Article(id=1153433636310340301, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433633999282214, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20241223001, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1734883200000, receivedDateStr=2024-12-23, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752929608657, onlineDateStr=2025-07-19, pubDate=1742832000000, pubDateStr=2025-03-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752929608657, onlineIssueDateStr=2025-07-19, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752929608657, creator=13701087609, updateTime=1752929608657, updator=13701087609, issue=Issue{id=1153433633999282214, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='6', pageStart='1', pageEnd='322', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1752929608105, creator=13701087609, updateTime=1758086445549, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1175062977960096080, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433633999282214, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1175062977960096081, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433633999282214, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=309, endPage=315, ext={EN=ArticleExt(id=1153433636763325143, articleId=1153433636310340301, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Analysis of pathogenic characteristics of Vibrio parahaemolyticus isolated in the markets and catering of Yangzhou in 2023, columnId=1151895321388347923, journalTitle=Journal of Food Safety & Quality, columnName=Food Analysis and Detection, runingTitle=null, highlight=null, articleAbstract=

Objective To investigate the prevalence, virulence phenotypes, and drug resistance phenotype of Vibrio parahaemolyticus in freshwater food, seafood, and ready-to-eat food in Yangzhou. Methods Freshwater food, seafood and ready-to-eat food samples were collected from 9 sampling points in Yangzhou markets for isolation and identification of Vibrio parahaemolyticus. The motility, hemolytic activity, and drug resistance phenotype of the isolates were analyzed. Results In 2023, a total of 289 samples were collected in the Yangzhou market, from which 63 strains of Vibrio parahaemolyticus were isolated, with an isolation rate of 21.80%. Among them, the isolation rate of freshwater food was 45.35% (39/86), seafood 28.75% (23/80), and ready-to-eat samples 0.81% (1/123), indicating a higher isolation rate in freshwater food compared to seafood in the markets of Yangzhou. The proportion of highly motility strains in freshwater food isolates was 30.77%, higher than that of the seafood isolates (17.39%). However, the proportion of strains with high hemolytic activity was 53.85% in freshwater food isolates, slightly lower than the 60.87% in seafood isolates. Antibiotic resistance profiling showed that all Vibrio parahaemolyticus isolates were resistant to ampicillin and cefazolin, while they were resistant to ceftazidime, tetracycline, ciprofloxacin, chloramphenicol, amikacin, meropenem and gentamicin. Conclusion This study highlights a high detection rate of Vibrio parahaemolyticus in Yangzhou markets and reveals differences in virulence and antibiotic resistance among isolates from different sources, offering valuable insights for monitoring and controlling the pathogen.

, correspAuthors=Dan GU, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Mei-Ling DU, Qiang-Qiang ZHU, Lei-Zhu GAO, Dan GU), CN=ArticleExt(id=1153433661010596422, articleId=1153433636310340301, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=2023年扬州市市场和餐饮中副溶血弧菌的病原特征分析, columnId=1151895321958773274, journalTitle=食品安全质量检测学报, columnName=食品分析与检测, runingTitle=null, highlight=null, articleAbstract=

目的 了解扬州市淡水产品、海产品及熟食餐饮中副溶血弧菌的流行情况、毒力表型和耐药表型。方法 在扬州市场9个采样点采集淡水产品、海产品和熟食样品, 进行副溶血弧菌分离鉴定, 并测定分离株的运动性、溶血活性和耐药表型。结果 2023年在扬州市共采集289份样品, 分离到63株副溶血弧菌, 总分离率为21.80%, 其中, 淡水产品分离率为45.35% (39/86), 海产品分离率为28.75% (23/80), 熟食样品分离率为0.81% (1/123), 表明扬州地区淡水产品分离率高于海水市场。淡水产品分离株中具有较强运动能力的菌株比例为30.77%, 高于海产品的17.39%; 而具有较强溶血活性的菌株比例在淡水产品为53.85%, 略低于海产品的60.87%。耐药表型的结果显示, 副溶血弧菌分离株对氨苄西林和头孢唑林的耐药率为100.00%, 对头孢他啶、四环素、环丙沙星、氯霉素、阿米卡星、美罗培南、庆大霉素7种抗生素均不耐药。结论 扬州市场副溶血弧菌检出率较高, 揭示了不同来源分离株在毒力和耐药性方面的差异, 为相关部门的监管和防治提供科学依据。

, correspAuthors=顾丹, authorNote=null, correspAuthorsNote=
* 顾丹(1988—), 女, 副教授, 主要研究方向为食源性病原菌流行传播规律。E-mail:
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杜美玲(1990—), 女, 主要研究方向为食源性病原菌流行传播规律。E-mail:

