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Article(id=1215670313682129603, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1215670311140381365, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20250321001, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1742486400000, receivedDateStr=2025-03-21, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1767767988843, onlineDateStr=2026-01-07, pubDate=1753372800000, pubDateStr=2025-07-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1767767988843, onlineIssueDateStr=2026-01-07, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1767767988843, creator=13701087609, updateTime=1767767988843, updator=13701087609, issue=Issue{id=1215670311140381365, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='14', pageStart='1', pageEnd='326', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1767767988237, creator=13701087609, updateTime=1767970098618, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1216518023599538606, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1215670311140381365, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1216518023599538607, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1215670311140381365, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=285, endPage=290, ext={EN=ArticleExt(id=1215670314017673927, articleId=1215670313682129603, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Comparative study on the resistance of different species strains to 3 kinds of food disinfectants, columnId=1153433739154678309, journalTitle=Journal of Food Safety & Quality, columnName=Food Safety Supervision and Management, runingTitle=null, highlight=null, articleAbstract=

Objective To compare the resistance differences of 18 kinds of bacterial strains and 2 kinds of biological indicators to chlorine-containing disinfectants, hydrogen peroxide and quaternary ammonium compounds. Methods Strains were selected from food, environment and human sources, including gram-negative bacteria, gram-positive bacteria and spore-forming bacteria (6 kinds of strains per category). Species identification was performed using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) and biochemical assays. The 2 kinds of biological indicators, namely Staphylococcus aureus ATCC6538 and Escherichia coli 8099 were used as reference strains. Bacterial suspensions (2.0×108-10.0×108 CFU/mL) were prepared to test their tolerance to sodium hypochlorite (available chlorine 0.00400%-0.00499%), hydrogen peroxide (7.5%), and benzalkonium bromide (3.996 mg/mL). All bacterial strains were treated with disinfectants for 3 minutes. By calculating the killing log value (KL) of disinfectants against different bacterial strains, the differences in resistance of different species of bacterial strains to 3 kinds of disinfectants were compared. Results The bactericidal efficacy ranked as sodium hypochlorite, hydrogen peroxide, benzalkonium bromide. Different species showed significant variations in resistance to disinfectants. Among 3 categories of bacterial strains (gram-negative bacteria, gram-positive bacteria and spore-forming bacteria), there were strains within each category that exhibit higher resistance to disinfectants than 2 kinds of biological indicators. Conclusion Sodium hypochlorite and hydrogen peroxide exhibit superior bactericidal effects compared to benzalkonium bromide. The resistance to disinfectant of bacteria is species-specific and cannot be predicted solely by gram classification. The screening scope for biological indicators should be expanded based on actual needs.

, correspAuthors=Sheng-Hui CUI, Jing-Yun LI, 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=Na LIU, Ya-Ping WANG, Xiu REN, Shuai ZHAO, Xue-Shuo WANG, Yue DOU, Sheng-Hui CUI, Jing-Yun LI), CN=ArticleExt(id=1215670315229827834, articleId=1215670313682129603, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=不同种属菌株对3种食品用消毒剂的抗性比对研究, columnId=1153433739318256173, journalTitle=食品安全质量检测学报, columnName=食品安全监管, runingTitle=null, highlight=null, articleAbstract=

目的 比较18种菌株和2种指示微生物对含氯、过氧化氢及季铵盐类消毒剂的抗性差异。方法 选取食品、环境和人体来源的革兰氏阴性菌、革兰氏阳性菌及芽孢菌3类菌株(每类6种), 经基质辅助激光解吸电离飞行时间质谱仪(matrix-assisted laser desorption ionization time of flight mass spectrometry, MALDI-TOF MS)和生化鉴定种属。以金黄色葡萄球菌ATCC6538和大肠杆菌8099 2种指示微生物为参考菌株, 制备2.0×108~10.0×108 CFU/mL菌悬液, 检测其对次氯酸钠(有效氯0.00400%~0.00499%)、过氧化氢(7.5%)及苯扎溴铵(3.996 mg/mL)的耐受性, 作用时间均为3 min。通过计算消毒剂对不同菌株的杀灭对数值(killing log value, KL), 比较不同种属菌株对3种消毒剂抗性差异。结果 消毒剂杀菌效力从强到弱为: 次氯酸钠、过氧化氢、苯扎溴铵; 不同菌种抗性存在显著差异, 且常见的3类菌株(革兰氏阴性菌、革兰氏阳性菌及芽孢菌)中, 均有对消毒剂抗性强于2种指示微生物的菌株。结论 次氯酸钠与过氧化氢的杀菌效果优于苯扎溴铵; 菌株抗性具有种属特异性, 不能单纯通过革兰氏分类预判; 指示微生物应依据实际需求扩大筛选范围。

