环境胁迫下沙门氏菌应激反应及抗性机制研究进展

细菌为了生存能够适应多种不利于生长的环境条件，因此许多食源性致病菌能够在某些食品保鲜处理中存活下来。沙门氏菌（Salmonella）是世界范围内常见的食源性致病细菌，在食品保鲜中可承受极端pH、温度、渗透压、寡营养和高压等胁迫。在与宿主相互作用过程中，沙门氏菌应对不同环境胁迫，可产生应激蛋白、进行系统自我调节和表达相关基因等，进而影响其致病能力，产生交叉抗性从而更好的生存和繁殖。因此，沙门氏菌能在遗传和表型水平上适应不同的极端环境。本文综述了沙门氏菌耐受环境胁迫，暴露于这些环境胁迫时发生的适应性变化，对细菌致病性的影响，以及产生的交叉保护机制。这些方面将有助于了解沙门氏菌的发病机制，对于制定抗击沙门氏菌感染的新策略，保障食品安全具有重要意义。     

The physiology of Campylobacter species and its relevance to their role as foodborne pathogens

Campylobacter jejuni and C. coli are recognised as the leading causes of bacterial foodborne diarrhoeal disease throughout the development world. While most foodborne bacterial pathogens are considered to be relatively robust organisms, as a consequence of the necessity to survive the inimical conditions imposed by food processing and preservation, Campylobacter species have uniquely fastidious growth requirements and an unusual sensitivity to environmental stress. Campylobacters also lack many of the well characterised adaptive responses that can be collated with resistance to stress in other bacteria. The aim of this review is to outline the unusual physiology of campylobacters (C. jejuni and C. coli) and to describe how this influences their role as foodborne pathogens.     

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.     

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."     

SURVIVAL AND GROWTH OF ACID ADAPTED ESCHERICHIA COLI STRAINS IN BROTH AT DIFFERENT PH LEVELS

Acid resistance of Escherichia coli O157:H7 strain UT 10 and Escherichia coli ATCC 25922 was determined in brain heart infusion broth at pH 7.4, 4.5 and 2.5. Variations due to acid stress in the counts of both strains were also determined. Acid adaptation enhanced the survival of both strains at pH 4.5, but neither strain could survive after 4 h at pH 2.5. At optimum growth conditions (pH 7.4), E. coli ATCC 25922 exhibited increased viability over E. coli UT 10. At pH 4.5, E. coli UT 10 was more tolerant to low pH than E. coli ATCC 25922. An increase in saturated fatty acids of both AA strains was observed, indicating the importance of lipid modification in enhancing survival at low pH. The results of this study indicated that the food industry should therefore adapt their processing/preservation procedures by taking the most acid tolerant pathogenic E. coli strains into consideration in order to ensure the safety of their products.

PRACTICAL APPLICATIONS

The study will enlighten the industry on the survival of acid adapted pathogens such as E. coli O157:H7 at low pH. It also indicates that current measures used in the preservation of low pH foods can trigger acid adaptation. For this reason, the effectiveness of current preservation measures in controlling foodborne pathogens should be reassessed. The importance of using cells adapted to different pH values in food challenge studies is also highlighted. Since fermented foods, which are generally regarded as safe, have been implicated in foodborne disease outbreaks, more attention should be given to the prevention of contamination of foods with pathogens such as E. coli O157:H7 since they may survive in low pH foods and cause disease.     

生物膜状态对食源性致病菌耐药与毒力的影响研究进展

生物膜是细菌抵御不利环境维持群体稳定性的一种常见的群落形态。生物膜状态增强了食源性致病菌的抗逆性和持久存活力,促进了细菌间的信息传导和物质交换。特别是生物膜状态显著增强了菌株对抗生素的耐受能力,提升了可移动元件在细菌间的转移效率,并且生物膜状态下的细菌具备更强的入侵和感染能力,成为食品安全和人类健康的重要危害。基于此,本文将从生物膜的结构特征与异质性、生物膜对基因突变和基因水平转移的影响、以及群体感应调控等多个角度,简述生物膜状态影响食源性致病菌耐药和毒力的研究进展,以期为深入研究生物膜的生物功能与危害防控提供新的思路。     

