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Salmon aquaculture is the fastest growing food production system in the world. Deficiencies in the quality or safety of salmon can have global repercussions. Controlling food safety aspects during production is therefore essential. Here, we investigate the state of hygiene in a salmon processing plant using next generation sequencing and classical culture-dependent methods to characterize the surface microbiota before and after cleaning and disinfection (C&D) at ten surface sampling points. Total aerobic counts revealed an average reduction in the bacterial loads of 1.1 log CFU/cm 2 by C&D. The highest relative abundance in the core microbiota before C&D was assigned to Acinetobacter, Mycoplasmataceae, Pseudomonas and Enterobacteriaceae in descending order. After C&D, we observed a significant increase in the relative abundance of Pseudomonas (p < 0.05). However, variations were found between conveyors, processing machines and drains. To assess the efficacy of commercial disinfectants, we performed susceptibility assays using advanced robotic high-throughput technologies and included foodborne bacteria which may affect food safety and spoilage. These included 128 Pseudomonas isolates, 46 Aeromonas isolates and 59 Enterobacterales isolates sampled from the salmon processing plant. Generally, minimum inhibitory concentrations (MICs) of the disinfectants were below the user concentration recommended by the producer for most isolates. BacTiter-Glo biofilm assays revealed that 30 min exposure to six out of eight commercial disinfectants resulted in an average reduction of relative luminescence >95 % in 59 single-species biofilms selected for screening. However, disinfection alone may not always be sufficient to eradicate biofilms completely. C&D routines must therefore be continuously assessed to maintain food safety and quality. The results from this study can contribute to understand and improve the state of hygiene in salmon processing environments., Competing Interests: Declaration of competing interest None to declare., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Sanitizers are widely incorporated in commercial apple dump tank systems to mitigate the cross-contamination of foodborne pathogens. This study validated the suitability of Enterococcus faecium NRRL B-2354 as a surrogate for Listeria monocytogenes during sanitizer interventions in dump tank water systems. E. faecium NRRL B-2354 inoculated on apples exhibited statistically equivalent susceptibility to L. monocytogenes when exposed to chlorine-based sanitizers (25-100 ppm free chlorine (FC)) and peroxyacetic acid (PAA, 20-80 ppm) in simulated dump tank water (SDTW) with 1000 ppm chemical oxygen demand (COD), resulting in 0.2-0.9 and 1.1-1.7 log CFU/apple reduction, respectively. Increasing the contact time did not affect sanitizer efficacies against E. faecium NRRL B-2354 and L. monocytogenes on apples. Chlorine and PAA interventions demonstrated statistically similar efficacies against both bacteria inoculated in SDTW. Chlorine at 25 and 100 ppm FC for 0.5-5 min contact yielded ~37.68-78.25 % and > 99.85 % inactivation, respectively, in water with 1000-4000 ppm COD, while ~51.55-99.86 % and > 99.97 % inactivation was observed for PAA at 20 and 80 ppm, respectively. No statistically significant difference was observed between the transference of E. faecium NRRL B-2354 and L. monocytogenes from inoculated apples to uninoculated apples and water, and from water to uninoculated apples during chlorine- or PAA-treated SDTW exposure. The data suggest E. faecium NRRL B-2354 is a viable surrogate for L. monocytogenes in dump tank washing systems, which could be used to predict the anti-Listeria efficacy of chlorine and PAA interventions during commercial apple processing. Further investigations are recommended to assess the suitability of E. faecium NRRL B-2354 as a surrogate for L. monocytogenes, when using different sanitizers and different types of produce to ensure reliable and comprehensive results., Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Vegetables are globally associated with a considerable number of foodborne outbreaks caused by viral infections, specifically human norovirus. In fresh produce industry, washing represents a critical step for food safety as process wash water (PWW) needs to be maintained at appropriate microbial quality to prevent water-mediated cross-contamination. This study aimed to explore the disinfection efficacy of chlorine (free chlorine, FC), chlorine dioxide (ClO 2 ) and peracetic acid (PAA) in PWW against infectious human norovirus and Tulane virus (TV). First, we tested the extent of TV inactivation in baby leaf, bell pepper, and vegetables mix PWW and monitored the viral decay by cell culture. Then, inactivation kinetics were defined for infectious human norovirus exposed to FC, ClO 2 and PAA in baby leaves PWW using the human intestinal enteroids (HIE) system. Finally, kinetic inactivation models were fitted to TV reduction and decay of sanitizers to aid the implementation of disinfection strategies. Results showed that >8 log 10 human norovirus and 3.9 log 10 TV were inactivated by 20 ppm FC within 1 min; and by 3 ppm ClO 2 in 1 min (TV) or 5 min (norovirus). PAA treatment at 80 ppm reduced ca. 2 log 10 TV but not completely inactivated the virus even after 20 min exposure, while 5 min treatment prevented norovirus replication in HIE. TV inactivation in PWWs was described using an exponential decay model. Taking these data together, we demonstrated the value of applying the HIE model to validate current operational limits for the most commonly used sanitizers. The inactivation kinetics for human norovirus and TV, along with the predictive model described in this study expand the current knowledge to implement post-harvest produce safety procedures in industry settings., 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 © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

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 [H 2 O 2 ], 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/cm 2 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 H 2 O 2  + P. These results suggested that combination of disinfectants (NaOCl, H 2 O 2 , 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., Competing Interests: Declaration of competing interest No potential conflict of interest was reported by the authors., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Salmonella has presented increasingly alarming rates of antimicrobial resistance believed to be a result of a high prevalence of integrons. It is speculated that disinfectant-resistant isolates are due to the expression of qacEΔ1, an efflux pump located in the 3' conserved sequence (3'CS) of class 1 integrons. With this concern, we tested the antibiotic and disinfectant resistance of 581 Salmonella strains collected from different sources, and characterized their integron structures. Gene expression and induction experiments were also performed. Results showed that Salmonella have high resistance to antimicrobials, especially to sulfonamides (SAs, 78.83 %), tetracyclines (TCs, 75.04 %) and benzalkonium chloride (BC, 87.26 %). The multi-drug resistance (MDR) frequency reached up to 63.17 %, and the prevalence of intI1 was 45.78 %. Molecular characterization of class 1 integrons exhibited nine different gene cassette arrays, of these, dfrA12-orf-aadA2 (n = 75), EstX (n = 25) and aadA2 (n = 14) were the most frequent. Importantly, 74.06 % of intI1-positive isolates were carrying qacEΔ1-sul1 genes in the 3'CS. This study also demonstrated that phenotypic resistance to both antibiotics and disinfectants was significantly correlated with the emergence of intI1 (p < 0.05). 91.37 % of qacEΔ1-sul1 positive Salmonella were found with disinfectant resistance. Additionally, expression of qacEΔ1 gene in Escherichia coli confirmed qacEΔ1 is predominantly involved in conferring disinfectant resistance. Disinfectant induction experiments further implicated qacEΔ1 in disinfectant resistance. RT-qPCR revealed a disinfectant-mediated increase in the relative expression of antibiotic-resistant genes (ARGs), aadA2 and dfrA12 on the integron, and efflux pump genes (mdtH and acrD) indicating that disinfectant could trigger co or cross-resistance. Therefore, our study confirmed that using disinfectant could provide selection pressure for strains with acquired resistance to antibiotics, providing new insights into the public health impact of Salmonella and guide continued efforts in antimicrobial stewardship and prevention of antibiotic resistance., 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 © 2023. Published by Elsevier B.V.)

The removal of C. difficile inoculated on fresh spinach leaves washed with antimicrobial solutions was investigated. In addition, the effect of washing solutions on the total aerobic mesophilic bacteria (TAMB) and Enterobacteriaceae in the fresh spinach was examined. The fresh spinach was washed through immersion in different concentrations (MIC, 2xMIC, and 4xMIC) of the natural disinfectant solution (NDS) consisting of EDTA, borax, and epigallocatechin gallate (EGCG) content developed in our laboratory and green tea extract-acetic acid (GTE-AA) for varying contact times (5 and 15 min). Different concentrations (50, 100, and 200 ppm) of sodium hypochlorite (NaOCl) and tap water as the control group were used to compare the effectiveness of the NDS. In addition, the effects of washing on the color, texture, and total phenol content of the spinach were determined. No statistical difference was observed in the washing of the spinach leaves with NDS prepared at 2xMIC and 4xMIC concentrations, while inhibition of C. difficile ranged between 2.11 and 2.32 logs. The highest inhibition was observed in the application of 50 ppm NaOCl for 15 min with a decrease of 2.88 logs in C. difficile spores. The GTE-AA and NDS decreased the number of TAMB by 2.27-3.08 log and, 3.21-3.66 log, respectively. Washing spinach leaves with natural disinfectant for 5 min caused a decrease of 2.58 logs in Enterobacteriaceae load, while washing with 50 ppm NaOCl for 15 min reduced Enterobacteriaceae load by 4 logs. Tap water was ineffective in reducing any microbial load. No difference was detected in the color parameters of the spinach through all washes. Although all antimicrobial washes made a difference in the texture of the spinach, the greatest loss in firmness was observed in the spinach washed with NaOCl. Washing spinach with epigallocatechin-based wash solutions can remove C. difficile in possible C. difficile contamination, thereby reducing the environmental load of C. difficile. Epigallocatechin-based disinfectants can be an alternative to chlorine-based disinfectants in improving the microbial quality of vegetables., Competing Interests: Declaration of competing interest There is no conflict of interest between the article's authors and other persons and/or institutions., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Fruits irrigated with contaminated water can transmit various pathogens. High sugar content in fruits such as black cherry (BC) fruit encourages microbial proliferation. A novel water-soluble decanoic acid (WSDA) was evaluated as a fruit sanitizer and compared with other traditional fruit sanitizers such as ethanol, bleach, or dishwasher surfactants. WSDA sanitizer killed yeasts, molds and bacteria including E. coli microbes effectively as other sanitizers with (4 log cycle reduction) of microbial load. Furthermore, the bacterial sanitization mechanism i.e. bactericidal or bacteriostatic was evaluated for alcohol, bleaching and WASDA solutions. E. coli was selected as the model pathogen used for such comparison. Results indicated that the mechanism of action for the three sanitizer solutions against E. coli was bactericidal. The problem with most used fruit sanitizers is their negative influence on fruit quality in terms of physical, mechanical and taste properties. In addition, some led to toxicological and ecological concerns. Thus, studies were conducted to explore the changes in the exocarp cell structure of BC fruit upon exposure to WSDA and other sanitizers using microscopic investigation. WSDA could have a very mild or gentle effect on the BC fruit cells compared to other sanitizers. Alcohol, bleaching and dishwasher surfactant changed the cellular structures and the intercellular spaces. Sanitizers may also affect fruit swelling. WSDA showed an increase in percent weight gain but it was significantly (p < 0.05) much lower than dishwasher surfactant and bleaching solution. BC Fruit flesh firmness and hardness were investigated upon exposure to different sanitizer solutions. BC fruit treated with WSDA showed the highest firmness values. Some liquid sanitizers could affect fruit quality in terms of fruit taste. Sensory evaluation in terms of the sanitizer's smell, texture and hedonic of BC fruit after soaking in different sanitizers was carried out. All sensory parameters of BC fruit soaked with WSDA were similar with insignificant differences (p > 0.05) compared to BC fruit soaked in tap water. However, the sensory parameters were significantly different (p < 0.05) when compared with alcohol, bleach and dishwasher surfactant. This ensures that WSDA was superior to other evaluated sanitizers in terms of physical, mechanical and fruit 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 B.V. All rights reserved.)