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杜美玲(1990—), 女, 主要研究方向为食源性病原菌流行传播规律。E-mail:

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3.扬州大学农业农村部农产品质量安全生物性危害因子(动物源)控制重点实验室, 扬州 225009
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2. Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China
3. Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture and Rural Affairs, Yangzhou University, Yangzhou 225009, China
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3.扬州大学农业农村部农产品质量安全生物性危害因子(动物源)控制重点实验室, 扬州 225009
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3. Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture and Rural Affairs, Yangzhou University, Yangzhou 225009, China
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Capital Journal of Public Health, 2020, 14(6): 285-290., articleTitle=Analysis on the etiological and molecular epidemiological characteristics of Vibrio parahaemolyticus in Beijing from 2010 to 2019, refAbstract=null), Reference(id=1175086852894765186, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433636310340301, doi=null, pmid=null, pmcid=null, year=2011, volume=26, issue=7, pageStart=539, pageEnd=542, url=null, language=null, rfNumber=[33], rfOrder=50, authorNames=傅慧琴, 苏靖华, 章红红, journalName=疾病监测, refType=null, unstructuredReference=傅慧琴, 苏靖华, 章红红. 318株副溶血性弧菌的血清分型和耐药性分析[J]. 疾病监测, 2011, 26(7): 539-542., articleTitle=318株副溶血性弧菌的血清分型和耐药性分析, refAbstract=null), Reference(id=1175086853012205699, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433636310340301, doi=null, pmid=null, pmcid=null, year=2011, volume=26, issue=7, pageStart=539, pageEnd=542, url=null, language=null, rfNumber=[33], rfOrder=51, authorNames=FU HQ, SU JH, ZHANG HH, journalName=Disease Surveillance, refType=null, unstructuredReference=FU HQ, SU JH, ZHANG HH. 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Drug resistance and molecular typing of Vibrio parahaemolyticus isolated from diarrhea patients in Songjiang District of Shanghai from 2016 to 2020[J]. Disease Surveillance, 2022, 37(2): 245-250., articleTitle=Drug resistance and molecular typing of Vibrio parahaemolyticus isolated from diarrhea patients in Songjiang District of Shanghai from 2016 to 2020, refAbstract=null)], funds=null, companyList=[AuthorCompany(id=1175086845097554938, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433636310340301, xref=null, ext=[AuthorCompanyExt(id=1175086845105943548, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433636310340301, companyId=1175086845097554938, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1. 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注: a. 低运动性(直径≤3 cm); b. 中等运动性(3 cm<直径≤6 cm); c. 高运动性(直径>6 cm)。

, figureFileSmall=Q5tzrD64x2V4srV3aGZQSQ==, figureFileBig=I3xOSUBlEq4yAp46aEJiww==, tableContent=null), ArticleFig(id=1175086847576387654, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433636310340301, language=EN, label=Fig.2, caption=Motility of Vibrio parahaemolyticus isolates from different sources, figureFileSmall=6lzBBC0WkKZVZk3oq3DIgg==, figureFileBig=EjxMnLmpL3/KEXNlgfZcsg==, tableContent=null), ArticleFig(id=1175086847639302215, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433636310340301, language=CN, label=图2, caption=不同来源副溶血弧菌分离株的运动性, figureFileSmall=6lzBBC0WkKZVZk3oq3DIgg==, figureFileBig=EjxMnLmpL3/KEXNlgfZcsg==, tableContent=null), ArticleFig(id=1175086847735771208, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433636310340301, language=EN, label=Fig.3, caption=Corresponding hemolytic bacterial circles of Vibrio parahaemolyticus with low, medium and high hemolysis, figureFileSmall=aCADRTAhtELmYs0iAhxREg==, figureFileBig=lqVCdhAcqABrVAxLeHuWHQ==, tableContent=null), ArticleFig(id=1175086847807074377, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433636310340301, language=CN, label=图3, caption=副溶血弧菌低、中、高溶血活性对应溶血菌圈

注: a. 低溶血活性(直径≤1.0 cm); b. 中等溶血活性(1.0 cm<直径≤1.3 cm); c. 高溶血活性(直径>1.3 cm)。

, figureFileSmall=aCADRTAhtELmYs0iAhxREg==, figureFileBig=lqVCdhAcqABrVAxLeHuWHQ==, tableContent=null), ArticleFig(id=1175086847878377546, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433636310340301, language=EN, label=Fig.4, caption=Hemolytic activity of Vibrio parahaemolyticus isolates from different sources, figureFileSmall=YjkMKCmmmA+RnOZ+HMBaOQ==, figureFileBig=AG1miAsHB3MY3/MeK0Eeww==, tableContent=null), ArticleFig(id=1175086847932903499, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433636310340301, language=CN, label=图4, caption=不同来源副溶血弧菌分离株的溶血活性, figureFileSmall=YjkMKCmmmA+RnOZ+HMBaOQ==, figureFileBig=AG1miAsHB3MY3/MeK0Eeww==, tableContent=null), ArticleFig(id=1175086848000012364, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433636310340301, language=EN, label=Table 1, caption=

