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Article(id=1153433748990316621, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433737141412332, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20250214002, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1739462400000, receivedDateStr=2025-02-14, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752929635522, onlineDateStr=2025-07-19, pubDate=1745510400000, pubDateStr=2025-04-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752929635522, onlineIssueDateStr=2025-07-19, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752929635522, creator=13701087609, updateTime=1752929635522, updator=13701087609, issue=Issue{id=1153433737141412332, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='8', pageStart='1', pageEnd='316', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1752929632696, creator=13701087609, updateTime=1757293087150, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1171735391666225233, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433737141412332, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1171735391666225234, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433737141412332, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=115, endPage=121, ext={EN=ArticleExt(id=1153433749959200862, articleId=1153433748990316621, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Effects of sodium diacetate on the growth and bongkrekic acid production of Burkholderia gladioli in wet rice noodles, columnId=1151895322692776479, journalTitle=Journal of Food Safety & Quality, columnName=Special Topic: Analysis and Monitoring of Toxic and Harmful Substances in Food, runingTitle=null, highlight=null, articleAbstract=

Objective To understand the effect of sodium diacetate on the growth and toxin production of Burkholderia gladioli in fresh wet rice noodles medium. Methods A total of 10 Burkholderia gladioli strains isolated from wet rice noodles samples and rice raw materials and purchased by the strain preservation center were used as research objects. The strains were inoculated into the medium supplemented with different proportions of sodium diacetate to study the growth and toxin production of the strains in the medium. Results The growth of 10 strains of Burkholderia gladioli in BHI broth was consistent. Sodium diacetate had consistent inhibitory effects on different bacterial strains. When the addition of sodium diacetate in BHI broth was more than 0.1%, the growth of 10 strains of Burkholderia gladioli was significantly inhibited, and there was no bacterial growth when the addition reached 0.2%. In BHI semi-solid medium and wet rice noodles medium, when sodium diacetate was added more than 0.1%, the growth of Burkholderia gladioli was significantly weakened or even completely inhibited, and the production of bongkrekic acid was also greatly reduced. Conclusions Sodium diacetate can effectively inhibit the growth of Burkholderia gladioli and reduce the production of bongkrekic acid. It is expected to become a safe and effective preservative or compound preservative that can act on Burkholderia gladioli, reducing the risk of contamination by Burkholderia gladioli at a more affordable cost in the industry.

, correspAuthors=Jing CHEN, 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=Jing-Min HUANG, Jing CHEN, Yan-Hui WU, Le-Shi LI, Mei-Zhu LI, Yu-Zhen JIANG), CN=ArticleExt(id=1153433767671747197, articleId=1153433748990316621, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=双乙酸钠在湿米粉中对唐菖蒲伯克霍尔德氏菌生长及产毒的影响, columnId=1151895322898297380, journalTitle=食品安全质量检测学报, columnName=本期专题：食品中有毒有害物质分析与监测, runingTitle=null, highlight=null, articleAbstract=

目的 了解双乙酸钠对唐菖蒲伯克霍尔德氏菌在实验室培养体系及湿米粉培养体系中的生长及产毒的影响。方法 以10株自湿米粉样品及其大米原料中分离得到的和菌种保藏中心购买的唐菖蒲伯克霍尔德氏菌菌株为研究对象, 接种至添加了不同比例的双乙酸钠的培养基中, 研究其在培养基中生长及产毒情况。结果 10株唐菖蒲伯克霍尔德氏菌在BHI肉汤中生长情况一致。双乙酸钠对不同菌株抑制作用一致, 当BHI肉汤中双乙酸钠添加量>0.1%时, 10株唐菖蒲伯克霍尔德氏菌的生长均明显被抑制, 添加量达到0.2%时, 没有菌生长。在BHI半固体培养基及湿米粉培养基中, 双乙酸钠添加量超过0.1%时, 唐菖蒲伯克霍尔德氏菌的生长显著减弱甚至被完全抑制, 米酵菌酸的产量也大大减少。结论 双乙酸钠能有效抑制唐菖蒲伯克霍尔德氏菌的生长和降低米酵菌酸的产生, 有望成为安全有效且可作用于该菌的防腐剂或复配防腐剂之一, 以行业更容易接受的成本来降低唐菖蒲伯克霍尔德氏菌污染风险。

