,13(34):16-20Han Rui,Kan Wangchen,Song Qian,et al.The antibiotic resistance of halophilic bacteria and the antibacterial activity of the halocin that secreted by halophilic bacteria separated from Ejnoor in Inner Mongolia[J].Science Technology and Engineering,2013,13(34):16-20
5 Kennedy D.Time to deal with antibiotics[J].Science,2016,14(9):563-575
6 Lu W F,Iang H Q,Yu L.Detection of sulfanilamide sensitivity and sulfonamide resistant related genes in clinical escherichia coli isolates[J].Journal of Jilin Agricultural University,2010,32(3):340-344
7 Chen B W,Liang X M,Huang X P,et al.Differentiating anthropogenic impacts on ARG s in the pearl River estuary by using suitable gene indicators[J].Water Research,2013,47:2811-2820
8 Wang Y T.The usage and dosage of poultry commonly used medicine[J].Technical Advisor for Animal Husbandry,2010,28(1):143-143
9 Heuer H,Solehati Q,Zimmerling U,et al.Accumulation of sulfonamide resistance genes in arable soils due to repeated application of manure containing sulfadiazine[J].Appl Environ Microbiol,2011,77:2527-2530
10 Yu S,Wang M,Hong Y W.Antibiotics in environmental matrices and their effects on microbial ecosystems[J].Acta Ecologica Sinica,2011,31(15):4437-4446
11 Clinical and Laboratory Standards Institute.Method for broth dilution susceptibility testing of bacteria isolated:M7-A8[S].Wayne,Pennsylvania,USA:Clinical and Laboratory Standards Institute,2006
12 National Committee for Clinical Laboratory Standards.Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals:M31-A2[S].Wayne,Pennsylvania USA:National Committee for Clinical Laboratory Standards,2001
13 Biro2ováL,Mackul’ak T,Bodík I,et al.Pilot study of seasonal occurrence and distribution of antibiotics and drug resistant bacteria in wastewater treatment plants in Slovakia[J].Science of the Total Environment,2014,490:440-444
14 Lopes G V,Pissetti C,Pellegrini D da Cruz Pay珓ao,et al.Resistance phenotypes and genotypes of salmonella enterica subsp.enterica isolates from feed,pigs,and carcasses in Brazil[J].J Food Prot,2015,78(2):407-413
15 Gao P,Mao D Q,Luo Y,et al.Occurrence of sulfonamide and tetracycline resistant bacteria and resistance genes in aquaculture environment[J].Water Research,2012,46:2355-2364
16 Radhouani H,Poeta P,Igrejas G,et al.Antimicrobial resistance and phylogenetic groups in isolates of Escherichia coli from seagulls at the berlengas nature reserve[J].Veterinary Record,2009,165:138-142
17 Xu B J,Luo Y,Zhou Q X,et al.Sources,dissemination,and ecological risks of antibiotic resistance genes(ARGs)in the environment[J].Environmental Chemistry,2010,29(2):169-178
18 Chen H,Shu W,Chang X,et al.The profile of antibiotics resistance and integrons of extended-spectrum beta-lactamase producing thermotolerant coliforms isolated from the Yangtze River basin in Chongqing[J].Environmental Pollution,2010,158:2459-2464
19 Looft T,Johnson T A,Allen H K,et al.In-feed antibiotic effects on the swine intestinal microbiome[J].Proc Natl Acad Sci USA,2012,109(5):1691-1696
20 张崇淼,庄凯,巨欣,等.紫外线消毒对3种大肠杆菌的灭活效果和耐药性影响[J].环境工程学报,2015,9(9):4097-4101Zhang Chongmiao,Zhuang Kai,Ju Xin,et al.Inactivation and antibiotic resistance variation of three Escherichia coli under ultraviolet disinfection[J].Chinese Journal of Environmental Engineering,2015,9(9):4097-4101.
21 李静.紫外消毒致耐药基因水平转移规律和影响因素的研究[D].天津:天津大学,2016Li Jing.Effect of UV disinfection on horizontal gene transfer of antibiotic resistance genes[D].Tianjin:Tianjin University,2016
22 Wen L H,Xu Y B,Zhou Y,et al.Effects of heavy metal stress on antibiotics resistance of Pseudomonas alcaligenes LH7[J].Acta Ecologica Sinica,2015,35(10):1-12