中国水产品中副溶血性弧菌耐药性及遗传特征分析
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摘要
目的研究我国水产品中分离的副溶血性弧菌耐药性、产毒特性及遗传特征。方法对我国26个省、直辖市和自治区水产品中分离的1 137株副溶血性弧菌进行耐药性分析和毒力基因检测,并用脉冲场凝胶电泳方法比对产毒菌株的遗传特征。结果 1 137株副溶血性弧菌中有254株(22.3%)受试菌株对测定的12种抗生素全部敏感,883株(77.7%)对测定的抗生素呈现不同程度耐药。所有受试菌株对左氧氟沙星和庆大霉素抗生素敏感,主要耐受的抗生素为氨苄西林(53.7%,611/1 137)、阿莫西林/克拉维酸(46.3%,526/1 137)和头孢唑林(24.2%,275/1 137)。1 137株副溶血性弧菌分离株全部检出tlh基因,有3株检出tdh基因,有19株检出trh基因,其中2株同时检出tdh和trh基因。20株产毒菌株遗传特征分散,与其食品来源和耐药谱之间也无密切关联,其中4株对测定的抗生素均敏感,其余菌株主要对青霉素类和头孢类抗生素耐药。结论我国水产品中副溶血性弧菌对青霉素类抗生素普遍耐药,毒力基因携带率较低,产毒菌株遗传特征多样。
Objective To evaluate antimicrobial resistance,virulence and genetic characteristics of Vibrio parahaemolyticus isolated from aquatic products in China. Methods The susceptibility of 1 137 isolates of Vibrio parahaemolyticus from 26 provinces was determined,and virulence genes including tlh,tdh and trh were detected. The genetic characteristics of Vibrio parahaemolyticus were analyzed by pulsed field gel electrophoresis method. Results Two hundred and fifty-four strains( 22. 3%) were susceptible to all the antibiotics tested and 883 strains( 77. 7%) presented resistance to single or multiple antibiotics. All strains were susceptible to gentamicin and levofloxacin. The highest resistance rates were ampicillin( 53. 7%,611/1 137),amoxicillin/clavulanic acid( 46. 3%,526/1 137) and cefazolin( 24. 2%,275/1 137). All the 1 137 isolates were positive for tlh gene,including 3 isolates were positive for tdh gene and 19 isolates were positive for trh gene. Twenty strains with virulence genes were resistance to mainly penicillins and cephalosporin and 4 strains of them were snsceptible to all the antibiotics tested. There was no certain relation between the source and drug-resistant spectrum in these strains with virulence genes,and they displayed different genetic characteristics. Conclusion Vibrio parahaemolyticus isolated from aquatic products were resistant to penicillins widely in China,but the rate of virulence genes of strains isolated from food was lower than clinical. The genetic characteristics of virulent strains were varied.
Objective To evaluate antimicrobial resistance,virulence and genetic characteristics of Vibrio parahaemolyticus isolated from aquatic products in China. Methods The susceptibility of 1 137 isolates of Vibrio parahaemolyticus from 26 provinces was determined,and virulence genes including tlh,tdh and trh were detected. The genetic characteristics of Vibrio parahaemolyticus were analyzed by pulsed field gel electrophoresis method. Results Two hundred and fifty-four strains( 22. 3%) were susceptible to all the antibiotics tested and 883 strains( 77. 7%) presented resistance to single or multiple antibiotics. All strains were susceptible to gentamicin and levofloxacin. The highest resistance rates were ampicillin( 53. 7%,611/1 137),amoxicillin/clavulanic acid( 46. 3%,526/1 137) and cefazolin( 24. 2%,275/1 137). All the 1 137 isolates were positive for tlh gene,including 3 isolates were positive for tdh gene and 19 isolates were positive for trh gene. Twenty strains with virulence genes were resistance to mainly penicillins and cephalosporin and 4 strains of them were snsceptible to all the antibiotics tested. There was no certain relation between the source and drug-resistant spectrum in these strains with virulence genes,and they displayed different genetic characteristics. Conclusion Vibrio parahaemolyticus isolated from aquatic products were resistant to penicillins widely in China,but the rate of virulence genes of strains isolated from food was lower than clinical. The genetic characteristics of virulent strains were varied.
