食品与发酵工业 >

2023 , Vol. 49 >Issue 4: 250 - 257

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.031964

分析与检测

上海市养殖水产品及环境中副溶血性弧菌的流行、毒力及耐药性分析

  • 刘军军 ,
  • 刘单阳 ,
  • 李达容 ,
  • 王慧 ,
  • 赵勇 ,
  • 孙晓红
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  • 1(上海海洋大学 食品学院,上海,201306)
    2(农业部水产品贮藏保鲜质量安全风险评估实验室,上海,201306)
    3(上海水产品加工及贮藏工程技术研究中心,上海,201306)
硕士研究生(孙晓红教授为通信作者,E-mail:xhsun@shou.edu.cn)

收稿日期: 2022-04-13

  修回日期: 2022-05-24

  网络出版日期: 2023-03-20

基金资助

上海市科技兴农重点攻关项目(2019-02-08-00-10-F01149);上海市科技创新行动计划(21N31900300)

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The prevalence, virulence, and antibiotic resistance of Vibrio parahemolyticus in aquatic products and their breeding environment in Shanghai

  • LIU Junjun ,
  • LIU Danyang ,
  • LI Darong ,
  • WANG Hui ,
  • ZHAO Yong ,
  • SUN Xiaohong
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  • 1(College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China)
    2(Laboratory of Quality and Safety Risk Assessment for Aquatic Products Storage and Preservation (Shanghai), Ministry of Agriculture, Shanghai 201306, China)
    3(Shanghai Engineering Research Center for Aquatic-Products Processing & Preservation, Shanghai 201306, China)

Received date: 2022-04-13

  Revised date: 2022-05-24

  Online published: 2023-03-20

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摘要

为了解上海市养殖水产品中副溶血性弧菌(Vibrio parahaemolyticus)污染、毒力、血清型、耐药性及遗传关系。2020—2021年,采集上海市不同养殖场中南美白对虾、草鱼、鲫鱼、河蟹及养殖水和底泥,参考GB 4789.7—2013分离V.parahaemolyticus,采用PCR法特异性扩增tlh、tdh、trh和pirA基因,多重PCR检测血清型,微量肉汤稀释法检测耐药性,肠杆菌基因间重复序列(enterobacterial repetitive intergenic consensus sequence, ERIC)-PCR分析V.parahaemolyticus遗传关系。结果表明,234份样本中V.parahaemolyticus总污染率为30.8%,南美白对虾污染最严重(78.4%)。V.parahaemolyticus污染受季节温度影响,夏季污染率最高(51.4%)。V.parahaemolyticus均未检出毒力基因,血清群呈多样分布,O1群占优势(66.7%)。72株V.parahaemolyticus对氨苄西林和磺胺甲恶唑100%耐药,对头孢唑啉和红霉素耐药率高于70%,对复方新诺明耐药率为18.1%。对亚胺培南、头孢噻肟、头孢吡肟、氯霉素和卡那霉素中介耐药/耐药率低于5%。ERIC-PCR聚类群组分散,遗传多样性高。该研究结果为上海市养殖水产品的质量安全提供数据支撑。

关键词： 副溶血性弧菌; 水产养殖; 血清型; 耐药性; 遗传多样性

本文引用格式

刘军军 , 刘单阳 , 李达容 , 王慧 , 赵勇 , 孙晓红 . 上海市养殖水产品及环境中副溶血性弧菌的流行、毒力及耐药性分析[J]. 食品与发酵工业, 2023 , 49(4) : 250 -257 . DOI: 10.13995/j.cnki.11-1802/ts.031964

Abstract

To investigate the prevalence, virulence, serotype, antibiotic resistance and genetic relationship of Vibrio parahaemolyticus in the aquaculture products in Shanghai, Penaeus vannamei, Ctenopharyngodon idellus, Carassius auratus and Eriocheir sinensis, as well as their aquaculture water and sediment, were collected from different farms in Shanghai inform 2020 to 2021. V. parahaemolyticus was isolated according to the National Standard GB 4789.7—2013 and PCR was used to detect thermolabile hemolysin gene (tlh), heat-resistant direct hemolysin gene (tdh), heat-resistant-related hemolysin gene (trh), Photorhabdus insect-related toxin gene (pirA) that associated with acute hepatopancreas necrosis. Multiplex PCR was used to detect serotypes. The micro-broth dilution and Enterobacter repeat intergenic consensus sequence PCR (ERIC-PCR) were used to detect antimicrobial susceptibility and genetic relationship, respectively. V. parahaemolyticus was detected in 72 of the total 234 samples (30.8%) and the pollution of P. vannamei was the most serious (78.4%). The seasonal temperature significantly affected the abundance of V. parahaemolyticus, and the pollution rate was 51.4% in summer. None of the isolates possessed pathogenic genes tdh, trh and pirA. The distribution of serotypes was diverse and the most prevalent serotype was O1 (66.7%). Seventy-two V. parahaemolyticus strains were all resistant to ampicillin and sulfamethoxazole, more than 70% strains were resistant to cefazolin and erythromycin, and 18.1% strains were resistant to sulfamethoxazole-trimoxazole. The rate of intermediate-resistance or resistance to imipenem, cefotaxime, cefepime, chloramphenicol and kanamycin was less than 5%. ERIC-PCR clusters were dispersed and showed high genetic diversity. The results of this study provide data support for the quality and safety of aquatic products in Shanghai.