Prevalence of Vibrio parahaemolyticus in marine products, freshwater aquatic products and catering in Yangzhou City

, figureFileSmall=null, figureFileBig=null, tableContent=
样品 海产品 淡水产品 熟食样品
数量 分离率/% 数量 分离率/% 数量 分离率/%
- - 24/42 57.14 - -
16/40 40.00 15/44 34.09 - -
7/40 17.50 - - - -
熟食 - - - - 1/123 0.81
总计 23/80 28.75 39/86 45.35 1/123 0.81
), ArticleFig(id=1175086848079704141, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433636310340301, language=CN, label=表1, caption=

扬州市场海产品和淡水产品以及熟食样品中副溶血弧菌的流行情况

, figureFileSmall=null, figureFileBig=null, tableContent=
样品 海产品 淡水产品 熟食样品
数量 分离率/% 数量 分离率/% 数量 分离率/%
- - 24/42 57.14 - -
16/40 40.00 15/44 34.09 - -
7/40 17.50 - - - -
熟食 - - - - 1/123 0.81
总计 23/80 28.75 39/86 45.35 1/123 0.81
), ArticleFig(id=1175086848163590222, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433636310340301, language=EN, label=Table 2, caption=

Resistance of Vibrio parahaemolyticus from different sources to different antibiotics

, figureFileSmall=null, figureFileBig=null, tableContent=
抗生素 稀释范围
/(mg/L)
MIC/(mg/L) 耐药菌株数/耐药率
S I R 海产品源
(n=23)
淡水产品源
(n=39)
熟食样品源
(n=1)
总计
(n=63)
数量 耐药率
/%
数量 耐药率
/%
数量 耐药率/% 数量 耐药率/%
氨苄西林 1~128 ≤8 16 ≥32 23 100.00 39 100.00 1 100.00 63 100.00
头孢唑林 0.5~128.0 ≤1 2 ≥4 23 100.00 39 100.00 1 100.00 63 100.00
四环素 0.25~128.00 ≤4 8 ≥16 0 0 0 0 0 0 0 0
环丙沙星 0.002~16.000 ≤1 2 ≥4 0 0 0 0 0 0 0 0
氯霉素 1~128 ≤8 16 ≥32 0 0 0 0 0 0 0 0
阿米卡星 0.25~256.00 ≤16 32 ≥64 0 0 0 0 0 0 0 0
美罗培南 0.004~32.000 ≤1 2 ≥4 0 0 0 0 0 0 0 0
庆大霉素 0.125~128.000 ≤4 8 ≥16 0 0 0 0 0 0 0 0
头孢他啶 0.5~256.0 ≤4 8 ≥16 0 0 0 0 0 0 0 0
), ArticleFig(id=1175086848285225039, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433636310340301, language=CN, label=表2, caption=

不同来源副溶血弧菌对不同抗生素的耐药性

, figureFileSmall=null, figureFileBig=null, tableContent=
抗生素 稀释范围
/(mg/L)
MIC/(mg/L) 耐药菌株数/耐药率
S I R 海产品源
(n=23)
淡水产品源
(n=39)
熟食样品源
(n=1)
总计
(n=63)
数量 耐药率
/%
数量 耐药率
/%
数量 耐药率/% 数量 耐药率/%
氨苄西林 1~128 ≤8 16 ≥32 23 100.00 39 100.00 1 100.00 63 100.00
头孢唑林 0.5~128.0 ≤1 2 ≥4 23 100.00 39 100.00 1 100.00 63 100.00
四环素 0.25~128.00 ≤4 8 ≥16 0 0 0 0 0 0 0 0
环丙沙星 0.002~16.000 ≤1 2 ≥4 0 0 0 0 0 0 0 0
氯霉素 1~128 ≤8 16 ≥32 0 0 0 0 0 0 0 0
阿米卡星 0.25~256.00 ≤16 32 ≥64 0 0 0 0 0 0 0 0
美罗培南 0.004~32.000 ≤1 2 ≥4 0 0 0 0 0 0 0 0
庆大霉素 0.125~128.000 ≤4 8 ≥16 0 0 0 0 0 0 0 0
头孢他啶 0.5~256.0 ≤4 8 ≥16 0 0 0 0 0 0 0 0
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2023年扬州市市场和餐饮中副溶血弧菌的病原特征分析
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杜美玲 1, 2, 3, 4 , 朱强强 1, 2, 3, 4 , 高镭铢 1, 2, 3, 4 , 顾丹 1, 2, 3, 4, *
食品安全质量检测学报 | 食品分析与检测 2025,16(6): 309-315
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食品安全质量检测学报 | 食品分析与检测 2025, 16(6): 309-315
2023年扬州市市场和餐饮中副溶血弧菌的病原特征分析
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杜美玲1, 2, 3, 4 , 朱强强1, 2, 3, 4, 高镭铢1, 2, 3, 4, 顾丹1, 2, 3, 4, *
作者信息
  • 1.扬州大学江苏省人兽共患病学重点实验室, 扬州 225009
  • 2.扬州大学江苏高校动物重要疫病与人兽共患病防控协同创新中心, 扬州 225009
  • 3.扬州大学农业农村部农产品质量安全生物性危害因子(动物源)控制重点实验室, 扬州 225009
  • 4.扬州大学教育部农业与农产品安全国际合作联合实验室, 扬州 225009
  • 杜美玲(1990—), 女, 主要研究方向为食源性病原菌流行传播规律。E-mail:

