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Organic matter (OM) accumulation is common in chill tanks used to decontaminate raw poultry parts during processing. OM negatively affects the antimicrobial activity of chlorine-based compounds, but its effect on the antimicrobial effectiveness of peroxyacetic acid (PAA) on poultry meat has not been described. Therefore, this study evaluated the effect of OM on the efficacy of PAA solutions in simulated postchill tanks to reduce Salmonella artificially inoculated onto chicken parts. Chicken thighs were inoculated with a five-strain cocktail of poultry-borne Salmonella enterica serovars at ca. 6 log CFU/mL. Then, the thighs were immersed for 30 or 45 s in PAA solutions (500 or 1,000 ppm) with chicken slurry to simulate OM accumulation (0, 15, or 30 g/L). The thighs were rinsed with neutralizing buffered peptone water (100 mL), and rinsates were plated onto xylose lysine desoxycholate agar. Experiments were performed in triplicate (three thighs per treatment per replicate). Chemical oxygen demand, total nitrogen, and pH were measured as the water quality parameters of the PAA solutions before and after use. Chemical oxygen demand ranged from 2,905 mg/L in unused 500-ppm solutions without added OM to 6,290 mg/L in used 1,000-ppm solutions with 30 g/L OM. Initial total nitrogen was 42.5 ± 2.0 and 60.9 ± 8.3 mg/L for 15 and 30 g/L OM, which increased by 27 ± 17 mg/L after use. The pH of solutions ranged from 3.16 ± 0.14 to 3.42 ± 0.09 for the 1,000-ppm solutions and from 3.59 ± 0.06 to 3.96 ± 0.06 for the 500-ppm solutions. Mean Salmonella reductions were 0.9 ± 0.1 log CFU/mL of rinsate for the 500-ppm PAA treatment and 1.1 ± 0.1 log CFU/mL of rinsate for 1,000-ppm PAA treatment. Exposure time did not have a significant effect on the logarithmic reductions. There was no significant effect of OM concentration (P0.05) on the reductions, indicating that the antimicrobial efficacy was not affected and that PAA solutions may continue to be reused as long as the PAA concentration is actively monitored.

Salmonella is a common cause of foodborne illness in the United States, and several strains of Salmonella have been identified as resistant to antibiotics. It is not known whether strains that are antibiotic resistant (ABR) and that have some tolerance to antimicrobial compounds are also able to resist the inactivation effects of antimicrobial interventions used in fresh meat processing. Sixty-eight Salmonella isolates (non-ABR and ABR strains) were treated with half concentrations of lactic acid (LA), peracetic acid (PAA), and cetylpyridinium chloride (CPC), which are used in beef processing plants to screen for tolerant strains. Six strains each from non-ABR and ABR Salmonella that were most tolerant of LA (2%), PAA (200 ppm), and CPC (0.4%) were selected. Selected strains were inoculated on surfaces of fresh beef and subjected to spray wash treatment with 4% LA, 400 ppm PAA, or 0.8% CPC for the challenge study. Tissue samples were collected before and after each antimicrobial treatment for enumeration of survivors. Spray treatment with LA, PAA, or CPC significantly reduced non-ABR Salmonella and ABR Salmonella on surfaces of fresh beef by 1.95, 1.22, and 1.33 log CFU/cm2, and 2.14, 1.45, and 1.43 log CFU/cm2, respectively. The order of effectiveness was LAPAA = CPC. The findings also indicated that LA, PAA, and CPC were equally (P ≤ 0.05) effective against non-ABR and ABR Salmonella on surfaces of fresh beef. These data contribute to the body of work that indicates that foodborne pathogens that have acquired both antibiotic resistance and antimicrobial tolerance are still equally susceptible to meat processing antimicrobial intervention treatments.

