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Article(id=1154123920300036919, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986579971429187, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20241109001, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1731081600000, receivedDateStr=2024-11-09, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1753094185185, onlineDateStr=2025-07-21, pubDate=1740412800000, pubDateStr=2025-02-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1753094185185, onlineIssueDateStr=2025-07-21, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1753094185185, creator=13701087609, updateTime=1753094185185, updator=13701087609, issue=Issue{id=1153986579971429187, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='4', pageStart='1', pageEnd='320', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1753061440699, creator=13701087609, updateTime=1758783495950, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1177986619249406427, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986579971429187, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1177986619249406428, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986579971429187, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=315, endPage=320, ext={EN=ArticleExt(id=1154123920757216056, articleId=1154123920300036919, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Study on the contamination status of Vibrio parahaemolyticus in the production chain of Liuzhou river snails rice noodles, columnId=1153433739154678309, journalTitle=Journal of Food Safety & Quality, columnName=Food Safety Supervision and Management, runingTitle=null, highlight=null, articleAbstract=

Objective To investigate the contamination level of Vibrio parahaemolyticus (VP) in the production chain of the Liuzhou river snails rice noodles, promptly assess the risk of VP in the production chain. Methods From 2023 to 2024, a total of 487 simlpes, including snails and snail meat, semi-finished soup packets, and pre-packed Liuzhou river snails rice noodles final products, were randomly collected from Liuzhou agricultural markets and manufacturing enterprises in Liuzhou. A conbination of culture-based methods and real-time fluorescent polymerase chain reaction (PCR) was used to detect VP. Results Among 70 snails samples from agricultural market with a detection rate of 38.6% and an average contamination level of 16.1 MPN/g in 44 snails samples. In the 41 snails and snail meat sample from manufacturing enterprises showing the detection rate of 2.4%; no VP was detected in the semi-finished soup package or final product. Conclusion Although VP contamination is relatively common in snails at the retail trade, the contamination rate of snails and snail meat in the enterprise is low. Furthermore, the repeated boiling in Liuzhou river snails rice noodles production can effectively control VP. Therefore, the risk of VP contamination in the Liuzhou river snails rice noodles production chain is relatively low.

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目的 调查预包装柳州螺蛳粉生产链中副溶血性弧菌的污染水平, 及时了解揭示生产链中污染副溶血性弧菌(Vibrio parahaemolyticus, VP)风险大小。方法 2023—2024年采用随机的方式收集柳州市农贸市场和柳州螺蛳粉生产企业的螺蛳及螺肉、半成品汤料包和预包装螺蛳粉终产品共487份, 利用培养法结合实时荧光聚合酶链式反应(polymerase chain reaction, PCR)法对VP进行精准检测。结果 农贸市场的70份螺蛳中VP的检出率为38.6%, 对其中的44份进行定量检测, 平均污染量为16.1 MPN/g; 生产企业的41份螺蛳及螺肉中VP的检出率为2.4%; 半成品汤料包和终产品中均未检出VP。结论 尽管零售端螺蛳中VP污染较为普遍, 但在生产端的螺蛳及螺肉中VP污染率较低。此外, 螺蛳粉生产工艺中的熬煮过程也能有效杀灭VP。总体而言, 螺蛳粉生产链中污染VP的风险较小。

, correspAuthors=刘兴, authorNote=null, correspAuthorsNote=
* 刘兴(1972—), 男, 高级工程师, 主要研究方向为食品质量与安全。E-mail:
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谢雨龙(1989—), 男, 硕士, 高级工程师, 主要研究方向为食品药品微生物检测。E-mail:

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谢雨龙(1989—), 男, 硕士, 高级工程师, 主要研究方向为食品药品微生物检测。E-mail:

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谢雨龙(1989—), 男, 硕士, 高级工程师, 主要研究方向为食品药品微生物检测。E-mail:

