Isolation, characterization, and application of bacteriophage on Vibrio parahaemolyticus biofilm to control seafood contamination.

This study intended to isolate a Vibrio-particular phage from the natural environment, analyze its characteristics and genome sequence, and investigate its reduction effect on V. parahaemolyticus biofilm as a biocontrol agent in squid and mackerel. 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), 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. Color 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, and luxS, as examples. 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.

Prophylactic efficacy of baicalin and carvacrol against Salmonella Typhimurium biofilm on food and food contact surfaces.

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.

Synergistic effects of ε-poly-l-lysine and lysozyme against Pseudomonas aeruginosa and Listeria monocytogenes biofilms on beef and food contact surfaces.

This study investigated the synergistic effects of ε-poly- L -lysine (ε-PL) and lysozyme against P. aeruginosa and L. monocytogenes biofilms. Single-culture biofilms of two bacteria were formed on silicone rubber (SR), stainless steel (SS), and beef surfaces and then treated with lysozyme (0.05-5 mg/mL) and ε-PL at minimum inhibitory concentrations (MICs) of 1 to 4 separately or in combination. On the SR surface, P. aeruginosa biofilm was reduced by 1.4 and 1.9 log CFU/cm2 within 2 h when treated with lysozyme (5 mg/mL) and ε-PL (4 MIC), respectively, but this reduction increased significantly to 4.1 log CFU/cm2 (P < 0.05) with the combined treatment. On beef surface, P. aeruginosa and L. monocytogenes biofilm was reduced by 4.2-5.0, and 3.3-4.2 log CFU/g when lysozyme was combined with 1, 2, and 4 MIC of ε-PL at 25 °C, respectively. Compared to 5 mg/mL lysozyme alone, the combined treatment with 1, 2, and 4 MIC of ε-PL on beef surface achieved additional reduction against P. aeruginosa biofilm of 0.5, 0.8, and 0.7 log CFU/g, respectively, at 25 °C. In addition, 0.25 mg/mL lysozyme and 0.5 MIC of ε-PL significantly (P < 0.05) suppressed the quorum-sensing (agrA) and virulence-associated (hlyA and prfA) genes of L. monocytogenes.

Efficacy of disinfectant and bacteriophage mixture against planktonic and biofilm state of Listeria monocytogenes to control in the food industry.

Fresh produce and animal-based products contaminated with Listeria monocytogenes have been the main cause of listeriosis outbreaks for many years. The present investigation explored the potential of combination treatment of disinfectants with a bacteriophage cocktail to control L. monocytogenes contamination in the food industry. A mixture of 1 minimal inhibitory concentration (MIC) of disinfectants (sodium hypochlorite [NaOCl], hydrogen peroxide [H2O2], and lactic acid [LA]) and multiplicity of infection (MOI) 100 of phage cocktail was applied to both planktonic cells in vitro and already-formed biofilm cells on food contact materials (FCMs; polyethylene, polypropylene, and stainless steel) and foods (celery and chicken meat). All the combinations significantly lowered the population, biofilm-forming ability, and the expression of flaA, motB, hlyA, prfA, actA, and sigB genes of L. monocytogenes. Additionally, in the antibiofilm test, approximately 4 log CFU/cm2 was eradicated by 6 h treatment on FCMs, and 3 log CFU/g was eradicated within 3 days on celery. However, <2 log CFU/g was eradicated in chicken meat, and regrowth of L. monocytogenes was observed on foods after 5 days. The biofilm eradication efficacy of the combination treatment was proven through visualization using scanning electron microscopy (SEM) and confocal microscopy. In the SEM images, the unusual behavior of L. monocytogenes invading from the surface to the inside was observed after treating celery with NaOCl+P or H2O2 + P. These results suggested that combination of disinfectants (NaOCl, H2O2, and LA) with Listeria-specific phage cocktail can be employed in the food industry as a novel antimicrobial and antibiofilm approach, and further research of L. monocytogenes behavior after disinfection is needed.

Control of Listeria monocytogenes in food industry by a combination treatment of natural aromatic compound with Listeria-specific bacteriophage cocktail.

