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。     

单核细胞增生李斯特菌对季铵盐类消毒剂的耐受机制及其防控研究进展

单核细胞增生李斯特菌（以下简称“单增李斯特菌”）是一种常见的食源性病原菌，可通过群体感应在不锈钢食品加工设备表面形成单一或多物种生物膜，成为食品潜在的污染源。季铵盐类消毒剂常用于食品企业加工设备表面的清洗消毒，而单增李斯特菌生物膜在主动外排泵系统、应激反应、基因调控等多种机制的介导下可对其产生耐受性。本文综述了单增李斯特菌生物膜的形成及耐受机制，汇总了近年来物理杀菌以及新型组合杀菌技术在食源性单增李斯特菌防控中的应用，为新型高效杀菌技术的开发提供思路，也为企业建立单增李斯特菌及其他微生物防控措施提供重要参考。     

基于群体感应的单增李斯特菌生物膜形成与控制研究进展

单增李斯特菌的生物膜结构可以赋予细菌对消毒剂更强的耐受性,这使得如何有效清除其生物膜成为了食品加工业的难题。群体感应与细菌生物膜的形成、毒力侵袭特性和应激响应等生理特性密切相关。因此,本文概述了单增李斯特菌生物膜的特性,重点从群体感应（呋喃糖基硼酸二酯系统和寡肽自诱导因子系统）角度阐述了单增李斯特菌生物膜的形成机制,并综述了当前控制单增李斯特菌生物膜形成的可行方法及植物提取物作为群体感应抑制剂的研究现状。以期为预防和控制单增李斯特菌生物膜的形成提供新思路。     

单增李斯特菌生物膜形成及其调控机制研究进展

单增李斯特菌是一种重要的食源性致病菌,极易在食品接触表面形成难以清除的生物膜,导致食品持续性的污染。细菌生物膜在形成量、活细菌数量、微观结构等方面受到环境因素及菌株自身特性的影响。在单增李斯特菌的生物膜形成过程中,多种调控机制发挥着重要的作用。该文介绍了细菌生物膜的形成过程及影响单增李斯特菌生物膜形成的重要因素,简述了与单增李斯特菌生物膜相关的基因调控、胞外聚合物质分泌与群体感应系统调节的研究进展,有助于更好地理解其生物膜形成的复杂生理过程,为单增李斯特菌生物膜的污染及防控提供一定参考。     

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。     

即食菜卷和肉汤中单增李斯特菌生长模型及控制研究

通过对单增李斯特菌在不同NaCl浓度、不同pH、不同温度下营养肉汤中生长情况的研究,确定了单增李斯特菌的最佳生长条件。建立了30℃下单增李斯特菌在营养肉汤和即食菜卷中的生长模型。以模型为依据,提出了即食菜卷加工中控制李斯特菌的措施。     

单核细胞增生李斯特菌对食品加工中的胁迫响应及其机制研究进展

单核细胞增生李斯特菌（简称单增李斯特菌）在食品加工过程中经常会遇到环境胁迫,如酸胁迫、热胁迫、冷胁迫、干燥和高渗透压胁迫以及交叉胁迫等,并产生应激反应,而经过环境胁迫的单增李斯特菌可显著增强其抵抗致死性环境胁迫的能力,给食品安全带来极大危害。单增李斯特菌的酸胁迫响应机制主要包括F0F1-ATPase系统、谷氨酸脱羧酶系统以及精氨酸脱亚胺酶系统。热休克蛋白、冷休克蛋白、相容性溶质在单增李斯特菌的热胁迫、冷胁迫和干燥及高渗透压胁迫反应中起重要作用。本文从以上几方面就单增李斯特菌对环境胁迫响应及其机制的研究现状进行了综述,并提出了今后可能的研究方向,为单增李斯特菌在食品加工过程中的胁迫响应研究及防控提供指导。     