引文
[1]刘秀梅,陈艳,樊永祥.2003年中国食源性疾病暴发的监测资料分析[J].卫生研究,2006,35(2):201-204.
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[3]THEETHAKAEW C,FEIL E J,CASTILLORAMREZ S,et al.Genetic relationships of Vibrio parahaemolyticus isolates from clinical,human carrier,and environmental sources in Thailand,determined by multilocus sequence analysis[J].Appl Environ Microbiol,2013,79(7):2358-2370.
[4]HONDA T,IIDA T.The pathogenicity of Vibrio parahaemolyticus and the role of thermostable direct hemolysin and related hemolysins[J].Rev Med Microbiol,1993,4(2):106-113.
[5]CECCARELLI D,HASAN N A,HUQ A,et al.Distribution and dynamics of epidemic and pandemic Vibrio parahaemolyticus virulence factors[J].Front Cell Infect Microbiol,2013,3(8):97.
[6]朱雪兰,陈艳,刘秀梅,等.副溶血性弧菌溶血素基因及其检测的研究进展[J].国外医学卫生学分册,2007,34(4):233-236.
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[8]JUN J W,KIM J H,CHORESCA J R,et al.Isolation,molecular characterization,and antibiotic susceptibility of Vibrio parahaemolyticus in Korean seafood[J].Foodborne Pathog Dis,2012,9(3):224-231.
[9]BEJ A K,PATTTERSON D P,BRASHER C W,et al.Detection of total and hemolysin-producing Vibrio parahaemolyticus in shellfish using multiplex PCR amplification of tlh,tdh and trh[J].J Microbiol Method,1999,36(3):215-225.
[10]Clinical Laboratory Standards Institute.Methods for antimicrobial dilution and disk susceptibility testing of infrequently isolated or fastidious bacteria;approved guideline[S].Wayne Pennsylvania:Clinical Laboratory Standards Institute,2015.
[11]KANG C H,SHIN Y,KIM W,et al.Prevalence and antimicrobial susceptibility of Vibrio parahaemolyticus isolated from oysters in Korea[J].Environ Sci Pollut Res,2016,23(1):1-9.
[12]HE Y,JIN L L,SUN F J,et al.Antibiotic and heavy-metal resistance of Vibrio parahaemolyticus isolated from fresh shrimps in Shanghai fish markets,China[J].Environ Sci Pollut Res,2016,23(15):1-8.
[13]韩毅,沙丹.2012—2014年无锡市不同来源副溶血性弧菌的病原学特征分析[J].中国食品卫生杂志,2016,28(6):795-799.
[14]ALOTHRUBI S M.Antibiotic resistance of Vibrio parahaemolyticus isolated from cockles and shrimp sea food marketed in Selangor,Malaysia[J].Clin Microbiol,2014,3(3):148.
[15]KANG C H,SHIN Y,JANG S,et al.Characterization of Vibrio parahaemolyticus isolated from oysters in Korea:resistance to various antibiotics and prevalence of virulence genes[J].Mar Pollut Bull,2017,118(1/2):261-266.
[16]XU X K,Cheng J H,WU Q P,et al.Prevalence,characterization,and antibiotic susceptibility of Vibrio parahaemolyticus isolated from retail aquatic products in North China[J].BMC Microbiol,2016,16(1):32.
[17]江艳华,姚琳,李风铃,等.青岛市售养殖海水虾中副溶血性弧菌的分离及耐药性分析[J].中国人兽共患病学报,2013,29(5):516-519.
[18]陈盛峰,陈佳,禤金彩,等.北海近岸养殖海域3种弧菌的分离鉴定及耐药性分析[J].动物医学进展,2016,37(6):29-34.
[19]NISHIBUCHI M,KAPER J B.Thermostable direct hemolysin gene of Vibrio parahaemolyticus:a virulence gene acquired by a marine bacterium[J].Infect Immun,1995,63(6):2093-2099.
[20]GUTIERREZ W C K,KLEIN S L,LOVELL C R.High frequency of virulence factor genes tdh,trh,and tlh in Vibrio parahaemolyticus strains isolated from a pristine estuary[J].Appl Environ Microbiol,2013,79(7):2247-2252.
