新疆、内蒙古部分地区乳房炎奶样中金黄色葡萄球菌的流行性与耐药性分析
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摘要
为确定患乳房炎奶牛乳中金黄色葡萄球菌的流行性和耐药性状况,试验自2018年4月至2018年5月采集了新疆地区70例和内蒙古地区50例患乳房炎奶牛生鲜乳样。通过对奶样中金黄色葡萄球菌进行增菌培养、分离纯化,KB纸片扩散法药敏试验及PCR耐药基因检测发现,新疆和内蒙古地区奶样中金黄色葡萄球菌分离率分别为28.6%和54.0%;新疆地区分离的金黄色葡萄球菌菌株对青霉素G的耐药性最高(75.0%),其次是磺胺异噁唑(60.0%)、林可霉素(55.0%);内蒙古地区的金黄色葡萄球菌菌株同样对青霉素G高度耐受,耐药率为70.4%,其次为林可霉素(63.0%)、克林霉素(51.9%);两地区所分离菌株中,88.9%以上携带多种耐药基因,呈现多重耐药性;两地区所检出的高频耐药基因分别是新疆地区的ermA(30.0%)、dfrS1(30.0%)基因和内蒙古地区的lnuA(46.2%)基因。
To evaluate the epidemicity and antimicrobial resistance of Staphylococcus aureus in the milk of mastitis cows,70 samples from Xinjiang and 50 samples from Inner Mongolia were collected from April to May in 2018.Staphylococcus aureus was cultivated,separated and purified.Antibiotic susceptibility test of purified isolates with KB paper and PCR analysis for the drug resistance gene were carried out.The results showed that the separation rate of Staphylococcus aureus in Xinjiang and Inner Mongolia was 28.6% and 54.0%,respectively.Penicillin G(75.0%) was the highest resistant antibiotic for Staphylococcus aureus isolated from Xinjiang,followed by sulfis oxazole(60.0%),lincomycin(55.0%).For the isolates from Inner Mongolia,penicillin G(70.4%) was also the highest resistant drug,followed by lincomycin(63.0%) and clindamycin(51.9%).The results of the resistance gene test showed that multi-resistance genes were detected in more than 88.9% of the isolates.The highest frequency resistance genes of isolated were ermA(30.0%) and dfrS1(30.0%) in Xinjiang and lnuA(46.2%) in Inner Mongolia,respectively.
To evaluate the epidemicity and antimicrobial resistance of Staphylococcus aureus in the milk of mastitis cows,70 samples from Xinjiang and 50 samples from Inner Mongolia were collected from April to May in 2018.Staphylococcus aureus was cultivated,separated and purified.Antibiotic susceptibility test of purified isolates with KB paper and PCR analysis for the drug resistance gene were carried out.The results showed that the separation rate of Staphylococcus aureus in Xinjiang and Inner Mongolia was 28.6% and 54.0%,respectively.Penicillin G(75.0%) was the highest resistant antibiotic for Staphylococcus aureus isolated from Xinjiang,followed by sulfis oxazole(60.0%),lincomycin(55.0%).For the isolates from Inner Mongolia,penicillin G(70.4%) was also the highest resistant drug,followed by lincomycin(63.0%) and clindamycin(51.9%).The results of the resistance gene test showed that multi-resistance genes were detected in more than 88.9% of the isolates.The highest frequency resistance genes of isolated were ermA(30.0%) and dfrS1(30.0%) in Xinjiang and lnuA(46.2%) in Inner Mongolia,respectively.
引文
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[3] MANO DE CARVALHO C B,MOREIRO J B,FERREIRA M C.Survey of Bacteroides fragilis group susceptibility patterns in Brazil[J].International Journal of Antimicrobial Agents,1997,8(2):115-120.
[4] 刘海燕,曹伟,谭玉林.98株金黄色葡萄球菌分布及耐药性分析[J].实用预防医学,2012,19(4):586-588. LIU H Y,CAO W,TAN Y L.Analysis on distribution and drug resistance of 98 strains of Staphylococcus aureus[J].Practical Preventive Medicine,2012,19(4):586-588.(in Chinese)
[5] EDWARDS R,GREENWOOD D.An investigation of beta-lactamases from clinical isolates of Bacteroides species[J].Journal of Medical Microbiology,1992,36(2):89-95.
