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Salmonella is one of the pathogens that cause many foodborne outbreaks throughout the world, representing an important global public health problem. Salmonella strains with biofilm-forming abilities have been frequently isolated from different food processing plants, especially in poultry industry. Biofilm formation of Salmonella on various surfaces can increase their viability, contributing to their persistence in food processing environments and cross-contamination of food products. In recent years, increasing concerns arise about the antimicrobial resistant and disinfectant tolerant Salmonella , while adaptation of Salmonella in biofilms to disinfectants exacerbate this problem. Facing difficulties to inhibit or remove Salmonella biofilms in food industry, eco-friendly and effective strategies based on chemical, biotechnological and physical methods are in urgent need. This review discusses biofilm formation of Salmonella in food industries, with emphasis on the current available knowledge related to antimicrobial resistance, together with an overview of promising antibiofilm strategies for controlling Salmonella in food production environments., Competing Interests: Conflict of interestThe authors declare no conflict of interest., (© The Korean Society of Food Science and Technology 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Oxidative stress and inflammation are basic pathogenic factors involved in tissue injury and pain, as well as acute and chronic diseases. Since long-term uses of synthetic steroids and non-steroidal anti-inflammatory drugs (NSAIDs) cause severe adverse effects, novel effective materials with minimal side effects are required. In this study, polyphenol content and antioxidative activity of rosebud extracts from 24 newly crossbred Korean roses were analyzed. Among them, Pretty Velvet rosebud extract (PVRE) was found to contain high polyphenols and to show in vitro antioxidative and anti-inflammatory activities. In RAW 264.7 cells stimulated with lipopolysaccharide (LPS), PVRE down-regulated mRNA expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and thereby decreased nitric oxide (NO) and prostaglandin E 2 (PGE 2 ) production. In a subcutaneous air-pouch inflammation model, treatment with PVRE decreased λ-carrageenan-induced tissue exudation, infiltration of inflammatory cells, and inflammatory cytokines such as tumor necrosis factor-α and interleukin-1β concentrations, as achieved with dexamethasone (a representative steroid). Notably, PVRE also inhibited PGE 2 , similar to dexamethasone and indomethacin (a representative NSAID). The anti-inflammatory effects of PVRE were confirmed by microscopic findings, attenuating tissue erythema, edema, and inflammatory cell infiltration. These results indicate that PVRE exhibits dual (steroid- and NSAID-like) anti-inflammatory activities by blocking both the iNOS-NO and COX-2-PG pathways, and that PVRE could be a potential candidate as an anti-inflammatory material for diverse tissue injuries.

This study evaluated the combined antibacterial effect of 460 nm LED illumination and chitosan on Escherichia coli O157:H7, Salmonella spp. and Listeria monocytogenes on fresh-cut melon surface and its impact on the quality of melon at a total dose of 2.4 kJ/cm 2 at 4 and 10 °C. Results showed that the antibacterial effect of LED illumination in combination with chitosan (0.5 and 1.0%) was much better than that of LED illumination alone, showing their synergistic effect. Among the pathogens, L. monocytogenes was the most susceptible pathogen to LED illumination. Although the color of melons became paler after LED illumination, there was little to no change in ascorbic acid content, total flavonoid content, or antioxidant capacity of the illuminated fruits compared with non-illuminated fruits. Thus, these results suggest that chitosan-mediated 460 nm LED illumination could be applied to inactivate foodborne pathogens on fresh-cut melons during storage at food establishments., Competing Interests: Conflict of interestThe authors declare no conflict of interest., (© The Korean Society of Food Science and Technology 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

We investigated the physiological and transcriptomic response of Escherichia coli at the early stationary phase to light-emitting diodes with different wavelengths. The growth and metabolic changes of E. coli O157:H7 were examined under the influence of 465, 520, and 625 nm illuminated light. Under 465 nm illumination, the growth of E. coli O157:H7 was significantly retarded compared to 520 nm and 625 nm illumination and non-illuminated control. Metabolic changes were examined under these illumination and non-illuminated conditions based on transcriptomic reads. Transcriptomic response under 520 nm and 625 nm remained almost similar to control except few up-and down-regulated genes. Carbohydrates metabolic transcriptomic reads were greatly down-regulated under 465 nm illumination compared to 520 nm and 625 nm illumination and non-illuminated control showing depletion of glucose as a sole energy source during the exponential phase. Fatty acid degradation such as fad regulon-related genes was up-regulated in cells under 465 nm illumination revealing the shifting of cells to use fatty acid as a new carbon energy source during the early stationary phase. Exposure of E. coli O157:H7 cells to 465 nm illuminated light down-regulated virulence factor genes such as hlyA, hlyB, hlyC, stx1A, stx2B, paa, and bdm. Under the stress of 465 nm illumination, expression of stress and flagellar motility-related genes were up-regulated causing consumption of energy and reduction in cell growth. Also, oxidative phosphorylated transcriptomic reads were up-regulated under 465 nm illumination probably due to the production of ROS that might involve in the reduction of cell growth during the early stationary phase. These results indicate that pathogenic E. coli O157:H7 respond differentially to a different wavelength of the light-emitting diodes used in this study., (© 2023. The Author(s).)

This study aimed to evaluate the influence of chitosan on the antibacterial efficacy of 405 nm LED illumination against Escherichia coli O157:H7, Salmonella spp., and Listeria monocytogenes on fresh-cut melons. The antibacterial efficacy of LED illumination (a total dose of 1.3 kJ/cm 2 ) with or without chitosan (0.5 and 1.0 %) against these three pathogens was determined at 4 and 10 °C, respectively. Non-illuminated and chitosan-treated fruits were stored in the dark for 36 h under the same temperature. Color changes, ascorbic acid content, and total flavonoid content of illuminated and non-illuminated fruits were also analyzed. The results showed that the populations of all three pathogens on the non-illuminated and chitosan-treated fruits remained unchanged during storage. Regardless of bacterial species and chitosan concentrations, LED illumination in combination with chitosan greatly reduced the bacterial populations by 1.5 - 3.5 log/cm 2 , which was greater than LED illumination alone. Among the three pathogens, L. monocytogenes was the most susceptible to chitosan-mediated LED illumination. However, the whiteness index of illuminated fruits significantly increased by 1.3-fold compared to that of non-illuminated fruits, regardless of the presence of chitosan. Unlike color, no significant difference was observed in ascorbic acid and total flavonoid contents between illuminated and non-illuminated fruits. Although the fruit color was changed by LED illumination, these results indicate that adding chitosan could enhance the antibacterial efficacy of 405 nm LED illumination against major foodborne pathogens on fresh-cut melons without changing nutritional quality., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Light-emitting diodes (LEDs) is an eco-friendly light source with broad-spectrum antimicrobial activity. Recent studies have extensively been conducted to evaluate its efficacy in microbiological safety and the potential as a preservation method to extend the shelf-life of foods. This review aims to present the latest update of recent studies on the basics (physical, biochemical and mechanical basics) and antimicrobial activity of LEDs, as well as its application in the food industry. The highlight will be focused on the effects of LEDs on different types (bacteria, yeast/molds, viruses) and forms (planktonic cells, biofilms, endospores, fungal toxin) of microorganisms. The antimicrobial activity of LEDs on various food matrices was also evaluated, together with further analysis on the food-related factors that lead to the differences in LEDs efficiency. Besides, the applications of LEDs on the food-related conditions, packaged food, and equipment that could enhance LEDs efficiency were discussed to explore the future trends of LEDs technology in the food industry. Overall, the present review provides important insights for future research and the application of LEDs in the food industry.

Most foodborne pathogens have biofilm-forming capacity and prefer to grow in the form of biofilms. Presence of biofilms on food contact surfaces can lead to persistence of pathogens and the recurrent cross-contamination of food products, resulting in serious problems associated with food safety and economic losses. Resistance of biofilm cells to conventional sanitizers urges the development of natural alternatives to effectively inhibit biofilm formation and eradicate preformed biofilms. Lactic acid bacteria (LAB) produce bacteriocins which are ribosomally synthesized antimicrobial peptides, providing a great source of nature antimicrobials with the advantages of green and safe properties. Studies on biofilm control by newly identified bacteriocins are increasing, targeting primarily onListeria monocytogenes, Staphylococcus aureus, Salmonella, and Escherichia coli. This review systematically complies and assesses the antibiofilm property of LAB bacteriocins in controlling foodborne bacterial-biofilms on food contact surfaces. The bacteriocin-producing LAB genera/species, test method (inhibition and eradication), activity spectrum and surfaces are discussed, and the antibiofilm mechanisms are also argued. The findings indicate that bacteriocins can effectively inhibit biofilm formation in a dose-dependent manner, but are difficult to disrupt preformed biofilms. Synergistic combination with other antimicrobials, incorporation in nanoconjugates and implementation of bioengineering can help to strengthen their antibiofilm activity. This review provides an overview of the potential and application of LAB bacteriocins in combating bacterial biofilms in food processing environments, assisting in the development and widespread use of bacteriocin as a promising antibiofilm-agent in food industries., (© 2022 Institute of Food Technologists®.)

