WEI Wenjuan, ZHAO Shu, WANG Yuan, ZHOU Junfang, LI Xincang, FANG Wenhong. Detection of antibiotics resistance and distribution of resistance genes in Vibrio parahaemolyticus from cultured shrimp[J]. South China Fisheries Science, 2020, 16(1): 9-16. DOI: 10.12131/20190165
Citation: WEI Wenjuan, ZHAO Shu, WANG Yuan, ZHOU Junfang, LI Xincang, FANG Wenhong. Detection of antibiotics resistance and distribution of resistance genes in Vibrio parahaemolyticus from cultured shrimp[J]. South China Fisheries Science, 2020, 16(1): 9-16. DOI: 10.12131/20190165

Detection of antibiotics resistance and distribution of resistance genes in Vibrio parahaemolyticus from cultured shrimp

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  • Received Date: August 21, 2019
  • Revised Date: November 21, 2019
  • Available Online: December 02, 2019
  • We used agar dilution method to detect the resistance of 36 strains of Vibrio parahaemolyticus to 16 drugs. And used PCR amplication and DNA sequencing to detect the antimicrobial resistance to quinolones (qnrA, qnrB, qnrS, qnrVC), phenicols (cat, optrA, floR, cfr), tetracyclines (tetA, tetB, tetM), sulfonamides (sul1, sul2, sul3), aminoglycosides (strA, strB, aadA, aacA), rifampicin (arr), β-lactams (carB) and macrolides (erm). The results indicate that the isolates exhibited high resistance to ampicillin (88.9%) and sulfamethoxazole (SMZ, 66.7%), all susceptible to neomycin sulfate, gentamicin, ceftriaxone and meropenem. In general, multi-drug resistance (MDR) was highly prevalent (61.1%), and one isolate was resistant to six antimicrobials. Furthermore, 72.2% and 58.3% of the isolates were primarily mediated by qnrVC and strB, respectively; and the macrolides and rifamycin resistant genes were not detected in all the isolates. Obvious mismatch was found between the antimicrobial resistance phenotypes and genotypes, revealing the complexity of resistance to V. parahaemolyticus.
  • [1]
    DARAMOLA B A, WILLIAMS R, DIXON R A. In vitro antibiotic susceptibility of Vibrio parahaemolyticus from environmental sources in northern England[J]. Int J Antimicrob Ag, 2009, 34(5): 499-500. doi: 10.1016/j.ijantimicag.2009.06.015
    [2]
    张德福, 付绪磊, 张明, 等. 副溶血弧菌毒力因子及致病机理的研究进展[J]. 食品科学, 2015, 36(7): 216-222. doi: 10.7506/spkx1002-6630-201507040
    [3]
    HU Q, CHEN L. Virulence and antibiotic and heavy metal resistance of Vibrio parahaemolyticus isolated from crustaceans and shellfish in Shanghai, China[J]. J Food Protect, 2016, 79(8): 1371-1377. doi: 10.4315/0362-028X.JFP-16-031
    [4]
    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 Infec, 2016, 22(3): 258.e9-258.16. doi: 10.1016/j.cmi.2015.11.003
    [5]
    陈旭凌, 黄志坚. 水生细菌耐药性的遗传机制[J]. 中国抗生素杂志, 2013, 38(8): 573-578. doi: 10.3969/j.issn.1001-8689.2013.08.003
    [6]
    曾德乾, 冯娟, 徐力文, 等. 海水养殖鱼哈维弧菌分离株的耐药谱型分析[J]. 中国水产科学, 2015, 22(1): 129-138.
    [7]
    RODRIGUEZ-BLANCO A, LEMOS M L, OSORIO C R. Integrating conjugative elements as vectors of antibiotic, mercury, and quaternary ammonium compound resistance in marine aquaculture environments[J]. Antimicrob Agents Ch, 2012, 56(5): 2619-2626. doi: 10.1128/AAC.05997-11
    [8]
    张德福, 安慧, 张健, 等. 副溶血弧菌耐药及其机制的研究进展[J]. 食品工业科技, 2018, 39(9): 311-317, 324.