Due to the dissemination of airborne conidia and spores, molds can contaminate various surfaces. In the food industry sector, their presence and development can have health and economic implications. In order to control these undesirable microorganisms, various approaches can be used but the main one relies on the use of disinfectants. The objective of this review is to report the existing studies on the effect of various disinfectant molecules (i.e., sodium hypochlorite, chlorine dioxide, ethanol and other alcohols, hydrogen peroxide, peracetic acid, and quaternary ammonium compounds) on the inactivation of fungal spores. These studies were sorted depending on the targeted fungal species. Noteworthy, in the food industry, four log and three log reductions are required to claim a fungicidal activity for suspension (European Standard 1650, 2019) and surface (European Standard 13697/IN1, 2019) treatments, respectively. Most of the presented studies concerned Penicillium and Aspergillus species (44 and 31% of the literature, respectively). In general, for a given disinfection procedure, ascospores were more resistant than conidia, and Aspergillus conidia were more resistant than Penicillium ones. However, the variability of encountered molds (e.g. species, strains, physiological state) and disinfection procedures (e.g. molecules, concentrations, contact time) affected the efficacy of disinfectants.

BACKGROUND Nontuberculous mycobacteria are commonly found pathogens; however, skin and soft tissue infections due to nontuberculous mycobacteria are often associated with surgical procedures, particularly after lipoplasty. Although nontuberculous mycobacteria are resistant to some chemical disinfectants, glutaraldehyde, peracetic acid, povidone iodine, alcohol, and chlorine are still used for the sterilization of medical instruments. This study investigated the efficacy of various disinfectants in a fatty environment with adipose and a bloody environment without adipose. In addition, this study was also used to identify the most effective disinfectant against nontuberculous mycobacteria. METHODS Three nontuberculous mycobacteria (Mycobacterium avium, M. abscessus, and M. fortuitum), pathogens frequently found in skin and soft tissue infections, were used. Seven chemical disinfectants were tested in both fatty and bloody environments. The disinfectants used were considered to have a sterilization effect when the log10 reduction factor exceeded 5. RESULTS Most disinfectants had some sterilizing effects against nontuberculous mycobacteria; however, glutaraldehyde was the most effective against all 3. Chlorhexidine and povidone iodine also displayed sterilizing effects. Of the disinfectants tested, only alkyldiaminoethylglycine hydrochloride showed a diminished effect with statistical significance, specifically against M. fortuitum in a fatty environment, whereas it had effective results in a bloody environment. CONCLUSIONS Glutaraldehyde showed the greatest sterilizing effect on nontuberculous mycobacteria with a log10 reduction factor >5 in both fatty and bloody environments. However, some chemical disinfectants did not show sufficient sterilizing effects in a fatty environment and, therefore, should be used with caution for the sterilization of nontuberculous mycobacteria. LEVEL OF EVIDENCE Level II.

During the severe acute respiratory syndrome coronavirus type 2 (SARS‐CoV‐2) pandemic, chlorine‐containing disinfectants have been widely used in nucleic acid amplification testing laboratories. Whether the use of disinfectants affect the results of viral nucleic acid amplification is unknown. We examined the impact of different hypochlorous acid (HOCl) concentrations on the quantitative results of SARS‐CoV‐2 by real‐time reverse‐transcription polymerase chain reaction (RT‐PCR). We also explored the mechanisms and models of action of chlorine‐containing disinfectants that affected the detection of SARS‐CoV‐2. The results showed that different HOCl concentrations and different action times had an impact on the SARS‐CoV‐2 results. High concentrations of ambient HOCl have a greater impact than low concentrations, and this effect will increase with the extension of the action time and with the increase in ambient humidity. Compared with the enzymes or the extracted RNA required for RT‐PCR, the impact of HOCl on the SARS‐CoV‐2 detection is more likely to be caused by damage to primers and probes in the PCR system. The false negative result still existed after changing the ambient disinfectant to ethanol but not peracetic acid. The use of HOCl in the environment will have an unpredictable impact on the nucleic acid test results of SARS‐CoV‐2. In order to reduce the possibility of false negative of SARS‐CoV‐2 nucleic acid test and prevent the spread of epidemic disease, environmental disinfectants should be used at the beginning and end of the experiment rather than during the experimental operation.