This study investigated the effect of LED illumination on the inactivation of Rhizopus stolonifer and Botrytis cinerea on strawberries and physicochemical properties of the strawberries. Twelve days of illumination resulted in an antifungal effect of 3.4 and 1.9 log CFU/g on R. stolonifer and B. cinerea respectively. The illumination caused no significant effect (P ≥ 0.05) on the mass, color and texture of strawberries. Furthermore, total phenolic content, trolox equivalent antioxidant capacity and anthocyanin content of the illuminated strawberries significantly increased (P < 0.05). Vitamin C content of illuminated strawberries was only significantly different (P < 0.05) from the control starting from Day 9. These results show that 405 nm LED illumination can potentially complement temperature and humidity control in preventing mold spoilage and preserving physicochemical quality of strawberries during refrigerated storage., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Acid stress is one of the most common stresses that foodborne pathogens encounter. It could occur naturally in foods as a by-product of anaerobic respiration (fermentation), or with the addition of acids. However, foodborne pathogens have managed to survive to acid conditions and consequently develop cross-protection to subsequent stresses, challenging the efficacy of hurdle technologies. Here, we cover the studies describing the cross-protection response following acid-adaptation, and the possible molecular mechanisms for cross-protection. The current and future prospective of this research topic with the knowledge gaps in the literature are also discussed. Exposure to acid conditions (pH 3.5 - 5.5) could induce cross-protection for foodborne pathogens against subsequent stress or multiple stresses such as heat, cold, osmosis, antibiotic, disinfectant, and non-thermal technology. So far, the known molecular mechanisms that might be involved in cross-protection include sigma factors, glutamate decarboxylase (GAD) system, protection or repair of molecules, and alteration of cell membrane. Cross-protection could pose a serious threat to food safety, as many hurdle technologies are believed to be effective in controlling foodborne pathogens. Thus, the exact mechanisms underlying cross-protection in a diversity of bacterial species, stress conditions, and food matrixes should be further studied to reduce potential food safety risks.HighlightsFoodborne pathogens have managed to survive to acid stress, which may provide protection to subsequent stresses, known as cross-protection.Acid-stress may induce cross-protection to many stresses such as heat, cold, osmotic, antibiotic, disinfectant, and non-thermal technology stress.At the molecular level, foodborne pathogens use different cross-protection mechanisms, which may correlate with each other.

In recent years, photodynamic inactivation (PDI) has emerged as a promising preservation method to complement refrigeration in the fresh produce supply chain. However, due to infrastructural limitations in the supply chain, fresh produce is often exposed to environmental conditions rather than recommended storage conditions. Hence, this study aimed to investigate the influence of two important environmental variables in the fresh produce supply chain - temperature and relative humidity (RH), on the PDI of fruit spoilage molds. It also aimed to demonstrate proof-of-concept of their inactivation on fruit surfaces. In the in vitro stage, Botrytis cinerea and Rhizopus stolonifer, the two molds selected for this study, were illuminated with 405 nm LEDs on Dichloran Rose-Bengal Chloramphenicol (DRBC) agar at three levels of temperature (7, 16 and 25 °C) and relative humidity (40, 60 and 80%). Illumination under these conditions caused reductions greater than 94% in the mold populations, at all temperatures and relative humidities. Even so, a temperature of 25 °C was observed to be marginally better for the inactivation as compared to 7 and 16 °C, as it necessitated the lowest dose (6-7 kJ) for the first log reduction of both the molds. Similarly, an RH of 40% worked slightly better for the inactivation of B. cinerea, as it induced inactivation without any lag phase and required the lowest dose (8.03 kJ) for the first log reduction. When the antifungal effect was investigated on fruit surfaces, it was discovered that the illumination reduced the populations of B. cinerea and R. stolonifer on strawberries by 67% and 19%, whereas on tomatoes, the respective inactivations were 79% and 70% respectively. These results demonstrate further promise of PDI as a postharvest technology for reducing the risk of fruit spoilage. This study is also the first to demonstrate the potential of PDI to add value to supply chains where compliance to ideal storage conditions is not feasible., (Copyright © 2021 Elsevier B.V. All rights reserved.)

This study investigated the antimicrobial activity of 405 nm light-emitting diode (LED) with and without riboflavin against Listeria monocytogenes in phosphate buffered saline (PBS) and on smoked salmon at different storage temperatures and evaluated its impact on food quality. The results show that riboflavin-mediated LED illumination in PBS 25 °C significantly inactivated L. monocytogenes cells by 6.2 log CFU/mL at 19.2 J/cm 2 , while illumination alone reduced 1.9 log CFU/mL of L. monocytogenes populations at 57.6 J/cm 2 . L. monocytogenes populations on illuminated smoked salmon decreased by 1.0-2.2 log CFU/cm 2 at 1.27-2.76 kJ/cm 2 at 4, 12, and 25 °C, regardless of the presence of riboflavin. Although illumination with and without riboflavin caused the lipid peroxidation and color change in smoked salmon, this study demonstrates the potential of a 405 nm LED to preserve the smoked salmon products, reducing the risk of listeriosis., Competing Interests: Conflict of interestThe authors declare no conflict of interest., (© The Korean Society of Food Science and Technology 2021.)

Although the anti-obesity effect of Korean red ginseng ( Panax ginseng Meyer) has been revealed, its underlying mechanisms are not clearly understood. Here, we demonstrate an involvement of gut microbiome in the inhibitory effect of Korean red ginseng on high-fat-diet (HFD)-induced mouse obesity, and further provides information on the effects of saponin-containing red ginseng extract (SGE) and saponin-depleted red ginseng extract (GE). Mice were fed with either SGE or GE every third day for one month, and their food intakes, fat weights, plasma glucose, and insulin and leptin levels were measured. Immunofluorescence assays were conducted to measure pancreatic islet size. Stools from the mice were subjected to metagenomic analysis. Both SGE and GE attenuated HFD-induced gain of body weight, reducing HFD-induced increase of food intakes and fat weights. They also reduced HFD-increased plasma glucose, insulin, and leptin levels, decreased both fasting and postprandial glucose concentrations, and improved both insulin resistance and glucose intolerance. Immunofluorescence assays revealed that they blocked HFD-induced increase of pancreatic islet size. Our pyrosequencing of the 16S rRNA gene V3 region from stools revealed that both SGE and GE modulated HFD-altered composition of gut microbiota. Therefore, we conclude that Korean red ginseng inhibits HFD-induced obesity and diabetes by altering gut microbiome.

Aims: Although a link between agricultural cephalosporin use and resistance in Salmonella has been demonstrated with the drug ceftiofur, the underlying mechanism of the correlation is unclear. This study investigated the impact of ceftiofur exposure in S. Saintpaul on ceftriaxone resistance, the gene expression and the conjugative transfer of the bla CTX-M-65 gene., Methods and Results: Prior ceftiofur exposure caused a twofold increase in MIC from 1024 to 2048 µg ml -1 towards ceftriaxone and increased the enzymatic activity of Bla CTX-M-65 2·2 folds from 3·46 to 7·67 nmol nitrocefin hydrolysed min -1 . A threefold upregulation in gene expression of the bla CTX-M-65 gene was also observed. Donors exposed to ceftiofur subsequently demonstrated a 2·5-fold decrease in transfer efficiency., Conclusions: Prior exposure of S. Saintpaul to ceftiofur led to increased phenotypic resistance towards ceftriaxone while its ability to spread the cephalosporin resistance through conjugation, conversely, was impaired., Significance and Impact of the Study: Findings from this study shed light on one possible mechanism in which agricultural cephalosporin exposure in Salmonella may subsequently impact clinical treatment. The finding that cephalosporin exposure in donors may hinder the subsequent spread of resistance instead of aiding it up was counter-intuitive., (© 2020 The Society for Applied Microbiology.)