, correspAuthors=崔生辉, 李景云, authorNote=null, correspAuthorsNote=
*崔生辉(1972—), 男, 博士, 研究员, 主要研究方向为食品安全检测。E-mail: ;
李景云(1972—), 男, 主任技师, 主要研究方向为食品安全检测。E-mail:
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刘娜(1988—), 女, 硕士, 助理研究员, 主要研究方向为食品安全检测。E-mail:

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刘娜(1988—), 女, 硕士, 助理研究员, 主要研究方向为食品安全检测。E-mail:

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刘娜(1988—), 女, 硕士, 助理研究员, 主要研究方向为食品安全检测。E-mail:

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Chinese Journal of Infectious Diseases, 2020, 38(9): 556-563., articleTitle=Visual analysis of the focuses and trends of research on microbial resistance to disinfectants in China and abroad, refAbstract=null)], funds=[Fund(id=1215670322586636417, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1215670313682129603, awardId=2022YFF1103100, language=CN, fundingSource=国家重点研发计划项目(2022YFF1103100), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1215670315506651917, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1215670313682129603, xref=null, ext=[AuthorCompanyExt(id=1215670315515040526, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1215670313682129603, companyId=1215670315506651917, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=National Institute for Food and Drug Control, Beijing 100050, China), AuthorCompanyExt(id=1215670315540206352, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1215670313682129603, companyId=1215670315506651917, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=中国食品药品检定研究院, 北京 100050)])], figs=[ArticleFig(id=1215670320606924871, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1215670313682129603, language=EN, label=Table 1, caption=

Information table of 18 species and strains of bacteria

, figureFileSmall=null, figureFileBig=null, tableContent=
序号 微生物名称 原始标号 来源 类别
1 阪崎克罗诺杆菌 FC2942 食品 革兰氏
阴性菌
2 弗氏柠檬酸杆菌 FC2251 市售鸽肉
3 产气肠杆菌 FC4541 病人
4 阴沟肠杆菌 FC1375 病人
5 溶藻弧菌 FC1621 海水
6 肺炎克雷伯菌 FC1382 病人
7 单核细胞增生李斯特氏菌 FC11109 食品 革兰氏
阳性菌
8 粪肠球菌 FC13911 食品
9 藤黄微球菌 FC1737 空气
10 鹑鸡肠球菌 FC6832 病人
11 玫瑰考克氏菌 FC1763 空气
12 纹带棒状杆菌 FC6831 病人
13 拟蕈状芽孢杆菌 FC10418 天坛公园泥土 芽孢菌
14 蜡样芽孢杆菌 FC1052 天津市大饼鸡蛋
15 枯草芽孢杆菌 FC3473 医院
16 人参土芽孢杆菌 FC3461 药品辅料
17 蕈状芽孢杆菌 FC3478 医院
18 地衣芽孢杆菌 FC14167 食品
), ArticleFig(id=1215670320745336910, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1215670313682129603, language=CN, label=表1, caption=

18个种属菌株信息表

, figureFileSmall=null, figureFileBig=null, tableContent=
序号 微生物名称 原始标号 来源 类别
1 阪崎克罗诺杆菌 FC2942 食品 革兰氏
阴性菌
2 弗氏柠檬酸杆菌 FC2251 市售鸽肉
3 产气肠杆菌 FC4541 病人
4 阴沟肠杆菌 FC1375 病人
5 溶藻弧菌 FC1621 海水
6 肺炎克雷伯菌 FC1382 病人
7 单核细胞增生李斯特氏菌 FC11109 食品 革兰氏
阳性菌
8 粪肠球菌 FC13911 食品
9 藤黄微球菌 FC1737 空气
10 鹑鸡肠球菌 FC6832 病人
11 玫瑰考克氏菌 FC1763 空气
12 纹带棒状杆菌 FC6831 病人
13 拟蕈状芽孢杆菌 FC10418 天坛公园泥土 芽孢菌
14 蜡样芽孢杆菌 FC1052 天津市大饼鸡蛋
15 枯草芽孢杆菌 FC3473 医院
16 人参土芽孢杆菌 FC3461 药品辅料
17 蕈状芽孢杆菌 FC3478 医院
18 地衣芽孢杆菌 FC14167 食品
), ArticleFig(id=1215670320866971736, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1215670313682129603, language=EN, label=Table 2, caption=