通讯作者:

* 顾丹(1988—), 女, 副教授, 主要研究方向为食源性病原菌流行传播规律。E-mail:
Analysis of pathogenic characteristics of Vibrio parahaemolyticus isolated in the markets and catering of Yangzhou in 2023
Mei-Ling DU1, 2, 3, 4 , Qiang-Qiang ZHU1, 2, 3, 4, Lei-Zhu GAO1, 2, 3, 4, Dan GU1, 2, 3, 4, *
Affiliations
  • 1. Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou 225009, China
  • 2. Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China
  • 3. Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture and Rural Affairs, Yangzhou University, Yangzhou 225009, China
  • 4. Joint International Research Laboratory of Agriculture and Agri-product Safety, Ministry of Education, Yangzhou University, Yangzhou 225009, China
出版时间: 2025-03-25 doi: 10.19812/j.cnki.jfsq11-5956/ts.20241223001
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目的 了解扬州市淡水产品、海产品及熟食餐饮中副溶血弧菌的流行情况、毒力表型和耐药表型。方法 在扬州市场9个采样点采集淡水产品、海产品和熟食样品, 进行副溶血弧菌分离鉴定, 并测定分离株的运动性、溶血活性和耐药表型。结果 2023年在扬州市共采集289份样品, 分离到63株副溶血弧菌, 总分离率为21.80%, 其中, 淡水产品分离率为45.35% (39/86), 海产品分离率为28.75% (23/80), 熟食样品分离率为0.81% (1/123), 表明扬州地区淡水产品分离率高于海水市场。淡水产品分离株中具有较强运动能力的菌株比例为30.77%, 高于海产品的17.39%; 而具有较强溶血活性的菌株比例在淡水产品为53.85%, 略低于海产品的60.87%。耐药表型的结果显示, 副溶血弧菌分离株对氨苄西林和头孢唑林的耐药率为100.00%, 对头孢他啶、四环素、环丙沙星、氯霉素、阿米卡星、美罗培南、庆大霉素7种抗生素均不耐药。结论 扬州市场副溶血弧菌检出率较高, 揭示了不同来源分离株在毒力和耐药性方面的差异, 为相关部门的监管和防治提供科学依据。

副溶血弧菌  /  分离鉴定  /  运动性  /  溶血活性  /  耐药表型

Objective To investigate the prevalence, virulence phenotypes, and drug resistance phenotype of Vibrio parahaemolyticus in freshwater food, seafood, and ready-to-eat food in Yangzhou. Methods Freshwater food, seafood and ready-to-eat food samples were collected from 9 sampling points in Yangzhou markets for isolation and identification of Vibrio parahaemolyticus. The motility, hemolytic activity, and drug resistance phenotype of the isolates were analyzed. Results In 2023, a total of 289 samples were collected in the Yangzhou market, from which 63 strains of Vibrio parahaemolyticus were isolated, with an isolation rate of 21.80%. Among them, the isolation rate of freshwater food was 45.35% (39/86), seafood 28.75% (23/80), and ready-to-eat samples 0.81% (1/123), indicating a higher isolation rate in freshwater food compared to seafood in the markets of Yangzhou. The proportion of highly motility strains in freshwater food isolates was 30.77%, higher than that of the seafood isolates (17.39%). However, the proportion of strains with high hemolytic activity was 53.85% in freshwater food isolates, slightly lower than the 60.87% in seafood isolates. Antibiotic resistance profiling showed that all Vibrio parahaemolyticus isolates were resistant to ampicillin and cefazolin, while they were resistant to ceftazidime, tetracycline, ciprofloxacin, chloramphenicol, amikacin, meropenem and gentamicin. Conclusion This study highlights a high detection rate of Vibrio parahaemolyticus in Yangzhou markets and reveals differences in virulence and antibiotic resistance among isolates from different sources, offering valuable insights for monitoring and controlling the pathogen.