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[14] RUEGG P L,OLIVEIRA L,JIN W,et al.Phenotypic antimicrobial susceptibility and occurrence of selected resistance genes in gram-positive mastitis pathogens isolated from Wisconsin dairy cows[J].Journal of Dairy Science,2015,98(7):4521-4534.
[15] UGWU C C,GOMEZ-SANZ E,AGBO I C,et al.Characterization of mannitol-fermenting methicillin-resistant Staphylococci isolated from pigs in Nigeria[J].Brazil Journal of Microbiology,2015,46(3):885-892.
[16] KEHRENBERG C,SCHWARZ S,SCHWARZ S.Distribution of florfenicol resistance genes fexA and cfr among chloramphenicol-resistant Staphylococcus isolates[J].Antimicrobial Agents and Chemotherapy,2006,50(4):1156-1163.
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[20] AVELAR K E,OTSUKI K,VICENTE A C,et al.Presence of the cfxA gene in Bacteroides distasonis[J].Research in Microbiology,2003,154(5):369-374.
[21] JAMALI H,PAYDAR M,RADMEHR B,et al.Prevalence and antimicrobial resistance of Staphylococcus aureus isolated from raw milk and dairy products[J].Food Control,2015,54:383-388.
[22] DAIGLE D M,HUGHES D W,WRIGHT G D.Prodigious substrate specificity of AAC(6′)-APH(2″),an aminoglycoside antibiotic resistance determinant in Enterococci and Staphylococci[J].Chemistry & Biology,1999,6(2):99-110.
[23] 田晓英,王军,于忠娜,等.天津地区乳房炎奶样中金黄色葡萄球菌耐药性研究[J].现代食品科技,2017,33(10):1-10. TIAN X Y,WANG J,YU Z N,et al.Staphylococcus aureus drug resistance of dairy cow mastitis in Tianjin region[J].Modern Food Science and Technology,2017,33(10):1-10.(in Chinese)
[24] 王秋东,刘琪,董志民,等.内蒙古部分地区致奶牛乳房炎金黄色葡萄球菌和产色葡萄球菌的分子流行病学及耐药性研究[J].中国预防兽医学报,2017,39(4):257-261. WANG Q D,LIU Q,DONG Z M,et al.Molecular epidemiology and antimicrobial resistance of Staphylococcus aureus and Staphylococcus chromogens from dairy cows in the Inner Mongolia[J].Chinese Journal of Preventive Veterinary Medicine,2017,39(4):257-261.(in Chinese)
[25] 张倩,屈勇刚,于会举,等.石河子地区奶牛乳房炎源金黄色葡萄球菌的分离鉴定及耐药性分析[J].黑龙江畜牧兽医,2017,1:102-105. ZHANG Q,QU Y G,YU H J,et al.Isolation,identification and analysis of drug resistance of Staphylococcus aureus from mastitis in dairy cows in Shihezi area[J].Heilongjiang Animal Science and Veterinary Medicine,2017,1:102-105.(in Chinese)
[26] WANG D,ZHANG L,ZHOU X,et al.Antimicrobial susceptibility,virulence genes,and randomly amplified polymorphic DNA analysis of Staphylococcus aureus recovered from bovine mastitis in Ningxia,China[J].Journal of Dairy Science,2016,99(12):1-10.
[27] LIU H,LI S,MENG L,et al.Prevalence,antimicrobial susceptibility,and molecular characterization of Staphylococcus aureus isolated from dairy herds in northern China[J].Journal of Dairy Science,2017,100(11):8796-8803.
[28] 殷国民,杨丹,顾常梅,等.烧伤科患者分离金黄色葡萄球菌耐药性和耐药基因研究[J].检验医学与临床,2018,15(18):2697-2701. YIN G M,YANG D,GU C M,et al.Drug resistance and drug resistance genes distribution of Staphylococcus aureus isolated from burn ward[J].Laboratory Medicine and Clinic,2018,15(18):2697-2701.(in Chinese)
[29] WATERS A E,CONTENTE-CUOMO T,BUCHHAGEN J,et al.Multidrug-resistant Staphylococcus aureus in US meat and poultry[J].Clinical Infectious Diseases,2011,52(10):1227-1230.
