Promising strategies to control persistent enemies: Some new technologies to combat biofilm in the food industry—A review

Biofilm is an advanced form of protection that allows bacterial cells to withstand adverse environmental conditions. The complex structure of biofilm results from genetic-related mechanisms besides other factors such as bacterial morphology or substratum properties. Inhibition of biofilm formation of harmful bacteria (spoilage and pathogenic bacteria) is a critical task in the food industry because of the enhanced resistance of biofilm bacteria to stress, such as cleaning and disinfection methods traditionally used in food processing plants, and the increased food safety risks threatening consumer health caused by recurrent contamination and rapid deterioration of food by biofilm cells. Therefore, it is urgent to find methods and strategies for effectively combating bacterial biofilm formation and eradicating mature biofilms. Innovative and promising approaches to control bacteria and their biofilms are emerging. These new approaches range from methods based on natural ingredients to the use of nanoparticles. This literature review aims to describe the efficacy of these strategies and provide an overview of recent promising biofilm control technologies in the food processing sector.     

Biofilms in the Spotlight: Detection,Quantification, and Removal Methods

Microorganisms can colonize and subsequently form biofilms on surfaces, which protect them from adverse conditions and make them more resistant than their planktonic free‐living counterparts. This is a major concern in the food industry because the presence of biofilms has significant implications for microbial food contamination and, therefore, for the transmission of foodborne diseases. Adequate hygienic conditions and various preventive and control strategies have consequently been developed to ensure the provision of safe, good‐quality food with an acceptable shelf‐life. This review focuses on the significance of biofilms in the food industry by describing the factors that favor their formation. The interconnected process among bacteria known as “quorum sensing,” which plays a significant role in biofilm development, is also described. Furthermore, we discuss recent strategic methods to detect, quantify, and remove biofilms formed by pathogenic bacteria associated with food processing environments, focusing on the complexity of these microbial communities.     

Perspectives and Trends in the Application of Photodynamic Inactivation for Microbiological Food Safety

Photodynamic inactivation is a phenomenon that has the potential to cause microbial inactivation using visible light. It works on the principle that photosensitizers within the microbial cell can be activated using specific wavelengths to trigger a series of cytotoxic reactions. In the last few years, efforts to apply this intervention technology for food safety have been on the rise. This review article offers a detailed commentary on this research. The mechanism of photodynamic inactivation has been discussed as have the factors that influence its efficacy in food. Efforts to inactivate bacteria, fungi, and viruses have been analyzed in dedicated sections and so has the application of this technology to specific product classes such as fresh produce, dry fruits, seafood, and poultry. The challenges and opportunities facing the application of this technology to food systems have been evaluated and future research directions proposed. Thus, this review will provide insights for researchers and industry personnel looking for a novel solution to combat microbial contamination and resistance.     

Stress resistance of biofilm and planktonic Lactobacillus plantarum subsp. plantarum JCM 1149

The purpose of this study was to investigate the change in resistance of biofilm and planktonic food spoilage lactic acid bacteria (LAB) to environmental stresses, which strongly inhibit bacterial growth and are important in food preservation or in disinfection. The stress responses of biofilm and planktonic cells of Lactobacillus plantarum subsp. plantarum JCM 1149, which was used as a model spoilage bacterium, in various organic acids (namely, acetic acid, citric acid, lactic acid, and malic acid), ethanol, and sodium hypochlorite, were investigated using survival tests. The bacterial cells in biofilms showed greater resistance to all treatments than the planktonic bacterial cells in either the stationary or logarithmic phase. The planktonic bacterial cells showed reduced resistance to acetic acid after the cell suspension was diluted; however, intriguingly, the bacterial cells in biofilms maintained their resistance to acetic acid even after they were suspended or the cell suspension was diluted. These findings suggested the risk for food spoilage due to LAB derived from biofilms and suspended or diluted in foods, and demonstrated the importance of controlling biofilms of LAB in the food industry.     