Poultry meat has been a vehicle of antibiotic resistant bacteria and genes. Yet, the diversity of selective pressures associated with their maintenance in the poultry-production chain remains poorly explored. We evaluated the susceptibility of Enterococcus spp. from chicken meat collected 20 years apart to antibiotics, metals, acidic pH and peracetic acid-PAA. Contemporary chicken-meat samples (n = 53 batches, each including a pool of neck skin from 10 single carcasses) were collected in a slaughterhouse facility using PAA as disinfectant (March-August 2018, North of Portugal). Broilers were raised in intensive farms (n = 29) using CuSO4 and organic acids as feed additives. Data were compared with that of 67 samples recovered in the same region during 1999-2001. All 2018 samples had multidrug resistant-MDR isolates, with >45 % carrying Enterococcus faecalis, Enterococcus faecium or Enterococcus gallinarum resistant to tetracycline, erythromycin, ampicillin, quinupristin-dalfopristin, ciprofloxa-cin, chloramphenicol or aminoglycosides. Resistance rates were similar (P > 0.05) to those of 1999-2001 samples for all but five antibiotics. The decrease of samples carrying vancomycin-resistant isolates from 46 % to % between 1999-2001 and 2018 was the most striking difference. Isolates from both periods were similarly susceptible to acid pH [minimum-growth pH (4.5-5.0), minimum-survival pH (3.0-4.0)] and to PAA (MIC90 = 100-120 mg/L/MBC90 = 140-160 mg/L; below concentrations used in slaughterhouse). Copper tolerance genes (tcrB and/or cueO) were respectively detected in 21 % and 4 % of 2018 and 1999-2001 samples. The tcrB gene was only detected in E. faecalis (MICCuSO4 > 12 mM), and their genomes were compared with other international ones of chicken origin (PATRIC database), revealing a polyclonal population and a plasmid or chromosomal location for tcrB. The tcrB plasmids shared diverse genetic modules, including multiple antimicrobial resistance genes (e.g. to tetracyclines, chloramphenicol, macrolide-lincosamide-streptogramin B-MLSB, aminoglycosides, bacitracin, coccidiostats). When in chromosome, the tcrB gene was co-located closely to merA (mercury) genes. Chicken meat remains an important vehicle of MDR Enterococcus spp. able to survive under diverse stresses (e.g. copper, acid) potentially contributing to these bacteria maintenance and flux among animal-environment -humans.

Since the first SARS-CoV-2 outbreak in Wuhan, China, there has been continued concern over the link between SARS-CoV-2 transmission and food. However, there are few studies on the viability and removal of SARS-CoV-2 contaminating food. This study aimed to evaluate the viability of SARS-CoV-2 on food matrices, depending on storage temperature, and inactivate the virus contaminating food using disinfectants. Two SARS-CoV-2 strains (L and S types) were used to contaminate lettuce, chicken, and salmon, which were then stored at 20,4 and -40 °C. The half-life of SARS-CoV-2 at 20 °C was 3-7 h but increased to 24-46 h at 4 °C and exceeded 100 h at -40 °C. SARS-CoV-2 persisted longer on chicken or salmon than on lettuce. Treatment with 70% ethanol for 1 min inactivated 3.25 log reduction of SARS-CoV-2 inoculated on lettuce but not on chicken and salmon. ClO2 inactivated up to 2 log reduction of SARS-CoV-2 on foods. Peracetic acid was able to eliminate SARS-CoV-2 from all foods. The virucidal effect of all disinfectants used in this study did not differ between the two SARS-CoV-2 strains; therefore, they could also be effective against other SARS-CoV-2 variants. This study demonstrated that the viability of SARS-CoV-2 can be extended at 4 and -40 °C and peracetic acid can inactivate SARS-CoV-2 on food matrices.