Identification results of 18 species and strains of bacteria

, figureFileSmall=null, figureFileBig=null, tableContent=
菌种名称 鉴定结果
生化方法 MALDI-TOF MS方法
阪崎克罗诺杆菌 阪崎克罗诺杆菌 阪崎克罗诺杆菌
弗氏柠檬酸杆菌 弗氏柠檬酸杆菌 弗氏柠檬酸杆菌
产气肠杆菌 产气肠杆菌 产气肠杆菌
阴沟肠杆菌 阴沟肠杆菌 阴沟肠杆菌
溶藻弧菌 溶藻弧菌 溶藻弧菌
肺炎克雷伯菌 肺炎克雷伯菌 肺炎克雷伯菌
单核细胞增生
李斯特氏菌		 单核细胞增生
李斯特氏菌		 单核细胞增生
李斯特氏菌
粪肠球菌 粪肠球菌 粪肠球菌
藤黄微球菌 藤黄微球菌 藤黄微球菌
鹑鸡肠球菌 鹑鸡肠球菌 鹑鸡肠球菌
玫瑰考克氏菌 玫瑰考克氏菌 玫瑰考克氏菌
纹带棒状杆菌 纹带棒状杆菌 纹带棒状杆菌
拟蕈状芽孢杆菌 拟蕈状芽孢杆菌 拟蕈状芽孢杆菌
蜡样芽孢杆菌 蜡样芽孢杆菌 蜡样芽孢杆菌
枯草芽孢杆菌 枯草芽孢杆菌 枯草芽孢杆菌
人参土芽孢杆菌 人参土芽孢杆菌 人参土芽孢杆菌
蕈状芽孢杆菌 蕈状芽孢杆菌 蕈状芽孢杆菌
地衣芽孢杆菌 地衣芽孢杆菌 地衣芽孢杆菌
), ArticleFig(id=1215670320988606565, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1215670313682129603, language=CN, label=表2, caption=

18个种属菌株的鉴定结果

, figureFileSmall=null, figureFileBig=null, tableContent=
菌种名称 鉴定结果
生化方法 MALDI-TOF MS方法
阪崎克罗诺杆菌 阪崎克罗诺杆菌 阪崎克罗诺杆菌
弗氏柠檬酸杆菌 弗氏柠檬酸杆菌 弗氏柠檬酸杆菌
产气肠杆菌 产气肠杆菌 产气肠杆菌
阴沟肠杆菌 阴沟肠杆菌 阴沟肠杆菌
溶藻弧菌 溶藻弧菌 溶藻弧菌
肺炎克雷伯菌 肺炎克雷伯菌 肺炎克雷伯菌
单核细胞增生
李斯特氏菌		 单核细胞增生
李斯特氏菌		 单核细胞增生
李斯特氏菌
粪肠球菌 粪肠球菌 粪肠球菌
藤黄微球菌 藤黄微球菌 藤黄微球菌
鹑鸡肠球菌 鹑鸡肠球菌 鹑鸡肠球菌
玫瑰考克氏菌 玫瑰考克氏菌 玫瑰考克氏菌
纹带棒状杆菌 纹带棒状杆菌 纹带棒状杆菌
拟蕈状芽孢杆菌 拟蕈状芽孢杆菌 拟蕈状芽孢杆菌
蜡样芽孢杆菌 蜡样芽孢杆菌 蜡样芽孢杆菌
枯草芽孢杆菌 枯草芽孢杆菌 枯草芽孢杆菌
人参土芽孢杆菌 人参土芽孢杆菌 人参土芽孢杆菌
蕈状芽孢杆菌 蕈状芽孢杆菌 蕈状芽孢杆菌
地衣芽孢杆菌 地衣芽孢杆菌 地衣芽孢杆菌
), ArticleFig(id=1215670322267869296, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1215670313682129603, language=EN, label=Table 3, caption=

Resistance test results of 18 species and strains of bacteria to 3 kinds of disinfectants

, figureFileSmall=null, figureFileBig=null, tableContent=
类别 菌种名称 KL
次氯酸钠 过氧化氢 苯扎溴铵
革兰氏
阴性菌
 克罗诺杆菌 ≥5.00 ≥5.00 4.10
弗氏柠檬酸杆菌 4.60 4.01 3.93
产气肠杆菌 4.46 ≥5.00 2.17
阴沟肠杆菌 ≥5.00 4.40 2.82
溶藻弧菌 ≥5.00 ≥5.00 4.49
肺炎克雷伯菌 ≥5.00 3.82 3.00
革兰氏
阳性菌
 单核细胞增生李斯特氏菌 ≥5.00 ≥5.00 1.77
粪肠球菌 ≥5.00 3.00 2.98
藤黄微球菌 4.19 ≥5.00 4.45
鹑鸡肠球菌 ≥5.00 ≥5.00 ≥5.00
玫瑰考克氏菌 ≥5.00 ≥5.00 2.40
纹带棒状杆菌 ≥5.00 ≥5.00 3.37
芽孢菌
 拟蕈状芽孢杆菌 2.70 ≥5.00 ≥5.00
蜡样芽孢杆菌 2.39 2.59 1.46
枯草芽孢杆菌 3.32 2.91 1.88
蕈状芽孢杆菌 2.71 ≥5.00 1.78
人参土芽孢杆菌 ≥5.00 ≥5.00 ≥5.00
地衣芽孢杆菌 ≥5.00 ≥5.00 2.48
参考
菌株		 大肠杆菌8099 3.15 4.37 2.82
金黄色葡萄球菌ATCC6538 2.90 3.13 3.78
), ArticleFig(id=1215670322360143989, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1215670313682129603, language=CN, label=表3, caption=