Antimicrobial activity of coriander oil and its effectiveness as food preservative

Foodborne illness represents a major economic burden worldwide and a serious public health threat, with around 48 million people affected and 3,000 death each year only in the USA. One of the possible strategies to reduce foodborne infections is the development of effective preservation strategies capable of eradicating microbial contamination of foods. Over the last years, new challenges for the food industry have arisen such as the increase of antimicrobial resistance of foodborne pathogens to common preservatives and consumers demand for naturally based products. In order to overcome this, new approaches using natural or bio-based products as food preservatives need to be investigated. Coriander (Coriandrum sativum L.) is a well-known herb widely used as spice, or in folk medicine, and in the pharmacy and food industries. Coriander seed oil is the world's second most relevant essential oil, exhibiting antimicrobial activity against Gram-positive and Gram-negative bacteria, some yeasts, dermatophytes and filamentous fungi. This review highlights coriander oil antimicrobial activity and possible mechanisms of action in microbial cells and discusses the ability of coriander oil usage as a food preservative, pointing out possible paths for the successful evolution for these strategies towards a successful development of a food preservation strategy using coriander oil.     

Plasma-activated liquids for mitigating biofilms on food and food contact surfaces

Plasma-activated liquids (PALs) are emerging and promising alternatives to traditional decontamination technologies and have evolved as a new technology for applications in food, agriculture, and medicine. Contamination caused by foodborne pathogens and their biofilms has posed challenges and concerns to the food industry in terms of safety and quality. The nature of the food and the food processing environment are major factors that contribute to the growth of various microorganisms, followed by the biofilm characteristics that ensure their survival in severe environmental conditions and against traditional chemical disinfectants. PALs show an efficient impact against microorganisms and their biofilms, with various reactive species (short- and long-lived ones), physiochemical properties, and plasma processing factors playing a crucial role in mitigating biofilms. Moreover, there is potential to improve and optimize disinfection strategies using a combination of PALs with other technologies for the inactivation of biofilms. The overarching aim of this study is to build a better understanding of the parameters that govern the liquid chemistry generated in a liquid exposed to plasma and how these translate into biological effects on biofilms. This review provides a current understanding of PALs-mediated mechanisms of action on biofilms; however, the precise inactivation mechanism is still not clear and is an important part of the research. Implementation of PALs in the food industry could help overcome the disinfection hurdles and can enhance biofilm inactivation efficacy. Future perspectives in this field to expand existing state of the art to seek breakthroughs for scale-up and implementation of PALs technology in the food industry are also discussed.     

Bacterial biofilms associated with food particles in the human large bowel

Bacteria within the gastro-intestinal tract affect host function via production of short-chain fatty acids and synthesis of vitamins. Additionally, the commensal enteric bacteria modulate the immune system and provide protection from potentially pathogenic bacteria. Only recently heterogeneous bacterial biofilms were found to be associated with food particles within the intestinal tract. There are a number of studies investigating the formation and function of pathogenic and single-species biofilms, though few studies have investigated the dynamics of multispecies biofilms, especially with regard to food/microbial/host interactions. The scope of this review is to discuss the current knowledge of bacterial biofilms associated with food particles in the human large bowel, examine the established mathematical models depicting bacterial attachment, and elucidate key areas for further research.     

食品工业中混合菌生物被膜的形成、相互作用与新型控制策略

细菌黏附在食品或食品接触表面并形成生物被膜可能导致设备损坏、食品变质甚至人类疾病。混合菌生物被膜作为细菌在食品工业中的主要存在形式,与单菌生物被膜相比,对消毒剂和抗菌素往往具有更强的抗性。然而,混合菌生物被膜的形成与种间相互作用十分复杂,其在食品工业中的潜在作用仍有待探索。本文总结了混合菌生物被膜的形成和种间相互作用以及近年来的新型控制策略,并对未来食品工业中混合菌生物被膜的污染防控进行了展望,旨在为混合菌生物被膜在食品工业中的深入研究以及制定高效的新型控制策略提供理论依据和参考,以期更好地保障食品安全与公众健康。     

Pulsed light for the inactivation of fungal biofilms of clinically important pathogenic Candida species