In the present study, bacterial-derived antimicrobial agents included 5 mM reuterin combined with either 103.91 mM lactic acid (RL) or 0.08 μM microcin J25 (RJ) were evaluated for their effects on the microbiota and sensory attributes of raw chicken legs. Peracetic acid (13.67 mM), a conventional chemical commonly used in the poultry industry, was used as a positive control to compare efficacy. The chicken legs were sprayed with antimicrobial solutions and aerobically stored at 4 °C for 10 days. The RL treatment maintained the total viable count below the limit of 7 log CFU/g until the 8th day. Therefore, compared to the nontreated group, shelf-life was extended by 3 days in the RL treated group. The RJ treatment extended the shelf-life to 7 days, which is similar to what was achieved with the use of peracetic acid. Based on culture-independent amplicon sequencing, the RL and RJ treatments affected the microbial community on the chicken legs, inducing a delay in the increase of Pseudomonas, Psychrobacter and Carnobacterium while decreasing of Shigella. Significant decreases in sensory scores occurred in the nontreated group, while slight changes occurred in the combinations treated groups over the same period. Overall, sensory property scores for chicken legs treated with RL and RJ remained higher (P < 0.05) than those treated with peracetic acid or without antimicrobial agents. The antimicrobial combinations delayed the deterioration of sensory attributes throughout the storage period. These results suggest that RL and RJ provide a promising natural-sourced antimicrobial approach to control the growth of spoilage microorganisms on chicken legs., 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 B.V. All rights reserved.)

Salmonella enterica is one of the leading causes of foodborne gastroenteritis worldwide. In the food production environment, many bacterial species co-exist on surfaces in biofilm structures, which can act as reservoirs of microbial contamination of food products. Polymicrobial biofilms have been shown to have greater tolerance to antimicrobials, such as disinfectants, however the mechanistic basis of this is poorly understood. In this study, S. enterica subsp. enterica serovar Liverpool was co-cultured in mixed-species biofilms with bacteria isolated from the food production environment and challenged with the cationic biocide disinfectant, benzalkonium chloride (BC). Co-culture with the common environmental bacterium Acinetobacter johnsonii resulted in >200-fold higher resistance of S. Liverpool to BC, compared to mono-culture biofilms. The transcriptional response of S. enterica to biofilm co-culture was determined using a dual RNA-seq strategy. Genes controlled by the PhoPQ and PmrAB two-component systems, involved in lipid A modification and associated with cationic antimicrobial peptide resistance (CAMP) of S. Liverpool, were significantly upregulated. Deletion of either the phoP or pmrA genes resulted in an increase in susceptibility to BC, suggesting that activation of their regulons during co-culture enhances BC resistance. S. Liverpool lipid A profiles changed significantly upon co-culturing, with greater incorporation of both phosphoethanolamine and palmitate, which was dependent upon activation of PhoPQ and PmrAB. We conclude that when grown in the presence of A. johnsonii, S. Liverpool increases its tolerance to cationic BC disinfection by remodelling its cell envelope including reducing the net negative charge of lipid A and increasing lipid A acyl density., Competing Interests: Declaration of competing interest None., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Biocidal compounds are frequently used as disinfectants in poultry industry and their widespread usage has risen concern due to the co-selection and persistence of antimicrobial resistance among bacteria. In this study, extended spectrum β-lactamase producing (ESBL) Escherichia coli isolates (n = 60) obtained from chicken meat were characterized by Pulsed Field Gel Electrophoresis (PFGE) and further tested for disinfectant and heavy metal resistance phenotypically and genotypically. Plasmid replicon types of these isolates were also determined. ESBL producing E. coli isolates were found to be resistant to ciprofloxacin (48.3 %) and gentamicin (15 %). The majority of these isolates (46.5 %) carried bla CTX-M-55 gene. The isolates showed higher minimal inhibitory concentrations to cetylpyridinium chloride (90 %), cetyltrimethylammonium bromide (50 %), hexadecyltrimethylammonium bromide (46.7 %), triclosan (38.3 %), benzalkonium chloride (28.3 %), chlorhexidine (21.7 %), acriflavine (3.3 %), benzethonium chloride (1.7 %) and N-alkyl dimethyl benzyl ammonium chloride (1.7 %), but 18.3 % of the isolates were resistant to triclosan. Of the quaternary ammonium compounds (QACs) tolerance genes, mdfA, sugE(c), ydgE and ydgF were most present in all isolates, but the qacE, qacG, oqxA and oqxB genes were not detected. Of genes mediating the heavy metal resistance, the zitB gene was detected in all isolates, whereas the copA and cueO genes were detected in 96.67 % and 95 % of isolates, respectively. The IncFIB plasmid was commonly present (93.3 %) in ESBL producing E. coli isolates. Consequently, given the detection of genes mediating disinfectant and heavy metal resistance commonly in ESBL producing E. coli isolates as well as high rate of MICs against disinfectant compounds, the use of QACs for decontamination of the facilities may not be as effective as expected in poultry sector in Turkey., (Copyright © 2022. Published by Elsevier B.V.)

Chlorine dioxide (ClO 2 ) is commonly generated by mixing sodium chlorite and acid. This study aimed to evaluate how acid affects the release kinetics and antimicrobial property of ClO 2 . Solutions made with weak acids released ClO 2 more slowly and had higher stability than those made with hydrochloric acid. Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes were treated with 1, 2.5, and 5 ppm ClO 2 for 3 or 5 min. Lettuce inoculated with the pathogenic bacteria were treated with 2.5 and 5 ppm ClO 2 for 5 min. The effects of peptone load at 0.01% and 0.02% on the antimicrobial efficacy of ClO 2 were investigated in S. Typhimurium cell suspensions. The contribution of acids alone at the pH of the ClO 2 solutions to bacterial reduction was also evaluated. The 2.5 ppm ClO 2 solutions made with citric acid, lactic acid, and malic acid showed higher reductions in all three bacteria than ClO 2 made with hydrochloric acid and sodium bisulfate. The 5 ppm ClO 2 solutions produced with organic acids reduced populations of all bacterial strains from 7 log CFU/mL to undetectable level in 3 min, except S. Typhimurium treated by ClO 2 produced with lactic acid. On inoculated Romaine lettuce model, 5 ppm ClO 2 produced with lactic acid and malic acid resulted in the highest reduction of E. coli O157:H7, S. Typhimurium, and L. monocytogenes of approximately 1.4, 1.7, and 2.4 log CFU/g, respectively. The antimicrobial efficacy of ClO 2 made with HCl and NaHSO 4 were affected by 0.01% and 0.02% peptone load, respectively. Food-grade organic acids produced aqueous ClO 2 solutions with stronger antimicrobial properties than inorganic acids. The acids alone at the pH of ClO 2 did not show significant bacterial reductions., (Copyright © 2022. Published by Elsevier B.V.)