The microbiology of minimally processed fresh fruits and vegetables

Minimally processed fresh (MPF) fruits and vegetables are good media for growth of microorganisms. They have been involved in outbreaks because of the consumption of products contaminated by pathogens. They are also sensitive to various spoilage microorganisms such as pectinolytic bacteria, saprophytic Gram‐negative bacteria, lactic acid bacteria, and yeasts. Contamination of MPF fruits and vegetables occurs at every stage of the food chain, from cultivation to processing. Polluted environments during cultivation or poor hygienic conditions in processing increase the risk of contamination with foodborne pathogens. Although MPF fruits and vegetables may harbor psychrotrophic microorganisms such as fluorescent pseudomonads or Listeria monocytogenes, good control of refrigeration temperature limits growth of spoilage and pathogenic microorganisms. Modified atmospheres are often efficient to maintain or improve visual and organoleptic quality of MPF fruits and vegetables, but their effects on microorganisms are inconsistent. Chemical disinfection can partially reduce the initial bacterial contamination; irradiation seems to be more efficient. The applications of legislations and quality assurance systems to control contamination, survival, and growth of foodborne pathogens in MPF fruits and vegetables are discussed.     

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.     

In vitro and in vivo characterization of the antibacterial activity and membrane damage mechanism of quinic acid against Staphylococcus aureus

In this study, we investigated the antibacterial activity of quinic acid (QA) and elucidated its membrane action mechanism against Staphylococcus aureus. In vitro antibacterial assay showed that QA effectively inhibited the growth of test bacteria. In model food systems of cabbage suspension and barley soup, QA possessed a great antibacterial capacity. The membrane hyperpolarization and a decrease of membrane fluidity demonstrated a significant effect of QA on damaging the normal functions of cell membrane, which was further validated by the observation of the transmission electron microscope. The interaction of QA with Phe residues in the membrane protein was revealed by fluorescence quenching. Overall, the present study illustrates QA as a potent antibacterial agent and its inhibition action on S. aureus by damaging the cell membrane.

Practical applications

In model food systems of cabbage suspension and barley soup, QA possessed a great antibacterial capacity to control the total counts of viable bacteria. This study provided the theoretical foundations for the application of QA in foods to prevent the growth of foodborne pathogens.     

The importance of lactic acid bacteria for the prevention of bacterial growth and their biogenic amines formation: A review

Foodborne pathogens (FBP) represent an important threat to the consumers' health as they are able to cause different foodborne diseases. In order to eliminate the potential risk of those pathogens, lactic acid bacteria (LAB) have received a great attention in the food biotechnology sector since they play an essential function to prevent bacterial growth and reduce the biogenic amines (BAs) formation. The foodborne illnesses (diarrhea, vomiting, and abdominal pain, etc.) caused by those microbial pathogens is due to various reasons, one of them is related to the decarboxylation of available amino acids that lead to BAs production. The formation of BAs by pathogens in foods can cause the deterioration of their nutritional and sensory qualities. BAs formation can also have toxicological impacts and lead to different types of intoxications. The growth of FBP and their BAs production should be monitored and prevented to avoid such problems. LAB is capable of improving food safety by preventing foods spoilage and extending their shelf-life. LAB are utilized by the food industries to produce fermented products with their antibacterial effects as bio-preservative agents to extent their storage period and preserve their nutritive and gustative characteristics. Besides their contribution to the flavor for fermented foods, LAB secretes various antimicrobial substances including organic acids, hydrogen peroxide, and bacteriocins. Consequently, in this paper, the impact of LAB on the growth of FBP and their BAs formation in food has been reviewed extensively.     

Effect of a fullerene water suspension on bacterial phospholipids and membrane phase behavior

Several fullerene-based nanomaterials generate reactive oxygen species that can damage cells. In this study, we investigated the effect of buckminsterfullerene (C60) introduced as colloidal aggregates in water (nC60) on bacterial membrane lipid composition and phase behavior. Pseudomonas putida (Gram-negative) and Bacillus subtilis (Gram-positive) responded to nC60 by altering membrane lipid composition, phase transition temperature, and membrane fluidity. P. putida decreased its levels of unsaturated fatty acids and increased the proportions of cyclopropane fatty acids in the presence of nC60, possibly to protect the bacterial membrane from oxidative stress. Fourier transform infrared spectroscopy measurement of intact bacterial cells showed slightly increased phase transition temperatures (Tm) and increased membrane fluidity for cells grown in the presence of high, growth-inhibiting concentrations (0.5 mg L(-1)) of nC60. B. subtilis responded to a low dose of nC6o (0.01 mg L(-1)) by significantly increasing the levels of iso- and anteiso-branched fatty acids (from 5.8 to 31.5% and 12.9 to 32.3% of total fatty acids, respectively) and to a high, growth-inhibiting concentration of nC60 (0.75 mg L-1) by increasing synthesis of monounsaturated fatty acids. In contrast to P. putida, B. subtilis response was a decrease in Tm and an increase in membrane fluidity. These findings represent the first demonstrated physiological adaptation response of bacteria to a manufactured nanomaterial, and they showthat response inlipid composition and membrane phase behavior depends on both the nC60 concentration and the cell wall morphology.     