Microorganisms are naturally found as biofilm communities more than planktonic free‐floating cells; however, planktonic culture remains the current model for microbiological studies, such as disinfection techniques. The presence of fungal biofilms in the clinical setting has a negative impact on patient mortality, as Candida biofilms have proved to be resistant to biocides in numerous in vitro studies; however, there is limited information on the effect of pulsed light on sessile communities. Here we report on the use of pulsed UV light for the effective inactivation of clinically relevant Candida species. Fungal biofilms were grown by use of a CDC reactor on clinically relevant surfaces. Following a maximal 72 h formation period, the densely populated biofilms were exposed to pulsed light at varying fluences to determine biofilm sensitivity to pulsed‐light inactivation. The results were then compared to planktonic cell inactivation. High levels of inactivation of C. albicans and C. parapsilosis biofilms were achieved with pulsed light for both 48 and 72 h biofilm structures. The findings suggest that pulsed light has the potential to provide a means of surface decontamination, subsequently reducing the risk of infection to patients. The research described herein deals with an important aspect of disease prevention and public health.     

噬菌体裂解酶结构特征、重组策略及其在控制食源性致病菌中的应用

持续的全球食源性疾病和耐药细菌的广泛流行,对食品安全和人类健康造成了极大的威胁,迫切需要研发新型杀菌、控菌技术。噬菌体裂解酶是大部分裂性噬菌体在裂解期释放一种活性蛋白,能够有效裂解宿主细胞壁,已被证明可应用于食品供应链的各个环节中控制食源性致病菌风险。天然噬菌体裂解酶具有高度的宿主特异性和强烈的裂解活性,能破坏细菌生物被膜,而且具备绿色安全、不易产生耐药等优势。同时,噬菌体裂解酶具有模块化结构特点,运用蛋白质工程技术将其重组,可增强其裂解活性、提高稳定性以及靶向性。本综述系统地描述了噬菌体裂解酶的模块化结构特征及作用位点,讨论了噬菌体裂解酶的重组策略和方法,总结了天然噬菌体裂解酶在控制食源性致病菌方面的应用进展,并对噬菌体裂解酶在食品工业中的应用进行了展望,以期为食源性致病菌及其耐药性的有效控制提供一种行之有效的新策略。     

食品中细菌生物被膜及其形成机制的研究进展

生物被膜是自然条件下细菌在固态表面生长时采取的一种较为独特的群体生活方式,该方式较浮游状态存在显著生物学特征差异,对食品的危害更为严重,但目前国内外食品领域对其研究较少.本文在收集、研究现有文献的基础上归纳介绍了细菌生物被膜的特点及其形成过程,概述了各种影响细菌生物被膜形成的因素与机制,旨在提高人们对细菌生物被膜的认识,防范和减少细菌生物被膜对食品质量与安全的危害,同时推动该领域的研究发展.     

Mercury methylation by planktonic and biofilm cultures of Desulfovibrio desulfuricans

While biofilms are now known to be the predominant form of microbial growth in nature, very little is yet known about their role in environmental mercury (Hg) methylation. Findings of Hg methylation in periphyton communities have indicated the importance of investigating how environmental biofilms affect Hg methylation, as periphyton can be the base of the food webs in aquatic ecosystems. Chemical speciation influences the microbial uptake and methylation of inorganic Hg by planktonic cultures of sulfate-reducing bacteria; however, the effect of speciation on Hg methylation by biofilm cultures of these organisms has previously not been studied. In the present study, Hg methylation rates in biofilm and planktonic cultures of two isolates of Desulfovibrio desulfuricans from a coastal wetland were compared. Notably, the specific Hg methylation rate found was approximately an order of magnitude higher (0.0018 vs. 0.0002 attomol cell(-1) day(-1)) in biofilm cells than in planktonic cells, suggesting an important role for environmental biofilms in Hg methylation. To investigate the role of chemical speciation of Hg, experiments were conducted at two levels of sulfide. Both biofilm and planktonic cultures produced methylmercury at roughly twice the rate at low sulfide, when HgS(0)(aq), rather than HgHS2-, was the dominant Hg species. This indicates that the presence of a biofilm does not alter the relative availability of the dominant Hg species in sulfidic medium, in accordance with our previous studies of Hg uptake by Escherichia coli along a chloride gradient.     