A carrier (stainless steel disc as a default carrier) testing method is very needed for use in the actual food-processing fields by following the standard guideline. Here, we aimed to compare the virucidal efficacy of four commercial liquid disinfectants, including sodium hypochlorite (NaOCl), chlorine dioxide (ClO 2 ), and peracetic acid (PAA) against hepatitis A virus (HAV) following the OECD guideline protocol based on the quantitative carrier testing method and compared carrier testing results with the suspension testing results. The OECD method specifies a test for establishing whether a chemical disinfectant or a microbicide has a virucidal activity on hard non-porous surfaces. The antiviral efficacy was evaluated by plaque assays, and disinfectants were considered effective if the virus reduction was greater than or equal to 3 log 10 (99.9% decrease) for carrier or 4 log 10 (99.99% decrease) for suspension tests. Results indicated that ClO 2 above 500 ppm and 50% ethanol were effective in the carrier test method. In contrast, more than 200 ppm NaOCl and 50 ppm ClO 2 for all exposure times and 70% ethanol with contact for more than 5 min were effective in suspension tests. Treatment with PAA (80-2500 ppm) were not effective in carrier or suspension tests. Therefore, we recommend the use of more than 500 ppm ClO 2 or 50% ethanol with exposure for 10 min to disinfect surfaces that may be contaminated with HAV. Thus, these results could be effective in establishing official antiviral efficacy testing methods and basic data., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Biofilm formation in food processing plants reduces the efficacy of sanitation. The presence of transmissible locus of stress tolerance (tLST) also enhances resistance of planktonic cells of Escherichia coli to sanitation chemicals but the role of tLST in resistance of biofilm-embedded cells remains unclear. This study investigated the link of tLST to biofilm formation and its contribution to resistance of biofilm-embedded E. coli to sanitation. Biofilms were formed as single-strain and as dual-strain biofilms in association with E. coli, Aeromonas australensis or Carnobacterium maltaromaticum. Biofilms on stainless steel were compared to floating biofilms formed at the air-liquid interface (pellicles). The resistance of biofilm-embedded tLST positive strains of E. coli to chlorine, hydrogen peroxide, and peroxyacetic acid was higher than the resistance of tLST negative strains. Higher biofilm density as measured by crystal violet staining was observed in tLST-positive strains of E. coli when compared to tLST negative strains. Biofilm density positively correlated to resistance to disinfectants. The use of confocal laser scanning microscopy detected more compact structure of pellicles compared to solid surface-attached biofilms, resulting in higher chlorine resistance despite the absence of tLST in strains of E. coli. Collectively, the findings of this study elucidated the impact of tLST in strains of E. coli on biofilm formation and sanitizer resistance. These findings may inform the development of improved sanitization protocols for food facilities., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Inactivation rate constant or inactivation coefficient (specific lethality) quantifies the rate at which a chemical sanitizer inactivates a microorganism. This study presents a modified disinfection kinetics model to evaluate the potential effect of organic content on the chlorine inactivation coefficient of Escherichia coli O157:H7 in fresh produce wash processes. Results show a significant decrease in the bactericidal efficacy of free chlorine (FC) in the presence of organic load compared to its absence. While the chlorine inactivation coefficient of Escherichia coli O157:H7 is 70.39 ± 3.19 L/mg/min in the absence of organic content, it drops by 73% for a chemical oxygen demand (COD) level of 600-800 mg/L. Results also indicate that the initial chlorine concentration and bacterial load have no effect on the chlorine inactivation coefficient. A second-order chemical reaction model for FC decay, which utilizes a proportion of COD as an indicator of organic content in fresh produce wash was employed, yielding an apparent reaction rate of (9.45 ± 0.22) × 10 -4 /μM/min. This model was validated by predicting FC concentration in multi-run continuous wash cycles with periodic replenishment of chlorine., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The ability of Listeria monocytogenes isolates to survive within the food production environment (FPE), as well as virulence, varies greatly between strains. There are specific genetic determinants that have been identified which can strongly influence a strains ability to survive in the FPE and/or within human hosts. In this study, we assessed the FPE fitness and virulence potential, including efficacy of selected hygiene or treatment intervention, against 52 L. monocytogenes strains isolated from various food and food environment sources. Phenotypic tests were performed to determine the minimum inhibitory concentration of cadmium chloride and benzalkonium chloride and the sensitivities to five clinically relevant antibiotics. A genomic analysis was also performed to identify resistance genes correlating to the observed phenotypic resistance profiles, along with genetic determinants of interest which may elude to the FPE fitness and virulence potential. A transposon element containing a novel cadmium resistance gene, cadA7, a Tn916 variant insert in the hypervariable Listeria genomic island 1 region and an LGI2 variant were identified. Resistance to cadmium and disinfectants was prevalent among isolates in this study, although no resistance to clinically important antimicrobials was observed. Potential hypervirulent strains containing full length inlA, LIPI-1 and LIPI-3 were also identified in this study. Cumulatively, the results of this study show a vast array of FPE survival and pathogenicity potential among food production-associated isolates, which may be of concern for food processing operators and clinicians regarding L. monocytogenes strains colonising and persisting within the FPE, and subsequently contaminating food products then causing disease in at risk population groups., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The foodborne pathogen L. monocytogenes can be present in food processing environments where it is exposed to various stressors. These antimicrobial factors, which aim to eliminate the pathogen, can induce sub-lethal injury to the bacterial cells. In the present study, we investigated the efficacy of different treatments (stresses) relevant to food processing and preservation as well as sanitation methods, in generating sub-lethal injury at 4 °C and 20 °C to two L. monocytogenes strains, ScottA and EGDe. Additionally, we evaluated the survival and extent of L. monocytogenes injury after exposure to commonly used disinfectants (peracetic acid and benzalkonium chloride), following habituation in nutrient-deprived, high-salinity medium. Each stress had a different impact on the survival and injury kinetics of L. monocytogenes. The highest injury levels were caused by peracetic acid which, at 4 °C, generated high populations of injured cells without loss of viability. Other injury-inducing stresses were lactic acid and heating. Long-term habituation in nutrient-limited and high salinity medium (4 °C) and subsequent exposure to disinfectants resulted in higher survival and injury in benzalkonium chloride and increased survival, yet with lower injury levels, in peracetic acid at 20 °C. Taken together, these results highlight the potential food safety risk emerging from the occurrence of injured cells by commonly used food processing methods. Consequently, in order to accurately assess the impact of an antimicrobial method, its potential of inducing sublethal injury needs to be considered along with lethality., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Glycerol monolaurate (GML) is a monoglycerol ester of the fatty lauric acids, which has a wide-spectrum antimicrobial capacity, but fails to inactivate Gram-negative bacteria, especial Salmonella. To enhance the population reduction rate of GML for Salmonella, this reagent was combined with three disinfectants: lactic acid (LA), cetylpyridinium chloride (CPC), and trisodium phosphate (TSP), which can present acid, neutral, and alkaline in solution, respectively. The results showed that the 1% GML and a complex disinfectant (0.5% GML-0.025% LA) could powerfully inactivate Salmonella. Their population reduction rates respectively were able to achieve 99.92% and 98.29% with the vortex treatment, indicating that the vortex treatment could improve GML to destruct the outer membrane of Salmonella. During the simulation test of the soaking and rinse processing of chicken, for a short time (0 h), the effect of 0.5% GML-0.025% LA compound was better and more suitable for instantaneous inactivation than others, while for a long time (4 h), 1% GML exhibited a better bactericidal effect, which indicated it to be more suitable for long-term bacteriostasis. The characterization of color and texture for chicken samples were determined using Colormeter Ci7600, TA.XT Plus and Hyper-spectral Imager, which demonstrated that all samples treated by these complex disinfectants were not significantly different from untreated group. In conclusion, GML is a potential and superior disinfectant for the chicken process., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Plasma-activated water (PAW) has good liquidity and uniformity and may be a promising candidate to inactivate Penicillium italicum and maintain the quality attributes of kumquat. In this study, the effect of plasma-activated water (PAW) on the viability of Penicillium italicum on kumquat and quality attributes of PAW-treated kumquats were then systematically investigated to elucidate the correlation between PAW and kumquat quality attributes. The effects of PAW on fruit decay, microbial loads, and firmness of postharvest kumquats during the 6-week storage were also investigated. The results showed that the viability of Penicillium italicum was notably inhibited by PAW on kumquats. Moreover, PAW did not significantly change the surface color of kumquats. No significant reductions in ascorbic acid, total flavonoid, and carotenoids were observed in kumquats after the PAW treatment. Results from nitrate and nitrite residue analyses showed that PAW did not leave serious nitrate and nitrite residues after treatment. The decay analysis results demonstrated that PAW has the potential to control kumquat decay and fungal contamination as well as maintain the firmness of postharvest kumquats throughout 6-week storage. Transmit electron microscope observation confirmed that PAW could cause the surface sculpturing in the skin cell wall of kumquat. The information obtained from this research may provide insight into the utilization of PAW to fight against fungal infection during the storage of citrus fruit., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The importance of leaf integrity, i.e. the effects of bruising (mechanical damage), and sanitisation with peroxyacetic acid (PAA) on bacterial communities of ready-to-eat baby spinach remains unclear. Two shelf-life studies were conducted at 4 °C to investigate the effect of bruising and sanitisation on the growth of spoilage microorganisms. In the first experiment, both bruising treatments (100% and 40% of leaves) halved shelf life to 12 d, whereas intact leaves had a shelf-life of 23 d. Bruising had no influence on bacterial diversity during shelf-life, though some differences in the relative abundance of minor genera were observed. Pseudomonas and Pantoea were the most dominant bacterial genera, regardless of bruising treatment. High throughput amplicon sequencing also identified other spoilage bacteria including Chryseobacterium, Stenotrophomonas, Bacillus, Sphingobacterium, Erwinia and Flavobacterium. In the second experiment, washing of intact baby spinach with a sanitiser (80 mg/L: PAA) reduced microbial load as determined by aerobic plate count but did not immediately affect the presence/relative abundance of most of the genera of spoilage bacteria observed. During shelf-life, the bacterial diversity of sanitised leaves was significantly lower than on water-washed leaves. Although sanitisation resulted in a higher initial log reduction in microbial load compared to control (portable tap water), sanitisation did not extend the shelf life of baby spinach (23 d). Sanitised spinach had reduced bacterial diversity however, by the end of shelf life, both sanitised and water-washed spinach was dominated by Pseudomonas and Pantoea spoilage bacteria. This study demonstrated for the first time that the shorter shelf life of bruised leaves was related to faster microbial growth rather than changes in bacterial diversity or community composition., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Clam jeotgal, called "jogaejeotgal," is a Korean fermented seafood product with, generally, a high amount of added salt to inhibit the growth of pathogenic microorganisms. This study aimed to evaluate the efficacy of chlorine dioxide (ClO 2 ) and sodium hypochlorite (NaOCl) against murine norovirus 1 (MNV-1), a surrogate for human norovirus, in salt-fermented clam, jogaejeotgal. The sequential effect of ClO 2 and electron-beam (e-beam) irradiation on the inactivation of MNV-1 was also investigated. Treatments of up to 300 ppm ClO 2 and 1000 ppm NaOCl were used to determine the disinfectant concentrations at which more than 1 log (90%) MNV-1 inactivation occurred. The sequential treatment of ClO 2 (50-300 ppm) and e-beam (1-5.5 kGy) was performed after storage at 4 °C for 7 days. There was a 1.9-log reduction of the virus in seasoned clams irradiated at 5.5 kGy after ClO 2 treatment at 300 ppm. No significant change (p > 0.05) in physicochemical quality was observed after the combined treatment, suggesting the potential for the use of a combined treatment using ClO 2 (300 ppm) and e-beam (5.5 kGy) in the jeotgal manufacturing industry for the reduction of norovirus., (Copyright © 2021 Elsevier B.V. All rights reserved.)