, correspAuthors=陈晶, authorNote=null, correspAuthorsNote=
* 陈晶(1976—), 女, 高级工程师, 主要研究方向为食品微生物。E-mail:
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黄静敏(1985—), 女, 主要研究方向为食品微生物。E-mail:

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黄静敏(1985—), 女, 主要研究方向为食品微生物。E-mail:

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注: *P<0.05。

, figureFileSmall=mBwBsB8L9+8Qp4+JG6oXWw==, figureFileBig=5/Y93/hm0mcWsTPljiaPWg==, tableContent=null), ArticleFig(id=1171733816805737302, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433748990316621, language=EN, label=Fig.3, caption=Growth and BA production patterns of BGC83756 and LMG1162 in BHI semisolids with varying sodium diacetate concentrations, figureFileSmall=vl9VQxJ+LQdWflbFRU1V5w==, figureFileBig=I4mKSQ44hbTFezndZ0LUxg==, tableContent=null), ArticleFig(id=1171733816868651863, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433748990316621, language=CN, label=图3, caption=分离株BGC83756和标准菌株LMG11626在添加了不同比例双乙酸钠的BHI半固体中的生长情况及BA产生情况, figureFileSmall=vl9VQxJ+LQdWflbFRU1V5w==, figureFileBig=I4mKSQ44hbTFezndZ0LUxg==, tableContent=null), ArticleFig(id=1171733816927372120, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433748990316621, language=EN, label=Table 1, caption=

Information of BGC strains used in the experiment

, figureFileSmall=null, figureFileBig=null, tableContent=
菌株编号 来源 产毒情况 备注
BGC79123 大米 产毒株 自分离, 已测序
BGC83756 湿米粉 产毒株 自分离, 已测序
BGC72074 混合米浆 产毒株 自分离, 已测序
BGC72073 米浆 产毒株 自分离, 已测序
BGC72069 大米 产毒株 自分离, 已测序
BGC15002 米浆 产毒株 自分离, 已测序
BG95947 米浆 非产毒株 自分离, 已测序
BG83139 湿米粉 非产毒株 自分离, 已测序
LMG11626 广东省微生物菌种保藏中心 产毒株 标准菌株
CICC10574 中国工业微生物菌种保藏管理中心 非产毒株 标准菌株
), ArticleFig(id=1171733817002869593, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433748990316621, language=CN, label=表1, caption=

实验用BGC菌株信息

, figureFileSmall=null, figureFileBig=null, tableContent=
菌株编号 来源 产毒情况 备注
BGC79123 大米 产毒株 自分离, 已测序
BGC83756 湿米粉 产毒株 自分离, 已测序
BGC72074 混合米浆 产毒株 自分离, 已测序
BGC72073 米浆 产毒株 自分离, 已测序
BGC72069 大米 产毒株 自分离, 已测序
BGC15002 米浆 产毒株 自分离, 已测序
BG95947 米浆 非产毒株 自分离, 已测序
BG83139 湿米粉 非产毒株 自分离, 已测序
LMG11626 广东省微生物菌种保藏中心 产毒株 标准菌株
CICC10574 中国工业微生物菌种保藏管理中心 非产毒株 标准菌株
), ArticleFig(id=1171733817069978458, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433748990316621, language=EN, label=Table 2, caption=

Results of the generalized estimating equation model for the growth of 10 strains of BGC

