宠物犬源沙门氏菌和葡萄球菌耐药性及耐药基因检测
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摘要
【目的】了解新疆乌鲁木齐市宠物犬源沙门氏菌和葡萄球菌的耐药性及耐药基因携带情况。【方法从77份宠物犬直肠粪便样品中分离沙门氏菌和葡萄球菌,采用琼脂稀释法对分离的菌株进行药敏试验,通过PCR方法对沙门氏菌6类23种耐药基因和葡萄球菌5类9种耐药基因进行检测。【结果】从77份宠物犬粪便样品中分离到沙门氏菌36株,葡萄球菌75株。除头孢噻呋、安普霉素、阿米卡星、硫酸庆大霉素和多黏菌素外,宠物犬源沙门氏菌对其他被捡抗菌药物的耐药率高达97.0%以上,多重耐药以8耐为主,tetB、blaTEM、blaOXA、qnrS、oqxA、oqxB、aac(6′)-Ib-cr、ant(3″)-Ia和floR基因检出率在97.0%以上。宠物犬源葡萄球菌对左氧氟沙星(93.5%)、苯唑西林(90.7%)和氟苯尼考(82.9%)耐药率在80.0%以上,多重耐药集中在4耐、6耐和9耐,主要检出的基因为fexA(40.0%,30/75)和femA(18.7%,14/75),其次为tetM(16.0%,12/75)、ermB(14.7%,11/75)和mecA(1.3%,1/75)。【结论】宠物犬源沙门氏菌和葡萄球菌耐药情况严重,耐药基因携带率高,使宠物犬的细菌性疾病治疗难度加大,应加强耐药菌的监测,降低宠物犬源耐药菌转移给人的潜在风险。
【Objective】This study investigated the resistance and resistant genes of Salmonella and Staphylococcus isolates from feces of pet dog against antibacterial drugs in Urumqi,Xinjiang.【Method】Salmonella and Staphylococci were isolated from 77 pet dog rectal feces samples.Agar dilution method was used to test the susceptibility of the isolates.The PCR method was used to detect 23 resistant genes of6 kinds of Salmonellaand 9 resistant genes of 5 kinds of Staphylococcus.【Result】A total of 36 strains of Salmonellaand 75 strains of Staphylococcus were isolated.Except ceftiofur,apramycin,amikacin,gentamicin and polymyxin,the resistant rates of Salmonellaisolates against other detected antimicrobial agents were above 97.0%.The multidrug resistance strains mainly focused on 8 kinds of drug resistance,and the detection rates of tetB,blaTEM,blaOXA,qnrS,oqxA,oqxB,aac(6′)-Ib-cr,ant(3″)-Ia and floR genes were larger than 97.0%.The resistant rates of Staphylococcus against levofloxacin(93.5%),oxacillin(90.7%)and florfenicol(82.9%)were higher than 80.0%.The multidrug resistance results of Staphylococcus were concentrated on 4 types,6 types and 9 types.The main detected genes were fexA(40.0%,30/75)and femA(18.7%,14/75),followed by tetM(16.0%,12/75),ermB(14.7%,11/75)and mecA(1.3%,1/75).【Conclusion】The drug resistance of Salmonella and Staphylococcus was severe,and the resistance genecarrying rates were high,resulting in difficult treatment of bacterial diseases in pet dogs.The monitoring of drug-resistant bacteria should be strengthened to reduce the potential risk of transfer of drug-resistant bacteria from pet dogs.
