食品加工环境胁迫因素对单核细胞增生李斯特菌生物膜形成的影响研究进展

单核细胞增生李斯特菌（以下简称单增李斯特菌）生物膜的生长可导致反复的食物污染。食品加工储藏常用的冷藏、干燥、酸处理以及消毒剂处理等使微生物长期处于胁迫环境下，对生物膜的形成产生影响。本文总结了常见的食品加工胁迫因素对单增李斯特菌生物膜形成的影响，其中重点介绍消毒剂处理对单增李斯特菌生物膜形成的影响，同时从膜流动性相关的适应策略、生物膜形成相关蛋白和基因调控表达的角度阐述胁迫条件下单增李斯特菌生物膜的形成机制。胁迫环境下单增李斯特菌生物膜形成的研究有助于揭示真实环境下其生物膜的形成及变化规律；充分考虑环境因素设定清洁、消毒标准有利于降低食源性致病菌传播的潜在风险。     

大肠杆菌在食品加工贮藏中胁迫响应机制的研究进展

大肠杆菌能够感受环境信号并对环境的变化迅速做出反应。因此,在食品加工贮藏中,大肠杆菌在面对物理、化学因子胁迫时会产生应激反应,使其仍然能够生存和保持毒力,给食品安全带来极大的威胁。本文总结了在常见的食品加工贮藏胁迫因子下,包括热激、冷激、干燥、高渗透压、抗菌肽和酸,大肠杆菌的分子及生理响应机制及其在食品工业中的应用,并对大肠杆菌胁迫响应的未来研究做出展望。     

单核细胞增生李斯特菌生物被膜交叉污染评估进展

单核细胞增生李斯特菌（Listeria monocytogenes）（以下简称单增李斯特菌）是一种引发李斯特菌病罹患者高住院率和高死亡率的食源性致病菌,其可在冷、热、干燥和消毒剂处理等不利条件下黏附于食品接触表面并进一步形成难以清除的生物被膜。交叉污染是单增李斯特菌传播的主要途径,生物被膜的形成提高了单增李斯特菌在工厂和厨房环境持续传播和污染的可能性,可引发相关食源性疾病暴发和食品召回等,从而造成健康和经济损失。本文首先介绍了单增李斯特菌生物被膜的胞外聚合物组分,并从外部生存环境和内部微生物自身因素两方面总结了影响单增李斯特菌生物被膜交叉污染转移的因素;进一步重点从研究类型和细菌收集两方面阐述了生物被膜交叉污染的相关研究进展;最后,归纳总结了针对单增李斯特菌生物被膜形成早期的防控策略,展望该领域的研究前景,以期为科学评估和早期精准防控单增李斯特菌生物被膜交叉污染的潜在风险提供理论依据。     

单核细胞增生李斯特菌营养胁迫表型特征及调控机制

单核细胞增生李斯特菌(单增李斯特菌)能引发侵袭性李斯特菌病，是一类可导致严重后果的食源性致病菌。单增李斯特菌广泛分布于环境复杂多变的食品供应链各环节中，其通过调整自身生理状态以应对营养缺乏等胁迫条件带来的生存挑战，继而对食品安全造成威胁。本文重点介绍营养胁迫条件下单增李斯特菌表型特征和可能的调控机制，包括生物膜形成能力、抗性和毒性，以及σB、(p)ppGpp和sRNA等相关调控因子，以便了解单增李斯特菌营养胁迫下的抗逆生存机制，为进一步开展相关精准防控工作提供参考。     

沙门氏菌和单增李斯特菌诱导性耐酸响应机制的研究进展

沙门氏菌和单增李斯特菌被认为是肉制品中最重要的食源性致病菌。它们在弱酸环境下会发生强烈的诱导性耐酸响应,同时诱导产生高毒、耐酸、耐渗透压的高危菌株,是影响消费者健康安全的重大隐患。本文主要从沙门氏菌和单增李斯特菌产生诱导耐酸的发现过程、诱导耐酸响应的危害、产生诱导耐酸的影响因素方面进行概述,进一步从pH值稳态系统、应激蛋白分子的调控及细胞膜组成和流动性调控的角度分析了产生诱导耐酸响应的分子机制。     

乳酸菌的热胁迫研究进展

乳酸菌和其他自由生活的微生物一样,在发酵及食品加工、贮藏过程中经常暴露于各种环境胁迫条件下,包括饥饿胁迫、渗透压胁迫以及热胁迫等,增加胁迫抗性是提高生物量及代谢产物积累的有效策略。热胁迫可能是自然界及工业应用中微生物所面临的最常见压力。目前研究表明来自不同生境的乳酸菌通过基因表达的快速变化对温度的突然升高做出反应,从而导致热休克蛋白的蛋白质水平升高。热休克蛋白在进化过程中具有高度保守性,在正常条件下,其有助于受体调节,细胞骨架稳定及蛋白质的折叠、组装、运输、降解。在热胁迫条件下,其功能变得尤为重要。综述了乳酸菌的热胁迫耐受机制及不同乳酸菌的热胁迫反应,为研究乳酸菌热胁迫应答机制提供一定的理论借鉴,有利于乳酸菌的工业化应用。     