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Liuzhou Quality Inspection and Testing Research Center, Liuzhou 545500, China
2. Liuzhou River Snails Rice Noodles Quality Inspection Center, Liuzhou 545500, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1177985455590093054, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, authorId=1177985455351017720, language=CN, stringName=刘兴, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=1, 2, *, address=1.柳州市质量检验检测研究中心, 柳州 545500
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Food Science, 2013, 34(21): 227-232., articleTitle=Sublethal thermal injury and resuscitation conditions of Vibrio parahaemolyticus, refAbstract=null), Reference(id=1177985464096141786, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, doi=null, pmid=null, pmcid=null, year=2020, volume=29, issue=3, pageStart=420, pageEnd=428, url=null, language=null, rfNumber=[30], rfOrder=54, authorNames=俞文英, 张昭寰, 钱慧, journalName=上海海洋大学学报, refType=null, unstructuredReference=俞文英, 张昭寰, 钱慧, 等. 巴氏消毒及冷藏温度作用下副溶血弧菌菌株失活异质性的比较[J]. 上海海洋大学学报, 2020, 29(3): 420-428., articleTitle=巴氏消毒及冷藏温度作用下副溶血弧菌菌株失活异质性的比较, refAbstract=null), Reference(id=1177985464184222174, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, doi=null, pmid=null, pmcid=null, year=2020, volume=29, issue=3, pageStart=420, pageEnd=428, url=null, language=null, rfNumber=[30], rfOrder=55, authorNames=YU WY, ZHANG SH, QIAN H, journalName=Journal of Shanghai Ocean University, refType=null, unstructuredReference=YU WY, ZHANG SH, QIAN H, et al. Comparison of inactivation heterogeneity of Vibrio parahaemolyticus strains under pasteurization and cold storage temperature[J]. Journal of Shanghai Ocean University, 2020, 29(3): 420-428., articleTitle=Comparison of inactivation heterogeneity of Vibrio parahaemolyticus strains under pasteurization and cold storage temperature, refAbstract=null)], funds=[Fund(id=1177985457934709054, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, awardId=2024SZ0505G004, language=CN, fundingSource=柳州市科技攻关计划项目(2024SZ0505G004), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1177985453220311225, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, xref=null, ext=[AuthorCompanyExt(id=1177985453232894138, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, companyId=1177985453220311225, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1. 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Liuzhou River Snails Rice Noodles Quality Inspection Center, Liuzhou 545500, China), AuthorCompanyExt(id=1177985453329363135, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, companyId=1177985453316780221, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.柳州螺蛳粉质量检验中心, 柳州 545500)])], figs=[ArticleFig(id=1177985456714166556, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, language=EN, label=Fig.1, caption=Partial real-time PCR results of Vibrio parahaemolyticus in snails sold in Liuzhou, figureFileSmall=EZufRA6qseOvqhnXNvd4Pg==, figureFileBig=IM3oMDZDT78Ni7K8jPr9Ag==, tableContent=null), ArticleFig(id=1177985456810635553, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, language=CN, label=图1, caption=柳州市市售螺蛳副溶血性弧菌实时荧光PCR部分检测结果, figureFileSmall=EZufRA6qseOvqhnXNvd4Pg==, figureFileBig=IM3oMDZDT78Ni7K8jPr9Ag==, tableContent=null), ArticleFig(id=1177985456869355812, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, language=EN, label=Fig.2, caption=Real-time PCR results of Vibrio parahaemolyticus in snails, snail meat of enterprises, figureFileSmall=mjaIO5Gf9TN9yUavhFZBHA==, figureFileBig=WWwzLUCmWy8/l89L/46ZDA==, tableContent=null), ArticleFig(id=1177985456999379239, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, language=CN, label=图2, caption=企业中螺蛳及螺肉副溶血性弧菌的实时荧光PCR检测结果, figureFileSmall=mjaIO5Gf9TN9yUavhFZBHA==, figureFileBig=WWwzLUCmWy8/l89L/46ZDA==, tableContent=null), ArticleFig(id=1177985457188122927, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, language=EN, label=Table 1, caption=