[21]李薇薇,王晓英,郭云昌.中国部分水产品副溶血性弧菌毒力基因的分布特征[J].中国食品卫生杂志,2010,22(3):239-243.
[22]VUDDHAKUL V,SOBOON S,SUNGHIRAN W,et al.Distribution of virulent and pandemic strains of Vibrio parahaemolyticus in three molluscan shellfish species(Meretrix meretrix,Perna viridis,and Anadara granosa)and their association with foodborne disease in southern Thailand[J].J Food Prot,2006,69(11):2615-2620.
[23]BHOOPONG P,PALITTAPONGARNPIM P R,KIATKITTIPONG A,et al.Variability of properties of Vibrio parahaemolyticus strains isolated from individual patients[J].J Clin Microbiol,2007,45(5):1544-1550.
[24]马聪,朱海明,严纪文,等.不同来源的副溶血性弧菌定性定量分析及毒素基因检测[J].中国食品卫生杂志,2009,21(5):402-405.
[25]吴青,韩海红,余东敏,等.北京市水产品污染与感染病例中副溶血性弧菌血清型和毒力基因型的比较研究[J].中国食品卫生杂志,2015,27(4):363-367.
[26]CHEN Y,CHEN X,YU F,et al.Serology,virulence,antimicrobial susceptibility and molecular characteristics of clinical Vibrio parahaemolyticus strains circulating in southeastern China from 2009 to 2013[J].Clin Microbiol Infect,2016,22(3):258-259.
[27]刘丽萍,陈盛霞,徐岚,等.江苏及周边地区副溶血性弧菌分子流行病学特征研究[J].中国食品卫生杂志,2012,24(2):106-109.
[28]陈洪友,陈敏,盛跃颖,等.副溶血性弧菌食源性疾病暴发分离株的血清型、核糖型及毒力基因研究[J].中国食品卫生杂志,2011,23(2):114-118.
[2]SU Y C,LIU C.Vibrio parahaemolyticus:a concern of seafood safety[J].Food Microbiol,2007,24(6):549-558.
[3]THEETHAKAEW C,FEIL E J,CASTILLORAMREZ S,et al.Genetic relationships of Vibrio parahaemolyticus isolates from clinical,human carrier,and environmental sources in Thailand,determined by multilocus sequence analysis[J].Appl Environ Microbiol,2013,79(7):2358-2370.
[4]HONDA T,IIDA T.The pathogenicity of Vibrio parahaemolyticus and the role of thermostable direct hemolysin and related hemolysins[J].Rev Med Microbiol,1993,4(2):106-113.
[5]CECCARELLI D,HASAN N A,HUQ A,et al.Distribution and dynamics of epidemic and pandemic Vibrio parahaemolyticus virulence factors[J].Front Cell Infect Microbiol,2013,3(8):97.
[6]朱雪兰,陈艳,刘秀梅,等.副溶血性弧菌溶血素基因及其检测的研究进展[J].国外医学卫生学分册,2007,34(4):233-236.
[7]OKOH A I,IGBINOSA E O.Antibiotic susceptibility profiles of some Vibrio strains isolated from wastewater final effluents in a rural community of the Eastern Cape Province of South Africa[J].BMC Microbiol,2010,10(1):143.
[8]JUN J W,KIM J H,CHORESCA J R,et al.Isolation,molecular characterization,and antibiotic susceptibility of Vibrio parahaemolyticus in Korean seafood[J].Foodborne Pathog Dis,2012,9(3):224-231.
[9]BEJ A K,PATTTERSON D P,BRASHER C W,et al.Detection of total and hemolysin-producing Vibrio parahaemolyticus in shellfish using multiplex PCR amplification of tlh,tdh and trh[J].J Microbiol Method,1999,36(3):215-225.
[10]Clinical Laboratory Standards Institute.Methods for antimicrobial dilution and disk susceptibility testing of infrequently isolated or fastidious bacteria;approved guideline[S].Wayne Pennsylvania:Clinical Laboratory Standards Institute,2015.
