Biofilm-forming pathogens such as Pseudomonas aeruginosa and Escherichia coli present major food safety risks due to their strong adhesion to processing surfaces and resistance to sanitization. This study evaluated the antibiofilm efficacy of Coenzyme Q0 (CoQ0) on stainless steel, aluminum, silicone rubber, polyethylene terephthalate, and shrimp shell surfaces, as well as in single- and mixed-species biofilms. CoQ0 exhibited minimum inhibitory concentration/minimum bactericidal concentration (MIC/MBC) values of 16/20 μg/mL for P. aeruginosa, 60/64 μg/mL for E. coli, and 100/200 μg/mL for mixed biofilms, with complete eradication achieved at 4 MIC. P. aeruginosa was more sensitive at lower concentrations, whereas E. coli showed a dose-dependent response. Mechanistic assays demonstrated adenosine triphosphate (ATP) depletion, increased reactive oxygen species (ROS) generation, reduced respiratory activity in the triphenyl tetrazolium chloride (TTC) assay, and membrane disruption confirmed by extracellular protein leakage. Nuclear magnetic resonance (NMR) spectroscopy revealed degradation of extracellular polymeric substances (EPS), while reverse transcription polymerase chain reaction (RT-PCR) showed downregulation of biofilm and quorum-sensing genes (algU, rhlR, exoS in P. aeruginosa; rpoS, luxS, csgA in E. coli). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in MA-104 cells confirmed good tolerance up to 60 μg/mL. Confocal laser scanning microscopy (CLSM) and field-emission scanning electron microscopy (FE-SEM) further validated biofilm disintegration and cell damage. Overall, CoQ0 exhibits a multifaceted antibiofilm mechanism with species- and surface-specific effects, supporting its potential as a natural antimicrobial for food safety applications.

S. Typhimurium biofilm formation on chicken and food-contact surfaces presents a major challenge to food safety by enhancing bacterial persistence and resistance to sanitation. This study investigated the efficacy and mechanisms of cinnamon leaf oil (CLO), a sustainable plant-based antimicrobial, in controlling S. Typhimurium (ATCC 14028) biofilm formation. CLO exhibited a minimum inhibitory concentration (MIC) of 0.09% (v/v) (equivalent to 0.9 μL/mL). At concentrations ranging from ¼× to 4× MIC, CLO significantly (p < 0.05) reduced biofilm formation on chicken meat and reduced bacterial populations within preformed biofilms on food-contact surfaces including low and high-density polyethylene, polyethylene terephthalate, rubber and stainless steel in a dose-dependent manner. Mechanistic analyses revealed that CLO delayed bacterial motility, reduced cell surface hydrophobicity by approximately 60% at 4× MIC and inhibited autoaggregation. Confocal laser scanning microscopy demonstrated concentration-dependent inhibition of biofilm formation, with biofilms formed in the presence of CLO appearing progressively thinner, less densely organized and containing a higher proportion of non-viable cells as the CLO concentration increased. Collectively, these findings indicate that CLO inhibits the early stages of biofilm formation by interfering with bacterial adhesion and colonization, while also reducing bacterial populations within pre-established biofilms on food-contact surfaces.

ABSTRACT: Cronobacter sakazakii, an opportunistic foodborne pathogen, primarily with the infections that can cause bacteremia, necrotizing enterocolitis, and neonatal meningitis. These pathogens are Gram-negative rods, motile with peritrichous flagella without spore-forming capability, can grow under aerobic and anaerobic conditions, and tolerance to acidic conditions and desiccations stress. Phenyllactic acid (PLA), a phenolic acid with broad-spectrum antibacterial activity, is a promising natural preservative. However, its effects on C. sakazakii biofilms remain unclear. This study evaluated the antibiofilm potential of PLA, a natural antimicrobial metabolite, against C. sakazakii and investigated its effects on cell membrane integrity. First, the minimum inhibitory concentration (MIC) of PLA was assessed at 4 mg/mL. The MIC (4 mg/mL) of PLA was observed to reduce C. sakazakii for planktonic cells and biofilm formation on food-contact surfaces (stainless steel, silicon rubber, and polyvinyl chloride) by about 3 log CFU/cm2, respectively. PLA at 4 mg/mL significantly altered ALP activity, intracellular and extracellular ATP, suggested membrane disruption, cell integrity, and leakage of intracellular components. Fourier-transform infrared spectroscopy (FTIR) analysis provided extracellular polymeric substances composition. As confirmed by confocal laser scanning microscopy and scanning electron microscopy revealed disrupted biofilm architecture and altered cell morphology. Mechanistic analyses demonstrated that PLA exposure increased membrane permeability, indicated by elevated extracellular ATP and protein leakage, while intracellular ATP levels decreased. All things considered, these results clarify PLA's effectiveness against biofilm of C. sakazakii. These results demonstrate that PLA inhibits C. sakazakii biofilm formation primarily by disrupting membrane integrity, suggesting its potential as a natural antimicrobial strategy for controlling biofilms and supports the potential application of PLA as an effective antimicrobial agent in food industry to enhancing food safety.

Biofilm-mediated contamination poses a persistent threat in the global meat industry, facilitating the survival of foodborne pathogens and reducing the effectiveness of conventional sanitation methods. This review highlights postbiotics, non-viable microbial derivatives and cellular components, including metabolic byproducts, peptides, organic acids, bacteriocins, biosurfactants, and exopolysaccharides which are emerging as multifunctional agents for biofilm control and meat preservation. Postbiotics exert their effects through diverse mechanisms including disrupting adhesion via surface conditioning and gene suppression, impairing EPS matrix formation by interfering with cyclic-di-GMP signaling, and inducing oxidative and metabolic stress in biofilm-forming cells. Unlike probiotics and prebiotics, postbiotics are stable under harsh processing conditions and deliver immediate antimicrobial action without requiring host interaction. Case studies demonstrate their efficacy in reducing Listeria monocytogenes, Salmonella spp., and Escherichia coli on meat surfaces, packaging materials, and equipment. In addition to microbial safety, postbiotics contribute to product quality by enhancing lipid stability, protein integrity, and color retention. A VOSviewer-based bibliometric analysis further maps global research trends, thematic evolution, and knowledge gaps in postbiotic applications within meat systems. By integrating bibliometric analysis with mechanistic insights, this review evaluates postbiotics as sustainable tools for biofilm control in the meat industry, while emphasizing the need to overcome challenges related to standardization, delivery systems, and regulatory approval to support their effective adoption in clean-label preservation strategies that safeguard meat safety, quality, and consumer acceptance

Bacteriophages are gaining increasing attention as targeted biological control agents against multidrug-resistant (MDR) foodborne pathogens. However, repeated exposure to bacteriophages can induce the emergence of bacteriophage-resistant mutant (BIM) strains, which may exhibit altered physiological characteristics that impair therapeutic efficacy. This study investigated the phenotypic and molecular adaptive mechanisms of multidrug-resistant Salmonella Typhimurium BIMs that acquired resistance to receptor-specific bacteriophages (STP-1 and STP-2) targeting O-antigen and flagella. Two representative BIM strains—MO-4 (O-antigen mutant) and MF-6 (flagella mutant)—were analyzed using phenotypic microarray, motility and biofilm experiments, confocal microscopy, and qRT-PCR. MO-4 exhibited extensive metabolic changes, including 10 alterations in carbon source utilization and increased resistance to 11 antibiotics, while MF-6 showed changes in sensitivity to osmotic and pH stress and increased susceptibility to 5 antibiotics. Both BIM strains exhibited reduced motility but maintained similar or enhanced biofilm formation ability on food contact surfaces. Confocal microscopy analysis revealed structurally distinct biofilms with reduced thickness and increased density. qRT-PCR analysis showed receptor-specific transcriptional changes: MO-4 lost rfaL expression, supporting O-antigen deficiency, while MF-6 showed increased fljB expression and decreased fliK expression, indicating changes in flagellar regulation. Both BIMs showed differences in the expression of tolC, rpoS, and luxS, suggesting changes in efflux, stress response, and quorum sensing. These results highlight receptor-dependent adaptation of phage-resistant Salmonella variants and emphasise the need to evaluate both genetic and phenotypic outcomes when designing effective phage-based control strategies for food safety.

Weissella is a fascinating genus that has garnered increasing attention for its probiotic potential and industrial versatility. In this study, we present a detailed genomic and phenotypic investigation of six Weissella strains isolated from kimchi, identified as W. cibaria (254, 256, 258, 260) and W. confusa (255, 257). Genomic analysis revealed species-specific differences, with W. cibaria linked to cell cycle regulation and replication, while W. confusa displayed adaptations for motility and metabolic diversity. Antibiotic resistance gene analysis revealed the presence of daunorubicin resistance ( drrA ), multidrug resistance determinants ( bmr3 , lmrA , ykkCD ), and species-specific resistance to bicyclomycin ( bcr1 , bcr2 ) and quaternary ammonium compounds ( qacC ). Phenotypic antibiotic susceptibility testing demonstrated intrinsic resistance to vancomycin alongside high sensitivity to ampicillin, erythromycin, chloramphenicol, and gentamicin. Probiotic assessments highlighted significant tolerance to acid, bile salts, and enzymatic stress, with W. cibaria 256 and W. confusa 255 showing superior survivability under simulated gastrointestinal conditions. High surface hydrophobicity observed in W. confusa 255 and 257 further underscores their adhesion capabilities, which are crucial for gut colonization. Exopolysaccharide (EPS) production was further characterized by FTIR analysis, revealing diverse functional groups associated with structural protection and probiotic functionality. Morphological characteristics as determined by field emission scanning electron microscopy (FE-SEM) revealed rod-shaped cells of 1.39–1.87 μm in length and 0.50–0.70 μm in width for W. cibaria strains versus 1.02–1.45 μm in length and 0.54–0.62 μm in width for W. confusa strains. These findings provide novel insights into the genomic and phenotypic underpinnings of Weissella species' probiotic properties, resistance mechanisms, and adaptability to fermented food environments, highlighting their promising applications in functional food development and human health enhancement.

Seafood plays a pivotal role in global nutrition and sustainability, yet its safety is frequently compromised by a wide array of hazards encountered throughout the aquatic food chain. This comprehensive review integrates bibliometric analysis of 1524 peer-reviewed publications (2000–2024) with an in-depth scientific discussion of physical, chemical, and biological threats affecting seafood safety. Keyword mapping using VOSviewer identified six major research clusters, revealing a clear temporal shift from contaminant detection toward exposure-based risk assessment, public health relevance, and regulatory concerns, with dominant contributions from North America, Europe, and East Asia. The scientific synthesis highlights key hazards, including foreign materials (e.g., bones, glass, metals), toxic environmental contaminants (e.g., heavy metals, microplastics, marine biotoxins), and microbial risks involving spoilage organisms, bacteria, viruses, fungi, and parasites. The review further outlines hazard-specific control strategies, emphasizing biofilm-associated microbial persistence as a cross-cutting challenge and evaluating emerging intervention technologies within a risk-based and One Health framework. Overall, this integrated approach advances seafood safety research by aligning global research trends with hazard prioritization and risk-informed management strategies.

Aquatic foods are essential sources of protein and micronutrients and play a critical role in global nutrition, trade, and livelihoods. However, their safety and sustainability are frequently compromised by microbial contamination and biofilm formation during farming, processing, storage, and retail. Biofilms persist on moist surfaces, resist conventional cleaning practices, and contribute to spoilage, cross-contamination, and economic loss. This article reviews emerging applications of artificial intelligence and Industry 4.0 technologies for biofilm prevention and control in aquaculture and seafood systems. Particular emphasis is placed on the use of continuous water quality sensing, imaging platforms for early detection and cleaning verification, genomic and omics tools for microbial trait-level insight, and digital twin frameworks for virtual simulation of sanitation strategies. Recent advances demonstrate that sensor telemetry can predict biofilm-favorable conditions, imaging can verify removal in real time, and genomic data can identify persistence traits and tolerance mechanisms. When integrated, these approaches enable facility-specific digital twins that anticipate surface-specific risks and recommend optimized interventions before implementation. The convergence of AI, sensor networks, imaging, and omics offers a shift from reactive to proactive biofilm management in aquatic food systems. Positioned within the transition to Industry 5.0, these innovations support earlier detection, targeted interventions, and measurable improvements in food safety, quality, sustainability, and resilience, while aligning production systems with human-centric goals.