A packaging system using gaseous chlorine dioxide generating film (CDGF) in a sealed container was developed to extend the shelf life of semi-dry longan pulp (moisture content 38.8 wt%; a w 0.8). The antimicrobial properties, formation of chloroxyanion residues and effects of CDGF on the quality of semi-dry longan pulp were investigated. CDGF was triggered by the moisture vapor from semi-dry longan pulp in the sealed container and released gaseous ClO 2 into the headspace of the container. The antifungal test showed that CDGF significantly inactivated artificially inoculated molds in semi-dry longan pulp and achieved reductions of over 3 log CFU/g after 28 days storage at room temperature (25 °C). CDGF reduced total aerobic bacterial populations by over 6.4 log CFU/g and maintained these population levels at around 2.0 log CFU/g throughout the 180-day storage period at room temperature. The residual concentrations of chloride, chlorate and perchlorate in longan pulp increased and then decreased during the 180-day storage. Residual chloride levels were maintained at 1.5 mg/g after Day 120 and residual chlorate and perchlorate levels were not detected after Day 120 and Day 180, respectively, in CDGF-treated samples. CDGF treatments reduced total polyphenol content but didn't have any significant impact on the levels of polysaccharides in samples. There were no significant differences between CDGF-treated and control samples in color changes during storage. The content of 5-hydroymethylfurfural (5-HMF) in both samples increased during storage, suggesting that the Maillard reaction occurred. This study demonstrated an effective approach to develop a new antimicrobial packaging system for semi-dry longan pulp., (Copyright © 2020 Elsevier B.V. All rights reserved.)

This study aimed to evaluate the sensitivity of heat-resistant molds isolated from spoiled thermally processed foods to antimicrobial compounds used for food industry sanitation. An ortho-phenylphenol-based smoke generator sanitizer, liquid chemical sanitizers (benzalkonium chloride, biguanide, iodine, peracetic acid, and sodium hypochlorite), and acidic and alkaline electrolyzed water were used against Aspergillus australensis (MB 2579; NFF 02), Aspergillus aureoluteus (NFC1), Paecilomyces fulvus (PFF 01), Paecilomyces niveus (PNT 01; PNDC 01; PNB1 01), and Paecilomyces variotii (PV 01; PV 01; PVCH 03). The fungal strains were exposed separately to liquid sanitizers and electrolyzed water in stainless steel discs for 15 min following the European Committee for Standardization (CEN) recommendations. Moreover, the fungal strains were exposed to the smoke generator sanitizer for 7 h following French protocol NF-T-72281. The best results of fungal inactivation were achieved when the highest concentration specified in the label of these sanitizers was tested. On the opposite, the lowest concentration specified in the label should be avoided since it was ineffective in most cases (94%). The ortho-phenyphenol-based smoke generator sanitizer and peracetic acid (1%) showed the best results of spore inactivation, while iodine and benzalkonium chloride achieved satisfactory results against the strains evaluated. Sodium hypochlorite and biguanide were ineffective against most of the fungi studied at all concentrations tested. Acidic and basic electrolyzed water was also ineffective to achieve the 3-log CFU reduction required in the concentrations tested. In general, Paecilomyces spp. was more sensitive than Aspergillus spp. against all sanitizers evaluated, whereas A. aureoluteus NFC1 was resistant to all agents and concentrations tested. The heat-resistant fungal strains showed varied sensitivity against the different agents. Notably, the two most effective commercial sanitizers against the heat-resistant strains were ineffective against the filamentous fungi recommended for sanitizer testing (A. brasiliensis ATCC 16404), which demonstrates the relevance of testing fungal isolates that cause spoilage to choose the most effective compound and obtain the best results of fungal control., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Disinfection of fruits is one of the most important steps since they are going to be eaten fresh-or minimally-processed. This step affects quality, safety, and shelf-life of the product. Despite being a common sanitizer in the fruit industry, chlorine may react with organic matter leading to the formation of toxic by-products. Alternative sustainable disinfection strategies to chlorine are under study to minimize environmental and human health impact. Water-assisted UV-C light (WUV-C) is proposed here as an alternative sanitizing method for strawberries. In this study, strawberries were washed for 1 or 5 min in a tank with 2 or 4 lamps on, each emitting UV-C light at 17.2 W/cm 2 , or in a chlorine solution (200 ppm, pH 6.5). Moreover, trials with 4 lamps on, together with a washing solution consisting on peracetic acid at 40 or 80 ppm, were carried out. Overall, quality and nutritional parameters of strawberries after treatments were maintained. Changes in color were not noticeable and fruits did not lose firmness. No major changes were observed in antioxidant activity, organic acid, anthocyanin, vitamin C, and total phenolic content. Yeasts and molds were not affected by the WUV-C treatment, and 5 min were needed to significantly reduce total aerobic mesophylls population. However, reductions of artificially inoculated Listeria innocua and Salmonella Typhimurium after WUV-C treatments were comparable to those obtained with chlorine-wash, which were 3.0 log CFU / g. Moreover, WUV-C light was effective to minimize microorganisms remaining in washing water, avoiding cross-contamination and thus, allowing water recirculation. This effect was improved when combining the action of UV-C light with peracetic acid, showing the suitability of this combined treatment, understood as an alternative to chlorine sanitation, for sanitizing strawberries and keeping the populations of pathogenic bacteria in washing water lower than 0.6 ± 0.1 log CFU / mL., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Strawberries are often consumed fresh or only receive minimal processing, inducing a significant health risk to the consumer if contamination occurs anywhere from farm to fork. Outbreaks of foodborne illness associated with strawberries often involve a broad range of microbiological agents, from viruses (human norovirus) to bacteria (Salmonella spp. and Listeria monocytogenes). The addition of sanitizers to water washes is one of the most commonly studied strategies to remove or inactivate pathogens on berries as well as avoid cross contamination due to reuse of process wash water. The risk posed with the safety issues of by-products from chlorine disinfection in the fruit industry has led to a search for alternative sanitizers. We evaluated the applicability of different chemical sanitizers (peracetic acid (PA), hydrogen peroxide (H 2 O 2 ), citric acid (CA), lactic acid (LA) and acetic acid (AA)) for the inactivation of S. enterica, L. monocytogenes and murine norovirus (MNV-1) on strawberries. A control treatment with chlorine (NaClO) (100 ppm) was included. For each sanitizer, different doses (40, 80 and 120 ppm for PA and 1, 2.5 and 5% for H 2 O 2 , LA, AA and CA) and time (2 and 5 min) were studied in order to optimize the decontamination washing step. The best concentrations were 80 ppm for PA, 5% for H 2 O 2 and 2.5% for organic acids (LA, AA and CA) after 2 min treatment. Results indicate that the sanitizers selected may be a feasible alternative to chlorine (100 ppm) for removing selected pathogenic microorganisms (P > 0.05), with reductions about ≥2 log for bacterial strains and ≥ 1.7 log for MNV-1. As the washing water may also increase the microbial counts by cross-contamination, we observed that no pathogenic bacteria were found in wash water after 5% H 2 O 2 and 80 ppm PA after 2 min treatment. On the other hand, we also reported reductions about total aerobic mesophyll (TAM) (0.0-1.4 log CFU/g) and molds and yeasts (M&Y) (0.3-1.8 log CFU/g) with all alternative sanitizers tested. Strawberries treated did not shown significant differences about physio-chemical parameters compared to the untreated samples (initial). For this study, the optimal sanitizer selected was PA, due to the low concentration and cost needed and its microbiocidal effect in wash water and fruit. Notwithstanding the results obtained, the effect of PA in combination with other non-thermal technologies such as water-assisted ultraviolet (UV-C) light should be studied in future research to improve the disinfection of strawberries., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Two decontamination methods were evaluated for inactivating a cocktail of Salmonella or Listeria monocytogenes inoculated onto model low moisture foods (LMFs; dried strawberry, dried apple, raisins, chocolate crumb, cornflakes, shell-on or deshelled pistachio nuts). One treatment was based on a peracetic acid-ethanol (PAA-ethanol) sanitizer combination with the other being an Advanced Oxidation Process (AOP) that simultaneously applied UV-C (254 nm), ozone and hydrogen peroxide. The low moisture food was spray inoculated then dried prior to treatment. With Salmonella it was found that a pre-incubation step in 1% w/v glycerol-tryptic soy broth for 1 h prior to plating, significantly increased recovery of the pathogen compared to TSB alone. However, no increased recovery of L. monocytogenes was observed using the TSB-glycerol pre-incubation step. No Salmonella was detected on cornflakes, chocolate crumb and strawberry using 1.25 parts per thousand (‰) PAA-ethanol. The inactivation of Salmonella on deshelled pistachio was significantly higher using 2.5‰ PAA-ethanol sanitizer compared to the AOP treatments tested. Only negligible reductions of Salmonella (<1 log cfu) were obtained with shell-on pistachio treated with PAA-ethanol sanitizer or AOP. Salmonella could be reduced on dried apple slices by >4 log CFU when 5.0‰ PAA-ethanol was applied. L. monocytogenes was more sensitive to PAA-ethanol compared to Salmonella and could be eliminated on all the LMFs apart from shell-on pistachio. An AOP treatment applied 10% v/v hydrogen peroxide, ozone and 54 mJ/cm 2 UV-C could significantly reduce Salmonella on dried apple slices compared to when the individual elements (hydrogen peroxide, ozone or UV-C) were applied. Salmonella was also eliminated by AOP on the other LMFs (apart from shell-on pistachio) although the same level of inactivation was achieved by spraying with 10% v/v hydrogen peroxide alone. L. monocytogenes was sensitive to hydrogen peroxide and AOP being eliminated from all the LMFs. Although this may suggest that hydrogen peroxide spray was equivalent to AOP treatment it was noted that no residual H 2 O 2 or changes in visual appearance was evident on samples treated with the latter process. The study has demonstrated that the two decontamination methods assessed can be applied to reduce Salmonella and L. monocytogenes on LMFs although efficacy is dependent on the pathogen and product type., Competing Interests: Declaration of competing interest The authors declare that the submitted work was conducted in the absence of any commercial or financial relationships that could be construed as a conflict of interest., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