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。     

两种抗菌剂对不同温度下形成的单增李斯特菌生物膜的清除作用

该研究考察单增李斯特菌在32℃和10℃下生物膜的形成,并以1/2、1、2、4 MIC(minimum inbibitory concentration,最低抑菌浓度)的薰衣草精油和84消毒液处理单增李斯特菌的生物膜,以结晶紫染色法、二甲氧唑黄法(XTT法)及叠氮溴化丙锭-荧光定量PCR(PMA-qPCR)等方法评估其清除效果。研究结果表明,单增李斯特菌在两种温度下均能形成稳定的生物膜,32℃下形成的生物膜量显著高于10℃; 4种浓度薰衣草精油处理3 h对32℃下形成的生物膜的清除率为62%～82%,对10℃的生物膜的清除率为25%～56%; 4种浓度84消毒液处理3 h对32℃和10℃下形成的生物膜的清除率分别为47%～78%和36%～56%; 4 MIC(6. 4%体积分数)的薰衣草精油处理3 h后10℃生物膜的残余活菌数为32℃生物膜的2倍以上。该研究阐明了单增李斯特菌的低温生物膜抗性更强,薰衣草精油对生物膜的清除效果优于84消毒液,对植物精油的应用以及控制单增李斯特菌残留具有积极的意义。     

散装熟肉制品中单增李斯特菌污染状况分析

目的分析散装熟肉制品中单增李斯特菌污染状况。方法根据GB 4789.30-2016《单核细胞增生李斯特氏菌检验》规定的方法对散装熟肉制品的加工用原料、生产环境、各加工环节中产品以及不同销售环境下产品中的单增李斯特菌进行定性和定量检测。结果原料肉的总体带菌率达到21%;生产环境中第三区(远离食品接触面的区域)检出率为2%,其余区域未检出;各加工环节中产品单增李斯特菌检测结果均小于10CFU/g;不同销售环境下产品中单增李斯特菌检测结果小于10 CFU/g的比例为93%,处于10～50 CFU/g之间的样本占比6%,大于50 CFU/g的占比为1%。结论原料散装熟肉制品中单增李斯特菌污染的主要来源,蒸煮等加工环节能有效地杀灭单增李斯特菌。销售环境也关系到散装熟肉制品单增李斯特菌的污染程度,对于未包装的产品,专卖店优于农产品市场。     

An Ecological Perspective of Listeria monocytogenes Biofilms in Food Processing Facilities

Listeria monocytogenes can enter the food chain at virtually any point. However, food processing environments seem to be of particular importance. From an ecological point of view, food processing facilities are microbial habitats that are constantly disturbed by cleaning and sanitizing procedures. Although L. monocytogenes is considered ubiquitous in nature, it is important to recognize that not all L. monocytogenes strains appear to be equally distributed; the distribution of the organism seems to be related to certain habitats. Currently, no direct evidence exists that L. monocytogenes-associated biofilms have played a role in food contamination or foodborne outbreaks, likely because biofilm isolation and identification are not part of an outbreak investigation, or the definition of biofilm is unclear. Because L. monocytogenes is known to colonize surfaces, we suggest that contamination patterns may be studied in the context of how biofilm formation is influenced by the environment within food processing facilities. In this review, direct and indirect epidemiological and phenotypic evidence of lineage-related biofilm formation capacity to specific ecological niches will be discussed. A critical view on the development of the biofilm concept, focused on the practical implications, strengths, and weaknesses of the current definitions also is discussed. The idea that biofilm formation may be an alternative surrogate for microbial fitness is proposed. Furthermore, current research on the influence of environmental factors on biofilm formation is discussed.     