Results of Vibrio parahaemolyticus in snail sold in Liuzhou

, figureFileSmall=null, figureFileBig=null, tableContent=
区域 样品 数量
/份		 定性阳性
结果/份		 总平均值
污染率/%		 平均污染量
(2024年)
/(MPN/g)
鱼峰区 田螺 / / 31.3 1.5
石螺 16 5
柳南区 田螺 7 4 56.3 14.0
石螺 9 5
柳北区 田螺 3 1 33.3 6.9
石螺 3 1
阳和新区 田螺 2 2 71.4 66.3
石螺 5 3
城中区 田螺 / / 50.0 1.5
石螺 2 1
柳江区 田螺 4 2 21.7 6.5
石螺 19 3
), ArticleFig(id=1177985457334923570, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, language=CN, label=表1, caption=

柳州市市售螺蛳副溶血性弧菌检测结果

, figureFileSmall=null, figureFileBig=null, tableContent=
区域 样品 数量
/份		 定性阳性
结果/份		 总平均值
污染率/%		 平均污染量
(2024年)
/(MPN/g)
鱼峰区 田螺 / / 31.3 1.5
石螺 16 5
柳南区 田螺 7 4 56.3 14.0
石螺 9 5
柳北区 田螺 3 1 33.3 6.9
石螺 3 1
阳和新区 田螺 2 2 71.4 66.3
石螺 5 3
城中区 田螺 / / 50.0 1.5
石螺 2 1
柳江区 田螺 4 2 21.7 6.5
石螺 19 3
), ArticleFig(id=1177985457435586868, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, language=EN, label=Table 2, caption=

Test results of Vibrio parahaemolyticus in snails, snail meat and semi-finished soup of enterprises

, figureFileSmall=null, figureFileBig=null, tableContent=
企业 样品 样品数量/份 定性检测阳性结果/份 定量检测结果/(MPN/g)
企业1 螺蛳及螺肉 5 0 /
汤料包(未灭菌) 15 0 /
企业2 螺蛳及螺肉 5 0 /
汤料包(未灭菌) 10 0 /
企业3 螺蛳及螺肉 9 0 /
汤料包(未灭菌) 6 0 /
企业4 螺蛳及螺肉 9 0 /
汤料包(未灭菌) 5 0 /
企业5 螺肉 2 0 /
企业6 螺蛳 6 0 /
汤料包(未灭菌) 10 0 /
企业7 螺蛳 3 0 /
汤料包(未灭菌) 10 0 /
企业8 螺蛳 2 1 1.5
汤料包(未灭菌) 5 0 /
企业9 汤料包(未灭菌) 5 0 /
), ArticleFig(id=1177985457582387510, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, language=CN, label=表2, caption=

企业中螺蛳和螺肉及半成品汤料副溶血性弧菌检测结果

, figureFileSmall=null, figureFileBig=null, tableContent=
企业 样品 样品数量/份 定性检测阳性结果/份 定量检测结果/(MPN/g)
企业1 螺蛳及螺肉 5 0 /
汤料包(未灭菌) 15 0 /
企业2 螺蛳及螺肉 5 0 /
汤料包(未灭菌) 10 0 /
企业3 螺蛳及螺肉 9 0 /
汤料包(未灭菌) 6 0 /
企业4 螺蛳及螺肉 9 0 /
汤料包(未灭菌) 5 0 /
企业5 螺肉 2 0 /
企业6 螺蛳 6 0 /
汤料包(未灭菌) 10 0 /
企业7 螺蛳 3 0 /
汤料包(未灭菌) 10 0 /
企业8 螺蛳 2 1 1.5
汤料包(未灭菌) 5 0 /
企业9 汤料包(未灭菌) 5 0 /
), ArticleFig(id=1177985457666273591, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, language=EN, label=Table 3, caption=