The aim of this study was to assess the impact of the physiological state and intraspecific variability on the efficacy of 70% ethanol to inactivate conidia of Penicillium commune, used as a representative species of dairy product contaminants. Four physiological states were obtained by modifying the water activity during the production of conidia (0.995 and 0.950) and the harvesting conditions (hydrated and non-hydrated). These conditions were applied to four different P. commune strains isolated from contaminated dairy products. Five minutes exposure to 70% ethanol at ambient temperature allowed total inactivation of conidia (>4 log 10 ) regardless of the physiological state or the strain. For 1 min exposure, regardless of the strains, only dry-harvested conidia produced at a w 0.950 exhibited survivors. Survival after 2 min exposure was observed for this physiological state for P. commune UBOCC-A-116003 only. For this strain, the impact of the physiological state was greater than 1.54 log 10 between dry-harvested conidia produced at a w 0.950 that exhibited survivors after 1 min treatment and the 3 other kinds of conidia that were all inactivated. For 1 min exposure, by comparing the more resistant strain to the three other strains, the impact of the intraspecific variability was 2.35 log 10 . These results demonstrated that the physiological state of the conidia, the representativeness of the tested species and strains should be taken into account to assess the efficacy of disinfectants in dairies., 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 © 2020 Elsevier B.V. All rights reserved.)

Salmonella (S.) Infantis is currently the most common serovar in broilers and boiler meat in the European Union. In the field, eradication of S. Infantis in affected poultry flocks is considered extremely difficult. Despite stringent cleaning and disinfection measures between the placement of flocks, recurrent infections are often reported. So far, the efficacy of disinfectants on S. Infantis has rarely been studied. Therefore, in the present in-vitro study the bacteriostatic and bactericidal efficacy of ten commercial disinfectants were tested against seven S. Infantis field isolates. Combinations of aldehyde and quarternary ammonium were the active compounds of five, peroxygen of three, cresol and alkylamines of one disinfectant, respectively. Investigations were performed according to standard protocols and regulations. Different concentrations of disinfectants were used to test the bacteriostatic efficacy. Different temperatures and low and high protein exposures were applied as variables to investigate the bactericidal efficacy. Following neutralization of the disinfectants an additional incubation step was introduced to investigate the revitalisation potential of S. Infantis. The bacteriostatic efficacy could be assessed for seven disinfectants. For three disinfectants a bacteriostatic effect was observed when the recommended concentration was used, whereas with four disinfectants only increased concentrations led to this effect. The bactericidal efficacy was not influenced by temperature, whereas high protein exposure decreased the efficacy of nine disinfectants. Furthermore, reactivation of S. Infantis was revealed after application of disinfectants for the majority of products. Interestingly, the strain of S. Infantis influenced the efficacy of the disinfectants. Overall, products based on aldehydes and quarternary ammonium compounds proved most efficient, followed by peroxgen, cresol and alkylamines., Competing Interests: Declaration of competing interest The authors declare that the present work was undertaken in the absence of any commercial or financial relationships that could be construed as conflict of interest.

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.)

Aims: The aim of this study was to evaluate the performance of sampling methods [contact plates, sponges, and swabs] in the recovery of biofilm Listeria monocytogenes populations considering the seafood environment conditions (nature of conditioning, of materials and bacterial species)., Methods and Results: Different materials (stainless steel, polyvinyl chloride, polyurethane) were conditioned with two fish filtrates, the ready-to-eat the most consumed in Europe (smoked salmon, cod). After, we added the suspension of Listeria monocytogenes, alone or with Pseudomonas fluorescens or Carnobacterium strains, and incubated for 48 h at 8 °C. Then, the 48 h-biofilms were sampled with different methods (contact plates, sponges, and swabs). The cultivable bacterial populations were enumerated on agar, while the L. monocytogenes total and viable populations were quantified by qPCR and propidium monoazide-qPCR (PMA-qPCR), respectively. The amount of L. monocytogenes in biofilms was affected only by the nature of the conditioning with lowest adherent bacteria with cod versus with smoked salmon conditioning. Considering the amount of total population, the swab displayed the lowest values versus the sponges and the contact plates. An explanation was that the observations of the swab by microscopy showed the bacteria trapped within it. The recovery of cultivable bacterial populations was not significantly different with the three sampling methods. On the contrary, we showed that the VBNC populations were only detached by two of three methods (contact plates, sponges) while for the dead populations, those were contact plates and swabs., Conclusions: The nature of the conditioning influenced the amount of the bacteria in biofilms. And the performance of the recovery of the bacterial populations (dead, VBNC, cultivable) was dependent on the methods used., Significance and Impact of the Study: This study showed that the seafood environmental conditions influenced the biofilm formation and the assessment of the efficiency of cleaning and disinfectant operations could be significantly affected by the used sampling methods., Competing Interests: Declaration of competing interest The authors have no conflict of interest to declare., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Fresh produce shall be thoroughly washed at the retail level prior to serving to the consumer with potable water. Foodborne pathogens if present on a product may transfer to the wash water and cross-contaminate other products immersed in the water. Typically, an entire carton of lettuce (24 heads) is washed together increasing the likelihood of cross-contamination due to the close contact between each head. This study aimed to compare the effects of two wash batch volumes - "low" (8 heads) and "high" (24 heads) on the efficacy of two commercial water antimicrobials and cross-contamination. Red leaf lettuce was spot-inoculated (~5.0 log CFU/g) with Shiga toxin-producing Escherichia coli (STEC) and Listeria monocytogenes. In the first batch of washing, inoculated lettuce was washed with non-inoculated lettuce, followed by reuse of the water/antimicrobials twice washing only non-inoculated lettuce. Samples of inoculated and non-inoculated lettuce were collected to determine aerobic plate count (APC) as well as the populations of STEC and L. monocytogenes. Microbiological analysis of the wash water was also conducted. Wash volume (8 versus 24 lettuce heads) had limited effects on the antimicrobial activities of the interventions evaluated. Instead, high-volume wash increased the rate of cross-contamination between non-inoculated and inoculated lettuce, and cross-contamination of non-inoculated lettuce through wash water reuse. Retailers should consider volume of product processed per batch, reuse of wash water, and use of an appropriate water antimicrobial in consideration of mitigating potential product cross-contamination., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The potential of using ultraviolet light (UV) in combination of peracetic acid (PAA) as an alternative to chlorine washing for lettuce was evaluated. Shredded iceberg lettuce was dip-inoculated with a four-strain Salmonella cocktail to final levels of 6-7.5 log CFU/g, following by air-drying and overnight cold storage. The inoculated lettuce (80 g) was then washed in turbid tap water containing 6% lettuce juice extract and silicon dioxide (turbidity of ~60 NTU; COD of ~2000 mg/L) while being treated with 1) 10 or 20 ppm free chlorine, 2) PAA solution (40 and 80 ppm), 3) UV (10, 20 and 30 mW/cm 2 ), 4) a combination of UV and PAA for 1, 2, and 5 min. Among all the single treatments, the 30 mW/cm 2 UV treatment achieved the highest Salmonella reduction on lettuce. For the 2-min treatment group, the 30 mW/cm 2 UV treatment achieved 1.98 log reduction, while the 80 ppm PAA and 20 ppm free chlorine resulted in 1.52 and 1.23 log reduction, respectively. The combined treatment of 30 mW/cm 2 UV and 80 ppm PAA achieved significantly higher (P < .05) Salmonella reduction than the 20 ppm free chlorine washing. For the 5-min treatment group, the combined treatment resulted in 3.24 log reduction, while the 20 ppm free chlorine washing only achieved 1.24 log reduction. The effect of the combined treatment of 30 mW/cm 2 UV and 80 ppm PAA was also compared with 20 ppm free chlorine washing on larger sample sizes of 200, 500, and 1000 g lettuce. The increase of sample size from 80 g to 1000 g did not significantly (P < .05) affect the inactivation of Salmonella on lettuce for the combined treatment. In addition, the combined treatment of 80 ppm PAA and 30 mW/cm 2 UV was able to maintain the Salmonella population in wash water under the detection limit of 0.3 log CFU/mL. It was therefore concluded that the combined treatment of 30 mW/cm 2 UV and 80 ppm PAA could be used as an alternative to chlorine washing for lettuce decontamination., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Fresh leafy greens like lettuce can be consumed raw and are susceptible to foodborne pathogens if they become contaminated. Recently, the number of reported pathogenic foodborne outbreaks related to leafy greens has increased. Therefore, it is important to try to alleviate the human health burden associated with these outbreaks. Processing of fresh-cut lettuce, including washing, is a step in the supply chain that needs to be well controlled to avoid cross-contamination. Current measures to control the quality of lettuce during washing include the use of chemicals like chlorine; however, questions regarding the safety of chlorine have prompted research for alternative solutions with peracetic acid (PAA). This study evaluates the effectiveness of a PAA (c.a. 75 mg/L) solution on the reduction of a commensal E. coli strain during the washing of fresh-cut lettuce. Experiments were performed at the laboratory scale and validated at the industrial scale. We observed that the use of PAA was not adversely affected by the organic load in the water. The contact time and dose of the PAA showed to be relevant factors, as observed by the approximately 5-log reduction of E. coli in the water. Results showed that once introduced during washing, E. coli remained attached to the lettuce, thus supporting the need to control for pathogenic bacteria earlier in the supply chain (e.g., during primary production) as well as during washing. Moreover, our results showed that the use of PAA during washing did not have an apparent effect on the levels of fluorescent pseudomonads (FP) and total heterotrophic bacteria (THB) in lettuce. Overall, our results at the laboratory and industrial scales confirmed that during the processing of fresh-cut produce, where the accumulation of soil, debris, and other plant exudates can negatively affect washing, the use of a PAA (c.a. 75 mg/L) solution was an effective and safe wash water disinfectant that can potentially be used at the industrial scale., (Copyright © 2020. Published by Elsevier B.V.)