Most Listeria monocytogenes found in the food industry are listeriosis-causing pathogens and possess the ability to form biofilms on food and food contact materials (FCMs). This study aims to evaluate the efficacy of the combination treatment of natural aromatic compounds (thymol, eugenol, carvacrol, and citral) with a Listeria-specific phage cocktail in mitigating the threat posed by L. monocytogenes in the food industry. In vitro combination treatment of 1 minimal inhibitory concentration (MIC) of natural aromatic compound with phage cocktail at multiplicity of infection (MOI) 100 reduced more than 4 log CFU/mL of L. monocytogenes planktonic cells and inhibited biofilm formation. In addition, the expression of virulence-related genes (flaA, motB, hlyA, prfA, and actA) and the stress response (sigB) gene were significantly downregulated. The combination of natural aromatic compound with phage cocktail reduced the biofilm cell population on contaminated celery by more than 2 log CFU/g and by more than 2 log CFU/cm2 on already-formed biofilm on FCMs, but it was less effective on chicken meat, with an approximate reduction of only 1 log CFU/g. The antibiofilm activity toward preformed L. monocytogenes biofilms was also observed using field-emission scanning electron microscopy (FESEM) and confocal laser scanning microscopy (CLSM). COMSTAT analysis of the structural change of biofilms revealed that major biofilm structure parameters (biovolume, thickness, diffusion distance, and microcolonies at substratum) were reduced after treatment. Our findings suggest that the combination of natural aromatic compounds with a phage cocktail has enormous potential as an antimicrobial and antibiofilm agent for controlling L. monocytogenes in the food industry.

Insights into the mechanisms and key factors influencing biofilm formation by Aeromonas hydrophila in the food industry: A comprehensive review and bibliometric analysis.

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.

Synergistic effects of sequential treatment using disinfectant and e-beam for inactivation of hepatitis a virus on fresh vegetables.

Hepatitis A virus (HAV) has adversely affected public health worldwide, causing an economic burden on many countries. Fresh vegetables are reported as a source of HAV infections during production, harvesting, and distribution, which cause the emergence of foodborne illnesses. Therefore, in this study, the synergistic effects of chemical (sodium hypochlorite [NaOCl] and chlorine dioxide [ClO2]) and physical (electron-beam [e-beam] irradiation) sequential treatment for HAV inactivation on fresh vegetables were investigated, and the physicochemical quality changes of vegetables were evaluated after each treatment. On bell pepper and cucumber sequentially treated with NaOCl (50-500 ppm) and e-beam (1-5 kGy), the HAV titer was reduced by 0.19-4.69 and 0.28-4.78 log10 TCID50/mL, respectively. Sequential treatment with ClO2 (10-250 ppm) and e-beam (1-5 kGy) reduced the HAV titer on bell pepper and cucumber by 0.41-4.78 and 0.26-4.80 log10 TCID50/mL, respectively. The sequential treatments steadily decreased the HAV titers on each food by a significant difference (p < 0.05) compared to the controls. The treatment combinations of 500 ppm NaOCl and 3 kGy (e-beam) on bell pepper and 150 ppm NaOCl and 1 kGy (e-beam) on cucumber provided maximum synergistic effects. It was also found that sequential treatment with 50 ppm ClO2 and 5 kGy (e-beam) on bell pepper and 10 ppm ClO2 and 5 kGy (e-beam) on cucumber most efficiently inactivated HAV. Additionally, bell pepper and cucumber showed no significant quality changes (p < 0.05) after the treatment. Therefore, the sequential treatment with NaOCl or ClO2 and e-beam is expected to effectively control HAV on fresh vegetables without changing the food quality compared to either treatment alone.

Isolation and characterization of multidrug-resistant Salmonella-specific bacteriophages and their antibacterial efficiency in chicken breast.

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.

Probiotics and biofilm interaction in aquaculture for sustainable food security: A review and bibliometric analysis.

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.

Inhibitory effects of vorinostat (SAHA) against food-borne pathogen Salmonella enterica serotype Kentucky mixed culture biofilm with virulence and quorum-sensing relative expression.

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.

Development and efficacy assessment of hand sanitizers and polylactic acid films incorporating caffeic acid and vanillin for enhanced antiviral properties against HCoV-229E.