This study aimed to develop alternative packaging materials by coating polyethylene terephthalate (PET) with antimicrobial substances (glucose oxidase and potassium sorbate) and confirming antimicrobial and antibiofilm activity of the coated PET. Before examining the antimicrobial activity of coated PET, attenuated total reflectance–Fourier-transform infrared spectroscopy was conducted to ensure that glucose oxidase and potassium sorbate were present and generously coated on the PET samples. The graph peaks indicated the conjugated interaction of glucose oxidase with potassium sorbate on the chitosan-based PET surface. At 4 × MIC, glucose oxidase–coated PET reduced E. coli biofilm on shrimp by 2.9 log CFU/g at 10 °C and 3.1 log CFU/g at 25 °C, and on beef by 2.6 log CFU/g at 10 °C and 1.5 log CFU/g at 25 °C, relative to untreated controls. Potassium sorbate–coated PET achieved reductions of 2.9 and 3.1 log CFU/g on shrimp, and 2.9 and 1.9 log CFU/g on beef at 10 and 25 °C, respectively. Notably, the combination-coated PET exhibited the greatest antibiofilm efficacy, reducing E. coli by 2.9 and 3.1 log CFU/g on shrimp and 3.7 and 3.8 log CFU/g on beef at 10 and 25 °C, respectively. The inhibitory effect of combination treatment with glucose oxidase and potassium sorbate against E. coli biofilm was visually confirmed using confocal laser scanning microscopy and field-emission scanning electron microscopy. Moreover, reverse transcription polymerase chain reaction analysis revealed that glucose oxidase and potassium sorbate could inhibit quorum sensing- and virulence-related gene (luxS, pdfs, and rpoS) expression in E. coli. Therefore, this study confirmed that combination treatment with glucose oxidase and potassium sorbate effectively reduces E. coli biofilms, thus increasing food safety.

Clostridium perfringens (C. perfringens) biofilms pose a persistent challenge in meat and poultry processing environments due to their structural resilience, spore-mediated survival and toxin-associated virulence. These biofilms readily develop on food-contact surfaces under typical processing conditions including organic residue accumulation, temperature fluctuations, and localized anaerobic niches, leading to increased tolerance to sanitation and thermal treatments. Mechanistically, biofilm resilience in C. perfringens is governed by the integration of sporulation processes, quorum sensing–regulated gene expression and extracellular polymeric substance (EPS) matrix formation, which collectively enhance stress tolerance, limit antimicrobial penetration, and facilitate persistence under fluctuating environmental conditions. The interaction between spore formation and EPS architecture further promotes survival during thermal processing and enables rapid re-establishment of biofilms following sanitation. This review synthesizes current knowledge on the formation and persistence of C. perfringens biofilms, key environmental drivers in meat and poultry processing systems and the mechanistic basis of their stress resistance and survival strategies. It also critically examines how these mechanisms influence the efficacy of existing intervention strategies. It further evaluates the limitations of conventional control strategies and highlights emerging approaches for biofilm prevention and control, including food-grade antimicrobials, surface engineering, enzymatic disruption, and microbiome-based interventions, with emphasis on their modes of action and applicability in industrial settings. Overall, this review provides a mechanistic and systems-level perspective to support the development of more effective biofilm control strategies in meat processing environments.

Fermentation research has traditionally concentrated on free-floating microorganisms. In contrast, biofilms are commonly seen as a contamination danger. However, many food fermentations, like kombucha, cheese rinds and koji, rely on surface-attached populations. This review defines sessile fermentation as the purposeful utilization of controlled biofilms as functional aids in food systems. We combine biofilm biology and process engineering to describe how natural biofilm fermentations maintain stability and safety. Key processes include diffusion reaction gradients, microenvironments and quorum sensing. Surface design, cell immobilization and reactor types are all parts of engineering techniques. We then focus on three practical contributions, namely continuous processing, bio-preservation and functional food production. We also handle real-world issues including scale-up, fouling, cleaning-in-place and process control. Overall, altering our perspective on biofilms from issue to platform provides a realistic pathway to more efficient and scalable food fermentation.

The authors regret after carefully reviewing the original dataset and the published table, we acknowledge that the values presented under the “chicken meat” column for the carvacrol treatment appear duplicated with those of the baicalin treatment at corresponding concentrations. This duplication was not intentional and occurred during the final stage of table preparation, where formatting and data consolidation errors led to incorrect insertion of values. Importantly, we would like to clarify that: (1) The raw experimental data for baicalin and carvacrol treatments are distinct and were generated independently.(2) The duplication is limited to the presentation in Table 2 and does not reflect the actual experimental outcomes.(3) The statistical analyses, interpretations, and conclusions of the study were conducted using the correct datasets.We have rechecked the original data files and prepared a corrected version of Table 2. In summary: The issue is a data transcription/formatting error during manuscript preparation. There is no intentional duplication of experimental results. The corrected table accurately represents the original experimental findings.The authors would like to apologise for any inconvenience caused.

Drug resistance is a serious problem for human health worldwide. Day by day this drug resistance is increasing and creating an anxious situation for the treatment of both cancer and infectious diseases caused by pathogenic microorganisms. Researchers are trying to solve this terrible situation to overcome drug resistance. Biosynthesized gold nanoparticles (AuNPs) could be a promising agent for controlling drug-resistant pathogenic microorganisms and cancer cells. AuNPs can be synthesized via chemical and physical approaches, carrying many threats to the ecosystem. Green synthesis of AuNPs using biological agents such as plants and microbes is the most fascinating and attractive alternative to physicochemical synthesis as it offers many advantages, such as simplicity, non-toxicity, cost-effectiveness, and eco-friendliness. Plant extracts contain numerous biomolecules, and microorganisms produce various metabolites that act as reducing, capping, and stabilizing agents during the synthesis of AuNPs. The characterization of green-synthesized AuNPs has been conducted using multiple instruments including UV–Vis spectrophotometry (UV–Vis), transmission electron microscopy (TEM), scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray diffraction (XRD), DLS, and Fourier transform infrared spectroscopy (FT-IR). AuNPs have detrimental effects on bacterial and cancer cells via the disruption of cell membranes, fragmentation of DNA, production of reactive oxygen species, and impairment of metabolism. The biocompatibility and biosafety of synthesized AuNPs must be investigated using a proper in vitro and in vivo screening model system. In this review, we have emphasized the green, facile, and eco-friendly synthesis of AuNPs using plants and microorganisms and their potential antimicrobial and anticancer applications and highlighted their antibacterial and anticancer mechanisms. This study demonstrates that green-synthesized AuNPs may potentially be used to control pathogenic bacteria as well as cancer cells.

The pervasive use of plastics in the food industry has led to significant microplastic contamination, heightening food safety concerns. Microplastics not only infiltrate food systems but also provide optimal substrates for biofilm formation due to their hydrophobic and rough surface properties, which enhance microbial attachment. These biofilm–microplastic complexes protect embedded bacteria from disinfection efforts and facilitate the dissemination of pathogens and antibiotic-resistance genes, posing substantial risks to human health. This review investigates the mechanisms of biofilm formation on various plastic materials and elucidates how these interactions contribute to antimicrobial resistance and contamination in food systems. It also evaluates preventive and remedial strategies, including the development of alternative packaging materials, advanced cleaning protocols, and detection techniques for monitoring biofilm–microplastic complexes. Understanding these interactions is crucial for developing targeted interventions to mitigate contamination risks and enhance food safety and security. By integrating recent findings and proposing innovative strategies, this review aims to guide future research and inform policy development, supporting safer and more sustainable food production practices.

Biofilm, complex structures formed by microorganisms within an extracellular polymeric matrix, pose significant challenges in the sector by harboring dangerous pathogens and complicating decontamination, thereby increasing the risk of foodborne illnesses. This article provides a comprehensive review of the sigma factor, rpoS’s role in biofilm development, specifically in gram-negative bacteria, and how the genetic, environmental, and regulatory elements influence rpoS activity with its critical role in bacterial stress responses. Our findings reveal that rpoS is a pivotal regulator of biofilm formation, enhancing bacterial survival in adverse conditions. Key factors affecting rpoS activity include oxidative and osmotic stress and nutrient availability. Understanding rpoS-mediated regulatory pathways is essential for developing targeted biofilm management strategies to improve food quality and safety. Furthermore, a bibliometric analysis highlights significant research trends and gaps in the literature, guiding future research directions. Future research should focus on detailed mechanistic studies of rpoS-mediated biofilm regulation, the development of specific rpoS inhibitors, and innovative approaches like biofilm-resistant surface coatings. This knowledge can lead to more effective contamination prevention and overall food safety enhancements.

Seafood is essential for achieving the United Nations Sustainable Development Goals (SDGs), particularly in enhancing food security and supporting sustainable livelihoods. However, the seafood industry faces challenges due to the short shelf life of its products, primarily caused by microbial contamination and biofilm formation, which increase resistance to conventional preservation methods. This review examines the microbial hazards associated with seafood, focusing on the complexities of biofilm formation on seafood and processing equipment. It explores advanced non-thermal decontamination technologies, such as electron beam irradiation, cold plasma, and ultrasonication, alongside natural antimicrobial strategies, including plant extracts, biosurfactants, and antimicrobial peptides. These innovative methods show potential for enhancing seafood safety, extending shelf life, and preserving nutritional quality. The review emphasizes the need for ongoing research into effective biofilm control, improved detection methods, and robust regulatory frameworks to ensure the production of safe, high-quality seafood. Additionally, a bibliometric analysis highlights current research trends and key focus areas within this field, providing a comprehensive overview of the evolving scientific landscape related to seafood safety and biofilm management, and identifying critical opportunities for future research and innovation.

This study provides a comprehensive analysis of biofilm formation, antibiotic resistance, motility, and gene expression in four Aeromonas hydrophila strains—ATCC 15467, ATCC 7966, KCTC 2358, and KCTC 11533—on stainless steel (SS), silicon rubber (SR), polyethylene terephthalate (PET), and high-density polyethylene (HDPE) surfaces over 24, 48, 72, and 96 h. Biofilm formation peaked at 72 h, with ATCC 7966 demonstrating the highest biofilm density on PET (6.50 ± 0.08 log CFU/cm²), underscoring PET's role as a favorable substrate for biofilm development. In contrast, HDPE consistently exhibited the lowest biofilm levels, reflecting its potential as a biofilm-resistant material. Antibiotic susceptibility profiling revealed multidrug resistance (MDR) in ATCC 15467 and KCTC 11533 (MARI = 0.80), particularly against beta-lactams, aminoglycosides, and fluoroquinolones while ATCC 7966 and KCTC 2358 displayed moderate resistance. Motility assays highlighted strain-specific capabilities, with KCTC 11533 exhibiting the highest swimming motility (76.0 ± 6.6 mm) and KCTC 2358 excelling in swarming (47.7 ± 3.5 mm). Genetic analysis confirmed the presence of luxS and ahyR in all strains, while csgA was exclusive to ATCC 7966, correlating with its superior biofilm formation. Confocal microscopy revealed biofilm maturation dynamics, with red fluorescence indicating cell death and aging at 96 h, while SEM images captured intricate surface-specific biofilm architectures. These findings elucidate the critical interplay between strain characteristics, surface properties, and incubation time, providing a foundation for developing targeted strategies to control A. hydrophila biofilms in food processing environments.