【Objective】This study investigated the resistance and resistant genes of Salmonella and Staphylococcus isolates from feces of pet dog against antibacterial drugs in Urumqi,Xinjiang.【Method】Salmonella and Staphylococci were isolated from 77 pet dog rectal feces samples.Agar dilution method was used to test the susceptibility of the isolates.The PCR method was used to detect 23 resistant genes of6 kinds of Salmonellaand 9 resistant genes of 5 kinds of Staphylococcus.【Result】A total of 36 strains of Salmonellaand 75 strains of Staphylococcus were isolated.Except ceftiofur,apramycin,amikacin,gentamicin and polymyxin,the resistant rates of Salmonellaisolates against other detected antimicrobial agents were above 97.0%.The multidrug resistance strains mainly focused on 8 kinds of drug resistance,and the detection rates of tetB,blaTEM,blaOXA,qnrS,oqxA,oqxB,aac(6′)-Ib-cr,ant(3″)-Ia and floR genes were larger than 97.0%.The resistant rates of Staphylococcus against levofloxacin(93.5%),oxacillin(90.7%)and florfenicol(82.9%)were higher than 80.0%.The multidrug resistance results of Staphylococcus were concentrated on 4 types,6 types and 9 types.The main detected genes were fexA(40.0%,30/75)and femA(18.7%,14/75),followed by tetM(16.0%,12/75),ermB(14.7%,11/75)and mecA(1.3%,1/75).【Conclusion】The drug resistance of Salmonella and Staphylococcus was severe,and the resistance genecarrying rates were high,resulting in difficult treatment of bacterial diseases in pet dogs.The monitoring of drug-resistant bacteria should be strengthened to reduce the potential risk of transfer of drug-resistant bacteria from pet dogs.
引文
[1]李鹏.藏猪源大肠杆菌和肠球菌耐药性调查及其耐药机制的研究[D].北京:中国农业大学,2015.Li P.Antimicrobial resistance characteristics and mechanisms of Escherichia coli&Enterococcus from Tibetan pigs[D].Beijing:China Agricultural University,2015.
[2]Zishiri O T,Mkhize N,Mukaratirwa S.Prevalence of virulence and antimicrobial resistance genes in Salmonella spp.isolated from commercial chickens and human clinical isolates from South Africa and Brazil[J].Onderstepoort J Vet Res,2016,83(1):1-11.
[3]Clinical and Laboratory Standards Institute.Performance standards for antimicrobial susceptibility testing,twenty-third informational supplement,M100-S23[S].Wayne,PA:Clinical and Laboratory Standards Institute,2013.
[4]Guerra B,Helmuth R,Thomas K,et al.Plasmid-mediated quinolone resistance determinants in Salmonellaspp.isolates from reptiles in Germany[J].The Journal of Antimicrobial Chemotherapy,2010,65(9):2043-2045.
[5]Gay K,Robicsek A,Strahilevitz J,et al.Plasmid-mediated quinolone resistance in non-typhi serotypes of Salmonella enterica[J].Clinical Infectious Diseases,2006,43(3):297-304.
[6]Wang M,Guo Q,Xu X,et al.New plasmid-mediated quinolone resistance gene,qnrC,found in a clinical isolate of Proteus mirabilis[J].Antimicrobial Agents and Chemotherapy,2009,53(5):1892-1897.
[7]刘海燕,张平.家禽沙门氏菌病的流行现状及防控方法[J].畜牧市场,2009(11):30-32.Liu H Y,Zhang P.Prevalence and control of Salmonella in poultry[J].Stockbreeding Market,2009(11):30-32.
[8]Rodríguez-Martínez J M,Díaz de Alba P,Briales A,et al.Contribution of oqxABefflux pumps to quinolone resistance in extended-spectrum-β-lactamase-producing Klebsiella pneumoniae[J].The Journal of Antimicrobial Chemotherapy,2013,68(1):68-73.
[9]Monstein H J,Ostholm-Balkhed A,Nilsson M V,et al.Multiplex PCR amplification assay for the detection of blaSHV,blaTEMand blaCTX-M genes in Enterobacteriaceae[J].APMIS,2007,115(12):1400-1408.
[10]Xia L N,Tao X Q,Shen J Z,et al.A survey ofβ-lactamase and16SrRNA methylase genes among fluoroquinolone-resistant Escherichia coli isolates and their horizontal transmission in Shandong,China[J].Foodborne Pathogens and Disease,2011,8(12):1241-1248.