BACKGROUND: Although three years after the outbreak of SARS-CoV-2, the virus is still having a significant impact on human health and the global economy. Infection through respiratory droplets is the main transmission route, but the transmission of the virus by surface contact cannot be ignored. Hand sanitizers and antiviral films can be applied to control SARS-CoV-2, but sanitizers and films show drawbacks such as resistance of the virus against ethanol and environmental problems including the overuse of plastics. Therefore, this study suggested applying natural substrates to hand sanitizers and antiviral films made of biodegradable plastic (PLA). This approach is expected to provide advantages for the easy control of SARS-CoV-2 through the application of natural substances. METHODS: Antiviral disinfectants and films were manufactured by adding caffeic acid and vanillin to ethanol, isopropyl alcohol, benzalkonium chloride, and PLA. Antiviral efficacies were evaluated with slightly modified international standard testing methods EN 14,476 and ISO 21,702. RESULTS: In suspension, all the hand sanitizers evaluated in this study showed a reduction of more than 4 log within 2 min against HCoV-229E. After natural substances were added to the hand sanitizers, the time needed to reach the detection limit of the viral titer was shortened both in suspension and porcine skin. However, no difference in the time needed to reach the detection limit of the viral titer was observed in benzalkonium chloride. In the case of antiviral films, those made using both PLA and natural substances showed a 1 log reduction of HCoV-229E compared to the neat PLA film for all treatment groups. Furthermore, the influence of the organic load was evaluated according to the number of contacts of the antiviral products with porcine skin. Ten rubs on the skin resulted in slightly higher antiviral activity than 50 rubs. CONCLUSION: This study revealed that caffeic acid and vanillin can be effectively used to control HCoV-229E for hand sanitizers and antiviral films. In addition, it is recommended to remove organic matter from the skin for maintaining the antiviral activity of hand sanitizer and antiviral film as the antiviral activity decreased as the organic load increased in this study.

Antibiofilm mechanism of peppermint essential oil to avert biofilm developed by foodborne and food spoilage pathogens on food contact surfaces.

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.

Innovative nonthermal technologies for inactivation of emerging foodborne viruses.

Various foodborne viruses have been associated with human health during the last decade, causing gastroenteritis and a huge economic burden worldwide. Furthermore, the emergence of new variants of infectious viruses is growing continuously. Inactivation of foodborne viruses in the food industry is a formidable task because although viruses cannot grow in foods, they can survive in the food matrix during food processing and storage environments. Conventional inactivation methods pose various drawbacks, necessitating more effective and environmentally friendly techniques for controlling foodborne viruses during food production and processing. Various inactivation approaches for controlling foodborne viruses have been attempted in the food industry. However, some traditionally used techniques, such as disinfectant-based or heat treatment, are not always efficient. Nonthermal techniques are considered a new platform for effective and safe treatment to inactivate foodborne viruses. This review focuses on foodborne viruses commonly associated with human gastroenteritis, including newly emerged viruses, such as sapovirus and Aichi virus. It also investigates the use of chemical and nonthermal physical treatments as effective technologies to inactivate foodborne viruses.

Current and future interventions for improving poultry health and poultry food safety and security: A comprehensive review.

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.

Prophylactic efficacy of Lactobacillus curvatus B67-derived postbiotic and quercetin, separately and combined, against Listeria monocytogenes and Salmonella enterica ser. Typhimurium on processed meat sausage.

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.

Stability and inactivation of SARS-CoV-2 on food contact surfaces.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected more than 269 million people and killed more than 5.3 million people worldwide. Although fomite transmission of SARS-CoV-2 has been continuously reported, few studies have been conducted on food contact surfaces. Therefore, this study aimed to investigate the viability of coronaviruses on food contact surfaces and to remove SARS-CoV-2 contaminated on food contact surfaces with disinfectants. At 20 °C, SARS-CoV-2 was inactivated within 48 h on all food contact surfaces. At 4 °C, it was inactivated at 48 h on kraft paper and 96 h on parchment paper, but it was viable up to 5 days in low-density polyethylene (LDPE). At -20 °C, SARS-CoV-2 did not decrease by even 1 log on all food contact surfaces until 5 days. Treatment with 70% ethanol or 1000 ppm sodium hypochlorite for 5 min was sufficient to completely remove SARS-CoV-2 from 6 food contact surfaces. Similarly, UV-C irradiation at 60 mJ/cm2 eliminated SARS-CoV-2 contaminated on food contact surfaces. Also, the wiping test showed that even wiping an area contaminated with SARS-CoV-2 with a cloth moistened with 70% ethanol or 1000 ppm sodium hypochlorite, it took 5 min to inactivate the virus. Our findings suggested that SARS-CoV-2 contaminated on food contact surfaces in local retail may be viable enough to be transported home. However, if the type and method of use of the disinfectant suggested in this study are followed, it is possible to sufficiently control the fomite transmission of SARS-CoV-2 through food contact surfaces at home.