Vibrio parahaemolyticus  /  isolation and identification  /  motility  /  hemolytic activity  /  drug resistance phenotype
杜美玲, 朱强强, 高镭铢, 顾丹. 2023年扬州市市场和餐饮中副溶血弧菌的病原特征分析. 食品安全质量检测学报, 2025 , 16 (6) : 309 -315 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241223001
Mei-Ling DU, Qiang-Qiang ZHU, Lei-Zhu GAO, Dan GU. Analysis of pathogenic characteristics of Vibrio parahaemolyticus isolated in the markets and catering of Yangzhou in 2023[J]. Journal of Food Safety & Quality, 2025 , 16 (6) : 309 -315 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241223001
副溶血弧菌(Vibrio parahaemolyticus)是一种革兰氏阴性嗜盐菌, 在适宜环境中生长迅速, 繁殖周期仅为8~9 min, 广泛分布于海洋底部以及寄居在鱼、虾、蟹类以及软体动物的体表。食用未煮熟或者污染副溶血弧菌的海产品可能会引发急性肠胃炎[1], 并伴有呕吐、恶心、腹泻和发烧等症状, 严重时还可以通过外露伤口引起败血症, 危及生命健康[2]。根据美国食品和药物管理局的统计, 每年因副溶血弧菌感染引发的胃肠炎病例约为4500例, 已成为美国乃至全球食用海鲜相关疾病的主要原因之一[35]。在我国, 2010至2020年期间, 共报告由副溶血弧菌引起的食源性疾病暴发事件1772起[6], 占细菌性食源性事件的50%以上, 已成为我国主要的食源性病原微生物之一[78]。此外, 副溶血弧菌也对水产动物的养殖造成严重危害, 可以导致凡纳滨虾红体病, 死亡率高达80%[9]; 引发南美对虾的急性肝胰腺坏死病, 可造成虾产量下降60%[10]; 还可引起棘皮动物和贝类等水产养殖物种的烂胃病、化板病等综合疾病[11]。作为全球范围内最常见的胃肠道感染病原菌之一, 副溶血弧菌主要通过被污染的海产品传播给人群[1214]。2019年, AMALINA等[15]马来西亚养殖石斑鱼中检测出副溶血弧菌, 检出率为25%。2024年, 范秋华等[16]在中国深圳采集海水样品314份, 分离出82株副溶血弧菌, 检出率为26.1%。这些流行病学监测结果表明, 副溶血弧菌已经成为国内外威胁人类生命健康和财产安全的重要病原菌。
副溶血弧菌在全球范围内的感染率持续居高不下, 这不仅与其强大的生存和繁殖能力有关, 还与防治过程中滥用抗生素导致的耐药性问题密切相关。目前, 用于防治副溶血弧菌的常用抗生素包括氨苄西林、头孢唑林、四环素、环丙沙星、庆大霉素等。然而, 随着抗生素的广泛使用, 副溶血弧菌的耐药性也逐渐增强。2021年, 墨西哥LVAREZ-CONTRERAS等[17]在不同种类的海鲜样品中发现, 大多数副溶血弧菌分离株对氨苄西林、头孢菌素类抗生素耐药。2024年, 范秋华等[16]在中国深圳分离出82株副溶血弧菌, 对青霉素类药物耐药率为100%, 氨苄西林耐药率为98.8%。李明珠[18]在江苏部分地区共采集了137份海水样品和159份淡水样品, 分离出副溶血弧菌49株和89株, 发现海水源和淡水源分离株对氨苄西林的耐药率为40.8%和61.8%, 对头孢唑林的耐药率为100.0%和96.6%。这些研究表明, 副溶血弧菌的耐药问题日益严重, 需要加强对副溶血弧菌流行情况和耐药性监测, 为科学有效防控副溶血弧菌提供理论依据。
扬州市位于江苏省中部, 地处长江和京杭大运河交汇处, 水产品除了上海、南通、盐城等沿海城市供应, 其地域内高邮湖、宝应湖、邵伯湖等水域也是淡水养殖的重要基地。副溶血弧菌是一种重要的人兽共患食源性病原菌, 其感染而导致食物中毒的事件时有发生。而部分副溶血弧菌分离株能够迅速适应不同的生长环境, 获得生长优势, 这也可能是导致副溶血弧菌感染的原因之一[19]。2019年扬州市食源性疾病监测结果显示, 副溶血弧菌是扬州地区主要的食源性致病微生物之一[20]。为了进一步探究副溶血弧菌在扬州市海产品、淡水产品和餐饮中的流行情况, 本研究于2023年夏秋两季对扬州市区9处菜场共289份样品进行收集处理, 分离副溶血弧菌, 并对其毒力表型和耐药表型进行测定, 为该地区副溶血弧菌的监测和防控提供科学理论依据。