, figureFileSmall=null, figureFileBig=null, tableContent=
参数 斜率 标准误差 95%瓦尔德置信区间 假设检验
下限 上限 瓦尔德卡方 自由度 显著性
BG83139 0.209 0.1238 -0.034 0.451 2.843 1 0.092
BG95947 0.139 0.1238 -0.103 0.382 1.268 1 0.260
BGC15002 0.174 0.1238 -0.069 0.416 1.966 1 0.161
BGC72069 0.185 0.1238 -0.058 0.427 2.226 1 0.136
BGC72073 0.087 0.1238 -0.156 0.330 0.493 1 0.483
BGC72074 0.158 0.1238 -0.084 0.401 1.638 1 0.201
BGC79123 0.226 0.1238 -0.017 0.468 3.320 1 0.068
BGC83756 0.124 0.1238 -0.119 0.367 1.004 1 0.316
CICC10574 0.096 0.1238 -0.146 0.339 0.603 1 0.437
LMG11626 0
), ArticleFig(id=1171733817162253147, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433748990316621, language=CN, label=表2, caption=

10株BGC生长情况的广义线性模型结果

, figureFileSmall=null, figureFileBig=null, tableContent=
参数 斜率 标准误差 95%瓦尔德置信区间 假设检验
下限 上限 瓦尔德卡方 自由度 显著性
BG83139 0.209 0.1238 -0.034 0.451 2.843 1 0.092
BG95947 0.139 0.1238 -0.103 0.382 1.268 1 0.260
BGC15002 0.174 0.1238 -0.069 0.416 1.966 1 0.161
BGC72069 0.185 0.1238 -0.058 0.427 2.226 1 0.136
BGC72073 0.087 0.1238 -0.156 0.330 0.493 1 0.483
BGC72074 0.158 0.1238 -0.084 0.401 1.638 1 0.201
BGC79123 0.226 0.1238 -0.017 0.468 3.320 1 0.068
BGC83756 0.124 0.1238 -0.119 0.367 1.004 1 0.316
CICC10574 0.096 0.1238 -0.146 0.339 0.603 1 0.437
LMG11626 0
), ArticleFig(id=1171733817262916444, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433748990316621, language=EN, label=Table 3, caption=

Growth and BA production patterns of BGC83756 and LMG1162 in wet rice noodles with varying sodium diacetate concentrations

, figureFileSmall=null, figureFileBig=null, tableContent=
样品添加量 菌株
BGC83756 LMG11626
菌落数/(CFU/g) BA含量/(μg/kg) 菌落数/(CFU/g) BA含量/(μg/kg)
湿米粉 3.0×108 30533±893 2.6×108 45039±920
0.1%双乙酸钠 <10 ND <10 ND
0.2%双乙酸钠 <10 ND <10 ND
0.3%双乙酸钠 <10 ND <10 ND
0.4%双乙酸钠 <10 ND <10 ND
), ArticleFig(id=1171733817334219613, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433748990316621, language=CN, label=表3, caption=

分离株BGC83756和标准菌株LMG11626在添加了不同比例双乙酸钠的湿米粉中的生长情况及BA产生情况

, figureFileSmall=null, figureFileBig=null, tableContent=
样品添加量 菌株
BGC83756 LMG11626
菌落数/(CFU/g) BA含量/(μg/kg) 菌落数/(CFU/g) BA含量/(μg/kg)
湿米粉 3.0×108 30533±893 2.6×108 45039±920
0.1%双乙酸钠 <10 ND <10 ND
0.2%双乙酸钠 <10 ND <10 ND
0.3%双乙酸钠 <10 ND <10 ND
0.4%双乙酸钠 <10 ND <10 ND
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双乙酸钠在湿米粉中对唐菖蒲伯克霍尔德氏菌生长及产毒的影响
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黄静敏 1 , 陈晶 1, * , 吴燕蕙 1 , 李乐诗 1 , 李媚竹 2 , 蒋玉珍 2
食品安全质量检测学报 | 本期专题：食品中有毒有害物质分析与监测 2025,16(8): 115-121
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食品安全质量检测学报 | 本期专题：食品中有毒有害物质分析与监测 2025, 16(8): 115-121
双乙酸钠在湿米粉中对唐菖蒲伯克霍尔德氏菌生长及产毒的影响
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黄静敏1 , 陈晶1, * , 吴燕蕙1, 李乐诗1, 李媚竹2, 蒋玉珍2
作者信息
  • 1.深圳市计量质量检测研究院, 深圳 518100
  • 2.华南农业大学食品学院, 广州 510642