Photodynamic inactivation of spoilers Pseudomonas lundensis and Brochothrix thermosphacta by food-grade curcumin and its application on ground beef

This study evaluated the bactericidal effects of curcumin-mediated photodynamic inactivation (PDI) combined with citric acid on meat spoilage bacteria, and their preservative effects of refrigerated beef. After treatment of PDI (60 μM curcumin and 33.01 J/cm²) combined with 0.5 mg/mL citric acid, the populations of Pseudomonas lundensis and Brochothrix thermosphacta in PBS solution were reduced by 3.07 and 6.14 log CFU/mL, respectively, compared to 0.31 and 1.39 log CFU/mL for PDI. Combined treatment enhanced reactive oxygen species (ROS) activity in the two strains, and damaged their cell morphology, especially B. thermosphacta. With increasing curcumin concentration, the growth rate of spoilage bacteria was significantly inhibited (P < 0.05), and the changes of dominant spoilage species in bacterial community were retarded in aerobic-packaged and vacuum-packaged ground beef at 4 °C. Compared to the control, trichloroacetic acid-soluble peptide and total volatile basic nitrogen production were greatly restrained, and changes in pH values and brown color were apparently reduced for the combined treatment, which could extend the shelf life by 2 days. These results suggested the synergistic potency of PDI combined with citric acid against spoilage bacteria in meat products.Industrial relevance: The proliferation and spread of some psychrotrophs are major causes of quality deterioration and cross-contamination in fresh meat during cold chain. This study revealed that photosensitized curcumin combined with citric acid significantly decreased the survival of two psychrotrophic spoilage bacteria and extended the shelf-life of ground beef. Thus, curcumin-based PDI provides a new insight for the development of efficient and economic preservation in the meat processing industry.     

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。     

LuxS/AI-2型群体感应系统调控细菌生物被膜形成研究进展

生物被膜是大多数细菌在自然状态下的一种生长方式,使菌体具有浮游态时不具有的优势。它的形成和发展受到群体感应系统的调控,该系统是细菌依赖于群体密度而调控其生理行为的一种机制。其中LuxS/2型自诱导物（autoinducer 2,AI-2）群体感应系统又称种间群体感应系统,广泛存在于G＋及G－菌中。其信号分子AI-2被认为是种间通用的信号分子,参与调控多种细菌的生物被膜。此外,目前已发现多种LuxS/AI-2型群体感应系统抑制剂,它们可以影响许多细菌生物被膜的形成。目前研究人员已开始致力于揭示LuxS/AI-2型群体感应系统调控生物被膜形成的分子机制,这对于进一步理解LuxS/AI-2型群体感应系统与生物被膜的关系具有重要意义。     

Current and Recent Advanced Strategies for Combating Biofilms

Biofilms are matrix‐enclosed microbial aggregates that adhere to a biological or nonbiological surface. Biofilm formation is a significant problem in the medical, food, and marine industries and can lead to substantial economic and health problems. The complex microbial community of a biofilm is highly resistant to antibiotics and sanitizers and confers persistent survival that is a challenge to overcome. There are several conventional approaches to combating biofilms, physical and/or mechanical removal, chemical removal, and the use of antimicrobials, sanitizers, or disinfectants to kill biofilm organisms. However, biofilms are highly resistant to these approaches as opposed to planktonic cells. Thus, novel approaches other than the conventional methods are urgently needed. In this review, we discuss current and new advanced antibiofilm strategies that are superior to the conventional method in terms of addressing the biofilm problem for the improvement of healthcare, food safety, and in industrial processes.     