    [9]
    SILVESTER R, ALEXANDER D, AMMANAMVEETIL M A. Prevalence, antibiotic resistance, virulence and plasmid profiles of Vibrio parahaemolyticus from a tropical estuary and adjoining traditional prawn farm along the southwest coast of India[J]. Annal Microbiol, 2015, 65(4): 2141-2149. doi: 10.1007/s13213-015-1053-x
    [10]
    YOU K G, BONG C W, LEE C W. Antibiotic resistance and plasmid profiling of Vibrio spp. in tropical waters of Peninsular Malaysia[J]. Environ Monit Assess, 2016, 188(3): 171-171. doi: 10.1007/s10661-016-5163-0
    [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 Poll Res Int, 2016, 23(1): 918-926. doi: 10.1007/s11356-015-5650-9
    [12]
    SHAH S Q, CABELLO F C, L'ABEE-LUND T M, et al. Antimicrobial resistance and antimicrobial resistance genes in marine bacteria from salmon aquaculture and non-aquaculture sites[J]. Environ Microbiol, 2014, 16(5): 1310-1320. doi: 10.1111/1462-2920.12421
    [13]
    LETCHUMANAN V, YIN W F, LEE L H, et al. Prevalence and antimicrobial susceptibility of Vibrio parahaemolyticus isolated from retail shrimps in Malaysia[J]. Front Microbiol, 2015, 6(1): 33-33.
    [14]
    黄伟德, 肖双燕, 黎姗梅, 等. 广西凡纳滨对虾源副溶血弧菌耐药性和耐药基因的检测[J]. 西南农业学报, 2018, 31(9): 1979-1988.
    [15]
    XIE T F, XU X K, WU Q P, et al. Prevalence, molecular characterization, and antibiotic susceptibility of Vibrio parahaemolyticus from ready-to-eat foods in China[J]. Front Microbiol, 2016, 7(97): 549.
    [16]
    刘英超, 孙静, 钟继红. 急性炎症性肠炎的病原菌分布与耐药性分析[J]. 中华医院感染学杂志, 2015(9): 1987-1988, 1996.
    [17]
    谢红意, 叶鸿雁, 周方满, 等. 宁波地区腹泻患者副溶血弧菌毒力基因及耐药性分析[J]. 检验医学, 2019, 34(2): 136-139. doi: 10.3969/j.issn.1673-8640.2019.02.009
    [18]
    罗鹏, 何香燕, 胡超群. 细菌整合性接合元件SXT/R391研究进展[J]. 微生物学报, 2014, 54(5): 471-479.
    [19]
    AEDO S, IVANOVA L, TOMOVA A, et al. Plasmid-related quinolone resistance determinants in epidemic Vibrio parahaemolyticus, uropathogenic Escherichia coli, and marine bacteria from an aquaculture area in Chile[J]. Microb Ecol, 2014, 68(2): 324-328. doi: 10.1007/s00248-014-0409-2
    [20]
    MATSUO T, NAKAMURA K, KODAMA T, et al. Characterization of all RND-type multidrug efflux transporters in Vibrio parahaemolyticus[J]. Microbiol Open, 2013, 2(5): 725-742.
    [21]
    MORITA Y, KODAMA K, SHIOTA S, et al. Nor M, a putative multidrug efflux protein of Vibrio parahaemolyticus and its homolog in Escherichia coli[J]. Antimicrob Agents Ch, 1998, 42(7): 1778-1782. doi: 10.1128/AAC.42.7.1778
    [22]
    CLSI. Methods for antimicrobial dilution and disk susceptibility testing of infrequently isolated or fastidious bacteria: approved guideline[M]. CLSI, 2010: 70-96.
    [23]
    刘旭. 海水养殖源弧菌耐药性调查及qnrVC基因在弧菌中的流行情况研究[D]. 上海: 上海海洋大学, 2016: 14-16.