Detection results of Vibrio parahaemolyticus in enterprise final products

, figureFileSmall=null, figureFileBig=null, tableContent=
品牌 生产企业 样品 样品数量
/份		 定性检测阳性结果/份
品牌1 企业8 预包装
螺蛳粉		 65 0
品牌2 企业1 70 0
品牌3 企业2 45 0
品牌4 企业3 35 0
品牌5 企业4 40 0
品牌6 企业6 35 0
品牌7 企业9 20 0
), ArticleFig(id=1177985457766936891, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1154123920300036919, language=CN, label=表3, caption=

企业终产品中副溶血性弧菌的检测结果

, figureFileSmall=null, figureFileBig=null, tableContent=
品牌 生产企业 样品 样品数量
/份		 定性检测阳性结果/份
品牌1 企业8 预包装
螺蛳粉		 65 0
品牌2 企业1 70 0
品牌3 企业2 45 0
品牌4 企业3 35 0
品牌5 企业4 40 0
品牌6 企业6 35 0
品牌7 企业9 20 0
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柳州螺蛳粉生产链中副溶血性弧菌的污染状况研究
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谢雨龙 1, 2 , 覃巧思 1 , 鄢航 1 , 覃海姣 1 , 吴燮昳 1 , 兰盈 1 , 刘兴 1, 2, *
食品安全质量检测学报 | 食品安全监管 2025,16(4): 315-320
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食品安全质量检测学报 | 食品安全监管 2025, 16(4): 315-320
柳州螺蛳粉生产链中副溶血性弧菌的污染状况研究
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谢雨龙1, 2 , 覃巧思1, 鄢航1, 覃海姣1, 吴燮昳1, 兰盈1, 刘兴1, 2, *
作者信息
  • 1.柳州市质量检验检测研究中心, 柳州 545500
  • 2.柳州螺蛳粉质量检验中心, 柳州 545500

    • 谢雨龙(1989—), 男, 硕士, 高级工程师, 主要研究方向为食品药品微生物检测。E-mail:

通讯作者:

* 刘兴(1972—), 男, 高级工程师, 主要研究方向为食品质量与安全。E-mail:
Study on the contamination status of Vibrio parahaemolyticus in the production chain of Liuzhou river snails rice noodles
Yu-Long XIE1, 2 , Qiao-Si QIN1, Hang YAN1, Hai-Jiao QIN1, Xie-Yi WU1, Ying LAN1, LIU-Xing1, 2, *
Affiliations
  • 1. Liuzhou Quality Inspection and Testing Research Center, Liuzhou 545500, China
  • 2. Liuzhou River Snails Rice Noodles Quality Inspection Center, Liuzhou 545500, China
出版时间: 2025-02-25 doi: 10.19812/j.cnki.jfsq11-5956/ts.20241109001
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摘要
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目的 调查预包装柳州螺蛳粉生产链中副溶血性弧菌的污染水平, 及时了解揭示生产链中污染副溶血性弧菌(Vibrio parahaemolyticus, VP)风险大小。方法 2023—2024年采用随机的方式收集柳州市农贸市场和柳州螺蛳粉生产企业的螺蛳及螺肉、半成品汤料包和预包装螺蛳粉终产品共487份, 利用培养法结合实时荧光聚合酶链式反应(polymerase chain reaction, PCR)法对VP进行精准检测。结果 农贸市场的70份螺蛳中VP的检出率为38.6%, 对其中的44份进行定量检测, 平均污染量为16.1 MPN/g; 生产企业的41份螺蛳及螺肉中VP的检出率为2.4%; 半成品汤料包和终产品中均未检出VP。结论 尽管零售端螺蛳中VP污染较为普遍, 但在生产端的螺蛳及螺肉中VP污染率较低。此外, 螺蛳粉生产工艺中的熬煮过程也能有效杀灭VP。总体而言, 螺蛳粉生产链中污染VP的风险较小。