Occurrence and distribution of listeria species in facilities producing ready-to-eat foods in British Columbia, Canada

In British Columbia (BC), Canada, food processing facilities licensed under provincial authority are not required to sample for Listeria monocytogenes in food products or processing environments. In 2009, we conducted a survey of dairy, fish, and meat facilities under BC authority to estimate the prevalence of Listeria spp. and L. monocytogenes in ready-to-eat (RTE) foods and production environments. From August to October, 250 RTE food samples and 258 swabs from the food processing environments of 43 facilities were collected. Standard culture methods were applied to both food samples and swabs. Of swabs collected from all 258 environmental surfaces, 15% were positive for Listeria spp. Significantly (P, 0.001) more fish facilities than dairy and meat facilities had food contact surfaces contaminated with Listeria spp. L. monocytogenes was found in RTE foods from fish facilities alone (5 of 12); in all five of the fish facilities with contaminated product, one or more environmental swabs were also positive for L. monocytogenes. The results suggest that while control of L. monocytogenes in BC-inspected dairy and meat facilities is effective in limiting food contamination, there is a need for provincial inspectors to initiate improved monitoring and management of contamination by L. monocytogenes in RTE fish processing facilities.     

食品环境中单核细胞增生李斯特菌菌膜形成、转移及防控措施研究进展

单核细胞增生李斯特菌（Listeria monocytogenes）是主要的食源性致病菌之一。单核细胞增生李斯特菌菌膜在食品环境中的形成与转移对食品的污染是目前广泛关注的难题。针对于单核细胞增生李斯特菌菌膜预防与控制措施的研究能够避免食品环境受到污染,从而保证食品的安全。本文整理了食品环境中检出的单核细胞增生李斯特菌菌膜形成能力,并且综述了单核细胞增生李斯特菌菌膜形成及转移因素的研究进展,最后从单核细胞增生李斯特菌菌膜的预防与控制两个方面进行较为全面的概述。加入更多菌株并且模拟更多实际情况进行单核细胞增生李斯特菌菌膜研究能够更好地了解菌膜的形成与转移情况,并为更便捷地清除菌膜提供理论依据。今后应研究更多实际情况下具有代表性的单核细胞增生李斯特菌菌株,从而更好地了解菌膜形成与转移,其次探究单核细胞增生李斯特菌菌膜形成机理及相关因素对菌膜的影响机制,为实施高效的菌膜防控措施提供依据。     

Survival of Listeria monocytogenes attached to stainless steel surfaces in the presence or absence of Flavobacterium spp

Contaminated surfaces of food processing equipment are believed to be a significant source of Listeria monocytogenes to foods. However, very little is known about the survival of Listeria in processing environments. In a mixed bacterial biofilm of L. monocytogenes and Flavobacterium spp., the number of L. monocytogenes cells attaching to stainless steel increased significantly compared to when L. monocytogenes was in a pure culture. The L. monocytogenes cells in the mixed biofilms were also recoverable for significantly longer exposure periods. On colonized coupons held at 15 degrees C and 75% humidity, decimal reduction times were 1.2 and 18.7 days for L. monocytogenes in pure and mixed biofilms, respectively. With increasing exposure time, the proportion of cells that were sublethally injured (defined as an inability to grow on selective agar) increased from 8.1% of the recoverable cell population at day 0 to 91.4% after 40 days' exposure. At 4 and -20 degrees C, decimal reduction times for L. monocytogenes in pure culture were 2.8 and 1.4 days, respectively, and in mixed culture, 10.5 and 14.4 days, respectively. The enhanced colonization and survival of L. monocytogenes on "unclean" surfaces increase the persistence of this pathogen in food processing environments, while the increase in the percentage of sublethally injured cells in the population with time may decrease the ability of enrichment regimes to detect it.     

Listeria: A foodborne pathogen that knows how to survive

The foodborne pathogen Listeria is the causative agent of listeriosis, a severe disease with high hospitalization and case fatality rates. Listeria monocytogenes can survive and grow over a wide range of environmental conditions such as refrigeration temperatures, low pH and high salt concentration. This allows the pathogen to overcome food preservation and safety barriers, and pose a potential risk to human health. This review focuses on the key issues such as survival of the pathogen in adverse environments, and the important adaptation and survival mechanisms such as biofilm formation, quorum sensing and antimicrobial resistance. Studies on the development of technologies to prevent and control L. monocytogenes contamination in foods and food processing facilities are also discussed.     

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.