Biofilm formation on dairy equipment is a persistent challenge in the dairy industry, contributing to product contamination, equipment inefficiency, and economic losses. Traditional methods such as manual cleaning and basic chemical sanitation are discussed as foundational approaches, followed by an in-depth investigation of cutting-edge technologies, including clean-in-place systems, high-pressure cleaning, foam cleaning, ultrasonic and electrochemical cleaning, dry ice blasting, robotics, nanotechnology-based agents, enzymatic cleaners, and oxidizing agents. Enhanced sanitation techniques, such as dry steam, pulsed light, acidic and alkaline electrolyzed water, hydrogen peroxide vapor, microbubble technology, and biodegradable biocides, are highlighted for their potential to achieve superior sanitation while promoting sustainability. The effectiveness, feasibility, and limitations of these methods are evaluated, emphasizing their role in maintaining dairy equipment hygiene and reducing biofilm-associated risks. Additionally, challenges, such as equipment compatibility, cost, and regulatory compliance, are addressed, along with insights into future directions and innovations, including automation, smart cleaning systems, and green cleaning solutions. This review provides a comprehensive resource for researchers, industry professionals, and policymakers aiming to tackle biofilm formation in dairy production systems and enhance food safety, operational efficiency, and sustainability.

The biofilm-mediated persistence of Pseudomonas aeruginosa in the food and biomedical sectors is currently a global concern. In light of this challenge, this study investigated a preventive approach against P. aeruginosa biofilm formation using Flavourzyme, a food-grade peptidase, considering its antibiofilm potential. The results revealed that a co-culture comprising 300 µL/mL (1 × minimum inhibitory concentration [MIC]) of Flavourzyme could kill P. aeruginosa. On the MBEC™ biofilm-forming device, 0.125 × MIC of Flavourzyme inhibited > 4.5 log CFU/peg of biofilm. Cell motilities and the biosynthesis of quorum sensing (QS) molecules such as N-acyl-homoserine lactones (AHLs), including C4-HSL, decreased significantly at 0.06 × MIC of Flavourzyme and became undetectable at 0.125 × MIC. Interestingly, while 0.03 × MIC of Flavourzyme elicited diverse expressions of QS and virulence-regulating genes, ≥ 0.06 × MIC of Flavourzyme remarkably suppressed the relative genomic expressions. This study proposes Flavourzyme as a potent antibiofilm agent against P. aeruginosa biofilms, recommending specific concentrations for effective use in food preservation.

Biofilms on food-contact surfaces pose persistent challenges to sanitation, safety, and product quality within food processing. Traditional cleaning methods and broad-spectrum antimicrobials often fail to disrupt the resilient matrix and multispecies communities characteristic of these biofilms. Clustered regularly interspaced short palindromic repeat (CRISPR)–CRISPR-associated (Cas) systems offer a transformative approach to enhancing food safety, enabling precise modulation of microbial gene networks with applications in diagnostics, programmable sanitation, and targeted microbial control. This review synthesizes recent advances in CRISPR–Cas technology, encompassing Cas9/Cas12-based gene editing, Cas13-mediated RNA targeting, and dead Cas9 (dCas9)-based transcriptional regulation (CRISPR interference/activation, CRISPRi/a), and evaluates their relevance to biofilm prevention and eradication in food environments. We critically assess delivery platforms, including plasmids, nanocarriers, phagemids, and conjugative systems, for their efficiency in complex biofilm settings. The review highlights innovations such as multiplexed repression of redundant pathways, activation of latent antibiofilm functions. These genetic strategies are increasingly being integrated with omics-based analytics (e.g., transcriptomics, proteomics, metabolomics) to reveal systems-level cellular responses and regulatory shifts triggered by biofilm-targeted interventions. We also address the practical limitations, such as delivery barriers, off-target effects, regulatory hurdles, and ethical considerations specific to food applications. Ultimately, we propose a framework for translating CRISPR–Cas technology into scalable, safety-compliant tools for precision control of biofilms in food processing environments. This review aims to guide future research and inform stakeholders on leveraging CRISPR–Cas technology for safe, sustainable, and targeted management of food-associated biofilms.

Background: Cultured meat, produced by cellular agriculture, offers a sustainable, ethical, and resource-efficient alternative to conventional meat. The production of cultured meat involves growing animal cells under optimal culture conditions in controlled bioreactors. However, microbial contamination, especially biofilm formation, poses a significant challenge. Biofilms are microbial communities encased in a matrix of extracellular polymeric substances (EPS) that adhere to production surfaces, scaffolds, and bioreactor walls, complicating cleaning, reducing production efficiency, and increasing food safety risks. Scope and approach: This review assesses biofilms as a potential food safety risk in cultured meat production. It explores critical production components—cells, culture media, scaffold, and bioreactors—and their role in biofilm formation. Bacterial penetration mechanisms are highlighted, such as scaffold defects and enzymatic matrix degradation, with a focus on key contaminants like Listeria monocytogenes, Salmonella spp., and Escherichia coli. In addition, the influence of surface materials, scaffold design, and bioreactor configuration on biofilm persistence is also discussed. Key findings and conclusions: While direct evidence of biofilm prevalence in cultured meat systems is limited, biofilm control remains essential. Effective strategies include surface modifications, enzymatic treatments, and quorum-sensing inhibitors. Advanced detection tools like biosensors, metagenomics, and artificial intelligence (AI)-driven monitoring offer proactive solutions. Regulatory alignment, hygiene protocols, and traceability are vital for commercialization, enhancing food safety, operational efficiency, and sustainability.

Biofilms formed by Pseudomonas aeruginosa and Escherichia coli on seafood and processing surfaces pose serious food safety risks. This study evaluates naringenin, a citrus-derived flavonoid, for its antibiofilm potential against both pathogens on biotic (shrimp) and abiotic surfaces (stainless steel, aluminum, silicone rubber, and polyethylene terephthalate). The minimum inhibitory concentrations (MICs) of naringenin were 64 μg/mL for P. aeruginosa and 128 μg/mL for E. coli. Naringenin significantly reduced 24- and 72-h biofilms in a dose-dependent manner, with complete removal at 4MIC. Mechanistic studies revealed inhibition of initial adhesion, suppressed adenosine triphosphate (ATP) production, increased reactive oxygen species (ROS), and disruption of cellular respiration (via TTC assay). Protein leakage, carbon-13 nuclear magnetic resonance (13C NMR), and reverse transcription-polymerase chain reaction (RT-PCR) confirmed membrane disruption and downregulation of biofilm-related genes (algU, rhlR, exoS, rpoS, csgA, and luxS). Confocal laser scanning microscopy (CLSM) and field-emission scanning electron microscopy (FE-SEM) further validated biofilm disruption. In conclusion, naringenin demonstrates strong multi-targeted antibiofilm activity and holds promise as a natural intervention for improving seafood hygiene and surface sanitation.

Biofilm formation in food processing environments significantly threatens food safety and quality due to its resistance to conventional cleaning and disinfection methods. These resilient microbial communities contribute to contamination, spoilage and foodborne illnesses, highlighting the need for innovative and technology-driven control strategies. Emerging digital tools, particularly blockchain technology and artificial intelligence (AI), offer new opportunities for enhancing biofilm management. Blockchain ensures secure, real-time traceability of hygiene records, contamination events and compliance activities across the supply chain. Complementing this, AI technologies such as machine learning and sensor-based analytics support early detection of microbial growth, anomaly identification and predictive risk assessment. Together, these tools promote data-driven decision-making and more proactive contamination prevention. While pilot applications show promise in improving transparency and sanitation outcomes, challenges remain, including data integration, implementation costs and regulatory barriers. Addressing these issues will require interdisciplinary collaboration and supportive policy frameworks. This review summarizes the current and potential roles of blockchain and AI in biofilm control and outlines future directions for research and industrial application.

Biofilm prevention is critical in the food industry due to its significant impact on food degradation, safety and quality. This review delves into the nature and formation of biofilms, identifying common biofilm-forming microorganisms and the factors influencing their adherence to food contact surfaces. It comprehensively analyzes the implications of biofilms for food spoilage, pathogen persistence, shelf life and consumer safety. Current strategies for biofilm prevention across various food sectors such as meat, dairy and seafood industries are evaluated with a focus on understanding how the surface properties (surface roughness, topography, surface free energy and chemistry) influence biofilm formation, optimizing the use of antimicrobial agents and developing biofilm-resistant materials. The challenges faced by the food industry, including microbial resistance and regulatory hurdles are discussed to highlight the complexity of effective biofilm control. Emerging technologies and innovations such as nanotechnology, advanced detection techniques and computational modeling, offer promising avenues for enhanced biofilm management. The review emphasizes the need for ongoing research and the adoption of cutting-edge strategies to mitigate food degradation and improve food safety. It concludes with key recommendations for industry and a call to action for sustained efforts in optimizing biofilm prevention methods.

Biofilms are silent but formidable threats in seafood processing, where Pseudomonas aeruginosa and Escherichia coli can quickly transform contact surfaces into reservoirs of contamination. This study explores the dynamic biofilm formation on aluminum, silicone rubber, stainless steel, and polyethylene terephthalate over 24 and 72 h. Quantitative assays including Colony Forming Unit (CFU), Crystal Violet (CV), 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) revealed a significant increase in biofilm density, particularly on aluminum and silicone rubber. Fourier-Transform Infrared Spectroscopy (FTIR) and 1H Nuclear Magnetic Resonance (NMR) analyses showed that biofilm EPS exhibits an evolving amphiphilic nature, with stable polysaccharides and increasing lipid content enhancing resilience. Confocal Laser Scanning Microscopy (CLSM), and Field Emission Scanning Electron Microscopy (FE-SEM) captured the shift from early attachment to mature, dense biofilms. These findings underscore the crucial impact of surface material on biofilm growth and the pressing need for tailored cleaning protocols to curb contamination risks in food processing environments.

This study investigated the synergistic mechanism of UV-C and postbiotics against Salmonella enterica ser. Thompson biofilm in the poultry industry. The synergistic activity of postbiotic with UV-C was estimated singly and in combination and observed by field-emission scanning electron microscopy (FE-SEM) and confocal laser scanning microscopy (CLSM). When applied as single treatment, J.27 postbiotic from Leuconostoc mesenteroides and UV-C treatment reduced the S. Thompson biofilm cells by about 1.23 log and 3.23 log CFU/cm2 on stainless steel [SS], 1.23 log and 4.74 log CFU/cm2 on silicon rubber [SR], 0.53 log and 0.72 log CFU/cm2 on chicken tenderloin [CT], and 1.28 log and 1.77 log CFU/cm2 on eggshell [ES], respectively. However, the combination treatment reduced the S. Thompson biofilm cells on SS, SR, CT, and ES surfaces by up to 4.89 log, 6.03 log, 2.08 log, and 3.66 log CFU/cm2, respectively. Especially when the combination of 2 × MIC J.27 postbiotic with 3 min UV-C on SS, 4 × MIC J.27 postbiotic with 3 min UV-C on SR, 4 × MIC J.27 postbiotic with 5 min UV-C on CT, and 1 × MIC J.27 postbiotic with 3 min UV-C on ES was treated to the biofilm formed on each surface, the synergistic value was highest, up to 0.91, 1.04, 0.83, and 0.83, respectively. This study found that LAB J.27 postbiotic and UV-C reduced the biofilm of a pathogenic bacterium on poultry and poultry-related products and showed an antibiofilm effect when treated in combination. The results of this study proposed that the combination of a biological control method with a physical method could be a promising environmentally friendly strategy for reducing food poisoning bacterial biofilm contamination during poultry processing.

Biofilm formation by Aeromonas hydrophila in the food industry poses significant challenges to food safety and quality. Therefore, this comprehensive review aimed to provide insights into the mechanisms and key factors influencing A. hydrophila biofilm formation. It explores the molecular processes involved in initial attachment, microcolony formation, and biofilm maturation; moreover, it concurrently examines the impact of intrinsic factors, including quorum sensing, cyclic-di-GMP, the efflux pump, and antibiotic resistance, as well as environmental conditions, such as temperature, nutrient availability, and osmotic pressure, on biofilm architecture and resilience. Furthermore, the article highlights the potential of bibliometric analysis as a promising method for conceptualizing the research landscape of and identifying knowledge gaps in A. hydrophila biofilm research. The findings underscore the requirement for focused interventions that prevent biofilm development and raise food sector safety. The consolidation of current information and incorporation of bibliometric analysis enhances existing understanding of A. hydrophila biofilm formation and offers insights for future research and control strategies within a food industry context.