柳州螺蛳粉  /  生产链  /  副溶血性弧菌  /  污染状况

Objective To investigate the contamination level of Vibrio parahaemolyticus (VP) in the production chain of the Liuzhou river snails rice noodles, promptly assess the risk of VP in the production chain. Methods From 2023 to 2024, a total of 487 simlpes, including snails and snail meat, semi-finished soup packets, and pre-packed Liuzhou river snails rice noodles final products, were randomly collected from Liuzhou agricultural markets and manufacturing enterprises in Liuzhou. A conbination of culture-based methods and real-time fluorescent polymerase chain reaction (PCR) was used to detect VP. Results Among 70 snails samples from agricultural market with a detection rate of 38.6% and an average contamination level of 16.1 MPN/g in 44 snails samples. In the 41 snails and snail meat sample from manufacturing enterprises showing the detection rate of 2.4%; no VP was detected in the semi-finished soup package or final product. Conclusion Although VP contamination is relatively common in snails at the retail trade, the contamination rate of snails and snail meat in the enterprise is low. Furthermore, the repeated boiling in Liuzhou river snails rice noodles production can effectively control VP. Therefore, the risk of VP contamination in the Liuzhou river snails rice noodles production chain is relatively low.

Liuzhou river snails rice noodles  /  production chain  /  Vibrio parahaemolyticus  /  contamination status
谢雨龙, 覃巧思, 鄢航, 覃海姣, 吴燮昳, 兰盈, 刘兴. 柳州螺蛳粉生产链中副溶血性弧菌的污染状况研究. 食品安全质量检测学报, 2025 , 16 (4) : 315 -320 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241109001
Yu-Long XIE, Qiao-Si QIN, Hang YAN, Hai-Jiao QIN, Xie-Yi WU, Ying LAN, LIU-Xing. Study on the contamination status of Vibrio parahaemolyticus in the production chain of Liuzhou river snails rice noodles[J]. Journal of Food Safety & Quality, 2025 , 16 (4) : 315 -320 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241109001