[11]Lim K T,Yeo C C,Yasin R M,et al.Characterization of multidrug-resistant and extended-spectrum beta-lactamase-producing Klebsiella pneumoniae strains from Malaysian hospitals[J].Journal of Medical Microbiology,2009,58(11):1463-1469.
[12]Prasertsee T,Khantaprab N,Yamsakul P,et al.Repetitive sequence-based PCR fingerprinting and the relationship of antimicrobial-resistance characteristics and corresponding genes among Salmonellastrains from pig production[J].Asian Pacific Journal of Tropical Disease,2016,6(5):390-395.
[13]Bacci C,Boni E,Alpiqiani I,et al.Phenotypic and genotypic features of antibiotic resistance in Salmonella entericaisolated from chicken meat and chicken and quail carcasses[J].International Journal Food Microbiology,2012,160(1):16-23.
[14]曹堃,王元兰,刘宗梁,等.实时荧光定量PCR检测沙门菌flor基因和sul2基因方法的建立[J].中国抗生素杂志,2015,40(7):531-537.Cao K,Wang Y L,Liu Z L,et al.Establish the detecting mothed of Salmonella flor and sul2 genes by Real-Time quantitative PCR[J].Chinese Journal of Antibiotics,2015,40(7):531-537.
[15]俞林锋.上海某猪场大肠杆菌中mcr-1基因的传播机制研究[D].广州:华南农业大学,2016.Yu L F.Transfer mechanism of colistin resistance gene mcr-1among Escherichia coli isolates from a livestock farm of Shanghai[D].Guangzhou:South China Agricultural University,2016.
[16]蔡建星.新疆猪源葡萄球菌cfr和fexA基因及其遗传元件的检测分析[D].乌鲁木齐:新疆农业大学,2017.Cai J X.Analysis of cfrand fexAgenes and their genetic elements Staphylococci from pigs in Xinjiang[D].Urumqi:Xinjiang Agricultural University,2017.
[17]杜伟伟.河北省乳源金黄色葡萄球菌耐药性检测及耐药基因的研究[D].保定:河北农业大学,2015.Du W W.Research on drug resistence and resistant genes of the Staphylococcus aureus from raw milk in province of Hebei[D].Baoding:Hebei Agricultural University,2015.
[18]杨守深,王晶,范克伟,等.福建地区猪源葡萄球菌耐药性分析及耐药基因检测[J].中国人兽共患病学报,2016,32(11):976-982.Yang S S,Wang J,Fan K W,et al.Antimicrobial resistance analysis and resistance genes detection in Staphylococcusisolated from swine in Fujian province,China[J].Chinese Journal of Zoonoses,2016,32(11):976-982.
[19]王元兰.合肥市宠物犬与犬粮携带沙门氏菌情况调查及耐药性研究[D].合肥:安徽农业大学,2013.Wang Y L.Investigation on the carrying situation of Salmanellafrom pet dogs and food in Hefei and analysis of their antibiotic resistance[D].Hefei:Anhui Agricultural University,2013.
[20]冯言言,田伟,罗倩怡,等.广东省宠物源伪中间型葡萄球菌的耐药性调查研究[J].动物医学进展,2012,33(6):6-10.Feng Y Y,Tian W,Luo Q Y,et al.Antimicrobial resistance of Staphylococcus pseudintermedius isolated from pets in Guangdong province[J].Progress in Veterinary Medicine,2012,33(6):6-10.
[21]周传铎,赵然,金艺鹏,等.北京地区警犬皮肤伪中间型葡萄球菌药敏试验及耐药基因筛查[J].中国兽医杂志,2016,52(11):100-103.Zhou C D,Zhao R,Jing Y P,et al.Isolation,identification and drug resistance test of Staphylococcus from the skin of police dogs in Beijing[J].Chinese Journal of Veterinary Medicine,2016,52(11):100-103.