Poultry processing establishments use antimicrobial aids on broiler parts to minimize Campylobacter contamination. A silver-stabilized hydrogen peroxide (SHP) product was assessed for use as an antimicrobial processing aid. In a series of experiments, wing segments with skin were inoculated with 103 to 107 cells of Campylobacter coli, followed by treatment with SHP at 15,000 or 30,000 mg/L, peroxyacetic acid (PAA) at 300 or 3,000 mg/L (parts per million), or water. Each treatment was applied by either dip or spray. Rinsates from each wing segment were analyzed for direct counts and prevalence of Campylobacter. Treatment with SHP or PAA significantly reduced Campylobacter levels compared with water controls by up to 2.22 log CFU/mL. At high inoculum levels (106 to 107), SHP and PAA applied by dip had up to 1.27 log CFU/mL further reductions of Campylobacter levels compared with spray-treated wing segments. Additionally, wing drumettes were observed to retain higher levels and prevalence of Campylobacter recovery compared with wing flats at a low inoculation level (103). The results indicated that there was no carryover effect of SHP (same day versus 24 h) and dip treatment with SHP or PAA decreased Campylobacter recovery on broiler chicken wing segments compared with a water control. Although a 2-log reduction was modest, SHP had similar efficacy as the commonly used processing aid PAA. SHP shows potential for further investigation as an antimicrobial processing aid for use on poultry parts. HIGHLIGHTS

Within the European Union, Salmonella is frequently reported in food and feed products. A major route of transmission is upon contact with contaminated surfaces. In nature, bacteria such as Salmonella are often encountered in biofilms, where they are protected against antibiotics and disinfectants. Therefore, the removal and inactivation of biofilms is essential to ensure hygienic conditions. Currently, recommendations for disinfectant usage are based on results of efficacy testing against planktonic bacteria. There are no biofilm-specific standards for the efficacy testing of disinfectants against Salmonella. Here, we assessed three models for disinfectant efficacy testing on Salmonella Typhimurium biofilms. Achievable bacterial counts per biofilm, repeatability, and intra-laboratory reproducibility were analyzed. Biofilms of two Salmonella strains were grown on different surfaces and treated with glutaraldehyde or peracetic acid. Disinfectant efficacy was compared with results for planktonic Salmonella. All methods resulted in highly repeatable cell numbers per biofilm, with one assay showing variations of less than 1 log10 CFU in all experiments for both strains tested. Disinfectant concentrations required to inactivate biofilms were higher compared to planktonic cells. Differences were found between the biofilm methods regarding maximal achievable cell numbers, repeatability, and intra-laboratory reproducibility of results, which may be used to identify the most appropriate method in relation to application context. Developing a standardized protocol for testing disinfectant efficacy on biofilms will help identify conditions that are effective against biofilms.

