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调味糖浆中防腐剂抗性及活的非可培养状态的形成

Resistance to preservatives and the viable but non-culturable state formation of in flavored syrups.

作者信息

Wen Xia, Chen Yiwen, Zhang Shuyao, Su Ai-Ting, Huang Di, Zhou Gang, Xie Xiaobao, Wang Jufang

机构信息

School of Biology and Biological Engineering, South China University of Technology, Guangzhou, China.

Guangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, South China University of Technology, Guangzhou, China.

出版信息

Front Microbiol. 2024 Feb 16;15:1345800. doi: 10.3389/fmicb.2024.1345800. eCollection 2024.

Abstract

Food security is a crucial issue that has caused extensive concern, and the use of food flavors has become prevalent over time. we used the molecular biological techniques, preservative susceptibility testing, viable but non-culturable (VBNC) state induction testing, and a transcriptome analysis to examine the bacterial contamination of favored syrup and identify the causes and develop effective control measures. The results showed that WLS1-1 is a microorganism that can spoil food and is a member of the acetic acid bacteria families. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) tests showed that WLS1-1 was susceptible to potassium sorbate (PS), sodium benzoate (SB), and sodium sulffte (SS) at pH 4.0. It revealed a progressive increase in resistance to these preservatives at increasing pH values. WLS1-1 was resistant to PS, SB and SS with an MIC of 4.0, 2.0 and 0.5  g/L at pH 5.0, respectively. The MIC values exceed the maximum permissible concentrations that can be added. The induction test of the VBNC state demonstrated that WLS1-1 lost its ability to grow after 321 days of PS induction, 229  days of SB induction and 52  days of SS induction combined with low temperature at 4°C. Additionally, laser confocal microscopy and a propidium monoazide-quantitative polymerase chain reaction (PMA-qPCR) assay showed that WLS1-1 was still alive after VBNC formation. There were 7.192 ± 0.081 (PS), 5.416 ± 0.149 (SB) and 2.837 ± 0.134 (SS) log(CFU/mL) of viable bacteria. An analysis of the transcriptome data suggests that Asaia lannensis can enter the VBNC state by regulating oxidative stress and decreasing protein synthesis and metabolic activity in response to low temperature and preservatives. The relative resistance of Asaia lannensis to preservatives and the induction of the VBNC state by preservatives are the primary factors that contribute to the contamination of favored syrup by this bacterium. To our knowledge, this study represents the first evidence of the ability of to enter the VBNC state and provides a theoretical foundation for the control of organisms with similar types of activity.

摘要

食品安全是一个引起广泛关注的关键问题,随着时间的推移,食用香料的使用已变得普遍。我们运用分子生物学技术、防腐剂敏感性测试、活的非可培养(VBNC)状态诱导测试以及转录组分析,来检测调味糖浆的细菌污染情况,确定其成因并制定有效的控制措施。结果表明,WLS1-1是一种能够 spoilage 食品的微生物,属于醋酸菌科。最低抑菌浓度(MIC)和最低杀菌浓度(MBC)测试表明,在pH 4.0时,WLS1-1对山梨酸钾(PS)、苯甲酸钠(SB)和亚硫酸钠(SS)敏感。随着pH值升高,其对这些防腐剂的抗性逐渐增加。在pH 5.0时,WLS1-1对PS、SB和SS具有抗性,MIC分别为4.0、2.0和0.5 g/L。这些MIC值超过了可添加的最大允许浓度。VBNC状态诱导测试表明,在4°C低温下,经PS诱导321天、SB诱导229天和SS诱导52天后,WLS1-1失去了生长能力。此外,激光共聚焦显微镜和单叠氮碘化丙啶定量聚合酶链反应(PMA-qPCR)检测表明,VBNC形成后WLS1-1仍然存活。活菌数分别为7.192±0.081(PS)、5.416±0.149(SB)和2.837±0.134(SS)log(CFU/mL)。转录组数据分析表明,罗伊氏醋酸菌可通过调节氧化应激、降低蛋白质合成和代谢活性,以应对低温和防腐剂从而进入VBNC状态。罗伊氏醋酸菌对防腐剂的相对抗性以及防腐剂对VBNC状态的诱导是导致该细菌污染调味糖浆的主要因素。据我们所知,本研究首次证明了其进入VBNC状态的能力,并为控制具有类似活性类型的生物体提供了理论基础。 (注:原文中“spoilage”有误,推测应为“spoil”;“favored syrup”推测应为“flavored syrup” ;“Asaia lannensis”推测应为“Acetobacter lannensis” ,已按推测修改后翻译,供参考。)

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d11/10904602/3fe71e4b8e5d/fmicb-15-1345800-g001.jpg

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