2023年, 在扬州海水市场、淡水市场和餐饮市场共采集289份样品, 其中海产品80份, 淡水产品86份, 熟食样品123份。按照GB 4789.7—2013《食品安全国家标准 食品微生物学检验 副溶血性弧菌》方法, 分离得到63株副溶血弧菌菌株。以副溶血弧菌标准株RIMD2210633和大肠埃希菌ATCC25922作为实验质控菌株。
T100TMThermal Cycler聚合酶链式反应(polymerase chain reaction, PCR)仪、Gel Doc TM XR+全自动数码凝胶图像成像仪(美国BIORAD公司); DK-600电热恒温水槽(上海精宏实验仪器设备有限公司); Centrifuge 5425小型离心机、Biophotometer Centrifuge 5425分光光度计、ThermoMixer C金属浴(德国Eppendorf公司); ZQZY-88BE震荡培养箱(上海知楚仪器有限公司); SPX-300BSH-II生化培养箱(上海新苗医疗器械制造有限公司); YT3004电子天平(精度0.1 mg, 昆山优科维特电子科技有限公司); FR-250电泳仪(上海复日科技有限公司)。
血琼脂平板、氯化钠结晶紫增菌液、MH液体培养基(青岛高科技工业园海博生物技术有限公司); NaCl、丙三醇(分析纯, 国药集团化学试剂有限公司); 蛋白胨、酵母提取物(英国OXOID公司); 琼脂粉、琼脂糖[生工生物工程(上海)股份有限公司]; 硫代硫酸盐柠檬酸盐胆盐蔗糖(thiosulfate citrate bile salts sucrose agar culture medium, TCBS)琼脂培养基、MH琼脂培养基(广东环凯微生物科技有限公司); 科玛嘉弧菌显色平板(上海科玛嘉微生物技术有限公司); 2x Taq Master mix(南京诺唯赞生物科技有限公司); DL2000 Marker[宝生物(大连)有限公司]; tlh引物(北京擎科生物科技有限公司); 绿如蓝核酸染料(北京大学生命科学华东产业研究院生物试剂中心); 抗生素: 氨苄西林(纯度90.0%)、头孢唑林(纯度95.0%)、四环素(效价>900 µg/mg)、环丙沙星(纯度86.5%)、氯霉素(效价> 900 µg/mg)、阿米卡星(纯度97.0%)、美罗培南(纯度98.0%)、庆大霉素(效价> 590 IU/mg)、头孢他啶(纯度99.0%)(中国食品药品检定所)。
LB肉汤培养基: 30 g NaCl, 10 g蛋白胨, 5 g酵母提取物, 蒸馏水定容至1000 mL。
LB琼脂培养基: 30 gNaCl, 10 g蛋白胨, 5 g酵母提取物, 15 g琼脂粉, 蒸馏水定容至1000 mL。
LB半固体培养基: 30 g NaCl, 10 g蛋白胨, 5 g酵母提取物, 3 g琼脂粉, 蒸馏水定容至1000 mL。
样品采集后用冰袋保温箱保存, 2 h内运至实验室处理。在无菌条件下将鱼类、虾类、贝类、熟食样品处理后放入50 mL氯化钠结晶紫增菌液中富集, 37 ℃、180 r/min孵育18~24 h。用无菌接种环蘸取增菌液在TCBS平板上三区划线, 置于37 ℃培养箱中培养24 h。用无菌接种环挑取典型的绿色单菌落, 在科玛嘉弧菌显色平板三区划线, 置于37 ℃培养箱中培养24 h。用无菌接种环挑取典型紫色单菌落在LB固体琼脂培养基上三区划线, 置于37 ℃培养箱中培养24 h。用无菌接种环在LB固体琼脂培养基上挑取单菌落, 接入至3 mL LB液体培养基内。37 ℃、180 r/min孵育3 h, 吸取100 μL菌悬液到EP管中, 100 ℃金属浴加热10 min, 细胞破裂释放出DNA。用副溶血弧菌特异性基因tlh作为靶向基因, 使用tlh-F (5’-AAAGCGGATTATG CAGAAGCACTG-3’)、tlh-R (5’-GCTACTTTCTAGCATTT TCTCTGC)的引物序列, 通过PCR扩增tlh基因, 以副溶血弧菌标准菌株RIMD2210633为阳性对照, 进行鉴定。