Aquaculture is one of the most significant food sources from the prehistoric period. As aquaculture intensifies globally, the prevalence and outbreaks of various pathogenic microorganisms cause fish disease and heavy mortality, leading to a drastic reduction in yield and substantial economic loss. With the modernization of the aquaculture system, a new challenge regarding biofilms or bacterial microenvironments arises worldwide, which facilitates pathogenic microorganisms to survive under unfavorable environmental conditions and withstand various treatments, especially antibiotics and other chemical disinfectants. However, we focus on the mechanistic association between those microbes which mainly form biofilm and probiotics in one of the major food production systems, aquaculture. In recent years, probiotics and their derivatives have attracted much attention in the fisheries sector to combat the survival strategy of pathogenic bacteria. Apart from this, Bibliometric analysis provides a comprehensive overview of the published literature, highlighting key research themes, emerging topics, and areas that require further investigation. This information is valuable for researchers, policymakers, and stakeholders in determining research priorities and allocating resources effectively.

The recalcitrance of microbial aggregation or biofilm in the food industry underpins the emerging antimicrobial resistance among foodborne pathogens, exacerbating the phenomena of food spoilage, processing and safety management failure, and the prevalence of foodborne illnesses. The challenges of growing tolerance to current chemical and disinfectant-based antibiofilm strategies have driven the urgency in finding a less vulnerable to bacterial resistance, effective alternative antibiofilm agent. To address these issues, various novel strategies are suggested in current days to combat bacterial biofilm. Among the innovative approaches, phytochemicals have already demonstrated their excellent performance in preventing biofilm formation and bactericidal actions against resident bacteria within biofilms. However, the diverse group of phytochemicals and their different modes of action become a barrier to applying them against specific pathogenic biofilm-formers. This phenomenon mandates the need to elucidate the multi-mechanistic actions of phytochemicals to design an effective novel antibiofilm strategy. Therefore, this review critically illustrates the structure − activity relationship, functional sites of actions, and target molecules of diverse phytochemicals regarding multiple major antibiofilm mechanisms and reversal mechanisms of antimicrobial resistance. The implementation of the in-depth knowledge will hopefully aid future studies for developing phytochemical-based next-generation antimicrobials.

This study examines the antimicrobial and antibiofilm effectiveness of baicalin and carvacrol against Salmonella enterica ser. Typhimurium on food contact surfaces and chicken meat. The minimum inhibitory concentrations (MIC) for baicalin and carvacrol were found to be 100 μg/mL and 200 μg/mL, respectively, which aligns with findings from previous studies. The compounds exhibited a concentration-dependent decrease in microbial populations and biofilm formation. When used together, they displayed a remarkable synergistic effect, greatly augmenting their antibacterial activity. The assessment of food quality demonstrated that these treatments have no negative impact on the sensory characteristics of chicken meat. The impact of the structure on biofilms was observed through the use of Field Emission Scanning Electron Microscopy (FE-SEM) and Confocal Laser Scanning Microscopy (CLSM), revealing disrupted biofilm architectures and decreased cell viability. Crucially, RT-PCR analysis revealed a marked downregulation of quorum sensing (luxS), virulence (hilA), and stress response (rpoS) genes, highlighting the multifaceted antimicrobial mechanism of action. This gene-specific suppression suggests a targeted disruption of bacterial communication and virulence pathways, offering insight into the comprehensive antibiofilm strategy. This provides further insight into the molecular mechanisms that contribute to their antibiofilm effects.

Objective: This study intended to isolate a Vibrio-particular phage from the natural environment, analyse its characteristics and genome sequence, and investigate its reduction effect on V. parahaemolyticus biofilm as a biocontrol agent in squid and mackerel. Methods: Among 21 phages, phage CAU_VPP01, isolated from beach mud, was chosen for further experiments based on host range and EOP tests. When examining the reduction effect of phage CAU_VPP01 against Vibrio parahaemolyticus biofilms on surfaces (stainless steel [SS] and polyethylene terephthalate [PET]) and food surfaces (squid and mackerel). Results: The phage showed the most excellent reduction effect at a multiplicity-of-infection (MOI) 10. Three-dimensional images acquired with confocal laser scanning microscopy (CLSM) analysis were quantified using COMSTAT, which showed that biomass, average thickness, and roughness coefficient decreased when treated with the phage. Colour and texture analysis confirmed that the quality of squid and mackerel was maintained after the phage treatment. Finally, a comparison of gene expression levels determined by qRT-PCR analysis showed that the phage treatment induced a decrease in the gene expression of flaA, vp0962, andluxS, as examples. Conclusion: This study indicated that Vibrio-specific phage CAU_VPP01 effectively controlled V. parahaemolyticus biofilms under various conditions and confirmed that the isolated phage could possibly be used as an effective biocontrol weapon in the seafood manufacturing industry.

The risk of foodborne disease outbreaks increases when the pathogenic bacteria are able to form biofilms, and this presents a major threat to public health. An emerging non-thermal cold plasma (CP) technology has proven a highly effective method for decontaminating meats and their products and extended their shelf life. CP treatments have ability to reduce microbial load and, biofilm formation with minimal change of color, pH value, and lipid oxidation of various meat and meat products. The CP technique offers many advantages over conventional processing techniques due to its layout flexibility, nonthermal behavior, affordability, and ecological sustainability. The technology is still in its infancy, and continuous research efforts are needed to realize its full potential in the meat industry. This review addresses the basic principles and the impact of CP technology on biofilm formation, meat quality (including microbiological, color, pH value, texture, and lipid oxidation), and microbial inactivation pathways and also the prospects of this technology.

Foodborne pathogen Escherichia coli frequently causes foodborne infections. In our study, we investigated the antibiofilm activity of orange terpene (OT) against E. coli biofilms on a food surface (beef) and different surfaces that come into touch with food, including stainless steel (SS), polyethylene terephthalate (PET), low-density polyethylene (LDPE), and rubber (SR). The study findings revealed that OT significantly (P < 0.05) eliminated 48-h-old biofilms from all food contact surfaces (SS: 2.09 log CFU/cm2, PET: 1.95 log CFU/cm2, LDPE: 1.94 log CFU/cm2, and SR: 1.4 log CFU/cm2). Additionally, on beef surfaces, OT at a minimum inhibitory concentration (MIC) of 0.13 % demonstrated the ability to inhibit biofilm development by approximately 1.5 log CFU/cm2 and reduced pre-formed biofilms by 2.02 log CFU/cm2. Our sensory evaluations showed that it had no adverse impacts on beef color and texture, although it slightly altered the natural odor of beef. Quantitative and qualitative assessments showed that OT has strong bactericidal actions on biofilm populations. It significantly altered cell surface hydrophobicity, reduced cellular ATP levels, and inhibited cell auto-aggregation in planktonic cells (P < 0.05). As a result, our findings emphasize the antibacterial potentiality of OT in reducing the biofilm of E. coli in the food sector.

Enzymes play a vital role in baking, providing significant benefits from dough development to extending shelf life, which enhances product quality and consistency. Acting as biological catalysts, enzymes such as proteases and amylases break down proteins and starches, modifying dough rheology and improving fermentation. Lipases and oxidases further refine dough texture through emulsification and oxidation, while lipases also produce fatty acid derivatives during fermentation, contributing to the flavor and aroma of baked goods. Xylanases and cellulases optimize dough handling by altering fiber structure, and amylases help maintain moisture and texture, extending the shelf life of baked products. Ensuring regulatory compliance is essential when incorporating enzymes into baking processes, as bakers must address enzyme stability and determine appropriate dosages for reliable outcomes. Ongoing research is exploring innovative enzyme applications, including customized enzyme blends that target specific product qualities, offering new possibilities for product differentiation and innovation. In summary, enzyme-driven advancements present bakers with opportunities to improve product quality, shelf life, and consistency, while meeting industry regulations. This review emphasizes the critical impact enzymes have on dough properties and finished product characteristics, highlighting their role in driving future innovations within the baking industry.

There has been an increasing interest in phytochemicals with antimicrobial properties in food safety management. However, the mechanisms of action of phytochemicals remain mostly unknown, preventing their selective applications against biofilms of specific pathogenic bacteria. The objectives of the present study were to understand the antibiofilm and antibacterial properties of luteolin (LUT) and its modes of action against two detrimental foodborne pathogens, Salmonella enterica ser. Typhimurium and enterohemorrhagic Escherichia coli. The focus encompasses biofilm inhibition, interference with cell adhesion, disruption of membrane integrity, and dysfunctions of membrane-linked bacterial electron transport systems and energy metabolism. Our findings evidence that LUT prevents pathogenic biofilm formation on both biotic (eggshell) and abiotic (stainless steel and silicon rubber) surfaces by resisting irreversible cell adhesion and interfering with hydrophobic interactions between bacteria and contact surfaces. Our comprehensive biochemical and visual analyses further reveal that LUT mainly exhibits bactericidal activities by imposing oxidative stress on cells, leading to severe cellular structure damage, inhibition of metabolic and respiratory activities, and dysfunctions of energy metabolism, ultimately resulting in bacterial cell death. This study suggests that LUT has the potential to serve as a solution for developing effective, novel antimicrobials for improving food safety management.

Salmonella is a food-borne microorganism that is also a zoonotic bacterial hazard in the food sector. This study determined how well a mixed culture of Salmonella Kentucky formed biofilms on plastic (PLA), silicon rubber (SR), rubber gloves (RG), chicken skin and eggshell surfaces. In vitro interactions between the histone deacetylase inhibitor-vorinostat (SAHA)-and S. enterica serotype Kentucky were examined utilizing biofilms. The minimum inhibitory concentration (MIC) of SAHA was 120 µg mL−1. The addition of sub-MIC (60 µg mL−1) of SAHA decreased biofilm formation for 24 h on PLA, SR, RG, Chicken skin, and eggshell by 3.98, 3.84, 4.11, 2.86 and 3.01 log (p < 0.05), respectively. In addition, the initial rate of bacterial biofilm formation was higher on chicken skin than on other surfaces, but the inhibitory effect was reduced. Consistent with this conclusion, virulence genes expression (avrA, rpoS and hilA) and quorum-sensing (QS) gene (luxS) was considerably downregulated at sub-MIC of SAHA. SAHA has potential as an anti-biofilm agent against S. enterica serotype Kentucky biofilm, mostly by inhibiting virulence and quorum-sensing gene expression, proving the histone deacetylase inhibitor could be used to control food-borne biofilms in the food industry.

Salmonella Thompson is a foodborne pathogen that has attracted attention in Korea since it was identified as the cause of a large-scale domestic food poisoning outbreak in 2018. The present study aims to isolate and characterize bacteriophages specific to Salmonella Thompson and evaluate their efficacy in reducing Salmonella Thompson biofilm on eggshell. In addition, advanced microscopic analysis using field-emission scanning electron microscopy and confocal laser scanning microscopy was conducted to confirm the anti-biofilm activity. The two isolated bacteriophages CAU-STP-1 and CAU-STP-2 showed very high specificity by targeting only Salmonella Thompson. They were determined to be stable at 60–65 °C and pH 4–11. Bacteriophage particles quickly adsorbed to the host cells, with a latent period and burst size of 30–40 min and 19–100 plaque-forming units per cell, respectively. When comparing the two bacteriophages individually, CAU-STP-1 exhibited the highest in vitro inhibition ability, but treatment with both bacteriophages together in a cocktail was significantly more effective than with CAU-STP-1 alone. This bacteriophage cocktail was consequently employed to treat Salmonella Thompson biofilm on eggshell at different storage temperatures. There was a significant reduction in the biofilm on the eggshell with bacteriophage treatment at 10 and 20 °C after 7 days. For example, After 7 days at 10 °C, the biofilm on eggshell was reduced by 0.26, 0.96, and 2.50 log CFU/cm2 for MOI 103, MOI 104, and MOI 105, respectively. On the other hand, at 20 °C, the biofilm on eggshells was reduced by 0.10, 0.24, and 0.33 log CFU/cm2 for MOI 103, MOI 104, and MOI 105, respectively This study thus provides promising information for the potential development of an effective biocontrol agent against Salmonella Thomson within the food industry.

