噬菌体在食品安全领域的研究进展

微生物致病菌引起的食源性疾病在全世界频频发生,对人类健康造成严重危害,由抗生素滥用导致的耐药菌株出现,以及抗生素滥用,使得人们对食源性细菌疾病的控制更加困难。噬菌体是细菌的天敌,具有感染并裂解细菌的功能,噬菌体及其编码裂解酶的发现为食源性致病菌的检测及生物防治开辟了新的途径。因此近年来有关噬菌体作为抗菌制剂的研究受到普遍关注。就噬菌体在食品中病原微生物检测和控制方面的研究作以综述。     

噬菌体裂解酶在食品安全领域的研究进展

食源性疾病引发的食品安全问题对人类健康造成严重危害, 其中微生物致病菌是引起食源性疾病的最主要因素,近年来国内外由微生物致病菌引起的食源性疾病事件频频发生,受到世界各国的高度关注。食品工业防治食源性致病微生物的传统方法中,化学防腐剂存在副作用、天然防腐剂较弱的抗微生物活性以及大规模抗生素使用带来的耐药性等一系列问题,使寻求新的抗菌药物或制剂迫在眉睫。噬菌体裂解酶是双链DNA噬菌体复制后期表达, 能够裂解细菌细胞壁释放子代噬菌体的一种蛋白水解酶。随着近些年针对噬菌体及其产物展开的研究不断深入,噬菌体裂解酶凭借高度特异性、不影响正常菌群等特性, 从治疗人类耐药感染到控制多个领域的细菌污染, 成为了包括微生物食品安全在内多种应用中有效的抗微生物制剂。     

沙门氏菌和副溶血性弧菌裂解性噬菌体及其裂解酶研究进展

由微生物引发的食品安全问题给人类健康带来严重危害,其中沙门氏菌和副溶血性弧菌是最主要的2种微生物病原。噬菌体及其裂解酶的发现与应用为食源性致病菌的控制提供了新的技术,而因其具有安全、高效、特异性强、不易产生抗性等特点,在食品安全领域具有良好的应用前景。本文综述了沙门氏菌和副溶血性弧菌裂解性噬菌体及其裂解酶的研究与应用进展,沙门氏菌和副溶血性弧菌这2种致病菌的裂解性噬菌体在环境中广泛存在,对畜禽肉、海产品及其他食品中的沙门氏菌和副溶血性弧菌具有显著的减菌作用,其噬菌体裂解酶的研究还处于初级阶段,仅有几种裂解酶的研究报道,也显示了较好的溶菌活性。本文旨在为利用噬菌体及其裂解酶对食源性疾病的防控和人类的健康提供保障措施。     

李斯特菌噬菌体裂解酶基因在大肠杆菌中的表达

单核细胞增生李斯特菌是一种重要的食源性致病菌,存在于多种环境中,造成牛奶、肉类、水产品等的食品安全问题。李斯特菌噬菌体编码的裂解酶能够特异性地裂解宿主细胞,释放胞内的子代噬菌体。本研究旨在构建李斯特菌噬菌体A500裂解酶的细胞壁结合结构域CBD500的异源表达系统,获得重组蛋白。首先通过体外扩增得到CBD500基因,克隆到表达载体p ET32a(+)中,转化表达菌株大肠杆菌BL21(DE3)。阳性重组菌在30℃,0.1 mmol/L IPTG中诱导4 h后,以可溶形式表达分子质量约35 ku的重组蛋白。利用镍离子亲和层析纯化目的蛋白,纯化产物质量浓度为0.62 mg/mL。间接ELISA试验证实,纯化的重组蛋白具有生物学活性,可用于进一步研究裂解酶的理化性质和作用机制。     

裂解酶对食源性病原菌的生物防治研究及应用

噬菌体裂解酶能够高效消化细菌细胞壁,对多重耐药病原菌表现出独特的裂解能力,是一种新型抗菌分子,具有重要的应用潜力和研究价值。本文对裂解酶在食品生产中的研究进展进行了概述,重点综述了裂解酶及其嵌合体对食源性病原菌的生物防治研究以及裂解酶作用于革兰氏阴性菌的研究进展。     

噬菌体防控食源性致病菌的研究进展

噬菌体因其高效特异识别并裂解宿主菌的特性被认为是最具潜力的抗生素替代品之一。越来越多的学者将其应用于临床、兽医和食品领域中致病菌的防控。该文就噬菌体的基本特性、抑菌机理、安全性以及在对各种食品中食源性致病菌控制的研究进展以及应用前景进行综述。     