Fresh produce, as a known or suspected source of multiple foodborne outbreaks, harbors large populations of diverse microorganisms, which are partially released into wash water during processing. However, the dynamics of bacterial communities in wash water during produce processing is poorly understood. In this study, we investigated the effect of chlorine (FC) and peracetic acid (PAA) on the microbiome dynamics in spinach and romaine lettuce rinse water. Treatments with increasing concentrations of sanitizers resulted in convergence of distinct microbiomes. The resultant sanitizer resistant microbiome showed dominant presence by Bacillus sp., Arthrobacter psychrolactophilus, Cupriavidus sp., and Ralstonia sp. Most of the FC and PAA resistant bacteria isolated from spinach and lettuce rinse water after sanitation were gram positive spore forming species including Bacillus, Paenibacillus, and Brevibacillus spp., while several PAA resistant Pseudomonas spp. were also isolated from lettuce rinse water. Inoculation of foodborne pathogens altered the microbiome shift in spinach rinse water under PAA treatment, but not in lettuce rinse water or FC treated samples. These inoculated foodborne pathogens were not isolated among the sanitizer resistant strains., 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., (Published by Elsevier B.V.)

The aim of this study was to compare the sporicidal effect of the disinfectants peracetic acid (PAA) or hydrogen peroxide (H 2 O 2 ) applied as a fog or as a liquid. The efficacy of fogging of the disinfectants was tested in a closed isolator cabinet using highly heat and chemical-resistant spores of Geobacillus stearothermophilus. Fogging of a 0.06% solution of PAA resulted in over 5-log reduction of spores in 10 min, whereas for PAA used in liquid form the same reduction was achieved in 4.5 min. The inactivation curves for fog and liquid were fitted using three different models (Linear with shoulder, Weibull, Gauss-Eyring). This showed a shoulder for the fog with an estimated length of 4.1 min, but the D values, calculated for the linear parts of the curves, were not significantly different (1.1 and 0.8 min for the PAA fog and solution, respectively). Similar results were obtained for a 12% H 2 O 2 solution, albeit that H 2 O 2 was less effective compared to PAA, requiring 60 min to reach 3-log reduction when applied as a fog, with an estimated shoulder of 18.5 min. Fogging of a 0.06% peracetic acid solution effectively inactivated G. stearothermophilus spores. Overall, the data show that fogging can be an effective method of applying disinfectants but that a shoulder in the inactivation curves should be considered in process design. This study provides inactivation kinetics for disinfection using PAA or H 2 O 2 -based fog, which can aid in selection and validation of process parameters for disinfection of contained areas by fogging., Competing Interests: Declaration of competing interest None., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Ultrasound (US) has a high capacity to increase food safety. Although high and/or moderate temperature in combination with US has been studied, the knowledge about cooling/low temperatures as well as its combined effect with chemical preservation methods is scarce. Therefore, the aim of this study was to describe the inactivation of Staphylococcus spp. (SA) present in the natural microbiota of sliced Brazilian dry-cured loin (Socol, BDL) using US (40 kHz and 5.40 W/g) at 1.6-17.9 kJ/g, temperature (T) between 6.4 and 73.6 °C and peracetic acid (PA) between 5.5 and 274.5 mg/L employing the Central Composite Rotatable Design. The model fully describes how the combination of US, T, and PA affects SA inactivation. In BDL, an increase in US acoustic energy density (kJ/g) allows the reduction of T necessary to inactivate SA because of the occurrence of synergistic effect. However, US applied at low T was inefficient. On the other hand, PA was more efficient at low T, since high T degraded this compound at different rates according to the holding T. Therefore, the data indicates a relation between the technologies used in the combined decontamination of sliced BDL improving dry-cured meat safety., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Listeria monocytogenes is a food-borne pathogen responsible for the disease listeriosis and is commonly isolated from food and food production facilities. Many L. monocytogenes strains contain plasmids, though the contributions of plasmids to survival in food production environments are unknown. Three L. monocytogenes ST5, ST8, and ST121 strains containing plasmids, which harbor putative stress response genes, were cured of their plasmids. Wildtype (WT) and plasmid-cured strains were exposed to disinfectant, oxidative, heat, acid, or salt stress. After stress exposure, cells were plated for colony forming unit (CFU) counts to determine survivors. L. monocytogenes WT strains exposed to 0.01% (vol/vol) H 2 O 2 , 1% (vol/vol) lactic acid, and 15% (wt/vol) NaCl, pH 5 showed significantly higher counts of survivors compared to the plasmid-cured strains. The number of survivors for the ST5 WT strain exposed to 10 μg/mL benzalkonium chloride (BC) was significantly higher than in the plasmid-cured strain. The ST8 and ST5 strains were exposed to elevated temperature (50° and 55 °C respectively); only the ST5 WT strain had significantly higher numbers of survivors than the plasmid-cured strains. Our data revealed that L. monocytogenes ST5, ST8, and ST121 plasmids contribute to tolerance against elevated temperature, salinity, acidic environments, oxidative stress and disinfectants., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The sanitising effects of combined lactic acid (LA, 2%, v/v) and low-concentration sodium hypochlorite (SH, 4 mg/L) on Listeria innocua were investigated. The disinfectant kinetics and percentage of injured cells showed that the combined treatment resulted in dramatic inactivation of L. innocua after 60 s. The Weibull model with a high R 2 (1.00) and low root mean squared error (RMSE) (0.19) and Akaike Information Criterion (AIC) (-7.58) was verified as the most superior model to describe effects of the combination treatment. Protein leakage and the fluorescence intensity ratio revealed that LA might disrupt the cytoplasmic membrane rapidly and that SH could synergistically promote the sanitising effect. Atomic force microscopy (AFM) images showed notable morphological changes in the combination treated cells, which presented lower width (0.73 μm) and height (0.29 μm), and increased roughness (10.37 nm). Moreover, massive leakage of intracellular components indicated severe cell collapse. The antimicrobial effects on organic broccoli sprouts showed that the combined treatment resulted in reductions of 0.82, 1.51, and 1.77 log colony forming units (CFU)/g fresh weight (FW) for aerobic bacteria, yeasts and moulds, and inoculated L. innocua at day 0. In addition, there were no significant differences of sensory quality results between treatment and control groups and they were all greater than the acceptance limit. Thus, the disinfection treatments had no negative effects on the storage quality of the organic broccoli sprouts. The combined treatment could be a potential sanitising strategy for organic sprouts., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In this study, the efficacy of household sanitizers application on reduction of Escherichia coli O157:H7, ascorbic acid, total phenolics, DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging and overall acceptability of inoculated fresh basil leaves (Ocimum basilicum), at temperature of 40 °C was investigated. Sanitizers containing lactic acid (LA), acetic acid (AA) and citric acid (CA) were used at concentration of 2%, individually or in combination with H 2 O 2 (1% or 2%), and NaCl (7%). Control a and b were unwashed and washed leaves with distilled, deionized and sterilized water, respectively. All sanitizing treatments, in comparison to the control a, reduced the numbers of E. coli