SUMMARY: Consumers are more conscious of both the quality and microbial safety of the food they eat. Therefore, their demand for safe food and food products has motivated researchers to develop non-conventional processing technologies. Numerous investigations have shown the effectiveness of high-pressure processing, ultraviolet, pulsed light, ultrasonic sterilisation, pulsed electric field, irradiation, and cold plasma to produce safe and high-quality products compared to conventional processes. This review aims to summarise the advanced applications of these promising non-conventional thermal technologies to provide safe and high-quality food products. Many innovative strategies have been proposed as an alternative to conventional technologies, normally applied to reduce pathogenic microbes. This study presents some potential non-conventional technologies that show emerging microbial decontamination abilities for improving finished food products’ freshness, safety and multiplication. It also summarises the inactivation mechanism of foodborne pathogens, which presents an outline of the effects on finished food product quality based on microbiological and physicochemical perspectives. The key advantage of these technologies is their potential to inactivate foodborne microorganisms without affecting the nutritional value of food products. These technologies are gradually entering the food industry due to technological advances and low investment costs. A comprehensive study is critical to keep abreast of these technologies’ merits, application scope and recent developments to ensure food products satisfy consumer demands while maintaining the highest quality and safety standards.

This study investigated the antimicrobial and antibiofilm efficacy of separate and combined treatments of Lactobacillus curvatus B67-produced postbiotic and the polyphenolic flavanol quercetin against Listeria monocytogenes and Salmonella enterica ser. Typhimurium. The antimicrobial potentiality of the postbiotic was chiefly associated with organic acids (e.g., lactic and acetic acids). At sub-minimum inhibitory concentration (1/2 MIC), the postbiotic and quercetin effectively reduced the pathogenic biofilm cells on processed pork sausage and meat-processing surfaces (e.g., stainless-steel and rubber). Moreover, the postbiotic exhibited strong residual antimicrobial efficacy over diverse pH and temperature ranges. In addition, the combination of postbiotic with quercetin increased the leakage of pathogenic intracellular metabolites (e.g., nucleic acids and protein) and inhibited pathogenic biofilm formation on both biotic and abiotic surfaces. Therefore, this study confirmed that lactic acid bacteria-derived postbiotic and plant-derived quercetin could be used as potential alternative bioprotective agents in the meat processing industry.

Poultry is thriving across the globe. Chicken meat is the most preferred poultry worldwide, and its popularity is increasing. However, poultry also threatens human hygiene, especially as a fomite of infectious diseases caused by the major foodborne pathogens (Campylobacter, Salmonella, and Listeria). Preventing pathogenic bacterial biofilm is crucial in the chicken industry due to increasing food safety hazards caused by recurring contamination and the rapid degradation of meat, as well as the increased resistance of bacteria to cleaning and disinfection procedures commonly used in chicken processing plants. To address this, various innovative and promising strategies to combat bacterial resistance and biofilm are emerging to improve food safety and quality and extend shelf-life. In particular, natural compounds are attractive because of their potential antimicrobial activities. Natural compounds can also boost the immune system and improve poultry health and performance. In addition to phytochemicals, bacteriophages, nanoparticles, coatings, enzymes, and probiotics represent unique and environmentally friendly strategies in the poultry processing industry to prevent foodborne pathogens from reaching the consumer. Lactoferrin, bacteriocin, antimicrobial peptides, cell-free supernatants, and biosurfactants are also of considerable interest for their prospective application as natural antimicrobials for improving the safety of raw poultry meat. This review aims to describe the feasibility of these proposed strategies and provide an overview of recent published evidences to control microorganisms in the poultry industry, considering the human health, food safety, and economic aspects of poultry production.

In recent years, biosynthesized zinc oxide nanoparticles (ZnONPs) have gained tremendous attention because of their safe and non-toxic nature and distinctive biomedical applications. A diverse range of microbes (bacteria, fungi and yeast) and various parts (leaf, root, fruit, flower, peel, stem, etc.) of plants have been exploited for the facile, rapid, cost-effective and non-toxic synthesis of ZnONPs. Plant extracts, microbial biomass or culture supernatant contain various biomolecules including enzymes, amino acids, proteins, vitamins, alkaloids, flavonoids, etc., which serve as reducing, capping and stabilizing agents during the biosynthesis of ZnONPs. The biosynthesized ZnONPs are generally characterized using UV-VIS spectroscopy, TEM, SEM, EDX, XRD, FTIR, etc. Antibiotic resistance is a serious problem for global public health. Due to mutation, shifting environmental circumstances and excessive drug use, the number of multidrug-resistant pathogenic microbes is continuously rising. To solve this issue, novel, safe and effective antimicrobial agents are needed urgently. Biosynthesized ZnONPs could be novel and effective antimicrobial agents because of their safe and non-toxic nature and powerful antimicrobial characteristics. It is proven that biosynthesized ZnONPs have strong antimicrobial activity against various pathogenic microorganisms including multidrug-resistant bacteria. The possible antimicrobial mechanisms of ZnONPs are the generation of reactive oxygen species, physical interactions, disruption of the cell walls and cell membranes, damage to DNA, enzyme inactivation, protein denaturation, ribosomal destabilization and mitochondrial dysfunction. In this review, the biosynthesis of ZnONPs using microbes and plants and their characterization have been reviewed comprehensively. Also, the antimicrobial applications and mechanisms of biosynthesized ZnONPs against various pathogenic microorganisms have been highlighted.

The use of phages as biocontrol agents against antibiotic-resistant strains of Salmonella spp. is gaining attention. This study aimed to isolate lytic bacteriophages specific for multidrug-resistant Salmonella enterica serovars Typhimurium; it also evaluated the bactericidal effect of isolated phages (STP-1, STP-2, STP-3, and STP-4) from sewage sample against S. Typhimurium as host strains. Moreover, a current study evaluated the efficacy of a bacteriophage cocktail against S. Typhimurium cocktail in chicken breast meat. The 4 phages were classified under the Caudoviricetes class by morphology characterization. On host range testing, they exhibited lytic activities against S. Typhimurium, S. Enteritidis, and S. Thompson. In the stability test, the phages exhibited resistance to heat (above 70°C for 1 h) and pH (strongly alkaline for 24 h). Additionally, the phages had comparable adsorption rates (approximately 80% adsorption in under 5 min). Additionally, the latent periods ranged from 30 to 50 min, with respective burst sizes of 31, 218, 197, and 218 PFU/CFU. In vitro, bacterial challenge demonstrated that at a multiplicity of infection (MOI) of 10, each phage consistently inhibited S. Typhimurium growth at 37°C for 24 h. In the food test, the phage cocktail (MOI = 1,000) reduced S. Typhimurium in artificially contaminated chicken breast meat stored at 4°C by 0.9 and 1.2 log CFU/g after 1 and 7 d, respectively. The results point toward a promising avenue for addressing the challenge of multidrug-resistant S. Typhimurium in the food industry through the use of recently discovered phages. Notably, the exploration of phage cocktails holds significant potential for combating S. Typhimurium in chicken breast products in the times ahead.

There is no doubt that ruminants have the capability to digest lignocellulosic compounds and to utilize them as an absorbable form of energy by tapping into enzymes produced by the microbial population in their rumens. Among the rumens of various ruminants, this study focused on Korean goat rumens because of their unique digestibility of lignocellulosic biomasses. Therefore, a novel Gene12 gene was screened and unmasked from the constructed rumen metagenomic library of a Korean black goat and expressed in a Bacillus megaterium system. The recombinant protein was distinguished as a novel α-L-arabinofuranosidase enzyme from glycosyl hydrolase family 43 (GH43) for its capability to hydrolyze the non-reducing end of α-1,5-L-arabinofuranose linkages in α-L-arabinofuranosyl groups. The enzyme can also break apart α-L-arabinofuranosidic linkages and act synergistically with other hemicellulolytic enzymes to release α-1,2- and α-1,3-L-arabinofuranosyl groups from L-arabinose-comprising polysaccharides. In silico, phylogenetic, and computational analyses proclaimed that the Gene12 gene encodes a novel carbohydrate-active enzyme possessing a V-shaped indentation of the GH43 catalytic and functional domain (carbohydrate-binding module 6). The recombinant Gene12 protein has shared 81% sequence homology with other members of the GH43 family. Enzymic synopses (optimal pH, temperatures, and stability studies) of the recombinant Gene12 enzyme and its substrate specificity (synthetic and natural substrates) profiling were considered. The recombinant Gene12 α-L-arabinofuranosidase works best at pH 6.0 and 40 °C, and it is stable at pH 4.0 to 7.0 at temperatures of 20 to 50 °C. Additionally, 5-blended β-sheets were identified through a tertiary (3D) structure analysis along with the high substrate specificity against p-nitrophenyl-D-arabinofuranoside (pNPA). The highest substrate specificity of pNPA for Gene12 α-L-arabinofuranosidase indicated its confirmation as an exo-type arabinofuronidase. The results thus propose using the Gene12 protein as an exo-mannered GH43 α-L-arabinofuranosidase (EC 3.2.1.55) enzyme.

Establishing efficient methods to combat bacterial biofilms is a major concern. Natural compounds, such as essential oils derived from plants, are among the favored and recommended strategies for combatting bacteria and their biofilm. Therefore, we evaluated the antibiofilm properties of peppermint oil as well as the activities by which it kills bacteria generally and particularly their biofilms. Peppermint oil antagonistic activities were investigated against Vibrio parahaemolyticus, Listeria monocytogenes, Pseudomonas aeruginosa, Escherichia coli O157:H7, and Salmonella Typhimurium on four food contact surfaces (stainless steel, rubber, high-density polyethylene, and polyethylene terephthalate). Biofilm formation on each studied surface, hydrophobicity, autoaggregation, metabolic activity, and adenosine triphosphate quantification were evaluated for each bacterium in the presence and absence (control) of peppermint oil. Real-time polymerase chain reaction, confocal laser scanning microscopy, and field-emission scanning electron microscopy were utilized to analyze the effects of peppermint oil treatment on the bacteria and their biofilm. Results showed that peppermint oil (1/2× minimum inhibitory concentration [MIC], MIC, and 2× MIC) substantially lessened biofilm formation, with high bactericidal properties. A minimum of 2.5-log to a maximum of around 5-log reduction was attained, with the highest sensitivity shown by V. parahaemolyticus. Morphological experiments revealed degradation of the biofilm structure, followed by some dead cells with broken membranes. Thus, this study established the possibility of using peppermint oil to combat key foodborne and food spoilage pathogens in the food processing environment.

Biosynthesized metal nanoparticles, especially silver and gold nanoparticles, and their conjugates with biopolymers have immense potential in various fields of science due to their enormous applications, including biomedical applications. Polymeric nanoparticles are particles of small sizes from 1 nm to 1000 nm. Among different polymeric nanoparticles, chitosan-coated silver and gold nanoparticles have gained significant interest from researchers due to their various biomedical applications, such as anti-cancer, antibacterial, antiviral, antifungal, anti-inflammatory technologies, as well as targeted drug delivery, etc. Multidrug-resistant pathogenic bacteria have become a serious threat to public health day by day. Novel, effective, and safe antibacterial agents are required to control these multidrug-resistant pathogenic microorganisms. Chitosan-coated silver and gold nanoparticles could be effective and safe agents for controlling these pathogens. It is proven that both chitosan and silver or gold nanoparticles have strong antibacterial activity. By the conjugation of biopolymer chitosan with silver or gold nanoparticles, the stability and antibacterial efficacy against multidrug-resistant pathogenic bacteria will be increased significantly, as well as their toxicity in humans being decreased. In recent years, chitosan-coated silver and gold nanoparticles have been increasingly investigated due to their potential applications in nanomedicine. This review discusses the biologically facile, rapid, and ecofriendly synthesis of chitosan-coated silver and gold nanoparticles; their characterization; and potential antibacterial applications against multidrug-resistant pathogenic bacteria.