    • 黄静敏(1985—), 女, 主要研究方向为食品微生物。E-mail:

通讯作者:

* 陈晶(1976—), 女, 高级工程师, 主要研究方向为食品微生物。E-mail:
Effects of sodium diacetate on the growth and bongkrekic acid production of Burkholderia gladioli in wet rice noodles
Jing-Min HUANG1 , Jing CHEN1, * , Yan-Hui WU1, Le-Shi LI1, Mei-Zhu LI2, Yu-Zhen JIANG2
Affiliations
  • 1. Shenzhen Academy of Metrology & Quality Inspection, Shenzhen 518100, China
  • 2. College of Food Science, South China Agricultural University, Guangzhou 510642, China
出版时间: 2025-04-25 doi: 10.19812/j.cnki.jfsq11-5956/ts.20250214002
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摘要
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目的 了解双乙酸钠对唐菖蒲伯克霍尔德氏菌在实验室培养体系及湿米粉培养体系中的生长及产毒的影响。方法 以10株自湿米粉样品及其大米原料中分离得到的和菌种保藏中心购买的唐菖蒲伯克霍尔德氏菌菌株为研究对象, 接种至添加了不同比例的双乙酸钠的培养基中, 研究其在培养基中生长及产毒情况。结果 10株唐菖蒲伯克霍尔德氏菌在BHI肉汤中生长情况一致。双乙酸钠对不同菌株抑制作用一致, 当BHI肉汤中双乙酸钠添加量>0.1%时, 10株唐菖蒲伯克霍尔德氏菌的生长均明显被抑制, 添加量达到0.2%时, 没有菌生长。在BHI半固体培养基及湿米粉培养基中, 双乙酸钠添加量超过0.1%时, 唐菖蒲伯克霍尔德氏菌的生长显著减弱甚至被完全抑制, 米酵菌酸的产量也大大减少。结论 双乙酸钠能有效抑制唐菖蒲伯克霍尔德氏菌的生长和降低米酵菌酸的产生, 有望成为安全有效且可作用于该菌的防腐剂或复配防腐剂之一, 以行业更容易接受的成本来降低唐菖蒲伯克霍尔德氏菌污染风险。

双乙酸钠  /  唐菖蒲伯克霍尔德氏菌  /  椰毒假单胞菌  /  湿米粉  /  米酵菌酸

Objective To understand the effect of sodium diacetate on the growth and toxin production of Burkholderia gladioli in fresh wet rice noodles medium. Methods A total of 10 Burkholderia gladioli strains isolated from wet rice noodles samples and rice raw materials and purchased by the strain preservation center were used as research objects. The strains were inoculated into the medium supplemented with different proportions of sodium diacetate to study the growth and toxin production of the strains in the medium. Results The growth of 10 strains of Burkholderia gladioli in BHI broth was consistent. Sodium diacetate had consistent inhibitory effects on different bacterial strains. When the addition of sodium diacetate in BHI broth was more than 0.1%, the growth of 10 strains of Burkholderia gladioli was significantly inhibited, and there was no bacterial growth when the addition reached 0.2%. In BHI semi-solid medium and wet rice noodles medium, when sodium diacetate was added more than 0.1%, the growth of Burkholderia gladioli was significantly weakened or even completely inhibited, and the production of bongkrekic acid was also greatly reduced. Conclusions Sodium diacetate can effectively inhibit the growth of Burkholderia gladioli and reduce the production of bongkrekic acid. It is expected to become a safe and effective preservative or compound preservative that can act on Burkholderia gladioli, reducing the risk of contamination by Burkholderia gladioli at a more affordable cost in the industry.