Key words： Vibrio parahaemolyticus; aquaculture; serotype; antibiotic resistance; genetic diversity

参考文献

[1] RYU A R, MOK J S, LEE D E, et al.Occurrence, virulence, and antimicrobial resistance of Vibrio parahaemolyticus isolated from bivalve shellfish farms along the southern coast of Korea[J].Environmental Science and Pollution Research, 2019, 26(20):21 034-21 043.
[2] JEAMSRIPONG S, KHANT W, CHUANCHUEN R.Distribution of phenotypic and genotypic antimicrobial resistance and virulence genes in Vibrio parahaemolyticus isolated from cultivated oysters and estuarine water[J].FEMS Microbiology Ecology, 2020, 96(8):fiaa081.
[3] Food and Agriculture Organization of the United Nations (FAO). Fishery and Aquaculture Statistics[EB/OL].[2022-04-07].https://www.fao.org/fishery/static/Yearbook/YB2019_USBcard/navigation/index_intro_e.htm.
[4] MOK J S, RYU A, KWON J Y, et al.Distribution of Vibrio species isolated from bivalves and bivalve culture environments along the Gyeongnam coast in Korea:Virulence and antimicrobial resistance of Vibrio parahaemolyticus isolates[J].Food Control, 2019, 106:106697.
[5] JIANG Y H, CHU Y B, XIE G S, et al.Antimicrobial resistance, virulence and genetic relationship of Vibrio parahaemolyticus in seafood from coasts of Bohai Sea and Yellow Sea, China[J].International Journal of Food Microbiology, 2019, 290:116-124.
[6] JIANG H, YU T, YANG Y T, et al. Co-occurrence of antibiotic and heavy metal resistance and sequence type diversity of Vibrio parahaemolyticus isolated from Penaeus vannamei at freshwater farms, seawater farms, and markets in Zhejiang province, China[J].Frontiers in Microbiology, 2020, 11:1294.
[7] ZAHER H A, NOFAL M I, HENDAM B M, et al.Prevalence and antibiogram of Vibrio parahaemolyticus and Aeromonas hydrophila in the flesh of Nile tilapia, with special reference to their virulence genes detected using multiplex PCR technique[J].Antibiotics(Basel,Switzerland), 2021, 10(6):654.
[8] YAN L, PEI X Y, ZHANG X L, et al.Occurrence of four pathogenic Vibrios in Chinese freshwater fish farms in 2016[J].Food Control, 2019, 95:85-89.
[9] BEJ A K, PATTERSON D P, BRASHER C W, et al.Detection of total and hemolysin-producing Vibrio parahaemolyticus in shellfish using multiplex PCR amplification of tl, tdh and trh[J].Journal of Microbiological Methods, 1999, 36(3):215-225.
[10] NARAYANAN S V, JOSEPH T C, PEERALIL S, et al.Tropical shrimp aquaculture farms harbour pathogenic Vibrio parahaemolyticus with high genetic diversity and Carbapenam resistance[J].Marine Pollution Bulletin, 2020, 160:111551.
[11] 郭书林, 陈信忠, 尤颖哲, 等.福建省南美白对虾幼体急性肝胰腺坏死病病原分离与PCR检测[J].中国动物检疫, 2019, 36(6):96-99.
GUO S L, CHEN X Z, YOU Y Z, et al.Pathogen isolation and PCR detection of AHPND in Penaeus vannamei larvae in Fujian Province[J].China Animal Health Inspection, 2019, 36(6):96-99.
[12] KRUMPERMAN P H.Multiple antibiotic resistance indexing of Escherichia coli to identify high-risk sources of fecal contamination of foods[J].Applied and Environmental Microbiology, 1983, 46(1):165-170.
[13] 谢腾飞. 中国食源性副溶血性弧菌遗传多样性分析与冷胁迫研究[D].广州:华南理工大学, 2018.
XIE T F.Study on genetic diversity and cold stress of foodborne Vibrio parahaemolyticus in China[D].Guangzhou:South China University, 2018.
[14] 洪锦春, 钟凌, 陈惠龙, 等.福建省2017—2018年淡水鱼养殖场嗜盐性弧菌污染状况分析[J].海峡预防医学杂志, 2019, 25(6):53-54.
HONG J C, ZHONG L, CHEN H L, et al. Analysis of pollution status of halophilic Vibrio in freshwater fish farms in Fujian Province from 2017 to 2018[J]. Strait Journal of Preventive Medicine, 2019, 25(6):53-54.
[15] 秦思, 乔昕, 沈赟, 等.2016年江苏省淡水养殖环节常见致病性弧菌污染状况调查[J].食品安全质量检测学报, 2019, 10(12):3 946-3 951.