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0 引言
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食源性致病菌是导致我国食源性疾病发生的主要病因, 其中副溶血性弧菌一直是最主要的食源性致病菌之一, 在一些地区监测腹泻病中副溶血性弧菌致病比例高居榜首[1-4]。副溶血性弧菌是一种革兰氏阴性菌, 在贝类、鱼类和甲壳类中检出率较高, 其污染状况与季节相关。不同于沙门氏菌等食源性致病菌可以通过人做传染源进行传播, 副溶血性弧菌主要通过食物之间的交叉污染进行传播[5-7]。
柳州螺蛳粉作为柳州市的特色产品, 凭借其独特的风味深受广大消费者喜爱。由于螺蛳是生产螺蛳粉汤料包的最主要原料, 因此副溶血性弧菌成为预包装柳州螺蛳粉重点监测的致病菌之一[8]。根据粉包的生产工艺不同, 预包装柳州螺蛳粉分为水煮型螺蛳粉和方便型螺蛳粉, 分别执行不同的标准。水煮型螺蛳粉执行地方标准DBS 45/034—2018《食品地方安全标准 柳州螺蛳粉》, 其中副溶血性弧菌是其重点监测对象; 而方便型螺蛳粉执行GB 17400—2016《方便面》, 该标准对副溶血性弧菌并没有提出相应的要求。另一方面, 由于熟制等水产品中的副溶血性弧菌污染风险比较低, 随着GB 29921—2021《食品安全国家标准 预包装食品中致病菌限量》的实施, 熟制等水产品中副溶血性弧菌的监测要求已被取消, 仅保留生食水产品的检测。考虑到两种螺蛳粉汤料包的生产工艺相同, 因此对方便型预包装螺蛳粉是否需要纳入副溶血性弧菌的检测范围, 目前仍然有一定争议。当前对副溶血弧菌的研究主要集中在致病性和检测方法等研究[9-10], 对食品生产链中的污染调查研究相对较少。同时, 尽管已有学者对螺蛳粉中理化项目[11]、沙门氏菌[12]、霉菌[13]、重金属[14]和感官体系[15]等进行相关研究, 但是对副溶血性弧菌的研究仍鲜有报道。
本研究通过对柳州市6个城区批发及农贸市场的螺蛳、9家预包装螺蛳粉生产企业的螺蛳及螺肉和半成品汤料包、7个预包装螺蛳粉品牌终产品进行采样, 通过国标法培养和分离出疑似副溶血性弧菌的菌落, 并采用实时荧光聚合酶链式反应(polymerase chain reaction, PCR)法进行确证, 旨在全面了解预包装柳州螺蛳粉生产链中副溶血性弧菌的污染状况, 为柳州螺蛳粉的监管工作提供可循依据, 并为地方标准的修订提供数据支持。
1 材料与方法
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1.1 样品来源
2023—2024年在柳州市6个城区的批发及农贸市场采集石螺和田螺样品共70份, 螺蛳粉生产企业采集螺蛳及螺肉共41份、半成品汤料包样品66份, 线上及线下采集7个螺蛳粉品牌共310份。
1.2 试剂与仪器
3%氯化钠碱性蛋白胨水(广东环凯微生物科技有限公司); 弧菌显色培养基(上海科玛嘉微生物技术有限公司); Premix Ex TaqTM、细菌基因组DNA提取试剂盒[宝生物工程(大连)有限公司]; 上游引物GCGACCTTTCTCTGA AATATTAATTGT; 下游引物CATTCGCGTGGCAAACATC; taqman探针CGCACAAGGCTCGACGGCTGA, 序列参照SN/T 1870—2016《出口食品中食源性致病菌检测方法 实时荧光PCR法》由华大基因合成。
电子天平(常熟市双杰测试仪器厂); 均质器(宁波新芝生物科技股份有限公司); ABI7500型实时荧光定量PCR(美国ABI公司); Sigma1-14型离心机(德国Sigma公司)。
1.3 实验方法
1.3.1 样品处理
(1)螺蛳
样品的处理参照GB 4789.20—2024《食品安全国家标准 食品微生物学检验 水产品及其制品采样和检样处理》中的方法。样品运回实验室后先用煮沸冷却后的三级水冲洗螺壳上的泥沙, 沥干水分, 用75%酒精消毒外表面, 然后用消毒的锤子将螺壳砸开, 取出内容物称25 g加入到225 mL无菌的3%氯化钠碱性蛋白胨水中, 使用均质器均质, 制备成1:10的样液。
(2)螺肉及汤料包
称取25 g加入到225 mL无菌的3%氯化钠碱性蛋白胨水中, 使用均质器均质, 制备成1:10的样液。
(3)预包装螺蛳粉终产品
样品处理参照DBS 45/034—2018中的规定, 将所有配料包混合后称取25 g加入到225 mL无菌的3%氯化钠碱性蛋白胨水中, 使用均质器均质, 制备成1:10的样液。