sodium diacetate  /  Burkholderia gladioli  /  Pseudomonas cocovenenans  /  wet rice noodles  /  bongkrekic acid
黄静敏, 陈晶, 吴燕蕙, 李乐诗, 李媚竹, 蒋玉珍. 双乙酸钠在湿米粉中对唐菖蒲伯克霍尔德氏菌生长及产毒的影响. 食品安全质量检测学报, 2025 , 16 (8) : 115 -121 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250214002
Jing-Min HUANG, Jing CHEN, Yan-Hui WU, Le-Shi LI, Mei-Zhu LI, Yu-Zhen JIANG. Effects of sodium diacetate on the growth and bongkrekic acid production of Burkholderia gladioli in wet rice noodles[J]. Journal of Food Safety & Quality, 2025 , 16 (8) : 115 -121 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250214002
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0 引言
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唐菖蒲伯克霍尔德氏菌[椰毒假单胞菌酵米面亚种, Burkholderia gladioli (Pseudomonas cocovenenans subsp. Farinofermentans, BGC)]为革兰氏阴性无芽胞短小杆菌, 兼性厌氧, 在自然界分布广泛, 大多来源于土壤, 可能会随食物原料而污染食品。BGC不耐高温, 很容易被杀死, 但其代谢产物米酵菌酸(bongkrekic acid, BA)耐热性很强, 120 ℃高温处理1 h仍可保持毒性[1], 可引起食物中毒甚至死亡, 是我国微生物因素食源性疾病的首要死亡原因[2-5]。
我国BA中毒事件之前主要集中在发酵米面制品及其他变质淀粉类制品等食品[6-8], 主要是家庭自制。然而, 2018年广东省首次报道了因食用鲜湿米粉类食品引起的BA中毒事件[9-10]。区别于以往的中毒事件, 鲜湿米粉是工厂标准化批量生产的加工食品, 是我国一种产销量巨大的大众食品, 这种风险的严重性引起了社会和监管部门的高度重视。
湿米粉中BGC污染溯源研究表明, 其主要传播途径呈现多环节特征: (1)原料污染。稻米等原料在种植、采收过程中接触含菌土壤[11], 未彻底清洗的原料成为初级污染源[12-13]; (2)生产污染。加工设备由其是冷却成型车间环境及设施(含风扇、风口)及操作人员交叉接触导致的二次污染; (3)储运增殖。产品在流通环节中室温或较高温度存放(大于26 ℃持续4 h)会显著加速菌体繁殖[14]。湿米粉含水量高与pH (4.0~8.5)恰好位于BGC最适生长区间[15], 为微生物增殖创造理想基质条件。
鲜湿米粉的品质控制关键在于防止BGC的生长, 研究表明, 低温保存可有效抑制BGC生长[16-17]。但目前湿米粉行业已经存在冷链运输和存储难以推广的行业现实, 显示情形中, 由于食品行业冷链物流尚不完善且成本较高, 因此河粉等湿米粉制品的生产经营者更倾向于使用防腐剂以达到常温状态下延长货架期的目的。为解决实际需求, 需寻找可作用于BGC的防腐剂, 探索多手段结合的控制措施, 以行业更容易接受的成本提高风险管控水平, 具有重要的现实意义。
双乙酸钠是一种新型的高效低毒防腐、防霉和保鲜剂, 在食品和饮料等领域广泛应用[18-19]。双乙酸钠是继富马酸酯类和丙酸盐类之后开发研制和应用的粮食、食品及饲料防霉剂的新品[20-23]。在欧美和日本等发达国家已得到普遍使用, 世界卫生组织(World Health Organization, WHO)和联合国粮农组织(Food and Agriculture Organization of the United Nations, FAO)已批准双乙酸钠在谷物食品中作防霉保鲜剂使用[24], 1989年我国正式批准将其作粮食、食品和饲料等物质的防腐剂、防霉剂和保鲜剂。根据GB 2760—2024《食品安全国家标准 食品添加剂使用标准》, 双乙酸钠在食品中的最大使用量因食品类别而异。例如, 在原粮中最大使用量为1.0 g/kg, 粉圆中为4.0 g/kg, 糕点中为2.5 g/kg。
