2015年我国食源性副溶血性弧菌毒力基因及耐药特征研究
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摘要
目的探究我国2015年食源性副溶血性弧菌毒力基因分布情况和耐药特征。方法通过聚合酶链式反应,研究1046株副溶血性弧菌携带毒力基因tlh、tdh和trh情况,并采用微量肉汤稀释法,测定副溶血性弧菌对氨苄西林、阿莫西林/克拉维酸、头孢唑林、头孢噻肟、头孢他啶、亚胺培南、庆大霉素、四环素、环丙沙星、左氧沙星、氯霉素、复方新诺明等抗生素的敏感性。结果 1046株食源性副溶血性弧菌中有965株(92.3%)对一种或多种抗生素耐药,总耐药率为92.2%。全部菌株对庆大霉素和左氧氟沙星敏感,对其余10种抗生素有不同程度耐药,耐药率最高的前3位抗生素分别为头孢唑林(85.4%)、氨苄西林(59.4%)、阿莫西林/克拉维酸(49.3%)。1046株副溶血性弧菌全部携带tlh基因,其中19株(1.8%)trh基因阳性,4株(0.4%)tdh基因阳性。毒力基因阳性的所有菌株均未发现多重耐药现象,部分毒力基因阳性菌株对青霉素类和第一代头孢类抗生素耐药。结论我国2015年食源性副溶血性弧菌毒力基因携带率较低,对第一代头孢类和青霉素类抗生素耐药率较高;水产养殖地区应在养殖环节加强抗生素的管理并规范使用。
Objective To evaluate virulence distribution and antimicrobial resistance of Vibrio parahaemolyticusisolated from food in China in 2015.Methods A polymerase chain reaction method was used to detect virulence genes of tlh,tdh and trh.The susceptibility of 1046 isolates of Vibrio parahaemolyticus to 12 antibiotics were determined by a broth microdilution susceptibility test.The antibiotics of ampicillin,amoxicillin-clavulanic acid,cefazolin,cefotaxime,ceftazidime,imipenem,gentamicin,tetracycline,ciprofloxacin,levofloxacin,chloramphenico and trimethoprim/sulfamethoxazole were chosen.Results Totally 965 strains(92.2%) presented resistance to single or multiple antibiotics.All strains were sensitive to gentamicin and levofloxacin,and presented resistance to other antibiotics.The highest resistance rates were cefazolin(85.4%),ampicillin(59.4%) and amoxicillin-clavulanic acid(49.3%).All the 1046 isolates were positive to tlh gene,including 19 isolates were positive to trh gene and 4 isolates were positive to tdh gene.All the positive strains were not resistance to multiple antibiotics,and some of them were resistance to penicillins and the first-generation cephalosporin.Conclusion There is a lower positive rate of virulence genes in Vibrio parahaemolyticus isolated from food in China in 2015,though with a higher resistance rate to penicillins and the first-generation cephalosporin antibiotics.There is an urgent need to strengthen the management and standardization of antibiotics in a breeding process in the farming areas.
Objective To evaluate virulence distribution and antimicrobial resistance of Vibrio parahaemolyticusisolated from food in China in 2015.Methods A polymerase chain reaction method was used to detect virulence genes of tlh,tdh and trh.The susceptibility of 1046 isolates of Vibrio parahaemolyticus to 12 antibiotics were determined by a broth microdilution susceptibility test.The antibiotics of ampicillin,amoxicillin-clavulanic acid,cefazolin,cefotaxime,ceftazidime,imipenem,gentamicin,tetracycline,ciprofloxacin,levofloxacin,chloramphenico and trimethoprim/sulfamethoxazole were chosen.Results Totally 965 strains(92.2%) presented resistance to single or multiple antibiotics.All strains were sensitive to gentamicin and levofloxacin,and presented resistance to other antibiotics.The highest resistance rates were cefazolin(85.4%),ampicillin(59.4%) and amoxicillin-clavulanic acid(49.3%).All the 1046 isolates were positive to tlh gene,including 19 isolates were positive to trh gene and 4 isolates were positive to tdh gene.All the positive strains were not resistance to multiple antibiotics,and some of them were resistance to penicillins and the first-generation cephalosporin.Conclusion There is a lower positive rate of virulence genes in Vibrio parahaemolyticus isolated from food in China in 2015,though with a higher resistance rate to penicillins and the first-generation cephalosporin antibiotics.There is an urgent need to strengthen the management and standardization of antibiotics in a breeding process in the farming areas.
引文
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[15]Kang CH,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.
[16]Xu X,Cheng J,Wu Q,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.Jiang YH,Yao L,Li FL,et al.Isolation and antimicrobial resistance of Vibro parehaemolyticus in farmed marine shrimps from Qingdao markets[J].Chin J Zoono,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.Chen SF,Chen J,Xuan JC,et al.Isolation,identification and drug resistance analysis of three vibrio species in aquaculture area of Beihai[J].Prog Vet Med 2016,37(6):29-34
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[20]Gutierrez WCK,Klein SL,Lovell CR.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.Li WW,Wang XY,Guo YC.Distribution of virulence genes in Vibrio parahaemolyticusisolated from seafood in China[J].Chin J Food Hyg,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.