18个种属菌株对3种消毒剂的抗性测试结果

, figureFileSmall=null, figureFileBig=null, tableContent=
类别 菌种名称 KL
次氯酸钠 过氧化氢 苯扎溴铵
革兰氏
阴性菌
 克罗诺杆菌 ≥5.00 ≥5.00 4.10
弗氏柠檬酸杆菌 4.60 4.01 3.93
产气肠杆菌 4.46 ≥5.00 2.17
阴沟肠杆菌 ≥5.00 4.40 2.82
溶藻弧菌 ≥5.00 ≥5.00 4.49
肺炎克雷伯菌 ≥5.00 3.82 3.00
革兰氏
阳性菌
 单核细胞增生李斯特氏菌 ≥5.00 ≥5.00 1.77
粪肠球菌 ≥5.00 3.00 2.98
藤黄微球菌 4.19 ≥5.00 4.45
鹑鸡肠球菌 ≥5.00 ≥5.00 ≥5.00
玫瑰考克氏菌 ≥5.00 ≥5.00 2.40
纹带棒状杆菌 ≥5.00 ≥5.00 3.37
芽孢菌
 拟蕈状芽孢杆菌 2.70 ≥5.00 ≥5.00
蜡样芽孢杆菌 2.39 2.59 1.46
枯草芽孢杆菌 3.32 2.91 1.88
蕈状芽孢杆菌 2.71 ≥5.00 1.78
人参土芽孢杆菌 ≥5.00 ≥5.00 ≥5.00
地衣芽孢杆菌 ≥5.00 ≥5.00 2.48
参考
菌株		 大肠杆菌8099 3.15 4.37 2.82
金黄色葡萄球菌ATCC6538 2.90 3.13 3.78
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不同种属菌株对3种食品用消毒剂的抗性比对研究
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刘娜 , 王亚萍 , 任秀 , 赵帅 , 王学硕 , 窦越 , 崔生辉 * , 李景云 *
食品安全质量检测学报 | 食品安全监管 2025,16(14): 285-290
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食品安全质量检测学报 | 食品安全监管 2025, 16(14): 285-290
不同种属菌株对3种食品用消毒剂的抗性比对研究
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刘娜 , 王亚萍, 任秀, 赵帅, 王学硕, 窦越, 崔生辉* , 李景云*
作者信息
  • 中国食品药品检定研究院, 北京 100050

    • 刘娜(1988—), 女, 硕士, 助理研究员, 主要研究方向为食品安全检测。E-mail:

通讯作者:

*崔生辉(1972—), 男, 博士, 研究员, 主要研究方向为食品安全检测。E-mail: ;
李景云(1972—), 男, 主任技师, 主要研究方向为食品安全检测。E-mail:
Comparative study on the resistance of different species strains to 3 kinds of food disinfectants
Na LIU , Ya-Ping WANG, Xiu REN, Shuai ZHAO, Xue-Shuo WANG, Yue DOU, Sheng-Hui CUI* , Jing-Yun LI*
Affiliations
  • National Institute for Food and Drug Control, Beijing 100050, China
出版时间: 2025-07-25 doi: 10.19812/j.cnki.jfsq11-5956/ts.20250321001
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摘要
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目的 比较18种菌株和2种指示微生物对含氯、过氧化氢及季铵盐类消毒剂的抗性差异。方法 选取食品、环境和人体来源的革兰氏阴性菌、革兰氏阳性菌及芽孢菌3类菌株(每类6种), 经基质辅助激光解吸电离飞行时间质谱仪(matrix-assisted laser desorption ionization time of flight mass spectrometry, MALDI-TOF MS)和生化鉴定种属。以金黄色葡萄球菌ATCC6538和大肠杆菌8099 2种指示微生物为参考菌株, 制备2.0×108~10.0×108 CFU/mL菌悬液, 检测其对次氯酸钠(有效氯0.00400%~0.00499%)、过氧化氢(7.5%)及苯扎溴铵(3.996 mg/mL)的耐受性, 作用时间均为3 min。通过计算消毒剂对不同菌株的杀灭对数值(killing log value, KL), 比较不同种属菌株对3种消毒剂抗性差异。结果 消毒剂杀菌效力从强到弱为: 次氯酸钠、过氧化氢、苯扎溴铵; 不同菌种抗性存在显著差异, 且常见的3类菌株(革兰氏阴性菌、革兰氏阳性菌及芽孢菌)中, 均有对消毒剂抗性强于2种指示微生物的菌株。结论 次氯酸钠与过氧化氢的杀菌效果优于苯扎溴铵; 菌株抗性具有种属特异性, 不能单纯通过革兰氏分类预判; 指示微生物应依据实际需求扩大筛选范围。