琼脂糖凝胶电泳鉴定PCR产物: 配制1.0%的琼脂糖凝胶, 加热煮沸稍作冷却后加入绿如蓝染料, 吸取5 μL PCR产物在凝胶点样孔处点样, 以Marker DL2000作基因大小参考, 100 V电泳45 min, 当溴酚蓝移动到胶2/3位置时, 断开电源, 在凝胶成像仪上观察结果。扩增条带大小与阳性对照管一致, 判定为阳性管, 每个产物重复一次。在LB固体平板上挑取对应阳性单菌落, 接入LB液体试管中扩大培养, 37 ℃ 180 r/min培养12 h, 分别吸取800 µL的菌液和800 µL 50%甘油, 混匀于冻存管中, 在‒70 ℃超低温冰箱中保藏菌株。
(1)运动性实验
对63株不同来源的副溶血弧菌分离株进行运动性实验。取过夜培养的副溶血弧菌菌液30 µL稀释到3 mL新鲜的LB液体培养基中, 培养至OD600约为1.0。在无菌条件下用移液枪吸取5 μL菌液滴入含有0.3%琼脂的LB半固培养基平板正中心, 平板正置于培养箱中37 ℃孵育7 h。观察菌株生长情况并测量菌圈直径。每个菌株重复3次, 数值以3次结果的平均值±标准偏差表示。
(2)溶血性活性测定
对63株不同来源的副溶血弧菌分离株进行溶血活性实验。取过夜培养的副溶血弧菌菌液3 µL稀释到3 mL新鲜的LB液体培养基中, 培养至OD600约为1.0。将5 μL菌悬液滴于血琼脂平板(5%体积的兔血红细胞)上, 倒置于培养基中37 ℃培养30 h。在透射光条件下观察溶血菌圈的大小, 测定溶血圈直径。每个菌株重复3次, 数值以3次结果的平均值±标准偏差表示。
根据CLSI 2014标准, 用大肠杆菌ATCC 25922作为标准质控菌株, 在LB固体培养基上进行副溶血弧菌菌株复苏[21]。用无菌接种环选取单个菌落, 接种到无菌MH液体培养基中, 在37 ℃、180 r/min的条件下孵育4 h, 培养至OD600值约为0.6。将9种抗生素氨苄西林、头孢唑林、四环素、环丙沙星、氯霉素、阿米卡星、美罗培南、庆大霉素、头孢他啶配制成溶液, 进行逐级稀释, 每个稀释步骤中, 在空白平板上加入1 mL的无菌水, 从最高浓度的抗生素溶液开始, 取1 mL混合后, 转移到下一个平板中, 如此重复稀释。在稀释平板的顶部和底部, 保留两个只含1 mL无菌水的空白对照平板。向每个平板中加入19 mL的MH培养基, 混匀并冷却至凝固。取3 µL的菌液, 稀释至297 µL的磷酸盐缓冲液中, 按1:100的比例稀释, 充分混匀。从低药物浓度开始, 取稀释后的菌液5 µL依次点印到准备好的MH平板上。培养皿置于37 ℃培养箱中培养18 h, 记录不同菌株菌落生长情况。
本研究采用Excel 2021进行数据统计处理和图片绘制。
从扬州市不同市场共采集样品289份, 其中海产品80份, 淡水产品86份, 熟食样品123份, 共分离到副溶血弧菌63株, 总分离率为21.80%(表1)。如表1所示, 其中海产品中副溶血弧菌的分离率为28.75% (23/80), 淡水产品为45.35% (39/86), 熟食样品为0.81% (1/123)。结果表明海产品、淡水产品和熟食样品中均能分离出副溶血弧菌。其中, 海产品和淡水产品中的副溶血弧菌流行水平较高, 而餐饮市场中相关样品的感染程度较低。值得指出的是淡水产品中副溶血弧菌分离率(45.35%)高于海产品(28.75%), 与朱强强[22]研究副溶血弧菌在淡水产品分离率高于海产品的结论一致, 与近几年研究的副溶血弧菌在内陆地区淡水产品中带菌率普遍升高结果一致[2324]。其中, 值得注意的是淡水产品中鱼的分离率是57.14%, 远远高出陈坤才等[25]监测到广东地区淡水鱼样品副溶血性弧菌检出率为11.35%, 由此推测扬州市场淡水鱼养殖-运输-销售等多个环节副溶血弧菌污染严重, 相关部门要重视和加强对淡水鱼的监管。
对海产品、淡水产品和熟食样品共63株副溶血弧菌进行运动性实验。如图1所示, 菌落直径≤3 cm为低运动性; 3 cm<菌落直径≤6 cm为中等运动性; 菌落直径>6 cm为高运动性。结果显示, 海产品中分离出的副溶血弧菌低运动性菌株比例为17.39%、中等运动性为65.22%、高运动性为17.39%。淡水产品中分离出的副溶血弧菌低运动性菌株比例为5.13%、中等运动性为64.10%, 而高运动性增加到了30.77%。熟食样品中分离的菌株为低运动性(图2)。运动性结果表明, 海产品源副溶血弧菌运动性强的菌株比例低于淡水产品源菌株, 这一结果与李明珠[18]的研究结果一致, 表明淡水来源的副溶血弧菌具有更强的运动性。