[22]罗倩怡,徐燕莉,邓玉婷,等.宠物源细菌耐药性研究进展[J].中国兽医杂志,2009,45(3):57-59.Luo Q Y,Xu Y L,Deng Y T,et al.Research progress in drug resistance of pet source bacteria[J].Chinese Journal of Veterinary Medicine,2009,45(3):57-59.
[23]黄凯,陈素娟,黄骏,等.动物源性沙门氏菌的耐药性分析及氟苯尼考类耐药基因的鉴定[J].中国畜牧兽医,2015,42(2):459-466.Huang K,Chen S J,Huang J,et al.Analysis of antibiotic resistance of Salmonellaisolated from animals and identification of its florfenicol resistant gene[J].China Animal Husbandry&.Veterinary Medicine,2015,42(2):459-466.
[24]曾婉秋,杨晓农,陈瑞,等.成都地区健康犬及其畜主鼻腔金黄色葡萄球菌和中间型葡萄球菌的分离及耐药性研究[J].西南民族大学学报(自然科学版),2017,43(1):13-19.Zeng W Q,Yang X N,Chen R,et al.Investigation of carrying rates and drug resistance of Staphylococcus aureus and Staphylococcus intermedius in healthy dogs and their owners in Chengdu city[J].Journal of Southwest University for Nationalities(Natural Science Edition),2017,43(1):13-19.
[25]孙洋,冯书章.宠物在病原菌耐药性形成过程中的作用[J].动物医学进展,2007,28(12):106-109.Sun Y,Feng S Z.The role of pets in the pathogenesis of pathogen resistance[J].Progress in Veterinary Medicine,2007,28(12):106-109.
[26]陈耀能,梁景涛,陈爱贞,等.佛山市食源性和人源沙门氏菌血清型分布与耐药性研究[J].热带医学杂志,2012,12(8):955-959.Chen Y N,Liang J T,Chen A Z,et al.Serotype distribution and drug resistance of food-borne and human origin Salmonellain Foshan[J].Journal of Tropical Medicine,2012,12(8):955-959.
[27]杨怀珍,牟亚,罗薇.食源性沙门氏菌的研究进展[J].黑龙江畜牧兽医,2016(7):69-71,75.Yang H Z,Mou Y,Luo W.Research progress of food-borne Salmonella[J].Heilongjiang Animal Science and Veterinary Medicine,2016(7):69-71,75.
[28]张万江.猪源葡萄球菌和芽孢杆菌cfr基因流行病学及传播机制研究[D].北京:中国农业大学,2011.Zhang W J.The epidemiological study on the multi-resistance gene cfr and its transmission mechanism among Staphylococcus and Bacillus sp.isolates from swine[D].Beijing:China Agricultural University,2011.
[2]Zishiri O T,Mkhize N,Mukaratirwa S.Prevalence of virulence and antimicrobial resistance genes in Salmonella spp.isolated from commercial chickens and human clinical isolates from South Africa and Brazil[J].Onderstepoort J Vet Res,2016,83(1):1-11.
[3]Clinical and Laboratory Standards Institute.Performance standards for antimicrobial susceptibility testing,twenty-third informational supplement,M100-S23[S].Wayne,PA:Clinical and Laboratory Standards Institute,2013.
[4]Guerra B,Helmuth R,Thomas K,et al.Plasmid-mediated quinolone resistance determinants in Salmonellaspp.isolates from reptiles in Germany[J].The Journal of Antimicrobial Chemotherapy,2010,65(9):2043-2045.
[5]Gay K,Robicsek A,Strahilevitz J,et al.Plasmid-mediated quinolone resistance in non-typhi serotypes of Salmonella enterica[J].Clinical Infectious Diseases,2006,43(3):297-304.
[6]Wang M,Guo Q,Xu X,et al.New plasmid-mediated quinolone resistance gene,qnrC,found in a clinical isolate of Proteus mirabilis[J].Antimicrobial Agents and Chemotherapy,2009,53(5):1892-1897.