Salmonellosis is a prevalent food poisoning disease caused by Salmonella spp. that affects millions of people throughout the world. Contamination of chicken meat and processing equipment with Salmonella is a major issue in the food industry. Therefore, it is necessary to study not only food but also food contact surfaces [plastic (PLA) and rubber gloves (RG)] that can cause cross-contamination during processing. The inhibitory activities of quercetin, an antioxidant and antibacterial compound, were tested against S. Typhimurium and S. Enteritidis on PLA, RG, and chicken skin during biofilm. When quercetin (0–125 μg/mL) was supplemented, the inhibitory effect was 1.50–2.61 log CFU/cm2 (125; 1/2, 62.5; 1/4, and 31.25 μg/mL; 1/8 MIC). Furthermore, the initial biofilm formation rate of both bacteria was greater on the chicken skin surface than on the food contact surfaces. The inhibitory impact was visually observed using field-emission scanning electron microscopy and confocal laser scanning microscopy. The swimming and swarming motility was also found to be inhibited from 1/8 MIC. As determined by qRT-PCR, quercetin downregulated the expression levels of virulence (avrA, and hilA), stress response (rpoS), and quorum-sensing (luxS) genes. Our findings suggest that plant-derived quercetin is a possible antibiofilm agent against Salmonella spp. biofilm formation, as well as biofilm caused by other bacteria.

Foodborne pathogen-mediated biofilms in food processing environments are severe threats to human lives. In the interest of human and environmental safety, natural substances with antimicrobial properties and generally regarded as safe (GRAS) status are the futuristic disinfectants of the food industry. In this study, the efficacy of bioactive, soluble products (metabolic by-products) from lactic acid bacteria (LAB) and plant-derived essential oils (EO) were investigated as biocidal agents. The postbiotic produced by kimchi-derived Leuconostoc mesenteroides J.27 isolate was analyzed for its metabolic components to reveal its antimicrobial potential against three pathogenic microorganisms (Vibrio parahaemolyticus, Pseudomonas aeruginosa, and Escherichia coli). Additionally, the efficacy of food-grade EO (eugenol and thymol, respectively) was also assessed in our study. Determination of the minimum inhibitory concentration (MIC) of postbiotic and EO against three tested pathogens revealed that the sub-MIC (0.5 MIC) of postbiotic and EO could efficiently inhibit the biofilm formation on both seafood (squid) and seafood-processing surfaces (rubber and low-density polyethylene plastic). Moreover, the polymerase chain reaction (PCR) analysis confirmed that the LAB J.27 isolate possesses bacteriocin- and enzyme-coding genes. The residual antibacterial activity of the produced postbiotic was maintained over a diverse pH range (pH 1–6) but was entirely abolished at neutral or higher pH values. However, the activity was unaffected by exposure to high temperatures (100 and 121 °C) and storage (30 days). Notably, the leakage of intracellular metabolites, damage to DNA, and the down-regulation of biofilm-associated gene expression in the pathogens increased significantly (p > 0.05) following the combination treatment of postbiotic with thymol compared to postbiotic with eugenol. Nonetheless, all in vitro results indicated the prospective use of combining Leu. mesenteroides J.27-derived postbiotic with both EO as a “green preservative” in the seafood industry to inhibit the formation of pathogenic microbial biofilms.

The risk of salmonellosis is expected to increase with the rise in the consumption of poultry meat. The aim of this study was to investigate the combination treatment of peroxyacetic acid (PAA) or lactic acid (LA) with UV-C against Salmonella Enteritidis biofilms formed on food contact surface (stainless steel [SS], silicone rubber [SR], and ultra-high molecular weight polyethylene [UHMWPE]) and chicken skin. The biofilm on food contact surface and chicken skin was significantly decreased (P < 0.05) by combination treatment of PAA or LA with UV-C. Combination treatment of PAA (50–500 μg/mL) with UV-C (5 and 10 min) reduced 3.10–6.41 log CFU/cm2 and LA (0.5–2.0%) with UV-C (5 and 10 min) reduced 3.35–6.41 log CFU/cm2 of S. Enteritidis biofilms on food contact surface. Salmonella Enteritidis biofilms on chicken skin was reduced around 2 log CFU/g with minor quality changes in color and texture by combination treatment of PAA (500 μg/mL) or LA (2.0%) with UV-C (10 min). Additional reduction occurred on SS and UHMWPE by PAA or LA with UV-C, while only LA with UV-C caused additional reduction on chicken skin. Also, it was visualized that the biofilm on food contact surface and chicken skin was removed through field emission scanning electron microscopy (FESEM) and death of cells constituting the biofilm was confirmed through confocal laser scanning microscopy (CLSM). These results indicating that the combination treatment of PAA or LA with UV-C could be used for S. Enteritidis biofilm control strategy in poultry industry.

Green synthesis of silver nanoparticles (AgNPs) using biological resources is the most facile, economical, rapid, and environmentally friendly method that mitigates the drawbacks of chemical and physical methods. Various biological resources such as plants and their different parts, bacteria, fungi, algae, etc. could be utilized for the green synthesis of bioactive AgNPs. In recent years, several green approaches for non-toxic, rapid, and facile synthesis of AgNPs using biological resources have been reported. Plant extract contains various biomolecules, including flavonoids, terpenoids, alkaloids, phenolic compounds, and vitamins that act as reducing and capping agents during the biosynthesis process. Similarly, microorganisms produce different pri-mary and secondary metabolites that play a crucial role as reducing and capping agents during synthesis. Biosynthesized AgNPs have gained significant attention from the researchers because of their potential applications in different fields of biomedical science. The widest application of AgNPs is their bactericidal activity. Due to the emergence of multidrug-resistant microorganisms, researchers are exploring the therapeutic abilities of AgNPs as potential antibacterial agents. Al-ready, various reports have suggested that biosynthesized AgNPs have exhibited significant antibacterial action against numerous human pathogens. Because of their small size and large surface area, AgNPs have the ability to easily penetrate bacterial cell walls, damage cell membranes, produce reactive oxygen species, and interfere with DNA replication as well as protein synthesis, and result in cell death. This paper provides an overview of the green, facile, and rapid synthesis of AgNPs using biological resources and antibacterial use of biosynthesized AgNPs, highlighting their antibacterial mechanisms.

Biofilm may influence bacterial susceptibility to traditional disinfectants but identifying effective natural disinfectants as alternatives remains a great challenge to researchers. The objective of this research was to examine antibiofilm activity of carvacrol against Pseudomonas aeruginosa and Listeria monocytogenes. It was found that different concentrations of carvacrol dramatically reduced the P. aeruginosa and L. monocytogenes biofilms developed on both the MBEC™ biofilm device and polypropylene surface. On the MBEC™ biofilm device, the biofilm reduction was 5.04 log CFU/peg at 0.06% carvacrol for P. aeruginosa, and 3.81 log CFU/peg at 0.12% carvacrol for L. monocytogenes. On the polypropylene surface, P. aeruginosa was reduced by 4.79 log CFU/cm2 at 0.06% carvacrol, and L. monocytogenes was reduced by 4.62 log CFU/cm2 at 0.12% carvacrol. Visual observations using advanced microscopy confirmed the biofilm reduction results for both bacteria. This study demonstrated that carvacrol has potential use as a natural compound to prevent P. aeruginosa and L. monocytogenes related contaminations of food contact surfaces. These findings suggest that carvacrol could be used to formulate new sanitizers able to inhibit P. aeruginosa and L. monocytogenes biofilms.

A Gram-stain-negative, aerobic and rod-shaped novel bacterial strain, designated MAH-26T, was isolated from rhizospheric soil of a pine tree. The colonies were orange coloured, smooth, spherical and 0.7–1.8 mm in diameter when grown on Reasoner's 2A (R2A) agar for 2 days. Strain MAH-26T was able to grow at 10–40 °C, at pH 6.0–9.0 and with 0–1.0 % NaCl. Cell growth occurred on nutrient agar, R2A agar, tryptone soya agar and Luria–Bertani agar. The strain gave positive results in oxidase and catalase tests. Strain MAH-26T was closely related to Flavihumibacter sediminis CJ663T and Parasegetibacter terrae SGM2-10T with a low 16S rRNA gene sequence similarity (92.8 and 92.9 %, respectively) and phylogenetic analysis indicated that the strain formed a distinct phylogenetic lineage from the members of the closely related genera of the family Chitinophagaceae. Strain MAH-26T has a draft genome size of 6 857 405 bp, annotated with 5173 protein-coding genes, 50 tRNA and two rRNA genes. The genomic DNA G+C content was 41.5 mol%. The predominant isoprenoid quinone was menaquinone 7. The major fatty acids were iden-tified as iso-C15:0, iso-C15:1 G and iso-C17:0 3OH. On the basis of phylogenetic inference and phenotypic, chemotaxonomic and molecular properties, strain MAH-26T represents a novel species of a novel genus of the family Chitinophagaceae, for which the name Pinibacter aurantiacus gen. nov., sp. nov. is proposed. The type strain of Pinibacter aurantiacus is MAH-26T (=KACC 19749T=CGMCC 1.13701T).

The goal was to identify the biofilm-forming ability of Cronobacter sakazakii on surfaces of stainless steel (SS) and silicone rubber (SR) in contact with infant formula milk. Two representative bacteriophages (PBES04 and PBES19) were used to control the growth of C. sakazakii as well as its biofilm forming ability on either SS or SR surfaces. Bacterial growth was confirmed at 20 °C when PBES04 and PBES19 were used, whereas C. sakazakii was not normally detected in infant formula milk treated with both bacteriophages for 6 h. In an additional biofilm reduction experiment, the biofilm on SS or SR surfaces were reduced by 3.07 and 1.92 log CFU cm−2, respectively after PBES04 treatment, and 3.06 and 2.14 log CFU cm−2, respectively, after PBES19 treatment. These results demonstrate that bacteriophages can be effective in inactivating C. sakazakii in biofilms which could potentially increase food safety in commercial facilities.

The contamination of seafood with Vibrio species can have severe repercussions in the seafood industry. Vibrio species can form mature biofilms and persist on the surface of several seafoods such as crabs, oysters, mussels, and shrimp, for extended duration. Several conventional approaches have been employed to inhibit the growth of planktonic cells and prevent the formation of Vibrio biofilms. Since Vibrio biofilms are mostly resistant to these control measures, novel alternative methods need to be urgently developed. In this review, we propose environmentally friendly approaches to suppress Vibrio biofilm formation using a hypothesized mechanism of action.

A novel bacterial strain, designated MAH-20T, was isolated from a soil sample of a tomato garden. Cells of strain MAH-20T were Gram-stain negative, aerobic, motile, and rod-shaped. The colonies were light brown colored, smooth, spherical, and 0.2–0.7 mm in diameter when grown on Luria–Bertani agar for 2 days. Strain MAH-20T grows at 15–40 °C (optimum growth temperature 30–32 °C), at pH 5.0–10.0 (optimum growth pH 7.0) and at 0–2.0% NaCl. The strain showed positive activity for both oxidase and catalase tests. Cells were able to hydrolyze starch, DNA, urea, gelatin, L-arginine, and Tween 20. According to the 16S rRNA gene sequence similarity, the strain MAH-20T was identified as a new member of the genus Sphingomonas and had the close sequence similarity with Sphingomonas changbaiensis V2M44T (98.9%) and Sphingomonas tabacisoli X1-8T (98.1%). The genomic ANI value between strain MAH-20T and S. changbaiensis NBRC 104936T was 84.4%. The novel strain MAH-20T has a draft genome size of 3,350,026 bp (25 contigs), annotated with 3210 protein-coding genes, 46 tRNA, and 3 rRNA genes. The genomic DNA G + C content of isolate was 67.3 mol%, the predominant quinone was ubiquinone 10 and the major fatty acids were C16:0, C17:1ω6c and summed feature 8 (comprising C18:1ω7c and/or C18:1ω6c). On the basis of DNA-DNA hybridization results, phenotypic, genotypic, and chemotaxonomic data, the isolated strain MAH-20T represents a novel species, for which the name Sphingomonas horti sp. nov. is proposed, with MAH-20T as the type strain (= KACC 19746T = CGMCC1.13658T).