Survival of human coronavirus 229E at different temperatures on various food-contact surfaces and food and under simulated digestive conditions.

The COVID-19 pandemic caused by SARS-CoV-2 has had a major impact on human health and the global economy. Various transmission possibilities of SARS-CoV-2 have been proposed, such as the surface of food in the cold chain and food packaging, as well as the fecal-oral route, although person-to-person contact via droplets and aerosols has been confirmed as the main route of transmission. This study evaluated the survivability of HCoV-229E, a SARS-CoV-2 surrogate, in suspension, on food-contact surfaces and on food at various temperatures, and in simulated digestive fluids by TCID50 assay. In suspension, HCoV-229E survived after 5 days at 20 °C with a 3.69 log reduction, after 28 days at 4 °C with a 3.07 log reduction, and after 12 weeks at -20 °C with a 1.18 log reduction. On food-contact surfaces, HCoV-229E was not detected on day 3 on stainless steel (SS), plastic (LDPE), and silicone rubber (SR) at 20 °C with a 3.28, 3.24 and 3.28 log reduction, respectively, and survived after 28 days on SS and LDPE at 4 °C with a 3.13 and 2.88 log reduction, respectively, and survived after 12 weeks on SS, LDPE, and SR at -20 °C with a 1.92, 1.32 and 1.99 log reduction, respectively. On food, HCoV-229E was not detected on day 3 on lettuce and day 4 on chicken breast and salmon at 20 °C with a 3.61, 3.26 and 3.08 log reduction, respectively, and on day 14 on lettuce and day 21 on chicken breast and salmon at 4 °C with a 3.88, 3.44 and 3.56 log reduction, respectively. The virus remained viable for 12 weeks in all foods at -20 °C with 2-2.47 log reduction. In addition, in simulated digestive fluid experiments, HCoV-229E was relatively resistant in simulated salivary fluid (SSF; pH 7, 5), fed state simulated gastric fluid (FeSSGF; pH 3, 5, 7), and fasted state simulated intestinal fluid (FaSSIF; pH 7). However, the virus was less tolerant in fasted state simulated gastric fluid (FaSSGF; pH 1.6) and fed state simulated intestinal fluid (FeSSIF; pH 5). Therefore, this study suggested that HCoV-229E remained infectious on various food-contact surfaces and foods; in particular, it survived longer at lower temperatures and survived depending on the pH of the simulated digestive fluid.

Insights into antibiofilm mechanisms of phytochemicals: Prospects in the food industry.

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.

Inactivation methods for human coronavirus 229E on various food-contact surfaces and foods.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the cause of the COVID-19 outbreaks, is transmitted by respiratory droplets and has become a life-threatening viral pandemic worldwide. The aim of this study was to evaluate the effects of different chemical (chlorine dioxide [ClO2] and peroxyacetic acid [PAA]) and physical (ultraviolet [UV]-C irradiation) inactivation methods on various food-contact surfaces (stainless steel [SS] and polypropylene [PP]) and foods (lettuce, chicken breast, and salmon) contaminated with human coronavirus 229E (HCoV-229E). Treatments with the maximum concentration of ClO2 (500 ppm) and PAA (200 ppm) for 5 min achieved >99.9% inactivation on SS and PP. At 200 ppm ClO2 for 1 min on lettuce, chicken breast, and salmon, the HCoV-229E titers were 1.19, 3.54, and 3.97 log10 TCID50/mL, respectively. Exposure (5 min) to 80 ppm PAA achieved 1.68 log10 reduction on lettuce, and 2.03 and 1.43 log10 reductions on chicken breast and salmon, respectively, treated with 1500 ppm PAA. In the carrier tests, HCoV-229E titers on food-contact surfaces were significantly decreased (p < 0.05) with increased doses of UV-C (0-60 mJ/cm2) and not detected at the maximum UV-C dose (Detection limit: 1.0 log10 TCID50/coupon). The UV-C dose of 900 mJ/cm2 proved more effective on chicken breast (>2 log10 reduction) than on lettuce and salmon (>1 log10 reduction). However, there were no quality changes (p > 0.05) in food samples after inactivation treatments except the maximum PAA concentration (5 min) and the UV-C dose (1800 mJ/cm2).

Antibiofilm efficacy of Leuconostoc mesenteroides J.27-derived postbiotic and food-grade essential oils against Vibrio parahaemolyticus, Pseudomonas aeruginosa, and Escherichia coli alone and in combination, and their application as a green preservative in the seafood industry.

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.