[7]刘海燕,张平.家禽沙门氏菌病的流行现状及防控方法[J].畜牧市场,2009(11):30-32.Liu H Y,Zhang P.Prevalence and control of Salmonella in poultry[J].Stockbreeding Market,2009(11):30-32.
[8]Rodríguez-Martínez J M,Díaz de Alba P,Briales A,et al.Contribution of oqxABefflux pumps to quinolone resistance in extended-spectrum-β-lactamase-producing Klebsiella pneumoniae[J].The Journal of Antimicrobial Chemotherapy,2013,68(1):68-73.
[9]Monstein H J,Ostholm-Balkhed A,Nilsson M V,et al.Multiplex PCR amplification assay for the detection of blaSHV,blaTEMand blaCTX-M genes in Enterobacteriaceae[J].APMIS,2007,115(12):1400-1408.
[10]Xia L N,Tao X Q,Shen J Z,et al.A survey ofβ-lactamase and16SrRNA methylase genes among fluoroquinolone-resistant Escherichia coli isolates and their horizontal transmission in Shandong,China[J].Foodborne Pathogens and Disease,2011,8(12):1241-1248.
[11]Lim K T,Yeo C C,Yasin R M,et al.Characterization of multidrug-resistant and extended-spectrum beta-lactamase-producing Klebsiella pneumoniae strains from Malaysian hospitals[J].Journal of Medical Microbiology,2009,58(11):1463-1469.
[12]Prasertsee T,Khantaprab N,Yamsakul P,et al.Repetitive sequence-based PCR fingerprinting and the relationship of antimicrobial-resistance characteristics and corresponding genes among Salmonellastrains from pig production[J].Asian Pacific Journal of Tropical Disease,2016,6(5):390-395.
[13]Bacci C,Boni E,Alpiqiani I,et al.Phenotypic and genotypic features of antibiotic resistance in Salmonella entericaisolated from chicken meat and chicken and quail carcasses[J].International Journal Food Microbiology,2012,160(1):16-23.
[14]曹堃,王元兰,刘宗梁,等.实时荧光定量PCR检测沙门菌flor基因和sul2基因方法的建立[J].中国抗生素杂志,2015,40(7):531-537.Cao K,Wang Y L,Liu Z L,et al.Establish the detecting mothed of Salmonella flor and sul2 genes by Real-Time quantitative PCR[J].Chinese Journal of Antibiotics,2015,40(7):531-537.
[15]俞林锋.上海某猪场大肠杆菌中mcr-1基因的传播机制研究[D].广州:华南农业大学,2016.Yu L F.Transfer mechanism of colistin resistance gene mcr-1among Escherichia coli isolates from a livestock farm of Shanghai[D].Guangzhou:South China Agricultural University,2016.
[16]蔡建星.新疆猪源葡萄球菌cfr和fexA基因及其遗传元件的检测分析[D].乌鲁木齐:新疆农业大学,2017.Cai J X.Analysis of cfrand fexAgenes and their genetic elements Staphylococci from pigs in Xinjiang[D].Urumqi:Xinjiang Agricultural University,2017.
[17]杜伟伟.河北省乳源金黄色葡萄球菌耐药性检测及耐药基因的研究[D].保定:河北农业大学,2015.Du W W.Research on drug resistence and resistant genes of the Staphylococcus aureus from raw milk in province of Hebei[D].Baoding:Hebei Agricultural University,2015.
[18]杨守深,王晶,范克伟,等.福建地区猪源葡萄球菌耐药性分析及耐药基因检测[J].中国人兽共患病学报,2016,32(11):976-982.Yang S S,Wang J,Fan K W,et al.Antimicrobial resistance analysis and resistance genes detection in Staphylococcusisolated from swine in Fujian province,China[J].Chinese Journal of Zoonoses,2016,32(11):976-982.