Due to rapidly falling costs, whole genome sequencing (WGS) is becoming an essential tool in the surveillance of antimicrobial resistance (AMR) in Salmonella enterica. Although there have been many recent works evaluating the accuracy of WGS in predicting AMR from a large number of Salmonella isolates, little attention has been devoted to deciphering the underlying causes of disagreement between the WGS genotype and experimentally determined AMR phenotype. This study analyzed the genomes of six S. enterica isolates previously obtained from raw chicken which exhibited disagreements between WGS genotype and AMR phenotype. A total of five WGS false negative predictions toward ampicillin, amoxicillin/clavulanate, colistin, and fosfomycin resistance were presented in conjunction with their corresponding empirical phenotypic and/or genetic evidence of heteroresistance. A further case study highlighting the inherent limitations of WGS to detect the underlying genetic mechanisms of colistin heteroresistance was presented. These findings implicate heteroresistance as an underlying cause for false negative WGS-based AMR predictions in S. enterica and suggest that widespread use of WGS in the surveillance of AMR in food isolates might severely underestimate true resistance rates., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

This study aimed to evaluate three standard enrichment broth preparations for the recovery of healthy and chlorine-injured E. coli O157:H7 cells in kimchi. The growth of healthy and chlorine-injured cells in kimchi was observed in three different broths for 24 h. Results showed that the three broths were equally effective for the growth of healthy cells, although the broth described by the International Organization for Standardization (ISO) showed better performance in terms of maximum growth rate when compared to the other two broths described by the Korea Food Code (KFC) and the Food and Drug Administration (FDA). In the case of chlorine-injured cells, similar growth patterns were observed in KFC and ISO broths, whereas inhibition or no growth was found in FDA broth. Thus, this study suggests that KFC and ISO broths were more suitable than FDA broth for the enrichment of E. coli O157:H7 cells in kimchi., (© The Korean Society of Food Science and Technology 2020.)

Biofilms formed on food contact surfaces are frequently exposed to disinfectants at different concentrations. This study was designed to evaluate how S. Enteritidis in single species and dual species biofilms with P. fluorescens respond to quaternary ammonium compounds (QAC) residues on food contact surfaces. The 48 h-biofilms of S. Enteritidis and P. fluorescens in single/dual species were continuously exposed to 20 ppm QAC for 5 days, followed by QAC challenge at 200 ppm and 100 ppm for attached and detached cells, respectively. Biofilm structures were observed by confocal laser scanning microscopy (CLSM) and extracellular polymeric substances (EPS)-related gene expression was also evaluated. Results showed that QAC stress led to one log lower cell counts of S. Enteritidis and P. fluorescens single species biofilms. More cellulose observed by CLSM images and increased transcript levels of cellulose-related genes (csgD, bcsA and ardA) of S. Enteritidis were induced by QAC stress. Nevertheless, high percentage of membrane damaged cells in QAC pre-exposed biofilms might contribute to the increased sensitivity of S. Enteritidis in both attached and detached cells. Previous QAC exposure did not influence S. Enteritidis viable cell counts in dual specie biofilms, in which S. Enteritidis showed strong resistance to QAC with <2 log CFU/cm 2 reductions. Decreased transcript levels of cellulose-related genes were observed of S. Enteritidis in dual species biofilms, but EPS-related gene expression of P. fluorescens was not affected by single/duals species. The dual species biofilm matrix which has big microcolonies extruding from bottom layers with great amounts of polysaccharides mainly produced by P. fluorescens could possibly protect S. Enteritidis against disinfection. Enhanced survival of S. Enteritidis in dual species biofilms was also found when they were detached from the coupons. Overall, our findings highlight that although repeated exposures to low dose of QAC sensitized S. Enteritidis, the presence of P. fluorescens in dual species biofilms could enhance QAC resistance of S. Enteritidis, probably contributing to survival of S. Enteritidis in food processing plants., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Abstract: The BAX system for pathogen detection has been highly accurate in a variety of food products. However, false-negative results have been reported for the detection of pathogens in liquid egg products because of failed pathogen resuscitation and the existence of inhibitory components. In this study, a short-time enrichment step was used to simultaneously resuscitate the target cells to the detection level and to dilute the inhibitory components to reduce detection interference. The MP medium (BAX system) enabled faster multiplication of healthy Salmonella cells than did buffered peptone water (BPW) in tested liquid whole egg and egg yolk. However, MP failed to resuscitate heat-injured cells even after 24 h of incubation. Therefore, MP was replaced with BPW as the enrichment broth for the BAX system. However, the use of BPW for a one-step enrichment was not effective for removal of PCR inhibitors in egg yolk, and unstable detection results were obtained. To improve detection accuracy, a second step of enrichment with brain heart infusion was added. This two-step enrichment process shortened the enrichment time to 14 h and greatly increased the number of samples in which the pathogen was detected during the same enrichment time, especially in the liquid egg yolk samples. The validation study revealed 100% diagnostic accuracy of the two-step enrichment process plus the BAX system. These results indicate that a two-step enrichment process added to the BAX system can improve the detection of pathogenic Salmonella in liquid egg products., (Copyright ©, International Association for Food Protection.)

The objective of this study was to evaluate the effect of yellow mealworm ( Tenebrio molitor L.) exuviae (ME) given as a prebiotic in 20% of the diet fed to BALB/c mice. Analysis of the ME revealed that it was mostly composed of crude protein (52.94%), crude fiber (10.70%), and moisture (10.54%). When ME was fed to mice for 8 weeks, the number of intestinal lactic acid bacteria increased, reaching similar numbers (4.50 ± 0.80 CFU/mL) to those (4.70 ± 0.80 CFU/mL) of the control group not fed ME. Microbiome analysis showed that 8 weeks feeding of ME promoted the growth of Bifidobacteriaceae and Lactobacillaceae compared to the POS group, indicating the positive effects of feeding 20% ME on the intestinal microbiota of mice. These results suggest that ME can be considered as a dietary prebiotics to improve human gut microbial population, but further application study to human is necessary., Competing Interests: Conflict of interestThe authors have no conflicts of interest related to this study to disclose., (© The Korean Society of Food Science and Technology 2019.)

This study evaluated how the colonization sequence of Listeria monocytogenes and Pseudomonas fluorescens affects biofilm formation and biofilm cell response to food-related stress (desiccation or disinfection) as well as the transferability of L. monocytogenes to salmon products. The results showed that the colonization sequence did not affect the population of dual species biofilms. Furthermore, survival number of L. monocytogenes was 0.8 log CFU/cm 2 higher when P. fluorescens was the first colonizer during desiccation or disinfectant treatment in comparison with dual-species biofilms with other colonization sequences. A lower transfer rate of L. monocytogenes biofilm cells from dual-species biofilms was observed as compared to single species biofilms. In particular, L. monocytogenes cells detached at a slower rate during transfer to 10 slices of salmon from dual-species biofilms first established by P. fluorescens. Confocal images revealed more exopolysaccharide production in dual-speciesbiofilms first established by P. fluorescens than in biofilms generated via other sequences. These results indicate that preexisting P. fluorescens biofilms on stainless steel can enhance resistance of L. monocytogenes to desiccation and disinfection, although this setup decreased the transfer rate of L. monocytogenes to salmon slices. Thus, this study highlights the risk of L. monocytogenes contamination in pre-formed Pseudomonas biofilms at salmon processing facilities., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Essential oils (EOs) have demonstrated wide-spectrum antimicrobial activities and have been actively studied for their application in foods as alternative natural preservatives. However, information regarding microbial adaptive responses and changes in virulence properties following sublethal EO exposure is still scarce. The present study investigated the effect of sublethal thymol (Thy), carvacrol (Car), or trans -cinnamaldehyde (TC) adaptation on virulence gene expression and virulence properties of Escherichia coli O157:H7. The results demonstrated that E. coli O157:H7 grown to the early stationary phase in the presence of sublethal EO showed significantly ( P  < 0.05) reduced motility (reversible after stress removal), biofilm-forming ability, and efflux pump activity, with no induction of antibiotic resistance and no significant changes to its adhesion and invasion ability on a human colon adenocarcinoma (Caco-2) cell line. Reverse transcription-quantitative PCR revealed reduced expression of relevant virulence genes, including those encoding flagellar biosynthesis and function, biofilm formation regulators, multidrug efflux pumps, and type III secretion system components. This study demonstrated that Thy, Car, and TC at sublethal concentrations did not potentiate virulence in adapted E. coli O157:H7, which could benefit to their application in the food industry. IMPORTANCE The present study was conducted to evaluate changes in virulence properties in Escherichia coli O157:H7 adapted to sublethal essential oils (EOs). The results demonstrated reduced motility, biofilm-forming ability, and efflux pump activities in EO-adapted E. coli O157:H7, with no induction of antibiotic resistance or infection (adhesion and invasion) on Caco-2 cells. Reverse transcription-quantitative PCR results revealed changes in the expression of related virulence genes. Thus, the present study provides new insights into microbial virulence behavior following EO adaptation and suggests that Thy, Car, and TC sublethal exposure did not constitute a significant risk in inducing microbial virulence., (Copyright © 2019 American Society for Microbiology.)