[23]Bhoopong P,Palittapongarnpim PR,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]严寒秋,张新,黄瑛,等.2011-2014年北京市副溶血性弧菌血清型及毒力基因特征分析[J].现代预防医学,2015,42(24):4468-4470.Yan HQ,Zhang X,Huang Y,et al.Analysis on the characteristics of serotype and virulence gene of Vibrioparahaemolyticus in Beijing during from 2011 to 2014[J].Mod Prev Med,2015,42(24):4468-4470.
[25]曲梅,张新,刘桂荣,等.北京市副溶血弧菌病原学和分子流行病学特征分析[J].中华流行病学杂志,2011,32(12):1255-1258.Qu M,Zhang X,Liu GR,et al.Analysis on the etiological and molecular epidemiological characteristics of Vibrio parahaemolyticusin Beijing[J].Chin J Epidemiol,2011,32(12):1255-1258.
[26]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.Chen HY,Chen M,Sheng YY,et al.Distribution of serotype,ribotype and virulence gene of Vibrio parahaemolyticus isolated from foodborne disease outbreaks[J].Chin J Food Hyg,2011,23(2):114-118.
[27]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 to2013[J].Clin Microbiol Infect,2016,22(3):258-259.
[28]Shirai H,Ito H,Hirayama T,et al.Molecular epidemiologic evidence for association of thermostable direct hemolysin(TDH)and TDH-related hemolysin of Vibrio parahaemolyticus with gastroenteritis[J].Infect Immun,1990,58(11):3568-3573.
[2]Newton A1,Kendall M,Vugia DJ,et al.Increasing rates of vibriosis in the United States,1996-2010:Review of surveillance data from 2 systems[J].Clin Infect Dis,2012,(54):391-395.
[3]Su YC,Liu C.Vibrio parahaemolyticus:a concern of seafood safety[J].Food Microbiol,2007,24(6):549-558.
[4]Theethakaew C,Feil EJ,Castilloramírez S,et al.Genetic relationships ofVibrio 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.
[5]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.
[6]Ceccarelli D,Hasan NA,Huq A,et al.Distribution and dynamics of epidemic and pandemic Vibrio parahaemolyticus virulence factors[J].Front Cell Infect Microbiol,2013,3(8):97.
[7]Okoh AI,Igbinosa EO.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 JW,Kim JH,Jr CC,et al.Isolation,molecular characterization,and antibiotic susceptibility of Vibrio parahaemolyticus in Korean seafood[J].Foodborne Pathog Dis,2012,9(3):224-231.
[9]Jorgensen JH.Methods for antimicrobial dilution and disk susceptibility testing of infrequently isolated or fastidious bacteria[D].Wayne:Clinical and Laboratory Standards Institute,2015.
[10]Bej AK,Patterson DP,Brasher CW,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.
[11]He Y,Jin L,Sun F,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.
[12]Alothrubi SM.Antibiotic Resistance of Vibrio parahaemolyticusisolated from cockles and shrimp sea food marketed in Selangor,Malaysia[J].Clin Microbiol,2014,3(3):148.
[13]Melo LM,Almeida D,Hofer E,et al.Antibiotic resistance of Vibrio parahaemolyticus isolated from pond-reared Litopenaeus vannamei marketed in natal,Brazil[J].Br J Microbiol,2011,42(4):1463-1469.
[14]Kang CH,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.
[15]Kang CH,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.
[16]Xu X,Cheng J,Wu Q,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.Jiang YH,Yao L,Li FL,et al.Isolation and antimicrobial resistance of Vibro parehaemolyticus in farmed marine shrimps from Qingdao markets[J].Chin J Zoono,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.Chen SF,Chen J,Xuan JC,et al.Isolation,identification and drug resistance analysis of three vibrio species in aquaculture area of Beihai[J].Prog Vet Med 2016,37(6):29-34
[19]Nishibuchi M,Kaper JB.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 WCK,Klein SL,Lovell CR.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.Li WW,Wang XY,Guo YC.Distribution of virulence genes in Vibrio parahaemolyticusisolated from seafood in China[J].Chin J Food Hyg,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.
[23]Bhoopong P,Palittapongarnpim PR,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]严寒秋,张新,黄瑛,等.2011-2014年北京市副溶血性弧菌血清型及毒力基因特征分析[J].现代预防医学,2015,42(24):4468-4470.Yan HQ,Zhang X,Huang Y,et al.Analysis on the characteristics of serotype and virulence gene of Vibrioparahaemolyticus in Beijing during from 2011 to 2014[J].Mod Prev Med,2015,42(24):4468-4470.
[25]曲梅,张新,刘桂荣,等.北京市副溶血弧菌病原学和分子流行病学特征分析[J].中华流行病学杂志,2011,32(12):1255-1258.Qu M,Zhang X,Liu GR,et al.Analysis on the etiological and molecular epidemiological characteristics of Vibrio parahaemolyticusin Beijing[J].Chin J Epidemiol,2011,32(12):1255-1258.
[26]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.Chen HY,Chen M,Sheng YY,et al.Distribution of serotype,ribotype and virulence gene of Vibrio parahaemolyticus isolated from foodborne disease outbreaks[J].Chin J Food Hyg,2011,23(2):114-118.
[27]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 to2013[J].Clin Microbiol Infect,2016,22(3):258-259.
[28]Shirai H,Ito H,Hirayama T,et al.Molecular epidemiologic evidence for association of thermostable direct hemolysin(TDH)and TDH-related hemolysin of Vibrio parahaemolyticus with gastroenteritis[J].Infect Immun,1990,58(11):3568-3573.