本研究以自大米原料和湿米粉样品中分离到的BGC为研究对象, 研究双乙酸钠对鲜湿粉在贮藏过程中BGC的抑制效果, 以期为降低鲜湿粉被BGC污染风险提供参考。
1 材料与方法
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1.1 材料与试剂
从大米原料和湿米粉样品中自行分离的6株产毒株、2株非产毒株和2株菌株BGC标准菌株, 见表1。
双乙酸钠(化学纯, 天津市大茂化学试剂厂); BHI营养肉汤(北京陆桥技术有限责任公司); 血平板(郑州安图生物工程股份有限公司); BA标准品(Lot: 2358211, 500 mg/L甲醇, 1.2 mL, ANPEL上海安谱璀世公司); 大米为市购。
1.2 仪器与设备
DENSIMAT比浊仪(法国梅里埃公司); QUINTIX313-1CN千分之一电子天平(德国赛多利斯公司); GER IRM恒温恒湿培养箱、SCIEXTQ4500液相色谱串联质谱仪(爱尔兰沃特斯中国有限公司)。
1.3 方法
1.3.1 菌液制备
取出菌株冻干粉置于10 mL营养肉汤中, 36 ℃活化12 h。再划线到血平板上, 36 ℃培养24 h。挑取单菌落, 用5 mL无菌生理盐水制成菌悬液, 调菌液浓度为1.0 MCF, 作为接种菌液。
1.3.2 培养基制备
实验液体培养基: 按比例分别在BHI肉汤中加入0.1%、0.2%、0.4%、0.6%的双乙酸钠, 灭菌备用。
BHI半固体: 每100 mL BHI肉汤中加入0.5 g琼脂粉制成。
湿米粉培养基: 大米浸泡磨浆后(1.5 kg大米/3 kg水), 按米浆的量分别称取0.1%、0.2%、0.3%、0.4%的双乙酸钠, 边搅拌边向培养皿中加入25 mL混合米浆, 于蒸锅中蒸煮10 min, 再置于灭菌锅中121 ℃灭菌15 min, 冷却备用。
1.3.3 液体培养
取1 mL 1.0 MCF(约107)的菌悬液分别接种至100 mL BHI肉汤及添加了不同比例双乙酸钠的实验液体培养基, 37 ℃培养48 h, 按不同培养时间点取出300 μL培养物置于酶标仪上测定菌液的OD600值。
1.3.4 半固体培养
取100 μL 1.0 MCF(约107)的菌悬液分别涂布到BHI半固体培养基及添加了不同比例双乙酸钠的实验半固体培养基(每个平板25 mL培养基), 26 ℃培养 5 d, 进行菌落总数及BA测定, 菌落总数的测定按照 GB 4789.2—2022《食品安全国家标准 食品微生物学检验 菌落总数测定》进行[15], BA的测定按照GB 5009.189—2023《食品安全国家标准 食品中米酵菌酸的测定》进行[16]。
1.3.5 湿米粉培养基培养
取100 μL 1.0 MCF(约107)的菌悬液分别涂布到湿米粉培养基及添加了不同比例双乙酸钠的湿米粉培养基, 26 ℃培养 5 d, 取样时间分别为1、2、3、4、5 d。进行菌落总数及BA测定。
1.4 数据处理
数据图片均采用Excel 2010进行处理。数据统计采用SPSS 26进行, 采用单因素方差分析和广义估计方程进行统计分析, 以校正时间相关性对结果的影响。
2 结果与分析
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2.1 不同菌株在BHI肉汤中的生长情况
对6株分离产毒、2株非产毒的BGC以及2株标准菌株进行生长曲线的测定, 结果见图1。不产毒菌株CICC10574、BG95947、BG83139与产毒菌株LMG11626、BGC79123、BGC72074、BGC72069、BGC15002在培养0~4 h内生长缓慢, 处于生长迟缓期。4~16 h为生长对数期, 生长迅速。16 h后进入稳定期并持续到30 h, 30 h后逐渐进入衰亡期。不产毒菌株CICC10574、BG95947、BG83139与产毒菌株BGC79123、BGC72074、BGC72069、BGC15002在生长趋势上无明显差异。但产毒株BGC83756、BGC72073 4 h后进入对数期, 一直持续到24 h, 24~42 h一直处于稳定期, 42 h后逐渐进入衰亡期。