Photodynamic inactivation is a phenomenon that has the potential to cause microbial inactivation using visible light. It works on the principle that photosensitizers within the microbial cell can be activated using specific wavelengths to trigger a series of cytotoxic reactions. In the last few years, efforts to apply this intervention technology for food safety have been on the rise. This review article offers a detailed commentary on this research. The mechanism of photodynamic inactivation has been discussed as have the factors that influence its efficacy in food. Efforts to inactivate bacteria, fungi, and viruses have been analyzed in dedicated sections and so has the application of this technology to specific product classes such as fresh produce, dry fruits, seafood, and poultry. The challenges and opportunities facing the application of this technology to food systems have been evaluated and future research directions proposed. Thus, this review will provide insights for researchers and industry personnel looking for a novel solution to combat microbial contamination and resistance., (© 2019 Institute of Food Technologists®.)

Group B Streptococcus (GBS) was previously not known to be transmitted through food, but an outbreak investigation in Singapore in 2015 documented for the first time an association between GBS Type III Sequence Type 283 infection and consumption of raw fish dishes. As very little is known about the survival of GBS during heat treatment and the stomach transit, its survival under simulated conditions was studied, in comparison with that of Escherichia coli O157:H7 and Listeria monocytogenes . The mean D -values of four GBS strains ranging from 0.72 to 0.88 min in neutral pH tryptone soy broth at 56.4 °C and 0.44-1.43 min at pH 2.35 at 37 °C in simulated gastric fluid, were significantly lower ( p  < 0.05) than those of E. coli O157:H7 and L. monocytogenes . This study suggests possible factors other than acid or heat resistance of GBS to be instrumental to its pathogenicity., Competing Interests: The authors declare no conflict of interest.This article does not contain any studies with human or animal subjects performed by the any of the authors.

This study evaluated the potential of blue light-emitting diodes (LED) of wavelength 405 and 460 nm in combination with sodium chlorophyllin to inactivate Listeria monocytogenes and Salmonella spp. on cantaloupe rind. A cocktail culture of L. monocytogenes or Salmonella spp. strains was surface inoculated onto cantaloupe rinds to reach a final concentration of 4 log CFU/cm 2 and dip-treated in a 100 μM sodium copper chlorophyllin solution. The cantaloupe samples were then exposed to 405 or 460 nm LEDs at a total dose of 1210 J/cm 2 and 5356 J/cm 2 , respectively, at 4 and 20 °C. Results showed that the antibacterial efficacy against both pathogens on cantaloupe rinds between LED alone and LED with the chlorophyllin were statistically similar with bacterial inactivation ranging from 1.1 to 3 log CFU/cm 2 in most of the cases or the difference was only minimal. A similar inactivation of 3 log CFU/cm 2 was obtained in the case of L. monocytogenes and Salmonella spp. when illuminated by 405 nm LEDs while the inactivation of L. monocytogenes was higher than Salmonella spp. when illuminated by 460 nm LED. The δ value, a modified Weibull model parameter defined as the time (h) taken to reduce the bacterial population by 90%, was computed to compare the inactivation rates of the conditions. It was inferred that illumination with 405 nm LED required a lower δ value than 460 nm LED illumination (P < 0.05) for the inactivation of L. monocytogenes at 20 °C and Salmonella spp. at 4 °C. Thus the findings of this study indicate a promising application of blue LEDs to inactivate these pathogens on the surface of cantaloupe, minimizing the risk of listeriosis and salmonellosis by consumption of cantaloupe., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Thymol, carvacrol, and trans -cinnamaldehyde are essential oil (EO) compounds with broad-spectrum antimicrobial activities against foodborne pathogens, including Escherichia coli O157:H7. However, little is known regarding direct resistance and cross-resistance development in E. coli O157:H7 after adaptation to sublethal levels of these compounds, and information is scarce on microbial adaptive responses at a molecular level. The present study demonstrated that E. coli O157:H7 was able to grow in the presence of sublethal thymol (1/2T), carvacrol (1/2C), or trans -cinnamaldehyde (1/2TC), displaying an extended lag phase duration and a lower maximum growth rate. EO-adapted cells developed direct resistance against lethal EO treatments and cross-resistance against heat (58°C) and oxidative (50 mM H 2 O 2 ) stresses. However, no induction of acid resistance (simulated gastric fluid, pH 1.5) was observed. RNA sequencing revealed a large number (310 to 338) of differentially expressed (adjusted P value [ P adj ], <0.05; fold change, ≥5) genes in 1/2T and 1/2C cells, while 1/2TC cells only showed 27 genes with altered expression. In accordance with resistance phenotypes, the genes related to membrane, heat, and oxidative stress responses and genes related to iron uptake and metabolism were upregulated. Conversely, virulence genes associated with motility, biofilm formation, and efflux pumps were repressed. This study demonstrated the development of direct resistance and cross-resistance and characterized whole-genome transcriptional responses in E. coli O157:H7 adapted to sublethal thymol, carvacrol, or trans -cinnamaldehyde. The data suggested that caution should be exercised when using EO compounds as food antimicrobials, due to the potential stress resistance development in E. coli O157:H7. IMPORTANCE The present study was designed to understand transcriptomic changes and the potential development of direct and cross-resistance in essential oil (EO)-adapted Escherichia coli O157:H7. The results demonstrated altered growth behaviors of E. coli O157:H7 during adaptation in sublethal thymol, carvacrol, and trans -cinnamaldehyde. Generally, EO-adapted bacteria showed enhanced resistance against subsequent lethal EO, heat, and oxidative stresses, with no induction of acid resistance in simulated gastric fluid. A transcriptomic analysis revealed the upregulation of related stress resistance genes and a downregulation of various virulence genes in EO-adapted cells. This study provides new insights into microbial EO adaptation behaviors and highlights the risk of resistance development in adapted bacteria., (Copyright © 2018 American Society for Microbiology.)

The objective of this study was to evaluate probiotic effects of two Lactobacillus plantarum strains (GBL16 and 17) isolated from black raspberry. Results revealed that the number of GBL16 was gradually decreased as bile salt concentration was increased from 0.3 to 1%. However, GBL17 did not show any difference when GBL17 was applied to 1% bile salt, and it indicates that GBL17 is more tolerant to bile salt than GBL16. GBL17 exhibited higher heat resistance and adhesion ability to Caco-2 cells than GBL16. Regarding gut microbiome, no significant change in the number of total bacteria in intestines of mice after treatment with GBLs was determined. However, the combination of GBL16 and GBL17 significantly increased the number of total bacteria in intestines of mice after they were orally administered. Therefore, the results suggest that both GBL16 and 17 strains could be one of major probiotics that can improve human gut health., Competing Interests: Compliance with ethical standardsThe authors have no conflicts of interest related to this study to disclose.

Background: Salmonella enterica subspecies enterica serovar Saintpaul ( S . Saintpaul) is an important gut pathogen which causes salmonellosis worldwide. Although intestinal salmonellosis is usually self-limiting, it can be life-threatening in children, the elderlies and immunocompromised patients. Appropriate antibiotic treatment is therefore required for these patients. However, the efficacy of many antibiotics on S. enterica infections has been greatly compromised due to spreading of multidrug resistance (MDR) plasmids, which poses serious threats on public health and needs to be closely monitored. In this study, we sequenced and fully characterized an S. enterica MDR plasmid pSGB23 isolated from chicken., Results: Complete genome sequence analysis revealed that S. Saintpaul strain SGB23 harbored a 254 kb megaplasmid pSGB23, which carries 11 antibiotic resistance genes responsible for resistance to 9 classes of antibiotics and quaternary ammonium compounds that are commonly used to disinfect food processing facilities. Furthermore, we found that pSGB23 carries multiple conjugative systems, which allow it to spread into other Enterobacteriaceae spp. by self-conjugation. It also harbors multiple types of replicons and plasmid maintenance and addictive systems, which explains its broad host range and stable inheritance., Conclusions: We report here a novel MDR plasmid pSGB23 harboured by S. enterica . To our knowledge, it carried the greatest number of antibiotic resistance genes with the broadest range of resistance spectrum among S. enterica MDR plasmids identified so far. The isolation of pSGB23 from food sources is worrisome, while surveillance on its further spreading will be carried out based on the findings reported in this study.