Listeria monocytogenes is a major foodborne pathogen that adversely affects the food industry. In this study, 6 anti-listerial lactic acid bacteria (LAB) isolates were screened. These anti-listerial LAB isolates were identified via 16S rRNA gene sequencing and analyzed via repetitive extragenic palindromic-PCR. Probiotic assessment of these isolates, comprising an evaluation of the antibiotic susceptibility, tolerance to lysozyme, simulated gastric and intestinal juices, and gut conditions (low pH, bile salts, and 0.4% phenol), was carried out. Most of the isolates were resistant to streptomycin, vancomycin, gentamycin, kanamycin, and ciprofloxacin. All of the isolates were negative for virulence genes, including agg, ccf, cylA, cylB, cylLL, cylLS, cylM, esp, and gelE, and hemolytic activity. Furthermore, autoinducer-2 (a quorum-sensing molecule) was detected and quantified via HPLC with fluorescence detection after derivatization with 2,3-diaminonaphthalene. Metabolites profiles of the Lactobacillus sakei D.7 and Lactobacillus plantarum I.60 were observed and presented various organic acids linked with antibacterial activity. Moreover, freeze-dried cell-free supernatants from Lb. sakei (55 mg/mL) and Lb. plantarum (40 mg/mL) showed different minimum effective concentration (MEC) against L. monocytogenes in the food model (whole milk). In summary, these anti-listerial LAB isolates do not pose a risk to consumer health, are eco-friendly, and may be promising candidates for future use as bioprotective cultures and new probiotics to control contamination by L. monocytogenes in the food and dairy industries.

Salmonella is a foodborne pathogen and an emerging zoonotic bacterial threat in the food industry. The aim of this study was to evaluate the biofilm formation by a cocktail culture of 3 wild isolates of Salmonella enterica serotype Kentucky on plastic (PLA), silicon rubber (SR), and chicken skin surfaces under various temperatures (4, 10, 25, 37, and 42°C) and pH values (4.0, 5.0, 6.0, 7.0, and 8.0). Then, at the optimum temperature and pH, the effects of supplementation with glucose (0, 0.025, 0.05, and 0.4% w/v) on biofilm formation were assessed on each of the surfaces. The results indicated that higher temperatures (25 to 42°C) and pH values (7.0 and 8.0) led to more robust biofilm formation than lower temperatures (4 and 10°C) and lower pH levels (4.0 to 6.0). Moreover, biofilm formation was induced by 0.025% glucose during incubation at the optimum temperature (37°C) and pH (7.0) but inhibited by 0.4% glucose. Consistent with this finding, virulence related gene (rpoS, rpoH, hilA, and avrA) expression was increased at 0.025% glucose and significantly reduced at 0.4% glucose. This results also confirmed by field emission scanning electron microscope, confocal laser scanning microscopy, and autoinducer-2 determination. This study concluded that optimum environmental conditions (temperature 37°C, pH 7.0, and 0.25% glucose) exhibited strong biofilm formation on food and food contract surfaces as well as increased the virulence gene expression levels, indicating that these environmental conditions might be threating conditions for food safety.

The presence of Salmonella serotypes is a major safety concern of the food industry and poultry farmers. This study aimed to isolate and identify Salmonella spp. from a chicken processing facility by PCR and pulsed-field gel electrophoresis (PFGE). In addition, the biofilm-forming abilities of the isolated bacteria on stainless steel, silicone rubber, plastic, and chicken skin were also investigated. PCR was used for the confirmation of Salmonella serotypes, and then gene similarity within the same serotype was analyzed by PFGE. As a result, 26 S. Enteritidis isolates were detected at a high rate from both food contact surfaces and chicken products during processing. All of them were 100% genetically identical to the same bacteria. The results indicated that the virulence factors and effective biofilm-forming ability of S. Enteritidis isolates could affect human health and economic revenue. It was also suggested that the visual observation of food and food contact surfaces could be a great concern in the future. The continuous monitoring of S. Enteritidis molecular and biofilm characteristics is needed to increase food safety.

Salmonella enterica and Shiga toxin-producing (or verotoxin-producing) Escherichia coli are major foodborne pathogens, posing substantial food safety risks. Due to the negative effects of chemical treatment against foodborne pathogens, the application of enzyme-based techniques is currently receiving great attention. Here, we evaluated the inhibitory properties of Flavourzyme, a commercial peptidase, against these two foodborne pathogens. We noticed 4.0 and 5.5 log inhibition of biofilm formation by S. Typhimurium and E. coli, respectively, while treated with sub-minimum inhibitory concentrations of Flavourzyme for 24 h. For both bacteria, the enzyme exhibited quorum-quenching activity, preventing autoinducer-2 production completely by E. coli. In addition, Flavourzyme significantly suppressed the relative expression levels of biofilm-forming, quorum sensing, and virulence regulatory genes as measured by qRT-PCR. Based on our results, we suggest the use of Flavourzyme as a preventive agent against foodborne pathogens that possibly acts by inhibiting bacterial self-defense mechanisms following disruption of cellular proteins. This finding may shed light on how enzymes can be applied as a novel weapon to control foodborne illnesses to ensure food safety and public health.

Vibrio parahaemolyticus is a biofilm-producing seafood-borne pathogen that causes gastrointestinal disease in humans. Therefore, our study aimed to assess the elimination effects (alone or in combination) of ultraviolet C (UV–C; 5–60 mW × s/cm2) irradiation, sodium hypochlorite (NaOCl; 50–300 ppm), and slightly acidic electrolyzed water (SAEW; 1–15 min) on V. parahaemolyticus biofilms on crab and shrimp surfaces. SAEW alone had more significant effects than NaOCl even though it contained lower free chlorine concentrations. The combined UV-C/NaOCl treatment exhibited a significant (P < 0.05) but low reduction of V. parahaemolyticus biofilms on shrimp and crab surfaces, whereas UV-C/SAEW acquired a significantly higher (P < 0.05) reduction on both surfaces. 60 mW × s/cm2 UV-C coupled with 300 ppm NaOCl rendered maximum reductions of 3.78 and 3.32 log CFU/cm2 on shrimp and crab surfaces, respectively, whereas 60 mW × s/cm2 UV-C and 15 min of SAEW resulted in 4.41 and 4.06 log CFU/cm2 reductions in shrimp and crab, respectively. Therefore, our findings indicate that both UV-C/NaOCl and UV-C/SAEW treatment could be employed to improve the microbiological safety of the seafood industry.

Owing to their preservative and antimicrobial effects, postbiotics (metabolic byproducts of probiotics) are promising natural components for the food industry. Therefore, the present study aimed to investigate the efficacy of postbiotics collected from isolated Lactobacillus curvatus B.67 and Lactobacillus plantarum M.2 against Listeria monocytogenes pathogens in planktonic cells, motility, and biofilm states. The analysis of the metabolite composition of the postbiotics revealed various organic acids, along with a few well-known bacteriocin-encoding genes with potential antimicrobial effects. Postbiotics maintained their residual antimicrobial activity over the pH range 1–6 but lost all activity at neutral pH (pH 7). Full antimicrobial activity (100%) was observed during heat treatment, even under the autoclaving condition. Minimum inhibitory concentration (MICs) of L. curvatus B.67 and L. plantarum M.2 against L. monocytogenes were 80 and 70 mg/mL, respectively. However, four sub-MICs of the postbiotics (1/2, 1/4, 1/8, and 1/16 MIC) were tested for inhibition efficacy against L. monocytogenes during different experiment in this study. Swimming motility, biofilm formation, and expression levels of target genes related to biofilm formation, virulence, and quorum-sensing were significantly inhibited with increasing postbiotics concentration. Postbiotics from L. plantarum M.2 exhibited a higher inhibitory effect than the postbiotics from L. curvatus B.67. Nonetheless, both these postbiotics from Lactobacillus spp. could be used as effective bio-interventions for controlling L. monocytogenes biofilm in the food industry.

Gastroenteritis infections in humans are mainly associated with consumption of Vibrio parahaemolyticus contaminated shellfish, which causes health and economic loss. Virulence factor production, antibiotic resistance profile, and biofilm-forming capacity of Vibrio parahaemolyticus isolates on food and food contact surfaces at 30 °C were investigated to evaluate the antibiotic sensitivity and pathogenic level. Strains of V. parahaemolyticus were isolated from shellfish (e.g., Crassostrea gigas, Venerupis philippinarum, Mytilus coruscus, Anadara kagoshimensis) in Korea. When examined for 17 virulence factor-encoding genes, 53.3, 73.1, 87.1, 87.9, and 90.9% of the isolates were positive for genes encoding TDH, T6SS, T3SS1, T3SS2, and Type I pilus, respectively. All isolates showed resistance to vancomycin, tetracyclines, penicillin, nalidixic acid, and doxycycline, among 26 antibiotics tested, with most isolates resistant to kanamycin (93.5%), ampicillin (96.8%), clindamycin (96.8%), tobramycin (88.7%), amikacin (83.97%), and minocycline (80.7%). Biofilm formation, cell–cell attachment, and motility were high in most isolates. These findings may assist in monitoring the epidemics of the pathogen. Continuous monitoring could help to decrease V. parahaemolyticus infections and improve seafood safety.

Lactic acid bacteria (LAB) with probiotic potential have become highly sought-after by global consumers due to their wide applicability. Here, two LAB strains (J.27 and M.21) were isolated from a traditional Korean fermented vegetable (kimchi) and identified via 16S rRNA gene sequence analyses. Afterward, the two LAB isolates were evaluated for their antibiofilm activity against three foodborne pathogens (Vibrio parahaemolyticus, Pseudomonas aeruginosa, and Escherichia coli) on a seafood model (squid) and food-contact surfaces (rubber and high-density polyethylene [HDPE] plastic). This study also evaluated the effect of co-culture with LAB isolates on the relative expression of different virulence- and quorum sensing (QS)-associated genes from the three aforementioned pathogens. Our antimicrobial and genetic analyses revealed that the combined LAB isolates exhibited higher planktonic and biofilm cell inhibition on both biotic and abiotic surfaces after co-culture with the three mentioned foodborne pathogens assessed for 24 h. However, the antimicrobial activities of both LAB isolates were higher on abiotic surfaces (HDPE) than on biotic surfaces (squid). Therefore, the results of our in vitro experiments highlighted the potential applicability of both LAB isolates as “green” bioprotective agents against pathogenic microorganisms in the seafood industry.

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.