1.3.2 定性检测培养
样品按1.3.1进行处理后, 36 ℃过夜培养。
1.3.3 定量检测培养
将1.3.1中制备的1:10样品匀液摇匀后, 参照GB 4789.7—2013《食品安全国家标准 食品微生物学检验 副溶血性弧菌检验》定量检测, 用无菌移液管吸取1 mL加入到含有9 mL 3%氯化钠碱性蛋白胨水的试管中, 充分混匀, 制备成1:100的样品稀释液。另取一支无菌移液管, 按照同样的方法依次吸取1 mL加入到含有9 mL 3%氯化钠碱性蛋白胨水的试管中混匀后, 制备成1:1000的连续3个稀释梯度。每个稀释梯度接种3支试管, 然后将试管置于36 ℃恒温培养箱中, 过夜培养。
1.3.4 划线分离
取出培养箱中的试管摇匀, 用接种环伸入液面取一环菌液, 在弧菌显色平板上划线, 然后置于36 ℃恒温过夜培养。培养结束后观察平板上生长的菌落, 典型的副溶血性弧菌在弧菌显色平板上呈淡紫色菌落形态, 挑取淡紫红色菌落进行实时荧光PCR扩增确证。
1.3.5 DNA模版制备
参照SN/T 1870—2016的水煮法, 用无菌接种环从弧菌显色平板上的疑似菌落中沾取少量菌体, 置于50 μL DNA提取液中用移液器吹打重悬, 水煮5 min后12000 r/min离心5 min, 吸取上清液放置-20 ℃备用。
1.3.6 实时荧光PCR反应体系
采用20 μL实时荧光反应体系, 探针终浓度为0.4 μmol/L, 体系中同时还包含10 μL Premix Ex Taq(2×)、引物终浓度为0.4 μmol/L、0.2 μL参比荧光、2 μL的基因组DNA, 分别用无菌水补足剩余的体积。实时荧光PCR仪反应程序参照Premix Ex TaqTM说明书为: 95 ℃预变性30 s, 随后是40个循环的95 ℃变性5 s以及60 ℃复性和延伸34 s。
1.4 数据处理
根据副溶血性弧菌最可能数(most probable number, MPN)的检索表和PCR鉴定的阳性管数, 进行MPN计数, 当MPN<3.0 MPN/g, 分别估算为1.5 MPN/g; 当MPN>1100 MPN/g, 假定往后一个稀释梯度的阳性管数均不生长, 即阳性管数为3.3.0估算为2400 MPN/g[16]。
2 结果与分析
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2.1 柳州市市售螺蛳副溶血性弧菌的检出情况
2023—2024年夏季采集了柳州6个城区零售环节螺蛳共70份。根据实验方法经培养、平板分离及实时荧光PCR鉴定, 最终得到70份样品中有27份样品扩增出荧光曲线, 结果如表1所示。夏季为副溶血性弧菌的高发季节, 柳州市各城区螺蛳中副溶血性弧菌的污染率各有差异, 含有大型批发市场的柳江区污染率最低为21.7%, 而最小的城区阳和新区污染率最高为71.4%, 全市总平均污染率为38.6%, 其中市场田螺及螺肉总数16份, 阳性9份; 石螺及螺肉总数54份, 阳性18份。部分菌株荧光图谱结果如图1所示。对2024年采集的44份样品进行定量检测, 最高污染量为66.3 MPN/g, 平均污染量为16.1 MPN/g。
2.2 生产企业螺蛳和螺肉及半成品汤料副溶血性弧菌的检出情况
2023—2024年夏季采集了柳州市螺蛳粉生产企业用于生产汤料包的螺蛳及螺肉41份, 采集未进行消毒或灭菌的半成品汤料包66份, 利用国标法和实时荧光PCR法对其进行定性检测, 结果如表2所示。结果显示有1份螺蛳样品检出副溶血性弧菌, 污染率为2.4%, 定量检测结果为1.5 MPN/g, 荧光曲线图谱见图2; 半成品汤料包均未检出副溶血性弧菌。
2.3 预包装螺蛳粉副溶血性弧菌的检出情况
本研究对2023—2024年7个螺蛳粉品牌共310份预包装螺蛳粉进行检测, 检测结果如表3所示。结果显示310份预包装螺蛳粉定性检测均为未检出, 均符合DBS 45/034—2018标准的要求。
3 讨论与结论
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副溶血性弧菌是一种嗜盐的微生物, 在近岸海域中广泛存在, 在海产品中有较高的污染率[17-18]。但是近年的研究发现, 其在淡水产品中的污染率也呈上升的趋势[19-21], 甚至某些情况下在淡水食品中的流行率高于海产品[22]。尽管市售淡水产品中副溶血性弧菌出现比较高的污染率, 但实际污染量相对较低。如赵力等[23]研究发现, 万州零售淡水鱼中副溶血性弧菌平均污染率为38.3%, 平均污染量20.8 MPN/g。本研究得出类似的结论, 在对柳州市批发及农贸市场中随机采样的70份螺蛳样品中, 有27份呈阳性, 总污染率为38.6%。在这些样品中, 城区平均污染量最高为66.3 MPN/g, 最低为1.5 MPN/g, 全市平均污染量为16.1 MPN/g, 远低于国家标准对散装食品和预包装食品致病菌限量中所设定的安全限值1000 MPN/g。