消毒剂  /  指示微生物  /  种属  /  抗性  /  差异

Objective To compare the resistance differences of 18 kinds of bacterial strains and 2 kinds of biological indicators to chlorine-containing disinfectants, hydrogen peroxide and quaternary ammonium compounds. Methods Strains were selected from food, environment and human sources, including gram-negative bacteria, gram-positive bacteria and spore-forming bacteria (6 kinds of strains per category). Species identification was performed using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) and biochemical assays. The 2 kinds of biological indicators, namely Staphylococcus aureus ATCC6538 and Escherichia coli 8099 were used as reference strains. Bacterial suspensions (2.0×108-10.0×108 CFU/mL) were prepared to test their tolerance to sodium hypochlorite (available chlorine 0.00400%-0.00499%), hydrogen peroxide (7.5%), and benzalkonium bromide (3.996 mg/mL). All bacterial strains were treated with disinfectants for 3 minutes. By calculating the killing log value (KL) of disinfectants against different bacterial strains, the differences in resistance of different species of bacterial strains to 3 kinds of disinfectants were compared. Results The bactericidal efficacy ranked as sodium hypochlorite, hydrogen peroxide, benzalkonium bromide. Different species showed significant variations in resistance to disinfectants. Among 3 categories of bacterial strains (gram-negative bacteria, gram-positive bacteria and spore-forming bacteria), there were strains within each category that exhibit higher resistance to disinfectants than 2 kinds of biological indicators. Conclusion Sodium hypochlorite and hydrogen peroxide exhibit superior bactericidal effects compared to benzalkonium bromide. The resistance to disinfectant of bacteria is species-specific and cannot be predicted solely by gram classification. The screening scope for biological indicators should be expanded based on actual needs.