食品中亚致死损伤单增李斯特菌的研究进展

单核细胞增生李斯特菌（Listeria monocytogenes）是一类人畜共患的食源性致病菌。食品加工过程中所产生的亚致死损伤单增李斯特菌是不容忽视的,在适宜的环境下,损伤菌会恢复至正常状态继续生长,对消费者的健康造成威胁,因此亚致死损伤菌的存在是食品安全的一大隐患。探究亚致死损伤菌的检测、修复是目前研究的热点之一,本文将综合相关研究对近年来食品中亚致死损伤单增李斯特菌的检测、修复方法以及发展趋势进行综述。     

双组分调控系统及其对细菌诱导性耐酸响应调控机理的研究进展

双组分调控系统（two-component regulatory system,TCS）是维持细菌在压力环境中存活的重要结构。食品加工过程极易产生高渗、弱酸等压力环境,双组分系统能够帮助细菌感受外部环境的胁迫,及时动员体内对抗机制,这一过程容易产生耐酸、耐渗透压、耐高温甚至是高毒性的菌株,威胁食品安全。本文主要对TCS的结构组成、识别信号及调控作用等进行概述,并与细菌诱导耐酸响应（acid tolerance response,ATR）中的酸休克蛋白、细胞膜系统和氨基酸代谢等产生机制进行联系,综述了细菌在酸性条件下通过TCS响应信号分子激发ATR的具体过程。     

乳酸菌细菌素Durancin GL对单增李斯特菌的抗菌活性及机制

单增李斯特菌广泛分布于肉类、禽类、蛋类、乳制品及蔬菜中,且适应能力强,即使在4 ℃的冷藏环境下仍可生长繁殖,是食品中主要的食源性致病菌之一。乳酸菌细菌素Durancin GL是由干酪中肠球菌产生的一种新型细菌素,对单增李斯特菌具有靶向抑制作用。本实验研究了Durancin GL对单增李斯特菌的抗菌活性及作用机制。通过最小抑菌浓度和抑菌动力学实验检测Durancin GL对单增李斯特菌的抑制作用,结合监测胞内物质泄漏、菌体存活情况以及形态学分析,探讨Durancin GL对单增李斯特菌的抑菌机制。Durancin GL对单增李斯特菌最小抑菌浓度为（2.5±0.4）mg/L,可引起李斯特菌细胞质泄漏,增加细胞外液电导率,导致菌体细胞死亡,从而发挥其抑菌活性。     

单增李斯特菌鞭毛蛋白提取方法的研究

本实验对单增李斯特菌鞭毛蛋白提取方法进行研究,分别在23℃和37℃的条件下对单增李斯特菌(血清型IVb)进行扩增培养,发现23℃下单增李斯特菌产鞭毛的能力更强。运用酸解法处理单增李斯特菌后,分别通过超速离心法和硫酸铵沉淀法对鞭毛蛋白进行纯化,SDS-PAGE 电泳以及电镜的实验结果表明,超速离心较硫酸铵沉淀法简便、耗时短、纯度高、产量高,适用于单增李斯特菌鞭毛蛋白的提取。     

Heat and acid tolerance of Listeria monocytogenes after exposure to single and multiple sublethal stresses

The majority of published studies on the adaptive heat or acid tolerance response of Listeria monocytogenes have been performed with a single strain exposed to a single adaptation treatment; however, in food ecosystems, microorganisms commonly exist as multi-species communities and encounter multiple stresses, which may result in "stress hardening". Therefore, the present study evaluated the adaptive responses to heat (52, 57 and 63 degrees C) or lactic acid (pH 3.5) of a 10-strain composite of L. monocytogenes meat and human isolates at stationary phase, following exposure to combinations of osmotic (10% NaCl), acidic (pH 5.0 with HCl) and thermal (T; 46 degrees C) stresses, sequentially or simultaneously within 1.5h, in tryptic soy broth with 0.6% yeast extract (TSBYE). All treatments induced adaptive responses on L. monocytogenes at 57 degrees C, while no such cross-protection was observed at 52 and 63 degrees C. Survivor curves at 57 degrees C appeared convex with profound shoulders determined by a Weibull model. The highest thermotolerance was observed after combined exposure to acid and heat shock (pH-T), followed by exposure to osmotic shock, and by the combination of osmotic with heat shock (NaCl-T). Regarding acid tolerance, prior exposure to low pH, pH-T, or a combination of NaCl, pH and T resulted in a marked increase of resistance to pH 3.5, showing concave inactivation curves with tails at higher levels of survivors (log(10)CFU ml(-1)) than the control cultures. The sequence of exposure to sublethal stresses did not affect the thermotolerance of L. monocytogenes, whereas simultaneous exposure to most multiple stresses (e.g., NaCl-pH-T, NaCl-T and NaCl-pH) resulted in higher survivors of L. monocytogenes at pH 3.5 than exposure to the same stresses sequentially. The results indicate that combinations and sequences of sublethal hurdles may affect L. monocytogenes acid and heat tolerance, especially in acidic environments with mild heating or in low moisture environments.     