在养殖、销售、加工等各环节中, 销售环节的各类致病弧菌污染率相对较高, 农贸菜市是污染率最高的场所之一[24]。螺蛳中副溶血性弧菌的污染率主要与其销售方式相关。本研究发现, 规模较大的批发市场, 如柳江区其副溶血性弧菌污染率最低, 为21.7% (5/23)。这可能是因为大的批发市场出售的产品集中而单一, 而小的市场同个摊位出售的产品种类比较多, 从而增加了产品之间交叉污染副溶血性弧菌的风险。李成伟等[25]对临沂市的致病弧菌污染状况研究后发现, 销售环境中副溶血性弧菌的污染率为26%, 而养殖环节的污染率仅为4.1%, 研究者们推测市售产品副溶血性弧菌污染率高但污染量较低的原因可能是淡水水体中该菌数量比较少, 其在淡水产品中的存在主要通过交叉污染, 特别是混合销售的取样工具是主要的传播途径。此外, 还有研究表明, 超市中副溶血性弧菌的污染率要远低于一般农贸市场, 这主要得益于超市的分区销售, 有效避免了交叉污染[26-28]。
值得注意的是, 对螺蛳粉生产企业的螺蛳进行检测发现其阳性率远低于农贸市场, 样品检出副溶血性弧菌污染率仅为2.4% (1/41), 定量检测结果为1.5 MPN/g。这可能是因为企业通常从螺蛳产地或者比较大的批发市场采购原料, 从而减少副溶血性弧菌交叉污染的可能性。其次, 低温环境可以抑制副溶血性弧菌活性[24], 企业购置原料后一般会马上进行低温冷冻保存, 这有助于控制副溶血性弧菌的污染。同时, 在未灭菌的中间产品以及市售的终产品中均未检出副溶血性弧菌。这可能是因为原料中该菌污染率较低, 同时副溶血性弧菌不耐热, 45 ℃就可以让其达到亚致死状态[29], 超过65 ℃可以对其进行有效杀灭[30]。因此, 生产工艺中的煮沸和熬制步骤都可以对其进行有效控制。
综上所述, 尽管柳州市零售端田螺和石螺副溶血性弧菌污染率较高, 但污染量相对较低, 这可能是由于交叉污染导致的。螺蛳粉生产企业由于采购渠道和低温储存措施得当, 其污染率远低于农贸市场。此外, 生产过程中熬煮以及巴氏消毒也能有效杀灭副溶血性弧菌。因此, 预包装螺蛳粉生产链中副溶血性弧菌污染风险相对较小。
基金
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  • 柳州市科技攻关计划项目(2024SZ0505G004)
文献
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参考文献 引证文献
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2025年第16卷第4期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20241109001
  • 接收时间：2024-11-09
  • 首发时间：2025-07-21
  • 出版时间：2025-02-25
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  • 收稿日期：2024-11-09
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柳州市科技攻关计划项目(2024SZ0505G004)
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    1.柳州市质量检验检测研究中心, 柳州 545500
    2.柳州螺蛳粉质量检验中心, 柳州 545500

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* 刘兴(1972—), 男, 高级工程师, 主要研究方向为食品质量与安全。E-mail:
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
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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