通过广义估计方程对不同菌株生长情况进行统计分析, 不同菌株在BHI肉汤中生长差异不具有统计学意义(P>0.05)(表2)。总体而言, 几株产毒株和非产毒株或标准菌株和分离株的生长曲线均符合微生物典型的生长趋势, 虽个别菌株对数生长期持续时间有不同, 稳定期也稍有长短, 但总体生长情况一致。
2.2 在添加了不同比例双乙酸钠的实验液体培养基的生长情况
把6株分离产毒、2株非产毒的BGC以及2株标准菌株接种到添加了不同比例双乙酸钠的实验液体培养基中, 37 ℃培养48 h, 观察菌株生长情况。
双乙酸钠对8株分离株以及2株标准菌株的生长菌均有明显的抑制作用, 如图2所示。0.1%添加量时, 10株BGC生长显著减少, OD值减少70%以上; 当添加量达到0.2%以上时, 10株BGC在BHI肉汤中基本都不生长。对双乙酸钠比例和不同菌株两个因素进行检验, 发现双乙酸钠对菌株的抑制作用具有统计学意义(P<0.05), 而不同菌株对双乙酸钠的响应没有差异, 由此可见, 双乙酸钠可有效抑制BG生长, 且对不同菌株抑制作用一致。
2.3 BGC在添加了不同比例双乙酸钠的半固体培养基中的生长情况及BA产生情况
由2.1和2.2结果可见, 无论是产毒株或非产毒株, 还是分离株或标准菌株, 10株BGC在同等培养条件下, 生长情况没有显著差异, 故选取1株分离株BGC83756和1株标准菌株LMG11626接种到添加了不同比例双乙酸钠的半固体培养基中, 26 ℃培养 5 d, 后续观察菌株生长情况及BA产生情况。
结果见图3, 两株菌在BHI半固体中均生长良好, 培养后可达到108 CFU/g以上, BGC83756可产生26950 μg/kg的BA, LMG11626 BA的量可达到30214 μg/kg。当在半固体培养基中添加0.1%双乙酸钠时, 两株菌的生长均受到抑制, 菌量减少了两个数量级, 但BA的产量显著减少, 仅约1000 μg/kg, 下降了95%以上, 说明双乙酸钠可有效抑制毒素的产生。当双乙酸钠添加量达到0.2%时, BGC83756生长微弱, LMG11626生长被完全抑制, 此时两株菌均检测不到BA的产生。当双乙酸钠添加量达到0.4%时, BGC83756和LMG11626都没有生长。
2.4 BGC在添加了不同比例双乙酸钠的湿米粉培养基中的生长情况及BA产生情况
选取分离株BGC83756和标准菌株LMG11626接种到添加了不同比例双乙酸钠的湿米粉培养基中, 26 ℃培养5 d, 观察菌株生长情况及BA产生情况。由表3可见, 未添加双乙酸钠的湿米粉培养基上, BGC生长良好, 肉眼可见大量菌落, 湿米粉培养基全部变黄, 最终菌落数可达108以上, BA的浓度均可达30000 μg/kg以上。当双乙酸钠添加量大于0.1%时, 湿米粉培养基上没有菌落生长, 颜色没有变化, 也没有检测出BA。
3 讨论与结论
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鲜湿米粉是以大米为主要原料, 营养丰富, 水分高达60%以上, 非常适合微生物生长, 研究表明在常温条件下鲜湿米粉的保质期只有1~2 d[25]。本研究结果显示, 10株BGC在同等培养条件下, 生长差异不具统计学意义, 一般4 h后就开始快速生长, 12 h后菌量接近最高值。由前期调查结果可知, 鲜湿米粉从生产, 获得出厂证明, 然后运输到销售或经营者手里, 大概就需要6~8 h, 要是跨市的话, 时间更长。由此可见, 要是储运条件不当, 湿米粉到达消费者手里时, 已存在较高风险。低温条件下储运对BGC生长的抑制作用明显, 但从产品品质角度来说, 企业普遍反应产品冷藏后容易断条, 影响产品品质, 商家会因为冷链的米粉感官变差而拒绝接受产品。因此, 为了延长鲜湿米粉的货架期, 生产企业更偏向于添加一些食品添加剂, 常见的是脱氢乙酸钠。然而关于河源BA中毒事件流行病学调研结果表明: 河粉因超范围使用了脱氢乙酸钠, 常温存放超过3 d仍无肉眼可见的霉变、无明显酸馊气味, 反而掩盖了河粉已变质的问题[9]。脱氢乙酸及其钠盐是我国使用较为广泛的防霉剂[26-27], 但已有研究表明, 脱氢乙酸钠不能有效抑制BGC的生长及BA的产生[16]。2025年实施的GB 2760—2024中规定大米制品及淀粉制品均不能添加脱氢乙酸钠, 但现有调查仍发现广州市河粉等大米制品超范围使用脱氢乙酸及其钠盐的问题十分突出[28]。可见, 不少生产企业法律意识薄弱或没有充分意识到BGC污染的风险, 在没有其他安全有效的保鲜剂可选择的情况下, 仍然选择违规添加脱氢乙酸钠。