[19]王元兰.合肥市宠物犬与犬粮携带沙门氏菌情况调查及耐药性研究[D].合肥:安徽农业大学,2013.Wang Y L.Investigation on the carrying situation of Salmanellafrom pet dogs and food in Hefei and analysis of their antibiotic resistance[D].Hefei:Anhui Agricultural University,2013.
[20]冯言言,田伟,罗倩怡,等.广东省宠物源伪中间型葡萄球菌的耐药性调查研究[J].动物医学进展,2012,33(6):6-10.Feng Y Y,Tian W,Luo Q Y,et al.Antimicrobial resistance of Staphylococcus pseudintermedius isolated from pets in Guangdong province[J].Progress in Veterinary Medicine,2012,33(6):6-10.
[21]周传铎,赵然,金艺鹏,等.北京地区警犬皮肤伪中间型葡萄球菌药敏试验及耐药基因筛查[J].中国兽医杂志,2016,52(11):100-103.Zhou C D,Zhao R,Jing Y P,et al.Isolation,identification and drug resistance test of Staphylococcus from the skin of police dogs in Beijing[J].Chinese Journal of Veterinary Medicine,2016,52(11):100-103.
[22]罗倩怡,徐燕莉,邓玉婷,等.宠物源细菌耐药性研究进展[J].中国兽医杂志,2009,45(3):57-59.Luo Q Y,Xu Y L,Deng Y T,et al.Research progress in drug resistance of pet source bacteria[J].Chinese Journal of Veterinary Medicine,2009,45(3):57-59.
[23]黄凯,陈素娟,黄骏,等.动物源性沙门氏菌的耐药性分析及氟苯尼考类耐药基因的鉴定[J].中国畜牧兽医,2015,42(2):459-466.Huang K,Chen S J,Huang J,et al.Analysis of antibiotic resistance of Salmonellaisolated from animals and identification of its florfenicol resistant gene[J].China Animal Husbandry&.Veterinary Medicine,2015,42(2):459-466.
[24]曾婉秋,杨晓农,陈瑞,等.成都地区健康犬及其畜主鼻腔金黄色葡萄球菌和中间型葡萄球菌的分离及耐药性研究[J].西南民族大学学报(自然科学版),2017,43(1):13-19.Zeng W Q,Yang X N,Chen R,et al.Investigation of carrying rates and drug resistance of Staphylococcus aureus and Staphylococcus intermedius in healthy dogs and their owners in Chengdu city[J].Journal of Southwest University for Nationalities(Natural Science Edition),2017,43(1):13-19.
[25]孙洋,冯书章.宠物在病原菌耐药性形成过程中的作用[J].动物医学进展,2007,28(12):106-109.Sun Y,Feng S Z.The role of pets in the pathogenesis of pathogen resistance[J].Progress in Veterinary Medicine,2007,28(12):106-109.
[26]陈耀能,梁景涛,陈爱贞,等.佛山市食源性和人源沙门氏菌血清型分布与耐药性研究[J].热带医学杂志,2012,12(8):955-959.Chen Y N,Liang J T,Chen A Z,et al.Serotype distribution and drug resistance of food-borne and human origin Salmonellain Foshan[J].Journal of Tropical Medicine,2012,12(8):955-959.
[27]杨怀珍,牟亚,罗薇.食源性沙门氏菌的研究进展[J].黑龙江畜牧兽医,2016(7):69-71,75.Yang H Z,Mou Y,Luo W.Research progress of food-borne Salmonella[J].Heilongjiang Animal Science and Veterinary Medicine,2016(7):69-71,75.
[28]张万江.猪源葡萄球菌和芽孢杆菌cfr基因流行病学及传播机制研究[D].北京:中国农业大学,2011.Zhang W J.The epidemiological study on the multi-resistance gene cfr and its transmission mechanism among Staphylococcus and Bacillus sp.isolates from swine[D].Beijing:China Agricultural University,2011.
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