Effects of environmental stresses on the activities of the uspA, grpE and rpoS promoters of Escherichia coli O157:H7

Heat shock proteins and RNA polymerase sigma factor play an important role in protecting cells against environmental stresses, including starvation, osmotic and oxidative stresses, and cold shock. In this study, the effect of environmental stresses on activity of the auto-fluorescent Escherichia coli O157:H7 generated by the fusion of gfp(uv) to E. coli uspA, grpE and rpoS promoters were examined. Osmotic shock caused about a 4-fold increase in green fluorescence of E. coli O157:H7 harboring uspA::gfp(uv) or rpoS::gfp(uv) at 37 degrees C and room temperature whereas osmotic shock at 5 degrees C did not induce green fluorescence. When starved, E. coli O157:H7 possessing grpE::gfp(uv) was more sensitive for evaluating stress at low temperature while uspA::gfp(uv) was better suited for detecting the stress response at higher temperature. The uspA, grpE and rpoS promoters were up-regulated to varying degrees by stresses commonly encountered during food processing.     

Cold stress tolerance of Listeria monocytogenes: A review of molecular adaptive mechanisms and food safety implications

The foodborne pathogen Listeria monocytogenes has many physiological adaptations that enable survival under a wide range of environmental conditions. The microbes overcome various types of stress, including the cold stress associated with low temperatures in food-production and storage environments. Cold stress adaptation mechanisms are therefore an important attribute of L. monocytogenes, enabling these food pathogens to survive and proliferate to reach minimal infectious levels on refrigerated foods. This phenomenon is a function of many molecular adaptation mechanisms. Therefore, an improved understanding of how cold stress is sensed and adaptation measures implemented by L. monocytogenes may facilitate the development of better ways of controlling these pathogens in food and related environments. Research over the past few years has highlighted some of the molecular aspects of cellular mechanisms behind cold stress adaptation in L. monocytogenes. This review provides an overview of the molecular and physiological constraints of cold stress and discusses the various cellular cold stress response mechanisms in L. monocytogenes, as well as their implications for food safety.     

Microbial stress response in minimal processing.

"Bacteria have evolved adaptive networks to face the challenges of changing environments and to survive under conditions of stress. Therefore, the efficiencies of inactivation and preservation methods need to be assessed, especially with regard to the enormous potential of food pathogens to adapt to a wide variety of stress conditions. All adaptive responses, whether to changing nutrients or to various stresses encountered in minimal processing, involve a series of genetic switches that control the metabolic changes taking place. A common regulatory mechanism involves the modification of sigma (sigma) factors whose primary role is to bind to core RNA polymerase conferring promoter specificity directing expression of specialty regulons involved in heat-shock response, the chemotactic response, sporulation, and general stress response. Examples of the latter regulon in Gram-positive bacteria (the sigmaB regulon) and in Gram-negative bacteria (the RpoS regulon) will be discussed in more detail. Cellular adaptive mechanisms to starvation, cold shock, heat shock, (weak) acids, high osmolarity and high hydrostatic pressure will be described and their significance in food preservation and safety will be discussed."     

Combined effects of NaCl, NaOH, and biocides (monolaurin or lauric acid) on inactivation of Listeria monocytogenes and Pseudomonas spp

This study highlighted combinations of chemical stresses that could decrease or eliminate Listeria monocytogenes and Pseudomonas spp. surviving in food processing plants. Strains of L. monocytogenes, Pseudomonas fragi, and Pseudomonas fluorescens isolated from processing environments (meat and milk) were grown at 20 degrees C up to the early stationary phase. The strains were then subjected to 30 min of physicochemical treatments. These treatments included individual or combined acid (acetic acid), alkaline (NaOH), osmotic (NaCl), and biocides (fatty acids) challenges. Survival of the strains was studied after individual or combined acid (acetic acid), alkaline (NaOH), osmotic (NaCl), and biocides (monolaurin, lauric acid) challenges. Individual pH shocks had lower efficiencies than those used in combinations with other parameters. The treatment pH 5.4 followed by pH 10.5 had a low efficiency against L. monocytogenes. The opposite combination, pH 10.5 followed by pH 5.4, led to a 3-log reduction of the L. monocytogenes population. Pseudomonas spp. strains were much more sensitive than L. monocytogenes, and population reductions of 5 and 8 log (total destruction), respectively, were observed after the same treatments. As for L. monocytogenes, the combination pH 10.5 followed by pH 5.4 is more deleterious than the opposite. Whatever the bacterial species, the most efficient treatments were combinations of alkaline, osmotic, and biocide shocks. For instance, the combination pH 10.5 and 10% NaCl plus biocides showed reductions of 5 to 8 log for both bacteria. The origins of the observed lethal effects are discussed.