Biofilm Formation Characteristics of Pseudomonas lundensis Isolated from Meat

Biofilms formations of spoilage and pathogenic bacteria on food or food contact surfaces have attracted increasing attention. These events may lead to a higher risk of food spoilage and foodborne disease transmission. While Pseudomonas lundensis is one of the most important bacteria that cause spoilage in chilled meat, its capability for biofilm formation has been seldom reported. Here, we investigated biofilm formation characteristics of P. lundensis mainly by using crystal violet staining, and confocal laser scanning microscopy (CLSM). The swarming and swimming motility, biofilm formation in different temperatures (30, 10, and 4 °C) and the protease activity of the target strain were also assessed. The results showed that P. lundensis showed a typical surface‐associated motility and was quite capable of forming biofilms in different temperatures (30, 10, and 4 °C). The strain began to adhere to the contact surfaces and form biofilms early in the 4 to 6 h. The biofilms began to be formed in massive amounts after 12 h at 30 °C, and the extracellular polysaccharides increased as the biofilm structure developed. Compared with at 30 °C, more biofilms were formed at 4 and 10 °C even by a low bacterial density. The protease activity in the biofilm was significantly correlated with the biofilm formation. Moreover, the protease activity in biofilm was significantly higher than that of the corresponding planktonic cultures after cultured 12 h at 30 °C.     

Biofilm Formation by Shiga Toxin-Producing Escherichia coli O157:H7 and Non-O157 Strains and Their Tolerance to Sanitizers Commonly Used in the Food Processing Environment

Shiga toxin-producing Escherichia coli (STEC) strains are important foodborne pathogens. Among these, E. coli O157:H7 is the most frequently isolated STEC serotype responsible for foodborne diseases. However, the non-O157 serotypes have been associated with serious outbreaks and sporadic diseases as well. It has been shown that various STEC serotypes are capable of forming biofilms on different food or food contact surfaces that, when detached, may lead to cross-contamination. Bacterial cells at biofilm stage also are more tolerant to sanitizers compared with their planktonic counterparts, which makes STEC biofilms a serious food safety concern. In the present study, we evaluated the potency of biofilm formation by a variety of STEC strains from serotypes O157:H7, O26:H11, and O111:H8; we also compared biofilm tolerance with two types of common sanitizers, a quaternary ammonium chloride-based sanitizer and chlorine. Our results demonstrated that biofilm formation by various STEC serotypes on a polystyrene surface was highly strain-dependent, whereas the two non-O157 serotypes showed a higher potency of pellicle formation at air-liquid interfaces on a glass surface compared with serotype O157:H7. Significant reductions of viable biofilm cells were achieved with sanitizer treatments. STEC biofilm tolerance to sanitization was strain-dependent regardless of the serotypes. Curli expression appeared to play a critical role in STEC biofilm formation and tolerance to sanitizers. Our data indicated that multiple factors, including bacterial serotype and strain, surface materials, and other environmental conditions, could significantly affect STEC biofilm formation. The high potential for biofilm formation by various STEC serotypes, especially the strong potency of pellicle formation by the curli-positive non-O157 strains with high sanitization tolerance, might contribute to bacterial colonization on food contact surfaces, which may result in downstream product contamination.     

等离子体活化水在食品工业中应用研究进展

等离子体活化水(Plasma-activated water,PAW)具有活性组分含量高、低pH和氧化还原电位较高等特点,具有杀菌、抗生物被膜、促进种子萌发和幼苗生长等功能。作为一种新型的环境友好型非热加工技术,PAW在食品工业中的潜在应用前景受到广泛关注。本文综述了PAW在食品杀菌保鲜、肉制品护色、细菌生物被膜控制等领域中的应用研究,为PAW技术在食品工业中的广泛应用提供参考。     

光动力杀菌机制及在食品应用中的优势与不足

有害微生物是引起食品安全问题的重要原因之一。广谱抗生素的使用虽然有效地抑制了有害微生物,但耐药性导致其无法得到有效控制,这加重了食品安全隐患,同时造成更严重的经济损失。因此,迫切需要开发其它替代杀菌技术,这也是保护食品安全的重要挑战。光动力杀菌技术是通过光激发光敏剂后,产生的活性氧物质破坏致病菌的形态结构、细胞膜、核酸和蛋白质等,同时触发多种细菌难以抵消的死亡机制,此机制不易造成细菌耐受性。光动力杀菌技术杀菌效果佳,但此杀菌技术对食品营养成分及品质的影响关注较少,限制了其在食品领域中的推广和应用。本文系统综述了光动力杀菌技术的杀菌机制及影响因素,并且介绍了光动力杀菌在食品中的研究应用,就杀菌技术在食品应用中的优势和不足提出新见解,以期为我国光动力杀菌技术的工业化应用提供参考。