For the past decades, there has been a growing demand for natural antimicrobials in the food industry. Plant extracts have attracted strong research interests due to their wide-spectrum antimicrobial activities, but only a limited number have been investigated thoroughly. The present study aimed at identifying a novel anti-staphylococcal plant extract, to validate its activity in a food model, and to investigate on its composition and antimicrobial mechanism. Four plant extracts were evaluated against Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) in vitro, with Syzygium antisepticum leaf extract showing the strongest antimicrobial activity (MIC = 0.125 mg/mL). Relatively high total phenolic content (276.3 mg GAE/g extract) and antioxidant activities (90.2-138.0 mg TE/g extract) were measured in S. antisepticum extract. Food validation study revealed that higher extract concentration (32 mg/mL) was able to inhibit or reduce staphylococcal growth in cooked chicken, but caused color change on meat surface. By GC-MS, β-caryophyllene (12.76 area%) was identified as the dominant volatile compound in extract. Both crude extract and pure β-caryophyllene induced membrane damages in S. aureus. These results suggested good anti-staphylococcal properties of S. antisepticum plant extract, identified its major volatile composition and its membrane-damaging antimicrobial mechanism., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Extracts from medicinal plants have been reported to possess good antimicrobial properties, but a majority of them remain unexplored. This study aimed at identifying a novel plant extract with antimicrobial activity, to validate its efficacy in food model, and to elucidate its composition and antimicrobial mechanism. A total of 125 plant extracts were screened, and Cinnamomum javanicum leaf and stem extract showed potential antimicrobial activity against Listeria monocytogenes (MIC=0.13mg/mL). Total phenolic content of the extract was 78.3mg GAE/g extract and its antioxidant activity was 57.2-326.5mg TE/g extract. When applied on cold smoked salmon, strong strain-dependent antimicrobial effectiveness was observed, with L. monocytogenes LM2 (serotype 4b) and LM8 (serotype 3a) being more resistant compared to SSA81 (serotype 1/2a). High extract concentration (16mg/mL) was needed to inhibit or reduce the growth of L. monocytogenes on smoked salmon, which resulted in surface color change. GC-MS revealed that eucalyptol (25.54 area%) was the most abundant compound in the crude extract. Both crude extract and eucalyptol induced significant membrane damages in treated L. monocytogenes. These results suggest anti-L. monocytogenes activity of C. javanicum plant extract, identified its major volatile components, and elucidated its membrane-damaging antimicrobial mechanisms., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Aims: This study aimed to evaluate the biofilm formation and disinfectant resistance of Salmonella cells in mono- and dual-species biofilms with Pseudomonas aeruginosa, and to investigate the role of extracellular polymeric substances (EPS) in the protection of biofilms against disinfection treatment., Methods and Results: The populations of Salmonella in mono- or dual-species biofilms with P. aeruginosa on stainless steel (SS) coupons were determined before and after exposure to commercial disinfectant, 50 μg ml -1 chlorine or 200 μg ml -1 Ecolab ® Whisper™ V (a blend of four effective quaternary ammonium compounds (QAC)). In addition, EPS amount from biofilms was quantified and biofilm structures were observed using scanning electron microscopy (SEM). Antagonistic interactions between Salmonella and P. aeruginosa resulted in lower planktonic population level of Salmonella, and lower density in dual-species biofilms compared to mono-species biofilms. The presence of P. aeruginosa significantly enhanced disinfectant resistance of S. Typhimurium and S. Enteritidis biofilm cells for 2 days, and led to an average of 50% increase in polysaccharides amount in dual-species biofilms than mono-species biofilms of Salmonella. Microscopy observation showed the presence of large microcolonies covered by EPS in dual-species biofilms but not in mono-species ones., Conclusion: The presence of P. aeruginosa in dual-species culture inhibited the growth of Salmonella cells in planktonic phase and in biofilms, but protected Salmonella cells in biofilms from disinfection treatment, by providing more production of EPS in dual-species biofilms than mono-species ones., Significance and Impact of the Study: This study provides insights into inter-species interaction, with regard to biofilm population dynamics and disinfectant resistance. Thus, a sanitation protocol should be designed considering the protective role of secondary species to pathogens in biofilms on SS surface which has been widely used at food surfaces and manufacturers., (© 2017 The Society for Applied Microbiology.)

The objective of this study was to investigate the effect of 460 nm light-emitting diode (LED) on the inactivation of foodborne bacteria. Additionally, the change in the endogenous metabolic profile of LED illuminated cells was analyzed to understand the bacterial response to the LED illumination. Six different species of bacteria (Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, Escherichia coli O157:H7, Pseudomonas aeruginosa and Salmonella Typhimurium) were illuminated with 460 nm LED to a maximum dose of 4080 J/cm 2 at 4, 10 and 25 °C. Inactivation curves were modeled using Hom model. Metabolic profiling of the non-illuminated and illuminated cells was performed using a Liquid chromatography-mass spectrometry system. Results indicate that the 460 nm LED significantly (p < 0.05) reduced the populations of all six bacterial species. For example, the population of S. aureus reached below detection limit within 7 h. B. cereus was most resistant to photo-inactivation and exhibited about 3-log reduction in 9 h. Metabolic profiling of the illuminated cells indicated that several metabolites e.g. 11-deoxycortisol, actinonin, coformycin, tyramine, chitobiose etc. were regulated during LED illumination. These results elucidate the effectiveness of 460 nm LED against foodborne bacteria and hence, its suitability as a novel antimicrobial control method to ensure food safety., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

This study evaluated the antibacterial effect of 405 ± 5 nm light emitting diode (LED) illumination against four Salmonella serovars on fresh-cut papaya and on fruit quality at various storage temperatures. To determine the antibacterial mechanism of LED illumination at 0.9 kJ/cm 2 , oxidative damage to DNA and membrane lipids of Salmonella in phosphate-buffered saline solution was measured. The populations of Salmonella on cut fruits were significantly (P < 0.05) reduced by 0.3-1.3 log CFU/cm 2 at chilling temperatures following LED illumination for 36-48 h (1.3-1.7 kJ/cm 2 ). However, at room temperature, bacterial populations increased rapidly to 6.3-7.0 log CFU/cm 2 following LED illumination for 24 h (0.9 kJ/cm 2 ), which was approximately 1.0 log lower than the number of colonies on non-illuminated fruits. Levels of bacterial DNA oxidation significantly increased, whereas lipid peroxidation in bacterial membrane was not observed, suggesting that DNA oxidation contributes to photodynamic inactivation by LED illumination. LED illumination did not adversely affect the physicochemical and nutritional qualities of cut papaya, regardless of storage temperature. These results indicate that a food chiller equipped with 405 ± 5 nm LEDs can preserve fresh-cut papayas in retail stores without deterioration, minimizing the risk of salmonellosis., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

To investigate a potential of 405±5nm light emitting diode (LED) as a novel technology for food preservation, the antibacterial effect of 405±5nm LED on Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella spp. on the surface of fresh-cut mango and its influence on fruit quality were evaluated at different storage temperatures. LED-illumination inactivated 1.0-1.6 logCFU/cm 2 of populations at 4 and 10°C for 36-48h (total dose, 2.6-3.5kJ/cm 2 ) regardless of bacterial species, while those on non-illuminated mange remained unchanged or slightly increased during storage. At 20°C for 24h (total dose, 1.7kJ/cm 2 ), non-illuminated E. coli O157:H7 and Salmonella gradually grew, whereas LED-illumination reduced 1.2 log of Salmonella and inhibited the growth of E. coli O157:H7. Unlike these, non-illuminated L. monocytogenes cells rapidly increased to 7.3 log, while illuminated cells reached 4.6 log, revealing that LED-illumination delayed their growth. There were no significant (P>0.05) differences in color, antioxidant capacity, ascorbic acid, β-carotene, and flavonoid between non-illuminated and illuminated cut mangoes, regardless of storage temperature. These results suggest that 405±5nm LEDs in combination with chilling temperatures could be applied to preserve fresh-cut fruits without deterioration of physicochemical quality of fruits at food establishments, minimizing the risk of foodborne disease., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: As the preparation of salads involves extensive handling and the use of uncooked ingredients, they are particularly vulnerable to microbial contamination. This study aimed to determine the microbial safety and quality of pre-packed salads and salad bar ingredients sold in Singapore, so as to identify public health risks that could arise from consuming salads and to determine areas for improvement in the management of food safety., Results: The most frequently encountered organism in pre-packed salad samples was B. cereus, particularly in pasta salads (33.3%, 10/30). The most commonly detected organism in salad bar ingredients was L. monocytogenes, in particular seafood ingredients (44.1%, 15/34), largely due to contaminated smoked salmon. Further investigation showed that 21.6% (37/171) of the pre-packed smoked salmon sold in supermarkets contained L. monocytogenes. Significantly higher prevalence of L. monocytogenes and higher Standard Plate Count were detected in smoked salmon at salad bars compared to pre-packed smoked salmon in supermarkets, which suggested multiplication of the organism as the products move down the supply chain. Further molecular analysis revealed that L. monocytogenes Sequence Type (ST) 2 and ST87 were present in a particular brand of pre-packed salmon products over a 4-year period, implying a potential persistent contamination problem at the manufacturing level., Conclusions: Our findings highlighted a need to improve manufacturing and retail hygiene processes as well as to educate vulnerable populations to avoid consuming food prone to L. monocytogenes contamination.