disinfectant  /  biological indicators  /  species  /  resistance  /  difference
刘娜, 王亚萍, 任秀, 赵帅, 王学硕, 窦越, 崔生辉, 李景云. 不同种属菌株对3种食品用消毒剂的抗性比对研究. 食品安全质量检测学报, 2025 , 16 (14) : 285 -290 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250321001
Na LIU, Ya-Ping WANG, Xiu REN, Shuai ZHAO, Xue-Shuo WANG, Yue DOU, Sheng-Hui CUI, Jing-Yun LI. Comparative study on the resistance of different species strains to 3 kinds of food disinfectants[J]. Journal of Food Safety & Quality, 2025 , 16 (14) : 285 -290 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250321001
正文
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0 引言
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自新冠病毒疫情暴发以来, 微生物的潜在危害已引发社会各界的高度关注[1]。在食品生产、储存及销售的全链条流程中, 微生物污染始终是威胁食品安全的关键风险因素[2-3]。科学应用消毒剂作为防控微生物污染的核心手段[4], 其有效性直接关系到食品全产业链的生物安全[5-6]。本研究聚焦食品领域常用消毒剂(含氯、过氧化物类及季铵盐类消毒剂), 选取了从食品、环境和人等分离的18种常见菌种, 涵盖革兰氏阴性菌(具有外膜屏障结构)、革兰氏阳性菌(含致密肽聚糖层)及芽孢菌(具备厚壁保护结构)3大类群, 基于我国现行标准WS/T 774—2021《新冠肺炎疫情期间现场消毒评价标准》, 系统评估次氯酸钠(含氯消毒剂)、过氧化氢(过氧化物类消毒剂)[7]与苯扎溴铵(季铵盐类消毒剂)[8]的杀菌效果。通过建立多种属菌株消毒剂抗性评价体系, 揭示不同种属菌株对上述3类消毒剂的耐受特征, 为精准选择消毒剂类型及优化使用浓度提供科学依据; 构建基于微生物抗性特征的消毒方案决策模型, 有效规避传统消毒实践中因配伍不当导致的杀菌效能衰减或化学残留超标风险; 建立消毒剂抗性预警机制, 为防控多重耐消毒剂菌株的扩散提供技术支撑[9-10]。同时, 以目前常用的2种指示微生物(金黄色葡萄球菌ATCC6538和大肠杆菌8099)为参考菌株, 通过计算比较消毒剂对不同种属菌株与指示微生物的杀灭对数值(killing log value, KL), 明确上述3种消毒剂在实际应用中的消杀微生物范围, 也为今后筛选应用范围更广的指示微生物提供数据支撑。研究结果不仅可为食品企业建立全过程消毒质控体系提供理论指导, 更能为监管部门完善行业标准、健全食品安全保障体系提供数据支持, 从而推动食品产业在生物安全可控[11]的基础上实现高质量发展。
1 材料与方法
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1.1 材料与试剂
平板计数琼脂(plate count agar, PCA)培养基、乳酸菌MRS琼脂培养基(De Man Rogosa Sharpe agar, MRS)、胰蛋白胨大豆琼脂(tryptic soy agar, TSA)(美国BD公司); Bruker标准溶剂(货号900666)、次氯酸钠溶液(货号239305)(美国Sigma-Aldrich公司); α-氰基-4-羟基肉桂酸(α-cyano-4- hydroxycinnamic acid, HCCA)(货号8255344, 德国Bruker公司); 甲酸(纯度98%, 北京百灵威科技有限公司); 细菌基因组DNA提取试剂盒[天根生化科技(北京)有限公司]; 革兰氏阳性(gram positive, GP)细菌鉴定卡、革兰氏阴性(gram negative, GN)细菌鉴定卡、需氧芽孢杆菌(Bacillus, BCL)鉴定卡(法国生物梅里埃公司); 30%过氧化氢溶液、吐温80(上海沪试实验室器材股份有限公司); 复方新洁尔灭消毒液(苯扎溴铵)(货号30701122, 山东利尔康消毒科技股份有限公司); 硫代硫酸钠(分析纯, 国药集团化学试剂有限公司)。
1.2 仪器与设备
Thermo1389生物安全柜、Thermo 205050GC恒温培养箱(美国Thermo Fisher Scientific公司); VITEK COMPACT 2全自动微生物分析系统(法国梅里埃公司); autoflex基质辅助激光解吸电离飞行时间质谱仪(matrix-assisted laser desorption ionization time of flight mass spectrometry, MALDI-TOF MS)(德国Bruker公司); HYC-940螺旋涂布仪(西班牙IUL公司)。