O157:H7 (0.24 ± 0.12-3.37 ± 0.48 log CFU/g) at day 1 (1 h after sanitizing). The lowest number of E. coli O157:H7 population (2.35 ± 0.26 log CFU/g) was observed by applying the LA + H 2 O 2 (%2) treatment at first day of chilled storage. The highest amount of ascorbic acid (27.77 ± 0.06 mg/100 g), total phenolic (112.2 ± 0.5 mg gallic acid equivalents/100 g) and DPPH radical scavenging activity (95.2 ± 0.5%) was observed in control a at first day (P < 0.05). The results showed that the amount of ascorbic acid, total phenolics, DPPH radical scavenging activity and overall acceptability of basil leaves decreased during chilled storage. On day 2 of storage, the scores of sensory attributes for the control group were less than the minimum score of acceptance (i.e. 5 points). The results of this study indicated that LA + H 2 O 2 (2%) treatment rendered the samples favorable in terms of overall appearance (≥5) up to 48 h., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Strategies to mitigate antibiotic-resistant bacteria (ARB), including human pathogens, on raw vegetables are needed to reduce incidence of resistant infections. The objective of this research was to determine the effectiveness of standard post-harvest interventions, sanitizer washing and cold storage, to reduce ARB, including antibiotic resistant strains of the human pathogen E. coli O15:H7 and a common spoilage bacterium Pseudomonas, on raw carrots. To provide a background inoculum representing potential pre-harvest carryover of ARB, carrots were dip-inoculated in dairy cow manure compost slurry and further inoculated with known ARB. Inoculated carrots were washed with one of three treatments: sodium hypochlorite (50 ppm free chlorine), peroxyacetic acid (40 ppm peroxyacetic acid; 11.2% hydrogen peroxide), tap water (no sanitizer), or no washing (control). Washed carrots were air dried, packaged and stored at 10 °C for 7d or 2 °C for up to 60 d. Enumeration was performed using total heterotrophic plate counts (HPC), HPCs on antibiotic-containing media ("ARBs"), E. coli O157:H7, and Pseudomonas sp. immediately after washing (0 d) and after 7 d of storage. In addition to the cultured bacteria, changes to the surficial carrot microbiota were profiled by sequencing bacterial 16S rRNA gene amplicons to determine the effect of sanitizer wash, storage temperature, and time of storage (0, 1, 7, 14 and 60 d). Storage temperature, addition of a sanitizer during wash, and duration of storage significantly affected the bacterial microbiota (Wilcoxon, p < 0.05). Inclusion of either sanitizer in the wash water significantly reduced the log CFU/g of E. coli O157:H7 and Pseudomonas sp., as well as HPCs enumerated on cefotaxime- (10 μg/ml), sulfamethoxazole- (100 μg/ml), or tetracycline (3 μg/ml) supplemented media compared to the unwashed control (ANOVA, p < 0.05). However, no significant reductions to bacteria resistant to vancomycin or clindamycin occurred after washing and storage. Members of the Proteobactetria, Firmicutes, Actinobacteria, Planctomycetes, and Acidobacteria comprised the bacterial carrot microbiota. The diversity of the carrot microbiota was significantly affected by the temperature of storage and by extended storage (60 d), when spoilage began to occur. There were no significant differences to the relative abundance of bacterial groups associated with the type of sanitizer used for washing. Results of this study indicate that inclusion of a sanitizer in wash water, followed by storage at 2 °C, might be an effective strategy to prevent re-growth of pathogenic E. coli O157:H7 and reduce levels of bacteria resistant to certain antibiotics on carrots., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Foodborne illnesses caused by norovirus contaminated fresh produce remain a food safety concern worldwide. In the present study, the impacts of commercial and home processing conditions of strawberries were evaluated for inactivation of the MS2 bacteriophage. MS2 was used as a surrogate of norovirus and was spot inoculated onto strawberries to achieve 6.6 log PFU/g. The inoculated strawberries were washed with tap water, electrolyzed water, or 50 ppm chlorine for 90 s prior to and after storage. After initial washing, the strawberries were separately stored at -20 °C and -80 °C for 30 days. Change in MS2 populations on strawberries was evaluated by plaque assay method on day 1, 15, and 30 for -20 °C and -80 °C groups. The results showed that washing strawberries prior to storage resulted in a significant decrease (approximately 1 log PFU/g) of MS2 population regardless of the treatment (p < 0.05). Frozen storage had minor effects on inactivating MS2, which resulted in approximately a 0.5 log PFU/g reduction at the end of storage. Washing frozen berries in electrolyzed water or 50 ppm chlorine on day 30 resulted in an additional 1 log PFU/g decrease in MS2 compared to water alone. These results suggest that washing strawberries with a chemical antimicrobial prior to and post frozen storage may enhance microbial safety., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Peracetic acid (PAA) is a common oxidative sanitizer that is used in the food industry against various microorganisms. Limited information on the response of bacteria to this biocide is available. This study investigates the molecular response of the prevalent seafood-borne pathogenic Vibrio parahaemolyticus to PAA using mutants of peroxide scavenging genes. Among katE1, katE2, katG1, katG2, ahpC1 and ahpC2, and their regulator oxyR gene mutants, oxyR and katE mutants were highly susceptible to PAA. The growth and lethality of V. parahaemolyticus were harmed by 15 ppm of PAA in the △katE1E2 double mutant, and were significantly ameliorated in the presence of the katE1 gene in the wild-type strain and the gene-complementary strains that were pre-adapted in 2 ppm of PAA or 100 μM hydrogen peroxide. The application of PAA to these strains induced the accumulation of reactive oxygen species. The reduction of the level of hydrogen peroxide and gene expression during this treatment was influenced by the presence of katE genes. This investigation confirmed the major role of katE1 and a compensatory role of katE2 in the resistance of V. parahaemolyticus to PAA, and demonstrated some minor differences in the responses of this bacterium against PAA and hydrogen peroxide., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Effective and environmentally-friendly alternatives to traditional disinfectants are necessary to reduce the pollution and the emergence of antimicrobial-resistant bacterial strains in food-related environments. In the present study, treatments based in single and combined applications of plant essential oils (EOs) were evaluated for control Staphylococcus aureus biofilms. EOs of Lippia sidoides, Thymus vulgaris and Pimenta pseudochariophyllus showed a higher efficacy than peracetic acid and sodium hypochlorite against S. aureus planktonic cells and 24-h-old biofilms formed on polystyrene and stainless steel under food-related conditions. High concentrations of thymol and chavibetol were detected in these EOs, as well as the presence of other antimicrobial compounds such as carvacrol, eugenol, p-cymene, limonene, α-pinene, α-terpineol, terpinen-4-oil and linalool. L. sidoides oil were particularly effective against S. aureus, but doses higher than 2.75% (v/v) were required to completely eradicate 24-h-old biofilms. Binary combinations of L. sidoides, T. vulgaris and P. pseudochariophyllus allowed decrease significantly doses required to reduce 99.99% the number of biofilm cells. Furthermore, peracetic acid increased its efficacy against S. aureus biofilms by the combined application with these EOs. The most effective treatments against S. aureus biofilms were those combining L. sidoides with T. vulgaris or peracetic acid. Therefore, these EO-based treatments can be considered as an effective and environmentally-friendly alternative to control S. aureus biofilms in food-contact surfaces., (Copyright © 2018 Elsevier B.V. All rights reserved.)