The aim of this study was to elucidate the antibacterial mechanism of 405 ± 5-nm light-emitting diode (LED) illumination against Salmonella at 4°C in phosphate-buffered saline (PBS) by determining endogenous coproporphyrin content, DNA oxidation, damage to membrane function, and morphological change. Gene expression levels, including of oxyR , recA , rpoS , sodA , and soxR , were also examined to understand the response of Salmonella to LED illumination. The results showed that Salmonella strains responded differently to LED illumination, revealing that S. enterica serovar Enteritidis (ATCC 13076) and S. enterica subsp. enterica serovar Saintpaul (ATCC 9712) were more susceptible and resistant, respectively, than the 16 other strains tested. There was no difference in the amounts of endogenous coproporphyrin in the two strains. Compared with that in nonilluminated cells, the DNA oxidation levels in illuminated cells increased. In illuminated cells, we observed a loss of efflux pump activity, damage to the glucose uptake system, and changes in membrane potential and integrity. Transmission electron microscopy revealed a disorganization of chromosomes and ribosomes due to LED illumination. The levels of the five genes measured in the nonilluminated and illuminated S Saintpaul cells were upregulated in PBS at a set temperature of 4°C, indicating that increased gene expression levels might be due to a temperature shift and nutrient deficiency rather than to LED illumination. In contrast, only oxyR in S Enteritidis cells was upregulated. Thus, different sensitivities of the two strains to LED illumination were attributed to differences in gene regulation. IMPORTANCE Bacterial inactivation using visible light has recently received attention as a safe and environmentally friendly technology, in contrast with UV light, which has detrimental effects on human health and the environment. This study was designed to understand how 405 ± 5-nm light-emitting diode (LED) illumination kills Salmonella strains at refrigeration temperature. The data clearly demonstrated that the effectiveness of LED illumination on Salmonella strains depended highly on the serotype and strain. Our findings also revealed that its antibacterial mechanism was mainly attributed to DNA oxidation and a loss of membrane functions rather than membrane lipid peroxidation, which has been proposed by other researchers who studied the antibacterial effect of LED illumination by adding exogenous photosensitizers, such as chlorophyllin and hypericin. Therefore, this study suggests that the detailed antibacterial mechanisms of 405-nm LED illumination without additional photosensitizers may differ from that by exogenous photosensitizers. Furthermore, a change in stress-related gene regulation may alter the susceptibility of Salmonella cells to LED illumination at refrigeration temperature. Thus, our study provides new insights into the antibacterial mechanism of 405 ± 5-nm LED illumination on Salmonella cells., (Copyright © 2017 American Society for Microbiology.)

We conducted a microbiological assessment of Indian Rojak, a popular deep fried food in Singapore to evaluate its overall microbial quality, assess the effectiveness of reheating and identify key food items that could contribute to the microbial load of the dish. In 2009, an outbreak of foodborne illness associated with this food led to 154 reported cases of acute gastroenteritis, 48 were hospitalized and 2 died. Vibrio parahaemolyticus was isolated from the patients. We evaluated 455 Indian Rojak ingredients from 35 stalls; no Salmonella spp, Vibrio cholerae/parahaemolyticus or Escherichia coli O157:H7 were recovered from the studied samples. The reheating by the food handlers significantly reduced the overall median Standard Plate Count (SPC) of food from 4.5 to 2.7 log colony forming units (CFU)/g (p<0.05). The cooked ingredients with the highest microbial loads were tofu and fish cake, with those purchased from wet markets having significantly higher bacterial loads than those purchased from supermarkets (p<0.05). The Rojak gravy had the lowest median bacterial load (1.9 log CFU/g). Raw, ready-to-eat vegetables, namely green chillis, cucumbers and onions had higher levels ranging from 5.9 to 6.1 log CFU/g. Contamination with E. coli, Staphylococcus aureus, and Bacillus cereus was seen with some of the ready-to-eat raw vegetables. Repeated education of food handlers with emphasis on good hygiene practices should be conducted to reduce the risk of foodborne illnesses.

Volatile organic compounds (VOCs) as chemical spoilage indexes (CSIs) of raw chicken breast stored aerobically at 4, 10, and 21 °C were identified and quantified using solid phase microextraction (SPME) combined with gas chromatography-mass spectrometry (GC-MS). The growth dynamics of total viable count (TVC), psychrotrophs, Pseudomonas spp., lactic acid bacteria (LAB), Brochothrix thermosphacta and H2 S producing bacteria were characterized based on maximum growth rates (μmax ), maximal microbial concentration (Nmax ) and at the moment of microbial shelf life (Svalues ), calculated from Gompertz-fitted growth curves. Pseudomonas spp. was predominant species, while B. thermosphacta was characterized by the highest μmax . The microbiological and sensory shelf lives were estimated based on TVC, Pseudomonas spp., and B. thermosphacta counts and sensory evaluation, respectively. Among 27 VOCs identified by GC-MS in spoiled chicken samples, ethanol (EtOH), 1-butanol-3-methyl (1But-3M), and acetic acid (C2 ) achieved the highest Pearson's correlation coefficients of 0.66, 0.61, and 0.59, respectively, with TVC, regardless of storage temperature. Partial least squares (PLS) regression revealed that the synthesis of 1But-3M and C2 was most likely induced by the metabolic activity of B. thermosphacta and LAB, while EtOH was attributed to Pseudomonas spp. The increase in concentration of selected volatile spoilage markers (EtOH, 1But-3M, and C2 ) in the headspace over spoiled chicken breast was found to be statistically significant (P < 0.05) with TVC growth. These findings highlight the possibility of analyzing the combination of 3 selected spoilage markers: EtOH, 1But-3M, and C2 as rapid evaluation for poultry quality testing using SPME-GC-MS., (© 2016 Institute of Food Technologists®)

Fresh mung bean sprouts have been identified as a source of many Salmonella outbreaks worldwide. The aim of this study was to develop a rapid and accurate detection methodology for low levels of healthy and sanitizer-injured Salmonella on mung bean sprouts using real-time PCR coupled with either immunomagnetic separation (PCR-IMS) or centrifugation (PCR-cen). Initially, three parameters of IMS; specificity/sensitivity, bacterial concentration and bead incubation time were optimized. Secondly, limit of detection (LOD) was determined for the optimized PCR-IMS and PCR-cen. These two methods were compared against PCR alone (PCR) and the standard culture method (ISO) for their ability to detect Salmonella using inoculated and uninoculated sprouts. Under optimum IMS conditions (10(5)CFU/ml for 30 min), capture efficiency of Salmonella in sprout suspensions was lower than 40%, most probably due to the non-specific binding of the background microbiota. PCR-IMS and PCR-cen had a similar LOD at 10(3)CFU/ml, which was one log unit lower than PCR. Enrichment of 10h was sufficient to detect 100% of the inoculated sprouts with both PCR-IMS and PCR-cen, which was significantly faster compared to PCR and the ISO method. Moreover, the validation study using uninoculated sprouts revealed that PCR-IMS and PCR-cen were equally effective on Salmonella detection, showing 98.3% accuracy. These results suggest that PCR-cen would be the effective and less costly method for the detection of both healthy and sanitizer-injured Salmonella on mung bean sprouts., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Blue light-emitting diodes (LEDs) have been known to produce an antibacterial effect on various pathogenic bacteria. To extend this application to foods, blue 460-nm LEDs were evaluated for their antibacterial effect on Salmonella in orange juice. A cocktail of Salmonella enterica serovars Gaminara, Montevideo, Newport, Typhimurium, and Saintpaul was inoculated into pasteurized orange juice and illuminated with 460-nm LEDs at irradiances of 92, 147.7, and 254.7 mW/cm(2) and temperatures of 4, 12, and 20°C. Subsequently, linear, Weibull, and Gompertz models were fitted to the resultant survival curves. The color of the orange juice during illumination was also monitored. It was observed that irradiance and temperature both influenced the inactivation of Salmonella, which ranged from 2 to 5 log CFU/ml. The inactivation kinetics was best described by the Weibull model. An irradiance of 92 mW/cm(2) and temperatures of 12 and 20°C were the most bactericidal combinations, with D-values of 1,580 and 2,013 J/cm(2), respectively. Significant color changes were also observed after illumination; these changes could be minimized by choosing appropriate irradiance and temperature. These results demonstrate the potential of 460-nm LEDs for the preservation of fruit juices in the retail markets and their utility in minimizing the risk of salmonellosis.