对63株不同来源的副溶血弧菌菌株进行溶血活性测定。如图3所示, 溶血圈直径≤1.0 cm为低溶血活性; 1.1 cm<溶血圈直径≤1.3 cm为中等溶血活性; 溶血圈直径>1.3 cm为高溶血活性。统计结果显示, 海产品菌株中等溶血活性比例为34.78%, 高溶血活性为60.87%; 淡水产品菌株中等溶血活性为46.15%, 高溶血活性为53.85%(图4)。熟食样品中分离的菌株具有高溶血活性。海产品源中副溶血弧菌溶血活性高的菌株比例高于淡水产品源。值得注意的是, 在研究过程中发现, 海产品源中运动性高的菌株其溶血活性也高, 表明海产品源的副溶血弧菌分离株的运动性和溶血活性呈正相关性。在淡水产品源中, 12株运动性强菌株中有6株溶血活性也高, 占高运动性菌株的50.00% (6/12)。
对63株副溶血弧菌分离株进行耐药表型测定, 如表2所示, 所有菌株对氨苄西林和头孢唑林耐药率为100.00%。所有菌株对头孢他啶、四环素、环丙沙星、氯霉素、阿米卡星、美罗培南、庆大霉素7种抗生素均不耐药。近年来, 随着交通和物流快速发展, 使得副溶血弧菌在全国范围内快速传播, 已经成为我国沿海城市细菌性食物中毒的首要病原菌[26]。目前, 抗生素仍是防治副溶血弧菌的主要手段, 但这也导致了副溶血弧菌对抗生素耐药性的增强, 其感染也呈现上升趋势[2729]。本研究发现扬州市分离的63株副溶血弧菌对氨苄西林全部耐药, 这与深圳、北京、上海等地的分离株对氨苄西林的耐药率均超过95.0%的结果一致[3033]。此外, 俞佳莉等[34]研究显示, 上海松江区腹泻患者的副溶血弧菌分离株对头孢唑林耐药率为56.25%, 而本次分离的63株菌株对头孢唑林的耐药率高达100.00%。这一现象可能与头孢唑林等抗生素的大量使用有关, 导致耐药菌株不断产生。
对扬州市场副溶血弧菌流行情况的调查结果显示, 海产品菌株的分离率是28.75%, 淡水产品菌株的分离率是45.35%, 淡水市场副溶血弧菌的污染程度远高于海水市场。熟食样品的分离率为0.81%, 说明目前扬州市场熟食中副溶血弧菌污染率处于较低水平。海产品源副溶血弧菌运动性强的菌株比例为17.39%低于淡水产品源的30.77%。海产品源和淡水产品源中副溶血弧菌溶血性高的菌株比例分别为60.87%和53.85%。这些结果表明淡水产品源副溶血弧菌分离株具有较强的毒力, 需要加强对淡水产品中副溶血弧菌的监测, 并评估其致病能力。耐药表型测定结果表明所有菌株对氨苄西林和头孢唑林耐药率为100.00%, 需要加强对细菌耐药性的持续监测, 为有效遏制耐药菌株的进一步扩散, 相关部门需科学引导抗生素的合理使用, 从而保障公共卫生安全。
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2025年第16卷第6期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20241223001
  • 接收时间:2024-12-23
  • 首发时间:2025-07-19
  • 出版时间:2025-03-25
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  • 收稿日期:2024-12-23
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    1.扬州大学江苏省人兽共患病学重点实验室, 扬州 225009
    2.扬州大学江苏高校动物重要疫病与人兽共患病防控协同创新中心, 扬州 225009
    3.扬州大学农业农村部农产品质量安全生物性危害因子(动物源)控制重点实验室, 扬州 225009
    4.扬州大学教育部农业与农产品安全国际合作联合实验室, 扬州 225009

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* 顾丹(1988—), 女, 副教授, 主要研究方向为食源性病原菌流行传播规律。E-mail:
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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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