1.3 方法
1.3.1 菌种信息
基于食品微生物安全研究的特殊性, 本研究菌株筛选遵循以下核心原则: (1)生物多样性; (2)食源性污染关联性; (3)符合GB 4789系列等国家标准要求且在食品接触表面高频检出; (4)对现行检测体系具有稳定响应特性。通过多源样本(食品原料、生产环境及从业人员)分离纯化, 成功构建包含18种特征菌株的实验菌种库(详见表1), 充分体现食品污染微生物的种群多样性特征。参照WS/T 774—2021技术要求, 以2种指示微生物(金黄色葡萄球菌ATCC 6538及大肠埃希氏菌8099)作为参考菌株, 通过建立实验组-对照组平行比对体系, 确保消毒剂效能评价结果的可比性与溯源性。
1.3.2 18个种属菌株鉴定
将各个菌株接种于TSA平板, 置于(36±1) ℃恒温培养箱中培养18~24 h。挑取单菌落进行划线分离, 再次接种于TSA平板, 在相同条件下培养18~24 h, 获得纯化菌株备用。
取上述纯化后的各菌株第二代新鲜培养物, 分别采用VITEK COMPACT 2全自动微生物鉴定系统和MALDI-TOF MS进行菌种鉴定分析, 以确定各菌株的种属分类。
1.3.3 18个种属菌株对3种消毒剂的抗性测试
用无菌生理盐水将各菌株的二代新鲜培养物制备成2.0×108~10.0×108 CFU/mL的菌悬液, 备用。
次氯酸钠抗性实验中, 取4支12 mL无菌培养管, 分别加入以下试剂各4.5 mL: 次氯酸钠溶液(有效氯0.00400%~0.00499%)、无菌生理盐水、中和剂(1%硫代硫酸钠溶液, 设2个平行管), 分别加入0.5 mL备用菌悬液。其中, 消毒剂管作为试验组、生理盐水管作为阴性对照组。混匀后静置3 min, 分别取0.5 mL混合液加入中和剂管中, 混匀后静置10 min。各取100 μL涂布于PCA平板, (36±1) ℃培养18~24 h后计数菌落形成单位(CFU)。
过氧化氢抗性实验中, 采用次氯酸钠相同实验步骤, 使用7.5%过氧化氢溶液作为实验组消毒剂, 中和剂仍为1%硫代硫酸钠溶液。
苯扎溴铵抗性实验中, 使用3.996 mg/mL苯扎溴铵作为实验组消毒剂, 中和剂更换为含3%吐温80和1%卵磷脂的磷酸盐缓冲生理盐水(phosphate buffer saline, PBS), 其余条件保持不变。
所有实验均设置2株指示微生物作为参考菌株, 与18株实验菌株同步进行抗性测试, 每组实验重复3次。
1.4 数据处理
计算18个种属菌株与2种指示微生物的KL值, 采用Microsoft Excel 2021进行数据分析, 每个实验重复3次。
KL=lg(N0)–lg(Nt)
注: N0为消毒前初始微生物数量; Nt为消毒后存活的微生物数量。
2 结果与分析
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2.1 18个种属菌株的鉴定结果
18个种属菌株经生化和MALDI-TOF-MS的鉴定结果见表2, 与预期结果一致。
2.2 18个种属菌株对3种消毒剂抗性测试结果
本研究通过不同种属菌株的KL值, 系统评估了各类菌株的消毒剂抗性特征, 具体结果见表3。
革兰氏阴性菌中, 次氯酸钠处理后, 所有菌株KL值均高于参考菌株, 表明其对次氯酸钠的抗性弱于参考菌株。过氧化氢处理的弗氏柠檬酸杆菌和肺炎克雷伯菌的KL值介于大肠杆菌8099与金黄色葡萄球菌ATCC 6538之间, 显示其抗性强于大肠杆菌但弱于金黄色葡萄球菌; 其余4株菌的KL值均超过参考菌株, 即抗性弱于对照菌株。苯扎溴铵处理的产气肠杆菌KL值低于两株参考菌, 呈现最强抗性; 阴沟肠杆菌与大肠杆菌8099抗性相当, 均显著强于金黄色葡萄球菌; 肺炎克雷伯菌抗性表现与过氧化氢处理后抗性一致; 克罗诺杆菌等3株菌抗性弱于参考菌。通过对比3类消毒剂对典型革兰氏阴性菌的杀灭效果发现, 次氯酸钠和过氧化氢表现出卓越的杀菌效能, 其对克罗诺杆菌、溶藻弧菌的KL≥5.00, 达到高水平消毒标准。相比之下, 苯扎溴铵的杀菌效能较低, 其对产气肠杆菌和阴沟肠杆菌的KL仅为2.17和2.82, 尚未达到高效消毒要求(表3)。
革兰氏阳性菌中, 经次氯酸钠溶液和过氧化氢溶液处理后, 所有菌株KL显著高于参考菌株(除粪肠球菌), 抗性整体弱于参考菌株。苯扎溴铵处理后, 单核细胞增生李斯特氏菌和玫瑰考克氏菌抗性最强(KL最小); 粪肠球菌和纹带棒状杆菌抗性介于两个参考菌之间; 藤黄微球菌和鹑鸡肠球菌抗性最弱(KL最大)。强氧化性消毒剂次氯酸钠与过氧化氢对单核细胞增生李斯特氏菌和鹑鸡肠球菌等典型菌株的KL≥5.00。而季铵盐类消毒剂苯扎溴铵的杀菌效能呈现显著分化: 对藤黄微球菌表现出良好杀灭效果(KL为4.45), 但对单核细胞增生李斯特氏菌(KL为1.77)和玫瑰考克氏菌(KL为2.40)明显弱于另外两种消毒剂。