The purpose of this investigation was to identify and quantify the volatile chemical spoilage indexes (CSIs) for raw Atlantic salmon (Salmo salar) fillets stored under aerobic storage conditions at 4, 10 and 21 °C in relation to microbial and sensory shelf lives. The volatile organic compounds (VOCs) were analyzed with SPME-GC-MS technique. Through multivariate chemometric method, hierarchical cluster analysis (HCA) and Pearson's correlations, the CSIs: trimethylamine (TMA), ethanol (EtOH), 3-methyl-1-butanol (3Met-1But), acetoin and acetic acid (C2) were selected from the group of 28 detected VOCs. At the moment of microbiological shelf life established at total viable count (TVC) of 7.0 log CFU/g, the CSIs achieved levels of 11.5, 38.3, 0.3, 24.0 and 90.7 μg/g of salmon for TMA, EtOH, 3M-1But, acetoin and C2, respectively. Pseudomonas spp. was found as major specific spoilage organism (SSOs), suitable for shelf life prediction using modified Gompertz model at the cut-off level of 6.5 log CFU/g. H2S producing bacteria and Brochothrix thermosphacta were considered as important spoilage microorganisms; however, they were not suitable for shelf life estimation. Partial least square (PLS) regression revealed possible associations between microorganisms and synthetized VOCs, showing correlations between Pseudomonas spp. and 3Met-1But and aldehydes synthesis, lactic acid bacteria were linked with EtOH, C2 and esters, and B. thermosphacta with acetoin formation., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Aims: Little information is available on a direct comparison of the antibacterial efficacy of light emitting diode (LEDs) of different peak wavelengths. Thus, the objective of this study was to evaluate the effect of LEDs of three different wavelengths on bacterial inactivation., Methods and Results: Lactobacillus plantarum, Staphylococcus aureus and Vibrio parahaemolyticus were illuminated with 405, 460 and 520 nm LEDs at 4, 10 and 25°C respectively. Inactivation curves were plotted and fitted using Gompertz Model. Illumination with 405 and 460 nm LED produced significant inactivation (P < 0·05) in the population of V. parahaemolyticus (>4 log) while Lact. plantarum and Staph. aureus showed relatively less susceptibility to the LED illumination. The 520 nm LED produced negligible inactivation., Conclusions: The 405 and 460 nm LEDs proved more effective in inactivating the selected foodborne bacteria in this study compared to 520 nm LED. The 405 nm LED showed the greatest antibacterial effect at the same level of energy dose., Significance and Impact of the Study: The results in this study demonstrated the antibacterial efficacy of 405 nm LED on Lact. plantarum and V. parahaemolyticus, suggesting its potential for use in food industry for the control of these micro-organisms., (© 2015 The Society for Applied Microbiology.)

The ability of nine commercial broths to enrich healthy and 90% sanitizer-injured Salmonella Typhimurium and Salmonella cocktail on mung bean sprouts was evaluated to select an optimum broth for detection. Results showed that S. Typhimurium multiplied faster and reached a higher population in buffered peptone water (BPW), Salmonella AD media (AD) and ONE broth-Salmonella (OB), compared with other broths. Healthy and 90% sanitizer-injured Salmonella at low concentrations increased by 4.0 log CFU/ml in these three broths. However, no Salmonella growth was observed in lactose broth (LB). Further investigation showed that during incubation, pH of LB dropped from 6.7 to 4.2, due to production of lactic (66 mM) and acetic acids (62 mM) by lactic acid bacteria that were identified as dominant microbiota in bean sprouts. Though no cell membrane damage was detected by propidium monoazide combined with real-time PCR, it was found that LB inhibited Salmonella growth, especially from low inoculum levels. This study suggests that in consideration of effectiveness and cost, BPW would be a suitable enrichment broth to use for isolating and detecting Salmonella on mung bean sprouts, while using LB might cause false negative results in Salmonella detection by either PCR or standard cultural method., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

This study investigated the antibacterial effect of 405 ± 5 nm light emitting diode (LED) on Bacillus cereus, Listeria monocytogenes, and Staphylococcus aureus, and examined its antibacterial mechanism by determining the bacterial membrane and DNA damages. A 405 ± 5 nm LED illuminated the Gram-positive pathogens until 486 J/cm(2) at 4 °C. Weibull model was used to calculate reliable life (tR) to compare bacterial sensitivities to LED illumination. The membrane damage was determined by NaCl and LIVE/DEAD® assay, while comet assay and DNA ladder analysis were conducted to determine DNA degradation. The illumination resulted in 1.9, 2.1, and 1.0 log reductions for B. cereus, L. monocytogenes, and S. aureus at 486 J/cm(2), respectively. The comparison of tR values revealed that L. monocytogenes was identified as the most susceptible strain to LED illumination. The percentage of the bacterial sensitivity to NaCl remarkably increased in LED-illuminated cells compared to non-illuminated cells. Moreover, loss of membrane integrity was confirmed for LED-illuminated cells by LIVE/DEAD® assay, whereas no DNA breakage was indicated by comet assay and DNA ladder analysis. Thus, these findings suggest that the antibacterial effect of 405 ± 5 nm LED illumination on these pathogens might be due to physical damage to bacterial membrane rather than DNA degradation., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Photodynamic inactivation studies of microbial pathogens have focused on the use of an external photosensitizer or a precursor compound to eliminate bacteria. The present study investigated the inactivation kinetics of six bacterial pathogens by a 405nm light emitting diode (LED) without the addition of any external compound. The role of endogenous coproporphyrin on the bacterial susceptibility to LEDs was also examined. Pathogens were illuminated with LEDs at 25, 10 and 4°C for 9h and the inactivation curves were modeled using six different equations. Endogenous coproporphyrin was quantified using an HPLC system equipped with a fluorescence detector. At a dose of 306J/cm(2), the 405nm LED brought about 4.0, 2.1 and 1.9 log reductions in the populations of Staphylococcus aureus at 25, 10 and 4°C, respectively. At all three temperatures, the population of Bacillus cereus and Listeria monocytogenes reduced by approximately 2.3 and 1.9 log respectively. Salmonella Typhimurium and Escherichia coli O157:H7 showed moderate susceptibility to 405nm LED while Pseudomonas aeruginosa was most resistant. Of the six models tested, Hom model proved most suitable. This study demonstrated that 405nm LEDs can be useful in the inactivation of bacterial pathogens with the aid of endogenous coproporphyrin alone., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The objective of this study was to evaluate performance of the commercial kit based on loop-mediated isothermal amplification (LAMP) in comparison with the International Organization for Standardization method for detecting uninjured and sublethally injured Salmonella cells artificially inoculated at levels of 10(0) and 10(1) CFU/25 g on raw duck wing, raw mung bean sprouts, and processed fishballs. Injured cells were prepared by a heat treatment for duck wings and fishball samples and a chlorine treatment for bean sprout samples. Additionally, a validation study was performed on naturally contaminated food samples sold in Singapore. A total of 110 samples of each commodity were analyzed in this study. Regardless of inoculum levels, the detection by the commercial LAMP kit showed 100% sensitivity and specificity for both inoculated and uninoculated samples compared with the International Organization for Standardization method, with the exception of bean sprout samples. Only 20% of bean sprout samples inoculated with 10(0) CFU/25 g injured Salmonella cells were positive by using the commercial LAMP-based kit. However, all negative samples became positive following a secondary enrichment in Rappaport-Vassiliadis medium with soy broth or after concentration by centrifugation. These results suggest that secondary enrichment or centrifugation should be considered as an additional step to increase the sensitivity of the commercial LAMP-based kit with low numbers of injured target cells in samples with high background microflora (such as mung bean sprouts). The validation study also showed that the commercial LAMP-based kit provided 91% sensitivity and 95% specificity for naturally contaminated samples. Thus, this study demonstrates that the commercial LAMP-based kit might be a cost-effective method, as this system could provide rapid, accurate detection of both uninjured and injured Salmonella cells on raw duck wings, raw mung bean sprouts, and processed fishballs in less than 26 h.