Environmental conditions and serotype affect Listeria monocytogenes susceptibility to phage treatment in a laboratory cheese model

Listeria monocytogenes can survive and grow in a variety of environments, including refrigeration, making it difficult to control and highlighting the importance of optimizing control strategies against this pathogen. Listeria phages are attractive biocontrol agents because phages bind to specific wall teichoic acids (WTA) on the bacterial cell wall, inhibiting pathogens without disrupting the normal microbiota or structure of the food. Common stresses found on dairy products can affect cell wall composition and structure and subsequently affect the efficiency of control strategies that target the cell wall. The goal of this study was to determine the effect of a range of pH and temperatures on the effectiveness of a commercial phage cocktail treatment against several strains of L. monocytogenes in a cheese matrix. We developed a laboratory-scale cheese model that was made at different pH, treated with phage, and then inoculated with L. monocytogenes. Cheeses were incubated at 6, 14, or 22°C for 14 d, and bacterial counts were determined on d 1, 7, and 14. Our data show that phage treatment has a limited ability to reduce L. monocytogenes counts at each temperature tested; however, it was more effective on specific strains of L. monocytogenes when cheese was stored at higher temperatures. More specifically, the average counts of L. monocytogenes on phage-treated cheese stored at 22°C were significantly lower than those on phage-treated cheese stored at 6 or 14°C. Similarly, phage treatment was significantly more effective at inhibiting L. monocytogenes on cheese made at higher pH (6 and 6.5) compared with counts on cheese made at pH 5.5, where L. monocytogenes did not grow. Furthermore, serotype was found to affect the susceptibility of L. monocytogenes to phage treatment; serotype 1/2 strains showed significantly higher susceptibility to phage treatment than serotype 4b strains. Overall, our results suggest the importance of considering the efficacy of phage under conditions (i.e., temperature and pH) specific to a given food matrix when applying interventions against this important foodborne pathogen.     

Pore-forming toxins of foodborne pathogens

Pore-forming toxins (PFTs) are water-soluble molecules that have been identified as the most crucial virulence factors during bacterial pathogenesis. PFTs disrupt the host cell membrane to internalize or to deliver other bacterial or virulence factors for establishing infections. Disruption of the host cell membrane by PFTs can lead to uncontrollable exchanges between the extracellular and the intracellular matrix, thereby disturbing the cellular homeostasis. Recent studies have provided insights into the molecular mechanism of PFTs during pathogenesis. Evidence also suggests the activation of several signal transduction pathways in the host cell on recognition of PFTs. Additionally, numerous distinctive host defense mechanisms as well as membrane repair mechanisms have been reported; however, studies reveal that PFTs aid in host immune evasion of the bacteria through numerous pathways. PFTs have been primarily associated with foodborne pathogens. Infection and death from diseases by consuming contaminated food are a constant threat to public health worldwide, affecting socioeconomic development. Moreover, the emergence of new foodborne pathogens has led to the rise of bacterial antimicrobial resistance affecting the population. Hence, this review focuses on the role of PFTs secreted by foodborne pathogens. The review highlights the molecular mechanism of foodborne bacterial PFTs, assisting bacterial survival from the host immune responses and understanding the downstream mechanism in the activation of various signaling pathways in the host upon PFT recognition. PFT research is a remarkable and an important field for exploring novel and broad applications of antimicrobial compounds as therapeutics.     

大肠杆菌耐酸分子机制研究进展

具有耐强酸能力的大肠杆菌在酸性胁迫条件下更易生存。这主要是由于其具有葡萄糖-阻碍耐酸系统、氨基酸依赖型耐酸系统、伴侣蛋白抗酸作用以及保持膜电荷稳定等耐酸机制。了解大肠杆菌的这些耐酸分子机制,可以为食品加工工业控制大肠杆菌的污染提供新的认识,也对一些食源性致病菌的临床预防和治疗具有积极的作用。本文就大肠杆菌耐酸分子机制的研究进行综述。     

Detection of Escherichia coli via VOC profiling using secondary electrospray ionization-mass spectrometry (SESI-MS)