QIN S, QIAO X, SHEN Y, et al.Contamination investigation of common pathogenic Vibrio species in freshwater aquaculture processing in Jiangsu Province in 2016[J].Journal of Food Safety & Quality, 2019, 10(12):3 946-3 951.
[16] HE Y, WANG S, ZHANG J P, et al.Integrative and conjugative elements-positive Vibrio parahaemolyticus isolated from aquaculture shrimp in Jiangsu, China[J].Frontiers in Microbiology, 2019, 10:1574.
[17] RAHMAN M S, CARRARO R, CARDAZZO B, et al.Molecular typing of Vibrio parahaemolyticus strains isolated from mollusks in the North Adriatic sea[J].Foodborne Pathogens and Disease, 2017, 14(8):454-464.
[18] SILVA I P, DE SOUZA CARNEIRO C, ALICE FONTES SARAIVA M, et al.Antimicrobial resistance and potential virulence of Vibrio parahaemolyticus isolated from water and bivalve mollusks from Bahia, Brazil[J].Marine Pollution Bulletin, 2018, 131:757-762.
[19] RODGERS C, PARVEEN S, CHIGBU P, et al.Prevalence of Vibrio parahaemolyticus, and Vibrio vulnificus in blue crabs (Callinectes sapidus), seawater and sediments of the Maryland Coastal Bays[J].Journal of Applied Microbiology, 2014, 117(4):1 198-1 209.
[20] 李艳, 张晓蕾, 李金平, 等.2013—2017年感染性腹泻的病原体特点和流行特征[J].中华医院感染学杂志, 2019, 29(11):1 732-1 736.
LI Y, ZHANG X L, LI J P, et al.Etiological and epidemiological characteristics of with infectious diarrhea in a hospital from 2013 to 2017[J].Chinese Journal of Nosocomiology, 2019, 29(11):1 732-1 736.
[21] 李平, 黄涵, 钟汶兵.海口市市售贝类海产品副溶血性弧菌污染调查及耐药和毒力基因研究[J].中国食品卫生杂志, 2019, 31(4):366-370.
LI P, HUANG H, ZHONG W B.Investigation of Vibrio parahaemolyticus contamination, drug resistance and virulence genes in shellfish products sold in Haikou[J].Chinese Journal of Food Hygiene, 2019, 31(4):366-370.
[22] CHEN X, ZHU Q Y, YU F, et al.Serology, virulence and molecular characteristics of Vibrio parahaemolyticus isolated from seafood in Zhejiang Province[J].PLoS One, 2018, 13(10):e0204892.
[23] 严伟, 沈月华, 徐德顺.2019年浙江省湖州市副溶血性弧菌病原学特征分析[J].中国食品卫生杂志, 2021, 33(1):74-78.
YAN W, SHEN Y H, XU D S.Etiologic characteristics of Vibrio parahaemolyticus stains in Huzhou of Zhejiang in 2019[J].Chinese Journal of Food Hygiene, 2021, 33(1):74-78.
[24] LALITHA M K, WALTER N M, JESUDASON M, et al.An outbreak of gastroenteritis due to Vibrio parahaemolyticus in Vellore[J].The Indian Journal of Medical Research, 1983, 78:611-615.
[25] 魏文娟. 养殖源弧菌耐药性及qnrVC基因介导喹诺酮药物耐药机制研究[D].上海:上海海洋大学,2020.
WEI W J.Antimicrobial resistance and molecular mechanism of qnrVC on fluoroquinolones resistance in Vibrio spp.isolated from mariculture[D].Shanghai:Shanghai Ocean University, 2020.
[26] 强鑫华, 蔡云祥.食源性腹泻患者分离副溶血性弧菌毒力基因及药敏分析[J].中国卫生检验杂志, 2021, 31(2):155-158.
QIANG X H, CAI Y X.Virulence and antimicrobial susceptibility of Vibrio parahaemolyticus in patients with foodborne diarrhea[J].Chinese Journal of Health Laboratory Technology, 2021, 31(2):155-158.
[27] XIE T F, WU Q P, ZHANG J M, et al.Comparison of Vibrio parahaemolyticus isolates from aquatic products and clinical by antibiotic susceptibility, virulence, and molecular characterisation[J].Food Control, 2017, 71:315-321.
[28] SU C L, CHEN L M.Virulence, resistance, and genetic diversity of Vibrio parahaemolyticus recovered from commonly consumed aquatic products in Shanghai, China[J].Marine Pollution Bulletin, 2020, 160:111554.
[29] XIE T F, YU Q, TANG X, et al.Prevalence, antibiotic susceptibility and characterization of Vibrio parahaemolyticus isolates in China[J].FEMS Microbiology Letters, 2020, 367(16):fnaa136.
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