Light emitting diodes (LEDs) with their antibacterial effect present a novel method for food preservation. This effect may be influenced by environmental conditions such as the pH of the food contaminated by the pathogen. Thus, it is necessary to investigate the influence of pH on the antibacterial effect of LEDs before their application to real food matrices. Escherichia coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes in trypticase soy broth were illuminated using 10-W 461 (22.1 mW/cm(2)) and 521 nm (16 mW/cm(2)) LEDs at pH values of 4.5, 6.0, 7.3, 8.0 and 9.5 for 7.5 h at 15 °C. Using the 461 nm LEDs, the populations of E. coli O157:H7 decreased by 2.1 ± 0.02, 1.2 ± 0.08 and 4.1 ± 0.42 log CFU/ml at pH 4.5, 7.3 and 9.5 respectively, after a dosage of 596.7 J/cm(2). For L. monocytogenes, approximately a 5.8 ± 0.03 log reduction was observed after 238.7 J/cm(2) at pH 4.5 using the 461 nm LEDs, while the bacterial concentration was reduced by 1.8 ± 0.01 log at pH 9.5 after 596.7 J/cm(2). Bacterial inactivation using the 521 nm LEDs showed similar trends to the 461 nm LEDs at both acidic and alkaline pH conditions but with lower (1-2 log CFU/ml) reductions after 432 J/cm(2). Lower D-values were observed for L. monocytogenes when exposed to LEDs at acidic pH values, while the sensitivity of E. coli O157:H7 and S. Typhimurium to LED was markedly increased at an alkaline pH. Regardless of the pH at which the cultures were illuminated, the percentage of sublethal injury increased with the treatment time. These results highlight the enhanced antibacterial effect of the 461 nm LED under acidic and alkaline pH conditions, proving its potential to preserve foods as well as to have synergistic effect with acidic and alkaline antimicrobials., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Exposure of Salmonella to environmental stress, prior to its adherence to a food contact surface, may change the cell surface properties and consequently affect its initial attachment and biofilm formation. This study investigated the influence of temperature and pH preacclimation on the initial attachment of Salmonella Enteritidis to acrylic and stainless steel. Besides, changes in physicochemical properties of cells were examined; and their surface attachment was modeled by xDLVO theory. Results showed that control cells pre-grown at 37°C had significantly (P<0.05) higher initial attachment, followed by those pre-grown at 25, 42, and 10°C. The initial attachment of cells pre-grown at pH 5.3 or 6.3 was not significantly (P>0.05) different from control cells pre-grown at pH 7.3, but they were significantly higher compared to cells pre-grown at pH 8.3 and 9.0. No significant difference was observed between cell attachment to acrylic and stainless steel, although they had different physicochemical properties. The xDLVO theory successfully explained higher attachment for cells pre-grown at optimal condition on both contact surfaces. However, the xDLVO theory could not explain the similar attachment of cells to acrylic and stainless steel. This study elucidates that commonly used intervention technologies including cold storage, thermal treatment, and alkaline antimicrobial agents might alter the physicochemical properties of S. Enteritidis cells and result in varied initial attachment levels., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This study evaluated the acid and heat resistance of Salmonella Enteritidis in simulated gastric fluid (pH 2.0) and during thermal treatment (54-60 °C), respectively, after adaptation to lactic acid (LA) or trisodium phosphate (TSP) at various pHs (pH 5.3-9.0). The changes in membrane lipid composition and expression levels of RpoS and RpoH were examined to elucidate their roles in bacterial stress resistance. Transcriptional profile of several virulence-related genes was also analyzed. Results showed that LA-adapted cells at pH 5.3 and 6.3 had higher acid and heat resistance than control cells and cells adapted to TSP at pH 8.3 and 9.0. LA-adapted cells had the lowest ratio of unsaturated to saturated fatty acids, indicating that they might possess a less fluid membrane. It was observed that the expression levels of RpoH and RpoS were upregulated in TSP-adapted cells but not in LA-adapted cells. Thus, these results indicate that the increased acid and heat resistance of LA-adapted S. Enteritidis was possibly due to the decreased membrane fluidity instead of the upregulation of RpoS and RpoH. About 6.0, 2.1, and 2.46-fold upregulation of spvR, avrA, and hilA were observed in cells adapted to TSP at pH 9.0, except sefA that had its highest expression level in the control cells, indicating that the expression of these virulence genes highly depends on environmental conditions. This is the first study to show that the alteration in the cytoplasmic membrane rather than RpoS and RpoH plays a more crucial role in conferring greater acid and heat resistance on LA-adapted S. Enteritidis, thus providing a better understanding on the bacterial stress response to acidic conditions., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Conventional culture detection methods are time consuming and labor-intensive. For this reason, an alternative rapid method combining real-time PCR and immunomagnetic separation (IMS) was investigated in this study to detect both healthy and heat-injured Salmonella Typhimurium on raw duck wings. Firstly, the IMS method was optimized by determining the capture efficiency of Dynabeads(®) on Salmonella cells on raw duck wings with different bead incubation (10, 30 and 60 min) and magnetic separation (3, 10 and 30 min) times. Secondly, three Taqman primer sets, Sal, invA and ttr, were evaluated to optimize the real-time PCR protocol by comparing five parameters: inclusivity, exclusivity, PCR efficiency, detection probability and limit of detection (LOD). Thirdly, the optimized real-time PCR, in combination with IMS (PCR-IMS) assay, was compared with a standard ISO and a real-time PCR (PCR) method by analyzing artificially inoculated raw duck wings with healthy and heat-injured Salmonella cells at 10(1) and 10(0) CFU/25 g. Finally, the optimized PCR-IMS assay was validated for Salmonella detection in naturally contaminated raw duck wing samples. Under optimal IMS conditions (30 min bead incubation and 3 min magnetic separation times), approximately 85 and 64% of S. Typhimurium cells were captured by Dynabeads® from pure culture and inoculated raw duck wings, respectively. Although Sal and ttr primers exhibited 100% inclusivity and exclusivity for 16 Salmonella spp. and 36 non-Salmonella strains, the Sal primer showed lower LOD (10(3) CFU/ml) and higher PCR efficiency (94.1%) than the invA and ttr primers. Moreover, for Sal and invA primers, 100% detection probability on raw duck wings suspension was observed at 10(3) and 10(4) CFU/ml with and without IMS, respectively. Thus, the Sal primer was chosen for further experiments. The optimized PCR-IMS method was significantly (P=0.0011) better at detecting healthy Salmonella cells after 7-h enrichment than traditional PCR method. However there was no significant difference between the two methods with longer enrichment time (14 h). The diagnostic accuracy of PCR-IMS was shown to be 98.3% through the validation study. These results indicate that the optimized PCR-IMS method in this study could provide a sensitive, specific and rapid detection method for Salmonella on raw duck wings, enabling 10-h detection. However, a longer enrichment time could be needed for resuscitation and reliable detection of heat-injured cells., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Unlabelled: The aim of this study was to determine the growth kinetics of Listeria monocytogenes, with and without cold-adaption, on fresh-cut cantaloupe under different storage temperatures. Fresh-cut samples, spot inoculated with a 4-strain cocktail of L. monocytogenes (∼3.2 log CFU/g), were exposed to constant storage temperatures held at 10, 15, 20, 25, or 30 °C. All growth curves of L. monocytogenes were fitted to the Baranyi, modified Gompertz, and Huang models. Regardless of conditions under which cells grew, the time needed to reach 5 log CFU/g decreased with the elevated storage temperature. Experimental results showed that there were no significant differences (P > 0.05) in the maximum growth rate k (log CFU/g h(-1) ) and lag phase duration λ (h) between the cultures of L. monocytogenes with or without previous cold-adaption treatments. No distinct difference was observed in the growth pattern among 3 primary models at various storage temperatures. The growth curves of secondary modeling were fitted on an Arrhenius-type model for describing the relationship between k and temperature of the L. monocytogenes on fresh-cut cantaloupe from 10 to 30 °C. The root mean square error values of secondary models for non- and cold-adapted cells were 0.018, 0.021, and 0.024, and 0.039, 0.026, and 0.017 at the modified Gompertz, Baranyi, and Huang model, respectively, indicating that these 3 models presented the good statistical fit. This study may provide valuable information to predict the growth of L. monocytogenes on fresh-cut cantaloupes at different storage conditions., Practical Application: Listeriosis has occurred and increased along with the increased demand of fresh and fresh-cut fruits and vegetables. This study was conducted to predict the growth of non- and cold-adapted L. monocytogenes on fresh-cut cantaloupe at different temperature using mathematical model. These results can be helpful for risk assessments of L. monocytogenes in fresh-cut cantaloupe. This study provides valuable information to food handlers to choose proper storage temperatures for extending the shelf-life of fresh-cut cantaloupe., (© 2014 Institute of Food Technologists®)

The occurrence of Listeria monocytogenes in ready-to-eat (RTE) fish products is well documented and represents an important food safety concern. Contamination of this pathogen in vacuum-packed (VP) smoked fish products at levels greater than the RTE food limit (100 CFU/g) has been traced to factors such as poor sanitary practices, contaminated processing environments, and temperature abuse during prolonged storage in retail outlets. Intervention technologies including physical, biological, and chemical techniques have been studied to control transmission of L. monocytogenes to these products. High-pressure processing, irradiation, and pulsed UV-light treatment have shown promising results. Potential antilisterial effects of some sanitizers and combined chemical preservatives have also been demonstrated. Moreover, the concept of biopreservation, use of bioactive packaging, and a combination of different intervention technologies, as in the hurdle concept, are also under consideration. In this review, the prevalence, routes of contamination, and potential intervention technologies to control transmission of L. monocytogenes in VP smoked fish products are discussed., (© 2014 Institute of Food Technologists®.)