[2] HAN R W,ZHENG N,YU Z N,et al.Simultaneous determination of 38 veterinary antibiotic residues in raw milk by UPLC-MS/MS[J].Food Chemistry,2015,181:119-126.
[3] MANO DE CARVALHO C B,MOREIRO J B,FERREIRA M C.Survey of Bacteroides fragilis group susceptibility patterns in Brazil[J].International Journal of Antimicrobial Agents,1997,8(2):115-120.
[4] 刘海燕,曹伟,谭玉林.98株金黄色葡萄球菌分布及耐药性分析[J].实用预防医学,2012,19(4):586-588. LIU H Y,CAO W,TAN Y L.Analysis on distribution and drug resistance of 98 strains of Staphylococcus aureus[J].Practical Preventive Medicine,2012,19(4):586-588.(in Chinese)
[5] EDWARDS R,GREENWOOD D.An investigation of beta-lactamases from clinical isolates of Bacteroides species[J].Journal of Medical Microbiology,1992,36(2):89-95.
[6] BRAKSTAD O G,AASBAKK K,MAELAND J A.Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene[J].Journal of Clinical Microbiology,1992,30(7):1654-1660.
[7] ARGUDIN M A,VANDERHAEGHEN W,BUTAYE P.Diversity of antimicrobial resistance and virulence genes in methicillin-resistant non-Staphylococcus aureus Staphylococci from veal calves[J].Research in Veterinary Science,2015,99:10-16.
[8] GOMEZ-SANZ E,TORRES C,LOZANO C,et al.Detection,molecular characterization,and clonal diversity of methicillin-resistant Staphylococcus aureus CC398 and CC97 in Spanish slaughter pigs of different age groups[J].Foodborne Pathogens and Disease,2010,7(10):1269-1277.
[9] MARTINEAU F,PICARD F J,LANSAC N,et al.Correlation between the resistance genotype determined by multiplex PCR assays and the antibiotic susceptibility patterns of Staphylococcus aureus and Staphylococcus epidermidis[J].Antimicrobial Agents and Chemotherapy,2000,44(2):231-238.
[10] 李陇平.影响奶牛健康养殖的乳腺炎葡萄球菌研究[D].杨凌:西北农林科技大学,2015. LI L P.The Study of mastitis-inducing staphylococcal isolates for molecular typing and antibiotic resistance profiling[D].Yangling:Northwest A & F University,2015.(in Chinese)
[11] LINA G,QUAGLIA A,REVERDY M E,et al.Distribution of genes encoding resistance to macrolides,lincosamides,and streptogramins among Staphylo-cocci[J].Antimicrobial Agents and Chemotherapy,1999,43(5):1062-1066.
[12] JACKSON C R,FEDORKA-CRAY P J,BARRETT J B,et al.Prevalence of streptogramin resistance in Enterococci from animals:Identification of vatD from animal sources in the USA[J].International Journal of Antimicrobial Agents,2007,30(1):60-66.
[13] PATERSON G K,HARRISON E M,HOLMES M A.The emergence of mecC methicillin-resistant Staphylococcus aureus[J].Trends in Microbiology,2014,22(1):42-47.
[14] RUEGG P L,OLIVEIRA L,JIN W,et al.Phenotypic antimicrobial susceptibility and occurrence of selected resistance genes in gram-positive mastitis pathogens isolated from Wisconsin dairy cows[J].Journal of Dairy Science,2015,98(7):4521-4534.
[15] UGWU C C,GOMEZ-SANZ E,AGBO I C,et al.Characterization of mannitol-fermenting methicillin-resistant Staphylococci isolated from pigs in Nigeria[J].Brazil Journal of Microbiology,2015,46(3):885-892.
[16] KEHRENBERG C,SCHWARZ S,SCHWARZ S.Distribution of florfenicol resistance genes fexA and cfr among chloramphenicol-resistant Staphylococcus isolates[J].Antimicrobial Agents and Chemotherapy,2006,50(4):1156-1163.