噬菌体控制主要食源性致病菌的研究进展

食源性细菌疾病是当今世界上最广泛的公共卫生问题之一。由抗生素滥用导致的耐药菌株出现,以及抗生素禁用,使得人们对食源性细菌疾病的控制更加困难。噬菌体是细菌的天敌,具有感染并裂解细菌的功能,与抗生素相比,噬菌体制剂具有特异性强、自我增殖快、抗菌能力强、研发时间短等优点,因此近年来有关噬菌体作为抗菌制剂的研究受到普遍关注。本文就噬菌体在食源性病原微生物控制方面的应用研究进展作一综述,以期为噬菌体及其制剂在保障动物性食品安全领域的深入研究提供一定的参考。     

噬菌体防控沙门氏菌生物被膜的研究进展

沙门氏菌是重要的食源性致病菌，控制食品中沙门氏菌的污染，特别是防止其形成生物被膜交叉污染食品，对保障食品安全具有重要意义。现阶段传统的物理性、化学性控制措施都存在一定的局限性，难以有效将沙门氏菌生物被膜完全去除，因此，亟待开发针对沙门氏菌生物被膜的新型控制和清除策略。近年来噬菌体作为一种安全、有效、无残留的天然抑菌剂受到了普遍关注，与化学消毒剂相比，其具有特异性强、自我增殖快、安全性高、研发时间短等突出优势，在食源性致病菌生物被膜的控制和清除领域已表现出巨大潜力。该文综述了沙门氏菌生物被膜的结构成分与形成过程，并重点介绍了国内外用噬菌体破解沙门氏菌生物被膜的作用机制与应用现状。针对噬菌体的未来研究方向进行了展望，以期为食品加工过程中沙门氏菌生物被膜的有效控制提供新的技术与策略。     

噬菌体对鸡肉表面耐药沙门氏菌的抑制作用

沙门氏菌是一类典型的人畜共患食源性病原菌,其污染严重影响着生鲜畜禽肉类制品的质量和安全。裂解性噬菌体作为细菌的天然生物源抑菌剂,是防控沙门氏菌污染的有效方式之一。目的 为了更好的研究噬菌体对生鲜畜禽肉表面耐药沙门氏菌的抑制效果,方法 本研究以实验室前期筛选得到的沙门氏菌噬菌体D1-2和Pu20为研究材料,采用单独使用和1:1混合的方式对生鸡胸肉表面多重耐药沙门氏菌的抑制效果进行研究。结果 与单一噬菌体相比,噬菌体混合制剂对生鸡胸肉表面多重耐药沙门氏菌抑菌效果更显著。在4_^oC时,采用感染复数MOI值为10000的噬菌体混合制剂,其对肠炎沙门氏菌11561的抑菌效率达到98%,且可以完全抑制鼠伤寒沙门氏菌SJTUF13277。结论 该实验结果为生物制剂控制耐药沙门氏菌提供了实验数据与理论基础。     

基于噬菌体的食源性致病菌检测方法研究进展

全球每年有几百万人因食源性致病菌感染引发疾病，亟需开发快速且专一的食源性致病菌检测方法以保障食品安全。噬菌体具有反应效率高、特异性强以及容易制取等众多优势，被广泛用于食源性致病菌检测。文章首先介绍噬菌体并且阐述食源性致病菌检测现状，接着分析噬菌体与微生物学、免疫学、分子生物学、光学以及生物传感器等技术联合检测食源性致病菌的原理，重点总结了噬菌体联合这些技术的检测方法在实际应用中的最新进展，以期为噬菌体在食源性致病菌检测中的应用提供参考。     

Phages and their lysins: Toolkits in the battle against foodborne pathogens in the postantibiotic era

Worldwide, foods waste caused by putrefactive organisms and diseases caused by foodborne pathogens persist as public health problems even with a plethora of modern antimicrobials. Our over reliance on antimicrobials use in agriculture, medicine, and other fields will lead to a postantibiotic era where bacterial genotypic resistance, phenotypic adaptation, and other bacterial evolutionary strategies cause antimicrobial resistance (AMR). This AMR is evidenced by the emergence of multiple drug-resistant (MDR) bacteria and pan-resistant (PDR) bacteria, which produces cross-contamination in multiple fields and poses a more serious threat to food safety. A “red queen premise” surmises that the coevolution of phages and bacteria results in an evolutionary arms race that compels phages to adapt and survive bacterial antiphage strategies. Phages and their lysins are therefore useful toolkits in the design of novel antimicrobials in food protection and foodborne pathogens control, and the modality of using phages as a targeted vector against foodborne pathogens is gaining momentum based on many encouraging research outcomes. In this review, we discuss the rationale of using phages and their lysins as weapons against spoilage organisms and foodborne pathogens, and outline the targeted conquest or dodge mechanism of phages and the development of novel phage prospects. We also highlight the implementation of phages and their lysins to control foodborne pathogens in a farm–table–hospital domain in the postantibiotic era.     