双乙酸钠是一种新型的高效低毒防霉、防腐和保鲜剂。GB 2760—2024中规定双乙酸钠允许使用于原粮、粉圆和糕点等食品。现研究表明其显著的微生物抑制效果: 刘壮等[29]通过实验探究发现, 双乙酸钠可显著降低鲜湿米粉条霉菌总数, 并有效改善产品外观、色泽及风味特性; 张灿[30]对马铃薯鲜湿粉条的保鲜研究表明, 双乙酸钠对细菌和霉菌生长均具有显著抑制作用, 且不影响产品感官品质。本研究通过培养基体系实验发现, 当双乙酸钠添加量大于0.1%时, 在37 ℃条件下对10株BGC的生长有明显的抑制作用, 且各菌株抑制效果呈现高度一致性。在26 ℃模拟贮藏条件下, 同样有效抑制BA毒素的生物合成; 值得注意的是, 在模拟湿米粉基质体系中, 添加量大于0.1%双乙酸钠即可完全抑制BGC生长, 且BA生成量低于检出限。尽管现行国家标准尚未批准双乙酸钠应用于大米制品及淀粉制品(包括湿米粉), 但其对BGC菌株及毒素合成的双重抑制作用, 为开发安全高效的鲜湿米粉保质技术提供了理论依据。
然而需要指出的是, 本研究的实验体系基于培养基条件, 而实际湿米粉生产过程中涉及的复杂基质参数(如pH、水分活度及其他组分)可能对双乙酸钠的抑菌效能产生显著影响。虽然实验数据表明0.1%添加量即可实现完全抑制, 但该浓度对产品感官特性(包括质构特性、风味特征等)的影响仍需通过系统评价进行验证。双乙酸钠在湿米粉中的应用需解决两个关键问题: 一是法规限制(当前未获批用于大米制品); 二是添加后对产品品质的影响。未来研究可探索复配防腐剂(如与乳酸钠结合)以降低单一添加量, 或通过工艺优化减少对米粉质构的影响。此外, 推动双乙酸钠在大米制品中的合规性申报是实际应用的前提。
随着食品安全监管体系对BGC污染风险防控的持续强化, 以及消费者安全意识的不断提升, 预期将严格规范脱氢乙酸钠等添加剂的非法使用行为。在此背景下, 双乙酸钠凭借其特异性抑制BGC及控制BA毒素生成的特性, 有望发展成为符合行业成本控制需求的新型防腐解决方案, 为鲜湿米粉安全生产提供技术支撑。
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  • 深圳市科技计划项目(KCXFZ20211020165404007)
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2025年第16卷第8期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20250214002
  • 接收时间：2025-02-14
  • 首发时间：2025-07-19
  • 出版时间：2025-04-25
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  • 收稿日期：2025-02-14
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深圳市科技计划项目(KCXFZ20211020165404007)
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    1.深圳市计量质量检测研究院, 深圳 518100
    2.华南农业大学食品学院, 广州 510642

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* 陈晶(1976—), 女, 高级工程师, 主要研究方向为食品微生物。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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