[17] VESELY D L,GOWER W R,DIETZ J R,et al.Elevated atrial natriuretic peptides and early renal failure in type 2 diabetic Goto-Kakizaki rats[J].Metabolism,1999,48(6):771-778.
[18] SCHMITZ FJ,FLUIT A C,GONDOLF M,et al.The prevalence of aminoglycoside resistance and corresponding resistance genes in clinical isolates of Staphylococci from 19 European hospitals[J].The Journal of Antimicrobial Chemotherapy,1999,43(2):253-259.
[19] LIM K T,HANIFAH Y A,YUSOLF M,et al.ErmA,ermC,tetM and tetK are essential for erythromycin and tetracycline resistance among methicillin-resistant Staphylococcus aureus strains isolated from a tertiary hospital in Malaysia[J].Indian Journal of Medical Microbiology,2012,30(2):203-207.
[20] AVELAR K E,OTSUKI K,VICENTE A C,et al.Presence of the cfxA gene in Bacteroides distasonis[J].Research in Microbiology,2003,154(5):369-374.
[21] JAMALI H,PAYDAR M,RADMEHR B,et al.Prevalence and antimicrobial resistance of Staphylococcus aureus isolated from raw milk and dairy products[J].Food Control,2015,54:383-388.
[22] DAIGLE D M,HUGHES D W,WRIGHT G D.Prodigious substrate specificity of AAC(6′)-APH(2″),an aminoglycoside antibiotic resistance determinant in Enterococci and Staphylococci[J].Chemistry & Biology,1999,6(2):99-110.
[23] 田晓英,王军,于忠娜,等.天津地区乳房炎奶样中金黄色葡萄球菌耐药性研究[J].现代食品科技,2017,33(10):1-10. TIAN X Y,WANG J,YU Z N,et al.Staphylococcus aureus drug resistance of dairy cow mastitis in Tianjin region[J].Modern Food Science and Technology,2017,33(10):1-10.(in Chinese)
[24] 王秋东,刘琪,董志民,等.内蒙古部分地区致奶牛乳房炎金黄色葡萄球菌和产色葡萄球菌的分子流行病学及耐药性研究[J].中国预防兽医学报,2017,39(4):257-261. WANG Q D,LIU Q,DONG Z M,et al.Molecular epidemiology and antimicrobial resistance of Staphylococcus aureus and Staphylococcus chromogens from dairy cows in the Inner Mongolia[J].Chinese Journal of Preventive Veterinary Medicine,2017,39(4):257-261.(in Chinese)
[25] 张倩,屈勇刚,于会举,等.石河子地区奶牛乳房炎源金黄色葡萄球菌的分离鉴定及耐药性分析[J].黑龙江畜牧兽医,2017,1:102-105. ZHANG Q,QU Y G,YU H J,et al.Isolation,identification and analysis of drug resistance of Staphylococcus aureus from mastitis in dairy cows in Shihezi area[J].Heilongjiang Animal Science and Veterinary Medicine,2017,1:102-105.(in Chinese)
[26] WANG D,ZHANG L,ZHOU X,et al.Antimicrobial susceptibility,virulence genes,and randomly amplified polymorphic DNA analysis of Staphylococcus aureus recovered from bovine mastitis in Ningxia,China[J].Journal of Dairy Science,2016,99(12):1-10.
[27] LIU H,LI S,MENG L,et al.Prevalence,antimicrobial susceptibility,and molecular characterization of Staphylococcus aureus isolated from dairy herds in northern China[J].Journal of Dairy Science,2017,100(11):8796-8803.
[28] 殷国民,杨丹,顾常梅,等.烧伤科患者分离金黄色葡萄球菌耐药性和耐药基因研究[J].检验医学与临床,2018,15(18):2697-2701. YIN G M,YANG D,GU C M,et al.Drug resistance and drug resistance genes distribution of Staphylococcus aureus isolated from burn ward[J].Laboratory Medicine and Clinic,2018,15(18):2697-2701.(in Chinese)
[29] WATERS A E,CONTENTE-CUOMO T,BUCHHAGEN J,et al.Multidrug-resistant Staphylococcus aureus in US meat and poultry[J].Clinical Infectious Diseases,2011,52(10):1227-1230.