    [24]
    ROBICSEK A, STRAHILEVITZ J, SAHM D F, et al. Qnr prevalence in ceftazidime-resistant Enterobacteriaceae isolates from the United States[J]. Antimicrob Agents Ch, 2006, 50(8): 2872-2874. doi: 10.1128/AAC.01647-05
    [25]
    JACOBY G A, WALSH K E, MILLS D M, et al. QnrB, another plasmid-mediated gene for quinolone resistance[J]. Antimicrob Agents Ch, 2006, 50(4): 1178-1182. doi: 10.1128/AAC.50.4.1178-1182.2006
    [26]
    HATA M, SUZUKI M, MATSUMOTO M, et al. Cloning of a novel gene for quinolone resistance from a transferable plasmid in Shigella flexneri 2b[J]. Antimicrob Agents Ch, 2005, 49(2): 801-803. doi: 10.1128/AAC.49.2.801-803.2005
    [27]
    KITIYODOM S, KHEMTONG S, WONGTAVATCHAI J, et al. Characterization of antibiotic resistance in Vibrio spp. isolated from farmed marine shrimps (Penaeus monodon)[J]. FEMS Microbiol Ecol, 2010, 72(2): 219-227. doi: 10.1111/j.1574-6941.2010.00846.x
    [28]
    YOO M H, HUH M D, KIM E H. Characterization of chloramphenicol acetyltransferase gene by multiplex polymerase chain reaction in multidrug-resistant strains isolated from aquatic environments[J]. Aquaculture, 2003, 217(1/2/3/4): 1-21.
    [29]
    WANGY, LU Y. A novel gene, optr A, that confers transferable resistance to oxazolidinones and phenicols and its presence in entero-coccus faecalis and enterococcus faecium of human and animal origin[J]. J Antimicrob Ch, 2015, 70(8): 2182-2190. doi: 10.1093/jac/dkv116
    [30]
    KIM E H, AOKI T. Sequence analysis of the florfenicol resistance gene encoded in the transferable R-plasmid of a fish pathogen, Pasteurella piscicida[J]. Microbiol Immunol, 1996, 40(9): 665-669. doi: 10.1111/j.1348-0421.1996.tb01125.x
    [31]
    KEBRENBERG C, SCHWARZ S. Distribution of florfenicol resistance genes fex A and cfr among chloramphenicol-resistant Staph-yloccus isolates[J]. Antimicrob Agents Ch, 2006, 50(4): 1156-1163. doi: 10.1128/AAC.50.4.1156-1163.2006
    [32]
    CHIOU J C, LI R C, CHEN S. CARB-17 family of β-lactamases mediates intrinsic resistance to penicillins in Vibrio parahaemolyticus[J]. Antimicrob Agents Ch, 2015, 59(6): 3593-3595. doi: 10.1128/AAC.00047-15
    [33]
    汪建国. 我国渔药研究与应用综述(14)[J]. 渔业致富指南, 2013(14): 68-72.
    [34]
    汪涛, 杨再福, 陈勇航, 等. 磺胺类抗性基因的产生及演变研究进展[J]. 环境污染与防治, 2017, 39(11): 1251-1255.
    [35]
    JOSEPH S W, DEBELL R M, BROWN W P. In vitro response to chloramphenicol, tetracycline, ampicillin, gentamicin, and beta-lactamase production by halophilic vibrios from human and environmental sources[J]. Antimicrob Agents Ch, 1978, 13(2): 244-248. doi: 10.1128/AAC.13.2.244
    [36]
    CHARMAINE N G, CHEN H, GOH S G, et al. Microbial water quality and the detection of multidrug resistant E. coli and antibiotic resistance genes in aquaculture sites of Singapore[J]. Mar Pollut Bull, 2018, 135(7): 475-480.
    [37]
    JIANG Y, YAO L, LI F, et al. Characterization of antimicrobial resistance of Vibrio parahaemolyticus from cultured sea cucumbers (Apostichopus japonicas)[J]. Lett Appl Microbiol, 2014, 59(2): 147-154. doi: 10.1111/lam.12258
    [38]
    蒋魁, 徐力文, 苏友禄, 等. 2012年~2014年南海海水养殖鱼类病原菌哈维弧菌分离株的耐药性分析[J]. 南方水产科学, 2016, 12(6): 99-107. doi: 10.3969/j.issn.2095-0780.2016.06.013
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