[11]KANG C H,SHIN Y,KIM W,et al.Prevalence and antimicrobial susceptibility of Vibrio parahaemolyticus isolated from oysters in Korea[J].Environ Sci Pollut Res,2016,23(1):1-9.
[12]HE Y,JIN L L,SUN F J,et al.Antibiotic and heavy-metal resistance of Vibrio parahaemolyticus isolated from fresh shrimps in Shanghai fish markets,China[J].Environ Sci Pollut Res,2016,23(15):1-8.
[13]韩毅,沙丹.2012—2014年无锡市不同来源副溶血性弧菌的病原学特征分析[J].中国食品卫生杂志,2016,28(6):795-799.
[14]ALOTHRUBI S M.Antibiotic resistance of Vibrio parahaemolyticus isolated from cockles and shrimp sea food marketed in Selangor,Malaysia[J].Clin Microbiol,2014,3(3):148.
[15]KANG C H,SHIN Y,JANG S,et al.Characterization of Vibrio parahaemolyticus isolated from oysters in Korea:resistance to various antibiotics and prevalence of virulence genes[J].Mar Pollut Bull,2017,118(1/2):261-266.
[16]XU X K,Cheng J H,WU Q P,et al.Prevalence,characterization,and antibiotic susceptibility of Vibrio parahaemolyticus isolated from retail aquatic products in North China[J].BMC Microbiol,2016,16(1):32.
[17]江艳华,姚琳,李风铃,等.青岛市售养殖海水虾中副溶血性弧菌的分离及耐药性分析[J].中国人兽共患病学报,2013,29(5):516-519.
[18]陈盛峰,陈佳,禤金彩,等.北海近岸养殖海域3种弧菌的分离鉴定及耐药性分析[J].动物医学进展,2016,37(6):29-34.
[19]NISHIBUCHI M,KAPER J B.Thermostable direct hemolysin gene of Vibrio parahaemolyticus:a virulence gene acquired by a marine bacterium[J].Infect Immun,1995,63(6):2093-2099.
[20]GUTIERREZ W C K,KLEIN S L,LOVELL C R.High frequency of virulence factor genes tdh,trh,and tlh in Vibrio parahaemolyticus strains isolated from a pristine estuary[J].Appl Environ Microbiol,2013,79(7):2247-2252.
[21]李薇薇,王晓英,郭云昌.中国部分水产品副溶血性弧菌毒力基因的分布特征[J].中国食品卫生杂志,2010,22(3):239-243.
[22]VUDDHAKUL V,SOBOON S,SUNGHIRAN W,et al.Distribution of virulent and pandemic strains of Vibrio parahaemolyticus in three molluscan shellfish species(Meretrix meretrix,Perna viridis,and Anadara granosa)and their association with foodborne disease in southern Thailand[J].J Food Prot,2006,69(11):2615-2620.
[23]BHOOPONG P,PALITTAPONGARNPIM P R,KIATKITTIPONG A,et al.Variability of properties of Vibrio parahaemolyticus strains isolated from individual patients[J].J Clin Microbiol,2007,45(5):1544-1550.
[24]马聪,朱海明,严纪文,等.不同来源的副溶血性弧菌定性定量分析及毒素基因检测[J].中国食品卫生杂志,2009,21(5):402-405.
[25]吴青,韩海红,余东敏,等.北京市水产品污染与感染病例中副溶血性弧菌血清型和毒力基因型的比较研究[J].中国食品卫生杂志,2015,27(4):363-367.
[26]CHEN Y,CHEN X,YU F,et al.Serology,virulence,antimicrobial susceptibility and molecular characteristics of clinical Vibrio parahaemolyticus strains circulating in southeastern China from 2009 to 2013[J].Clin Microbiol Infect,2016,22(3):258-259.
[27]刘丽萍,陈盛霞,徐岚,等.江苏及周边地区副溶血性弧菌分子流行病学特征研究[J].中国食品卫生杂志,2012,24(2):106-109.
[28]陈洪友,陈敏,盛跃颖,等.副溶血性弧菌食源性疾病暴发分离株的血清型、核糖型及毒力基因研究[J].中国食品卫生杂志,2011,23(2):114-118.