Phages and engineered lysins as an effective tool to combat Gram-negative foodborne pathogens

One of the biggest challenges faced by food producers is ensuring microbiological safety. Despite strict criteria for food products, foodborne diseases are a global problem and pose a real risk to consumers. Therefore, it is necessary to identify new and more effective methods for eliminating pathogens from food and the food processing environment. According to the European Food Safety Authority (EFSA), the most common foodborne diseases are caused by Campylobacter, Salmonella, Yersinia, Escherichia coli, and Listeria. Out of the five listed, four are Gram-negative bacteria. Our review focuses on the use of bacteriophages, which are ubiquitous bacterial viruses, and bacteriophage endolysins to eliminate Gram-negative pathogens. Endolysins cleave specific bonds within the peptidoglycan (PG) of the bacterial cell, causing the cell to burst. Single phages or phage cocktails, which are, in some instances, commercially available products, eliminate pathogenic bacteria in livestock and various food matrices. Endolysins have matured as the most advanced class of antibacterial agents in the clinical sector, but their use in food protection is highly unexplored. Advanced molecular engineering techniques, different formulations, protein encapsulation, and the addition of outer membrane (OM) permeabilization agents enhance the activity of lysins against Gram-negative pathogens. This creates space for groundbreaking research on the use of lysins in the food sector.     

噬菌体在食品生产和加工中的生物防控应用及思考

噬菌体以其无以比拟的优势在生物防控和疾病治疗中扮演着重要角色,不仅有效抑制致病菌延长食品货架期,且具有特异无残留等优点。目前在农业、动物疾病防控、食品安全以及耐药性疾病治疗中的应用案例屡见不鲜,许多国家已有噬菌体产品应运而生。噬菌体在食品生产(“农场”)和加工后(“餐桌”)的干预措施,能够控制重要食源性病原,如沙门氏菌,李斯特菌和大肠杆菌等。在“农场-餐桌”的供应链中,噬菌体及其衍生物均能发挥重要作用,有效抑制致病菌来保障农产品质量安全。本文就噬菌体在食品生物防控中的应用进展以及应用中存在的问题进行综述。     

基于益生菌及其代谢物的食源性致病微生物控制技术研究进展

食源性致病微生物不仅会引起食品腐败,造成经济损失,也给人类健康带来很大威胁,因此亟需开发高效、安全、不影响食品品质的食源性致病微生物控制技术。益生菌是近年微生物和食品科研领域研究热点,益生菌除具有调节和改善健康功能,其本身及代谢物对微生物较强的清除和抑制作用,使其可应用于对食源性致病微生物的控制。明悉益生菌及其代谢物类型、潜在的抑制微生物机制以及应用现状对相关控制技术的研发至关重要。分析了现有益生菌及其代谢物对食源性致病微生物的抑制机制,主要为破坏细胞结构、影响遗传物质复制、阻断能量代谢途径、干扰群体感应系统、控制生物被膜形成、竞争关键性营养物质等方面的单机制或多机制联合作用;同时基于目前益生菌及其代谢物在食品安全和品质控制中的应用研究实例,探讨了其在果蔬、肉类等食品中的应用方式、应用条件,对货架期的延长效果等;分析了益生菌及其代谢物在抗菌效能提高、活性包装、与其他食品品质控制技术联用等方面研究的发展趋势。研究结果旨在为基于益生菌及其代谢物的食品安全和品质控制技术的开发和应用提供借鉴和参考。     

Phage-based technologies for highly sensitive luminescent detection of foodborne pathogens and microbial toxins: A review

Foodborne pathogens and microbial toxins are the main causes of foodborne illness. However, trace pathogens and toxins in foods are difficult to detect. Thus, techniques for their rapid and sensitive identification and quantification are urgently needed. Phages can specifically recognize and adhere to certain species of microbes or toxins due to molecular complementation between capsid proteins of phages and receptors on the host cell wall or toxins, and thus they have been successfully developed into a detection platform for pathogens and toxins. This review presents an update on phage-based luminescent detection technologies as well as their working principles and characteristics. Based on phage display techniques of temperate phages, reporter gene detection assays have been designed to sensitively detect trace pathogens by luminous intensity. By the host-specific lytic effects of virulent phages, enzyme-catalyzed chemiluminescent detection technologies for pathogens have been exploited. Notably, these phage-based luminescent detection technologies can discriminate viable versus dead microbes. Further, highly selective and sensitive immune-based assays have been developed to detect trace toxins qualitatively and quantitatively via antibody analogs displayed by phages, such as phage-ELISA (enzyme-linked immunosorbent assay) and phage-IPCR (immuno-polymerase chain reaction). This literature research may lead to novel and innocuous phage-based rapid detection technologies to ensure food safety.     