芽孢菌中, 次氯酸钠溶液处理后的拟蕈状芽孢杆菌、蜡样芽孢杆菌和蕈状芽孢杆菌抗性强于参考菌; 枯草芽孢杆菌、人参土芽孢杆菌和地衣芽孢杆菌3株抗性较弱。过氧化氢溶液处理后的蜡样芽孢杆菌和枯草芽孢杆菌呈现较强抗性; 拟蕈状芽孢杆菌、蕈状芽孢杆菌、人参土芽孢杆菌和地衣芽孢杆菌的抗性均弱于参考菌株。苯扎溴铵溶液处理后的蜡样芽孢杆菌、枯草芽孢杆菌、蕈状芽孢杆菌和地衣芽孢杆菌抗性强于参考菌, 拟蕈状芽孢杆菌和人参土芽孢杆菌抗性则弱于参考菌。
3 讨论与结论
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革兰氏阴性菌作为重要的致病菌及条件致病菌, 其有效灭活是公共卫生防控的关键环节[12]。革兰氏阴性菌对化学消毒剂的固有抗性源于其独特的细胞结构及防御机制: (1)外膜屏障系统。由脂多糖(lipopolysaccharide, LPS)构成的疏水性外膜可有效阻隔疏水性消毒剂的渗透[13], 外膜孔蛋白通过分子筛效应选择性排斥大分子及带电物质[14]; (2)动态防御体系。外排泵介导的消毒剂主动外排机制成分[14], 生物膜形成的物理性防护屏障[15-16]以及特异性酶介导的消毒剂降解系统[17]等; (3)遗传调控网络。基因表达调控影响外膜成分合成及抗性相关蛋白表达调控[18]。菌株内部通过多个方面共同调控对化学消毒剂抗性[19]。值得注意的是, 虽都属于革兰氏阴性菌, 不同种属间菌株对消毒剂的敏感性存在显著差异。这种差异性可能由于上述抗性机制的不同而造成。实验数据显示, 强氧化剂(次氯酸钠)通过破坏外膜完整性并氧化细胞内容物实现快速杀菌[20-21], 而过氧化氢则通过羟基自由基攻击DNA及酶系统发挥杀灭作用[22-23], 因此效果更优。季铵盐类消毒剂有效成分属于表面活性剂[24], 消毒效果不如强氧化剂类消毒剂。此外, 微生物生理状态或存在环境也可能影响其对消毒剂的抗性[25]。
革兰氏阳性菌对消毒剂的响应也呈现显著种属特异性, 其对消毒剂抗性机制与阴性菌类似: 包括(1)结构性屏障。厚肽聚糖层构成物理阻隔层, 其三维网状结构可阻滞大分子消毒剂渗透; 细胞壁磷壁酸通过静电作用吸附带正电荷季铵盐类物质, 显著降低其生物利用度; (2)动态防御系统。外排泵介导消毒剂主动外排; 过氧化氢酶等应激酶系可分解特定消毒剂成分; 形成生物膜通过物理阻隔和化学中和双重机制降低消毒剂渗透效率; (3)基因调控系统。而造成不同种属之间对消毒剂抗性差异的原因也是上述机制的不同[26]。
一般而言, 芽孢菌体(即营养细胞)对消毒剂的抗性弱于其对应芽孢, 但是强于非芽孢菌。这是由于芽孢菌的细胞壁通常含有更多交联程度高的肽聚糖层、细胞壁外可能有厚的荚膜或黏液层、细胞膜中有高含量能保持高强度流动性和稳定性的饱和脂肪酸、存在特殊的转运蛋白和泵系统、菌体内多种抗氧化酶、快速调整代谢途径等原因造成的[14,27]。但是由于细胞壁结构、生物膜形成能力、酶系统、外排泵系统以及基因调控等方面的不同, 也会造成不同芽孢菌对消毒剂抗性不同[14]。芽孢所独有的多层结构, 诸如皮质层、芽孢衣等, 如同坚实的堡垒, 能够有效抵御化学消毒剂的渗透。以次氯酸钠和过氧化氢为例, 需要调配更高的浓度, 或者延长其作用时间, 才有可能破坏芽孢的防御体系。
综合实验结果和分析, 本研究中消毒剂效果从强到弱为: 次氯酸钠、过氧化氢、苯扎溴铵, 且不同种属的菌株对消毒剂抗性存在明显差异; 在革兰氏阴性菌、革兰氏阳性菌和芽孢菌中, 均有对消毒剂抗性强于参考菌株的存在。在实际的医疗、卫生防疫与食品加工等诸多领域, 不能一概而论地使用同一种消毒剂及相同浓度、作用时间, 否则很可能出现对部分细菌消杀彻底[28], 却未除去另一些抗性强的细菌[29], 进而引发感染风险或产品污染隐患[30], 所以精准识别与区别对待不同菌株的抗性特点势在必行[31]。在筛选指示微生物时, 应该合理加大筛选范围。
基金
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  • 国家重点研发计划项目(2022YFF1103100)
文献
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2025年第16卷第14期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20250321001
  • 接收时间：2025-03-21
  • 首发时间：2026-01-07
  • 出版时间：2025-07-25
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  • 收稿日期：2025-03-21
基金
国家重点研发计划项目(2022YFF1103100)
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    中国食品药品检定研究院, 北京 100050

通讯作者:

*崔生辉(1972—), 男, 博士, 研究员, 主要研究方向为食品安全检测。E-mail: ;
李景云(1972—), 男, 主任技师, 主要研究方向为食品安全检测。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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