There has been an increasing demand for fresh fruit and vegetables in recent years. Along the processing line in fresh-cut vegetable production, disinfection is one of the most important processing steps affecting the quality and safety, and the shelf-life of the end produce. Although a range of antimicrobial compounds commonly termed biocides or disinfectants are available, chlorine has long been used to disinfect washing waters of fresh-cut vegetables. However, since chlorine reactions with organic matter lead to the production of by-products, alternative disinfectants to chlorine must be evaluated. A synthetic washing water formula has been developed to determine the antimicrobial efficiency of different families of potential disinfectants: quaternary ammonium compounds (QACs) as benzalkonium chloride (BZK), and didecyldimethylammonium chloride (DDAC); isothiazolinones (mixture of methylchloroisothiazolinone and methylisothiazolinone, CMIT:MIT 3:1 and 1:1); and essential oils (carvacrol, CAR). The twin configuration and higher length of the chains of alkyl groups of DDAC compared to BZK have led to a higher antimicrobial efficiency. In both cases, Gram-positive bacteria seemed to be much more sensitive to the QAC attack than Gram-negative. The opposite happened for CMIT:MIT. The chloro-substituted isothiazolinone (CMIT) has been proven to be much more effective than its unsubstituted form (MIT). In addition, in contrast to chlorine, its antimicrobial activity together with that of DDAC was not decreased when increasing the organic matter content of the water. Synergetic antimicrobial effects have been confirmed when combining BZK and CAR. MBC values were determined in SWW, during 90 s of contact time and Salmonella concentration of 10 3  CFU/mL, corresponding to: 100 (BZK), 30 (DDAC), 50 (CMIT:MIT 3:1), 100 (CMIT:MIT 1:1), 300 (CAR), 75 (BZK)-200 (CAR), and 9 (free chlorine) mg/L. MBC values for inactivating similar concentration of E. faecalis corresponded to: 50 (BZK), and 10 (DDAC) mg/L. Increasing contact times up to 5 min did not lead to higher antimicrobial efficiencies. CMIT:MIT 3:1 together with DDAC, and combinations of BZK-CAR seem to be a plausible alternative to chlorine., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This study was conducted to investigate the Salmonella inactivation effects of washing in combination with pulsed light (PL), ultrasound, and chlorine on lettuce shreds. First, the effect of washing combined with PL and chlorine on the inactivation of Salmonella on lettuce and in wash water was evaluated in a small-scale study with clear tap water and turbid tap water containing lettuce extract and silicon dioxide. In general, water wash combined with PL (PL wash) and chlorine wash combined with PL (PL-Cl) were significantly more effective on killing Salmonella on lettuce than the chlorine wash and water wash regardless the wash water quality and inoculation method. We then tested washing combined with PL, ultrasound and chlorine using a large-scale UV setup with turbid wash water. Increasing the sample size decreased the decontamination efficacy of all the treatments. All the treatments resulted in <2 log reductions of Salmonella on lettuce shreds. For both small- and large-scale studies, treatments involving chlorine could keep the Salmonella population in wash water under the detection limit of 2 CFU/mL for almost all the replicates. Taking everything into consideration, we concluded that the combined PL-Cl treatment could be a better alternative to the chlorine wash for lettuce decontamination since it was in general more effective on inactivating Salmonella on lettuce than chlorine wash and could maintain the Salmonella level in wash water under the detection limit of 2 CFU/mL regardless the inoculation method, water quality and sample size, preventing the potential cross contamination through wash water., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Different fungi, including the genera Aspergillus (Neosartorya), Paecilomyces (Byssochlamys) and Talaromyces, produce (asco)spores that survive pasteurization treatments and are regarded as the most stress-resistant eukaryotic cells. The sensitivity of the ascospores to treatments with industrial sanitizers containing chlorine dioxide and iodine (iodophors) has never been assessed before. Ascospores of 4 species of Eurotiales were tested and showed clear variations in sensitivity. The most resilient species, T. macrosporus and Pae. variotii (=B. spectabilis) survive 75, but not 200 ppm chlorine dioxide solution treatments. These species were able to survive 75 ppm iodine solution treatments, but relatively low amounts of ascospores (100-1000 spores) could be inactivated after 16 h of treatment. Inactivated spores did not show any sign of germination after 7 days following treatment on growth medium. As judged by microscopy, iodine inactivation resulted in visibly distorted ascospores. For the interpretation of results, the state of dormancy or activation of ascospores is highly important., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This study investigated the effect of a water-assisted ultraviolet system (WUV; samples were treated by UV while being immersed in agitated water) on the inactivation of Salmonella on baby spinach, iceberg lettuce, blueberry, grape tomato, and baby-cut carrot. The Salmonella inactivation effect of the WUV system was tested in two scales, and three disinfectants, chlorine, peroxyacetic acid (PAA) and hydrogen peroxide (H 2 O 2 ), were tested in combination with the system to see whether the Salmonella inactivation effect could be enhanced. The fresh produce samples were dip-inoculated with a Salmonella cocktail to final concentrations of 4.6-7.6 log CFU/g. To simulate the washing process in the industry, fresh produce extracts and/or silicon dioxide were added in the wash water to adjust chemical oxygen demand to ~2000 mg/L and turbidity to >60 NTU. In general, the decontamination efficacy of WUV treatments followed this order: Tomato > Carrot > Lettuce ≈ Blueberry > Spinach. In the small-scale study, WUV alone was able to achieve 0.9, 2.6, >3.6, 1.7, and 2.0 log CFU/g reductions of Salmonella on fresh produce for spinach, lettuce, tomato, blueberry, and carrot, respectively. For all fresh produce items, WUV combined with PAA could achieve significantly (P < 0.05) higher Salmonella reduction on fresh produce than chlorine wash and PAA wash. The WUV treatments combined with chlorine or PAA were able to keep residual Salmonella in wash water below the detection limit (2 CFU/mL) for almost all the replicates. Similar Salmonella reductions on fresh produce and in wash water were found in the large-scale study. Considering the decontamination efficacy on fresh produce, the ability to disinfect the wash water, and the cost, we recommend chlorine wash for baby spinach, WUV alone for grape tomato and WUV combined with PAA for iceberg lettuce, blueberry and baby-cut carrot., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The recovery at a range of incubation temperatures and pH of spores of Bacillus weihenstephanensis KBAB4 exposed to a peracetic acid-based disinfectant (PABD) or to pulsed light was estimated. Spores of B. weihenstephanensis were produced at 30 °C and pH 7.00, at 30 °C and pH 5.50, or at 12 °C and pH 7.00. The spores were treated with a commercial peracetic acid-based disinfectant at 80 mg·mL -1 for 0 to 200 min at 18 °C or by pulsed light at fluences ranging between 0.4 and 2.3 J·cm -2 for pulsed light treatment. After each treatment, the spores were incubated on nutrient agar at 12 °C, 30 °C or 37 °C, or at pH 5.10, 6.00 or 7.40. Incubation temperature during recovery had a significant impact only near the recovery limits, beyond which surviving spores previously exposed to a PABD or to pulsed light were not able to form colonies. In contrast, a decrease in pH of the recovery nutrient agar had a progressive impact on the ability of spores to form colonies. The time to first log reduction after PABD treatment was 29.5 ± 0.7 min with recovery at pH 7.40, and was tremendously shortened 5.1 ± 0.2 min with recovery at pH 5.10. Concerning the fluence necessary for the first log reduction, it was 1.5 times higher when the spores were recovered at pH 6.00 compared to a recovery at pH 5.10. The impact of recovery temperature and pH can be described with a mathematical model using cardinal temperature and pH as parameters. These effects of temperature and pH on recovery of Bacillus weihenstephanensis spores exposed to a disinfectant combining peracetic acid and hydrogen peroxide, or pulsed light are similar, although these treatments are of different natures. Sporulation temperature or pH did not impact resistance to the peracetic acid-based disinfectant or pulsed light., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This work investigated the effects of diamond-like carbon (DLC) coatings on the architecture and biocide reactivity of dual-species biofilms mimicking food processing contaminants. Biofilms were grown using industrial isolates of Escherichia coli and Pantoea agglomerans on bare stainless steel (SST) and on two DLC surface coatings (a-C:H:Si:O designated by SICON® and a-C:H:Si designated by SICAN) in order to evaluate their antifouling activities. Quantification and spatial organization in single- and dual-species biofilms were examined by confocal laser scanning microscopy (CLSM) using a strain specific labelling procedure. Those assays revealed that the E. coli isolate exhibited a higher adhesion to the modified surfaces and a decreased susceptibility to disinfectant in presence of P. agglomerans than alone in axenic culture. While SICON® reduced the short-term growth of E. coli in axenic conditions, both DLC surfaces increased the E. coli colonization in presence of P. agglomerans. However, both modified surfaces triggered a significantly higher log reduction of E. coli cells within mixed-species biofilms, thus the use of SICON® and SICAN surfaces may be a good approach to facilitate the disinfection process in critical areas of food processing plants. This study presents a new illustration of the importance of interspecies interactions in surface-associated community functions, and of the need to evaluate the effectiveness of hygienic strategies with relevant multi-species consortia., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Enterotoxigenic Clostridium perfringens, a leading foodborne pathogen can be cross-contaminated from food processing stainless steel (SS) surfaces to the finished food products. This is mostly due to the high resistance of C. perfringens spores adhered onto SS surfaces to various disinfectants commonly used in food industries. In this study, we aimed to investigate the survivability and adherence of C. perfringens spores onto SS surfaces and then validate the effectiveness of a simulated Clean-in-Place (CIP) regime on inactivation of spores adhered onto SS surfaces. Our results demonstrated that, 1) C. perfringens spores adhered firmly onto SS surfaces and survived for at-least 48 h, unlike their vegetative cells who died within 30 min, after aerobic incubation at refrigerated and ambient temperatures; 2) Spores exhibited higher levels of hydrophobicity than vegetative cells, suggesting a correlation between cell surface hydrophobicity and adhesion to solid surfaces; 3) Intact spores were more hydrophobic than the decoated spores, suggesting a positive role of spore coat components on spores' hydrophobicity and thus adhesion onto SS surfaces; and finally 4) The CIP regime (NaOH + HNO 3 ) successfully inactivated C. perfringens spores adhered onto SS surfaces, and most of the effect of CIP regime appeared to be due to the NaOH. Collectively, our current findings may well contribute towards developing a strategy to control cross-contamination of C. perfringens spores into food products, which should help reducing the risk of C. perfringens-associated food poisoning outbreaks., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The objective of this study was to evaluate how treatment temperature influences the solubility of ClO 2 gas and the antimicrobial effect of ClO 2 gas against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on produce and food contact surfaces. Produce and food contact surfaces inoculated with a combined culture cocktail of three strains each of the three foodborne pathogens were processed in a treatment chamber with 20 ppmv ClO 2 gas at 15 or 25 °C under the same conditions of absolute humidity (11.2-12.3 g/m 3 ) for up to 30 min. As treatment time increased, ClO 2 gas treatment at 15 °C caused significantly more (p < 0.05) inactivation of the three pathogens than treatment at 25 °C. ClO 2 gas treatment at 25 °C for 30 min resulted in 1.15 to 1.54, 1.53 to 1.88, and 1.00 to 1.78 log reductions of the three pathogens on spinach leaves, tomatoes, and stainless steel No.4, respectively. ClO 2 gas treatment at 15 °C for 30 min caused 2.53 to 2.88, 2.82 to 3.23, and 2.37 to 3.03 log reductions of the three pathogens on spinach leaves, tomatoes, and stainless steel No.4, respectively. Treatment with ClO 2 gas at 25 °C for 20 min resulted in 1.88 to 2.31 log reductions of the three pathogens on glass while >5.91 to 6.82 log reductions of these pathogens occurred after 20 min when treated at 15 °C. Residual ClO 2 levels after gas treatment at 15 °C were significantly (p < 0.05) higher than those at 25 °C. The results of this study can help the food processing industry establish optimum ClO 2 gas treatment conditions for maximizing the antimicrobial efficacy of ClO 2 gas., (Published by Elsevier B.V.)