Escherichia coli O157:H7 (EC O157:H7), as well as its recently emerging non-O157 relatives, are a notorious group of pathogenic bacteria associated with foodborne outbreaks. In this study, we demonstrated that secondary electrospray ionization mass spectrometry (SESI-MS) could be a rapid and accurate detection technology for foodborne pathogens. With SESI-MS volatile organic compound (VOC) profiling, we were able to detect and separate a group of eleven E. coli strains from two major foodborne bacteria, Staphylococcus aureus and Salmonella Typhimurium in three food modeling media. In addition, heatmap analysis of relative peak intensity show that there are six core peaks (m/z of 65, 91, 92, 117, 118 and 119) present and at a similar intensity in all eleven E. coli strains at the experimental conditions we tested. These peaks can be considered conserved VOC biomarkers for E. coli species (robustly produced after just 4 h of growth). Bacterial strain-level differentiation was also attempted via VOC profiling, and we found that EC O157:H7 and EC O145 were differentiable from all other EC strains under the conditions investigated.     

核酸交联剂在食源致病菌活菌检测中应用的 研究进展

近年来,由食源致病菌引起的食品安全问题越来越受到人们的重视,如何快速准确检测食品中是否存在食源致病菌是食品安全研究的热点问题。基于PCR检测食源致病菌的方法因快速且特异性强而被广泛应用,然而普通的PCR检测方法难以消除死菌残留DNA导致的假阳性结果,因而无法对致病菌进行准确检测。核酸交联剂是一种含有两个或两个以上烷基化官能团的烷基化试剂,目前,在食源致病菌检测中应用的核酸交联剂主要为EMA和PMA。核酸交联剂通过一定方式的诱导可选择性的透过死菌的细胞膜并与DNA产生共价交联,从而强有力的抑制死菌DNA的PCR扩增,达到鉴别死菌和活菌的效果。本文就核酸交联剂在食源致病菌活菌检测中应用的研究进展进行了综述,以期能为相关研究者开发食源致病菌活菌检测方法提供参考。     

Development of Wide-Spectrum Hybrid Bacteriocins for Food Biopreservation

The food industry demands new procedures and methods to produce minimally processed, ready to eat food with intact nutritional, taste, and flavor properties. The biopreservation and the use of both bacteriocins produced by lactic acid bacteria (LAB) and bacteriocinogenic strains as an alternative to substitute chemical antimicrobial for food preservation became increasingly important in the last two decades. When the new proposed natural preservatives techniques are applied, probiotics food can be obtained and, simultaneously, foodborne pathogens and spoilage contaminants can diminish. However, bacteriocins produced by LAB have a narrow antibacterial spectrum and are inactive against Gram-negative bacteria like Salmonella and the emergent enterohemorrhagic Escherichia coli. Knowing the mechanism of action and the structural features of microcins synthesized by Gram-negative bacteria and with potent antimicrobial activity against the mentioned microorganism, the proposal is to obtain hybrid peptides (microcin–bacteriocin) with broad antimicrobial spectrum. This review explains how the inability of bacteriocins to cross the outer membrane of Gram-negative bacteria unable them to act on the bacteria. It will also be discussed how a hybrid bacteriocin can be obtained.     

Microbial and safety implications of the use of modified atmospheres to extend the storage life of fresh meat and fish

Recently there has been a renewed interest in the use of modified and controlled atmospheres to extend the shelf-life of fresh muscle foods at reasonable cost. The use of vacuum packaging, hypobaric conditions and atmospheres based on gas blends (CO₂, O₂, N₂, CO) in fresh meat and fish preservation, recent trends in this area, and the possible food safety implications of the application of such technologies, are reviewed in this paper. Oxygen depletion and/or CO₂ addition into a package are effective in retarding the growth of the typical aerobic spoilage bacteria in muscle foods. Yet there is a possibility that when such foods are temperature abused they may become unsafe with respect to certain foodborne bacterial pathogens and especially with respect to Clostridium botulinum. The question of whether or not spoilage may precede toxigenesis and thus alarm the processor or the consumer has not been fully clarified. Lack of such knowledge has remained the limiting factor to greater commercial expansion, especially by the fish industry. Carbon dioxide enriched atmospheres have thus far received the greatest commercial usage for the bulk shipment of muscle food. Under strict temperature control atmospheric modification can extend the shelf-life of such foods safely. More studies are needed before we fully utilize modified atmosphere technology. Such studies should include investigations on the action and interactions of the various factors in modified atmospheres upon the potential growth of pathogens as well as development or use of additional antimicrobial ‘hurdles’ to assure a predictable 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...