It is essential to develop and discover alternative eco-friendly antibacterial agents due to the emergence of multi-drug-resistant microorganisms. In this study, we isolated and characterized a novel bacterium named Terrabacter humi MAHUQ-38T, utilized for the eco-friendly synthesis of silver nanoparticles (AgNPs) and the synthesized AgNPs were used to control multi-drug-resistant microorganisms. The novel strain was Gram stain positive, strictly aerobic, milky white colored, rod shaped and non-motile. The optimal growth temperature, pH and NaCl concentration were 30◦ C, 6.5 and 0%, respectively. Based on 16S rRNA gene sequence, strain MAHUQ-38T belongs to the genus Terrabacter and is most closely related to several Terrabacter type strains (98.2%–98.8%). Terrabacter humi MAHUQ-38T had a genome of 5,156,829 bp long (19 contigs) with 4555 protein-coding genes, 48 tRNA and 5 rRNA genes. The culture supernatant of strain MAHUQ-38T was used for the eco-friendly and facile synthesis of AgNPs. The transmission electron microscopy (TEM) image showed the spherical shape of AgNPs with a size of 6 to 24 nm, and the Fourier transform infrared (FTIR) analysis revealed the functional groups responsible for the synthesis of AgNPs. The synthesized AgNPs exhibited strong anti-bacterial activity against multi-drug-resistant pathogens, Escherichia coli and Pseudomonas aeruginosa. Minimal inhibitory/bactericidal concentrations against E. coli and P. aeruginosa were 6.25/50 and 12.5/50 µg/mL, respectively. The AgNPs altered the cell morphology and damaged the cell membrane of pathogens. This study encourages the use of Terrabacter humi for the ecofriendly synthesis of AgNPs to control multi-drug-resistant microorganisms.

The objective of this study was to investigate the antibacterial and antibiofilm activity of eugenol against V. parahaemolyticus planktonic and biofilm cells and the involved mechanisms as well. Atime-kill assay, a biofilm formation assay on the surface of crab shells, an assay to determine the reduction of virulence using eugenol at different concentrations, energy-filtered transmission electron microscope (EF-TEM), field emission scanning electron microscopy (FE-SEM), confocal laser scanning microscope (CLSM) and high-performance liquid chromatography (HPLC) were performed to evaluate the antibacterial and antibiofilm activity of eugenol. The results indicated that different concentrations of eugenol (0.1–0.6%) significantly reduced biofilm formation, metabolic activities, and secretion of extracellular polysaccharide (EPS), with effective antibacterial effect. Eugenol at 0.4% effectively eradicated the biofilms formed by clinical and environmental V. parahaemolyticus on crab surface by more than 4.5 and 4 log CFU/cm2, respectively. At 0.6% concentration, the reduction rates of metabolic activities for ATCC27969 and NIFS29 were 79% and 68%, respectively. Whereas, the reduction rates of EPS for ATCC27969 and NIFS29 were 78% and 71%, respectively. On visual evaluation, significant results were observed for biofilm reduction, live/dead cell detection, and quorum sensing (QS). This study demonstrated that eugenol can be used to control V. parahaemolyticus biofilms and biofilm-related infections and can be employed for the protection of seafood.

In this study, the effect of three essential oils (EOs)–clove oil (CO), thyme oil (TO), and garlic oil (GO), which are generally recognized as safe–on the planktonic growth, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), motility, biofilm formation, and quorum sensing (QS) of Vibrio parahaemolyticus was investigated. All three EOs showed bacteriostatic activity, with MICs in the range 0.02%–0.09% (v/v). CO and TO completely controlled planktonic growth at 0.28% and 0.08% (v/v), which is four times their MIC (4 × MIC), after 10 min, whereas GO completely controlled growth at 0.36% (v/v) (4 × MIC) after treatment for 20 min. V. parahaemolyticus motility was significantly reduced by all three EOs at 4 × MIC (0.28% for CO, 0.08% for TO, and 0.36% for GO), whereas QS was controlled and biofilm formation reduced by all three EOs at 8 × MIC (0.56% for CO, 0.16% for TO, and 0.72% for GO) after 30 min of treatment. These results suggest that CO, TO, and GO have a significant inhibitory effect on V. parahaemolyticus cells in biofilm sand thus represent a promising strategy for improving food safety. These results provide the evidence required to encourage further research into the practical use of the proposed EOs in food preparation processes.

The purpose of this research was to characterize Listeria monocytogenes from several environmental and clinical sources and assess the efficacy of single and combined physico-chemical treatments in reducing biofilm on lettuce leaves. PCR analysis of L. monocytogenes isolates collected from different clinical (10 strains) and environmental sources (12 strains) was used to look for the presence of one Listeria-specific gene and five virulence genes. Biofilms of L. monocytogenes were developed on lettuce leaves over 24 h. A 5-min ultrasound and a 300-ppm sodium hypochlorite (NaOCl) wash resulted in similar reductions in cell numbers of 0.82 log CFU cm−2. For chlorine dioxide (ClO2) at 60 ppm, the cell numbers were reduced by ∼5.45 log CFU cm−2. A combined treatment of 5 min of ultrasound plus 300 ppm NaOCl or 40 ppm ClO2, provided maximal efficacy, reducing the number of L. monocytogenes on the lettuce surface to non-detectable levels. Therefore, ClO2 has the potential to replace NaOCl for the disinfection of food products in the food industry.

Probiotics show great promise as alternative and environmentally friendly candidates to control microbial pathogens. Here, six isolated lactic acid bacteria (LAB) were chosen based on their anti-listerial activity. Anti-listerial LAB isolates were identified by 16S rRNA gene sequencing. The anti-listerial activities of these isolates were evaluated by Listeria monocytogenes biofilm inhibition assays on stainless-steel coupons (SS), lettuce, and a minimum biofilm eradication concentration (MBEC™) biofilm device. Results revealed that following co-culture with LAB for 24 h, L. monocytogenes biofilm cells were inhibited by up to 2.17 log CFU/cm2, 1.62 log CFU/cm2, and 1.09 log CFU/peg on SS, lettuce, and MBEC™, respectively. Although these LAB bacteria suppressed L. monocytogenes biofilm formation on both surfaces, the inhibitory effect on lettuce surfaces was lower than that on SS. These results support the potential use of LAB strains to inhibit biofilm formation by pathogenic bacteria on vegetable products and in the food industry, without associated risk to consumers.

A Gram-stain-negative, aerobic, non-motile and rod-or coccoid-shaped novel bacterial strain, designated MAH-25T, was isolated from soil sampled in a pine garden. The colonies were observed to be light pink-coloured, smooth, spherical and 1–2 mm in diameter when grown on nutrient agar for 2 days. Strain MAH-25T was found to be able to grow at 15–35 °C, at pH 5.0–8.0 and at 0–2.0% NaCl. Cell growth occurred on Reasoner's 2A agar and nutrient agar. The strain was found to be positive in both oxidase and catalase tests. According to 16S rRNA gene sequence comparisons, the isolate was identified as a member of the genus Ramlibacter and closely related to Ramlibacter solisilvae 5-10T (98.0% similarity), Ramlibacter henchirensis TMB834T (97.7%), Ramlibacter tataouinensis TTB310T (97.6%) and Ramlibacter rhizophilus YS3.2.7T (97.3%). The average nucleotide identity and digital DNA–DNA hybridization values between strain MAH-25T and the four closely related type strains were in the range of 78.8–81.3% and 22.3–24.1%, respectively. The novel strain MAH-25T has a draft genome size of 5505957 bp (11 contigs), annotated with 5210 protein-coding genes, 46 tRNA and three rRNA genes. The genomic DNA G+C content was determined to be 70.3 mol%. The predominant isoprenoid quinone was ubiquinone 8 (Q-8). The major fatty acids were identified as C16:0, summed feature 3 (C16:1 ω7c and/or C16:1 ω6c) and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c). The main polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. On the basis of DNA–DNA hybridization, genotypic analysis, chemotaxonomic and physiological data, strain MAH-25T represents a novel species within the genus Ramlibacter, for which the name Ramlibacter pinisoli sp. nov. is proposed, with MAH-25T (=KACC 19839T=CGMCC1.13660T) as the type strain.

The objective of this study was to investigate the virulence factors, genetic relationship, antibiotic resistance profile and the biofilm formation ability of Vibrio parahaemolyticus isolates on shrimp and mussel surfaces at 30?C. In this study, eight (n = 8) V. parahaemolyticus isolated from mussel were examined. We used the polymerase chain reaction (PCR) to examine the distribution of different genes, and Repetitive Extragenic Palindromic-PCR (REP-PCR) to compare the genetic relationship. Disk diffusion technique was used to assess antibiotic and multiple-antibiotic resistance. The biofilm formation assay, and field emission scanning electron microscopy (FE-SEM) were used to evaluate biofilm formation ability. Transmission Electron Microscope (TEM) was used to observe the morphological structure of bacterial cell. Our results indicated that the biofilm-associated genes, 16S rRNA, toxR, and tdh, were present in all the tested V. parahaemolyticus isolates (n = 8). Approximately, 62.5% (5 isolates among 8 isolates) isolates showed strong multiple-antibiotic resistance index with an average value of 0.56. All isolates (n = 8) showed strong genetic relationship and significant biofilm formation ability on shrimp and mussel surfaces. This study demonstrated that the presence of virulence factors, high multiple antibiotic resistance index (MARI) values, and effective biofilm formation ability of V. parahaemolyticus isolates could be a great threat to human health and economic values in future. It was also suggested that a high resistance rate to antibiotic could be ineffective for treating V. parahaemolyticus infections. The continuous monitoring of V. parahaemolyticus antibiotic, molecular and biofilm characteristics is needed to increase seafood safety.

The aim of this study was to evaluate the promotive and/or inhibitory effects of NaCl, glucose, their combinations on biofilm formation and quorum sensing (QS) autoinducer-2 (AI-2) production on black tiger shrimp surfaces, using three strains of Vibrio parahaemolyticus. Initially, six different NaCl concentrations (0.5, 1, 2, 3, 4, and 5%) were evaluated for a maximum biofilm formation (∼6.3 log CFU/cm2) at 30 °C. Inhibitory effects of glucose at six different levels (0.005, 0.01, 0.015, 0.02, 0.025, and 0.05%) were then investigated using the NaCl level previously found for a maximal biofilm formation. V. parahaemolyticus formed the best biofilm at 2% NaCl and the least biofilm at 5% NaCl, regardless of vibrio strain. In combination of 2% NaCl and glucose at different levels, the largest biofilm was observed at 0.015%, with the least biofilm seen at 0.05%, regardless of vibrio strain. Addition of salt and glucose more than the optimal level (2% salt and 0.015% glucose at 2% salt) inducted a stepwise inhibition of vibrio growth and biofilm formation in a continuous matter. In the visual evaluation, similar results were observed for vibrio growth, biofilm formation, live/dead cell detection, and quorum sensing.

Breast cancer was the commonest cancer among the female patients who attended Institute of Cancer Research & Hospital (NICRH), Dhaka, Bangladesh, which formed the second most common cancer worldwide. An expressive cross-sectional study was conceded out in NICRH. By convenience sampling, 322 female patients were selected. A design and well formatted pre-tested questionnaire was used to ask questions among the breast cancer patients. The objectives of the study was to investigate the distribution and determinants of breast cancer among the female patients admitted in NICRH. The period of the study was during July 2010 to June 2011. Majority (71 %) of the patients was in the age between 34 to 53 years and they were coming from rural areas. About 77 % respondents had experienced first menstruation in the age between 12 to 13 years. It was found that 42 % respondents gave their first child birth in between 21 to 25 years age and among all the respondent 64 % became pregnant up to 3 times. It also noted that 89 % respondents had practice to use oral contraceptives. While trying to get their history of contraceptive use; it was found that 42 % used contraception in between 4 to 6 years. In this study we found that 90 % of the respondent’s had the practice to breast fed their baby. But 24 % of the respondent’s have used HRT and among them 69 % had used it by ≤3 years. Around 56 % respondents had the parenchymal pattern of breast in mammography. Forty two percent respondent’s had the history of disease on breast cancer and 25 % had the history of benign breast cancer and also 2 % had the experiences to use radiation. This study explored the overall scenario of breast cancer among the patients admitted to NICRH in Bangladesh context; understanding the different type of determinants, risk factors to developing breast cancer and the distribution of patients in different area which will help to build up the national awareness. Measure should be taken immediately to detect the factors that were associated with breast cancer. This study was a very little one in respect of the national situation in our country and the result reflected only a fraction of the original condition. Broad based study in large population with sound methodology is required to find out real situation about this matter in Bangladesh.