乳酸菌拮抗食源性致病菌的研究及应用进展

食品在生产加工、物流运输和贮存消费过程中均可能出现被食源性致病菌污染的风险,造成安全隐患,目前通过热加工、辐照、电解水和等离子体等方法来控制食品中食源性致病菌,但在食品感官、规模化应用和安全生产等方面存在部分缺点.乳酸菌因其丰富的生物活性常作为生物发酵剂和保鲜剂广泛应用于食品中,并且可在食品中拮抗致病菌.本文主要从竞争...     

扩增型和非扩增型CRISPR/Cas系统在食源性致病菌快速检测领域应用的研究进展

食源性致病菌引起的食品安全事件频发已对公众健康和社会经济造成巨大的影响,构建针对其快速、高效的检测方法是保障食品安全的重要手段。成簇的间隔短回文重复序列（clustered regularly interspaced short palindromic repeats,CRISPR）及其相关蛋白（CRISPR-associated protein,Cas）组成的CRISPR/Cas是广泛存在于细菌和古细菌中的一种免疫系统,可以有效地识别并切割外源核酸。因此,可以利用CRISPR/Cas系统这一对外源核酸的识别及切割活性实现对食源性致病菌的快速高效检测。本文详细介绍基于核酸扩增和免核酸扩增的两种CRISPR/Cas系统,综述其在对各种食源性致病菌检测中的应用,讨论它们的优势、局限性以及未来的发展趋势,旨在为建立更高效的食源性致病菌检测方法提供技术参考,以期更有效地减少食源性致病菌造成的食品安全问题。     

Emerging foodborne pathogens

The broad spectrum of foodborne infections has changed dramatically over time, as well-established pathogens have been controlled or eliminated, and new ones have emerged. The burden of foodborne disease remains substantial: one in four Americans is estimated to have a significant foodborne illness each year. The majority of these illnesses are not accounted for by known pathogens, so more must remain to be discovered. Among the known foodborne pathogens, those more recently identified predominate, suggesting that as more and more is learned about pathogens, they come under control. In addition to the emergence or recognition of new pathogens, other trends include global pandemics of some foodborne pathogens, the emergence of antimicrobial resistance, the identification of pathogens that are highly opportunistic, affecting only the most high-risk subpopulations, and the increasing identification of large and dispersed outbreaks. New pathogens can emerge because of changing ecology or changing technology that connects a potential pathogen with the food chain. They also can emerge de novo by transfer of mobile virulence factors, often through bacteriophage. Though this is rarely observed, it can be reconstructed. Better understanding of the ecology and dynamics of phage transmission among bacteria will help us to understand the appearance of new pathogens in the future. One may look for emerging foodborne pathogens among the silent zoonoses, and among the severe infections affecting the immunocompromised humans. We should expect the unexpected. In the past, separating human sewage and animal manure from human food and water supplies was critical to improving public health. Now, our health depends increasingly on the safety of the feed and water supplies for the animals themselves. The successes of the 20th century and the new challenges we face mean that public health vigilance, careful investigation of new problems, responsible attention to food safety from farm to table, and partnerships to bring about new foodborne disease control measures will be needed for the foreseeable future.     

代谢组学在食品质量安全领域的应用进展

食品质量安全是保障现代经济社会和谐稳定发展的基础与前提，目前，食源性致病微生物、兽药残留、转基因食品及掺假等问题均会造成或可能造成食品安全隐患。代谢组学作为新兴技术不断发展，通过研究生物体受外界干扰前后小分子代谢产物的变化，进而探究机体内代谢机制，适用于食品安全多种微量危害因子的鉴别和监测。此外，危害因子在食品分解过程中产生的代谢物可成为特定的潜在生物标志物，为致病菌作用机制和品质安全控制的研究提供参考。本文阐述靶向代谢组学和非靶向代谢组学，介绍代谢组学技术常用的数据采集和数理统计方法，总结代谢组学技术在食源性致病菌、兽药残留、转基因食品、生鲜食品品质和肉制品掺假等食品品质和安全领域的研究进展，并对多组学联用技术提出展望，以期推动该技术在食品质量安全领域更广泛的应用。