As demand for fresh-cut produce increases, minimizing the risk of salmonellosis becomes critical for the produce industry. Sanitizers are routinely used during commercial flume washing of fresh-cut produce to minimize cross-contamination from the wash water. This study assessed the efficacy of a novel sanitizer blend consisting of peracetic acid (PAA; OxypHresh 15) with a sulfuric acid-surfactant (SS) antimicrobial (PAA-SS; ProduceShield Plus) against Salmonella during simulated commercial washing of diced tomatoes. Triplicate 9.1-kg batches of Roma tomatoes were dip inoculated in a two-strain avirulent Salmonella cocktail (Salmonella Typhimurium LT2 and MHM112) to achieve 5 to 6 log CFU per tomato and air dried for 2 h. After mechanical dicing, the tomatoes were washed in a pilot-scale processing line for 60 s with or without an added organic load in 90 ppm of PAA-SS (pH 1.8), SS at pH 1.8, 90 ppm of PAA, 5 or 10 ppm of free chlorine or sanitizer-free water as the control. Overall, PAA-SS (1.75 ± 0.75 log CFU/g) was significantly (P ≤ 0.05) more effective than water (0.69 ± 0.42 log CFU/g), chlorine (0.35 ± 0.36 log CFU/g), or SS (0.36 ± 0.19 log CFU/g) in reducing Salmonella. After washing for 20 s, PAA-SS was the only sanitizer to show a significant (P ≤ 0.05) reduction (1.93 ±0.59 log CFU/g) in Salmonella. All wash water samples were negative for Salmonella, except for 5 and 10 ppm of chlorine and the water control. Using PAA-SS with an organic load, yeast and mold populations were below the limit of detection (1.40 log CFU/g) and significantly (P ≤ 0.05) lower on diced tomatoes after 14 days of refrigerated storage compared with the other treatments (8.37 ± 0.08 log CFU/g), with SS at pH 1.8 (3.91 ± 0.93 log CFU/g) most effective against yeast and mold in the absence of an organic load. On the basis of these findings, the safety and shelf life of commercially washed diced tomatoes can be improved with PAA-SS. HIGHLIGHTS

The effect of bactericide Sekusept active (B SA), a peracetic acid-based preparation, on microbial strains, isolated from patients with severe infectious diseases who were treated in a regional children’s multi-specialty hospital, was studied. Based on the biochemical identification, the strains were classified as gram-negative non-fermenting bacteria (22 strains), Enterobacteriaceae family (18 strains), and bacilli - 3 strains. The biocidal activity of B SA was evaluated by the degree of inhibition of the growth of bacterial cells, existing in the planktonic form and in the form of biofilm (on a flat-bottomed plastic immunological tablet). It was shown that all the studied strains had the ability to biofilm formation, most of them (67,4%) formed moderately pronounced biofilms, and non-fermenting bacteria had a particularly pronounced coefficient of biofilm formation. The selected concentrations of B CA inhibited the growth of planktonic cells, and the ability of bactericide to prevent the formation of biofilms depended on the concentration (the most effective concentrations were 0,8 and 3,0%). Sensitivity of the strains existed in the aged biofilm to the bactericide was significantly lower, especially resistant to this effect were biofilms formed by non-fermenting bacteria and representatives of fam. Enterobacteriaceae. Our results confirm the importance of testing the effectiveness of biocides not only in accordance with standard methods developed for microorganisms in planktonic form, but also for biofilms.

Biofilms on the surface of food processing equipment act as potential reservoirs of microbial contamination. Bacterial interactions are believed to play key roles in both biofilm formation and antimicrobial tolerance. In this study, Aeromonas hydrophila, Chryseobacterium oncorhynchi, and Pseudomonas libanensis, which were previously isolated from Chinese raw milk samples, were selected to establish two dual-species biofilm models (P. libanensis plus A. hydrophila and P. libanensis plus C. oncorhynchi) on stainless steel at 7°C. Subsequently, three disinfectants, hydrogen peroxide (100 ppm), peracetic acid (100 ppm), and sodium hypochlorite (100 ppm), were used to treat the developed sessile communities for 10 min. Structural changes after exposure to disinfectants were analyzed with confocal laser scanning microscopy. The cell numbers of both A. hydrophila and C. oncorhynchi recovered from surfaces increased when grown as dual species biofilms with P. libanensis. Dual-species biofilms were more tolerant of disinfectants than were each single-species biofilm. Peracetic acid was the most effective disinfectant for removing biofilms, followed by hydrogen peroxide and sodium hypochlorite. The results expand the knowledge of mixed-species biofilms formed by psychrotrophic bacteria and will be helpful for developing effective strategies to eliminate bacterial mixed-species biofilms. HIGHLIGHTS

Salmonella can enter on the viable but non-culturable state (VBNC), characterized by the loss of ability to grow in routine culture media hindering detection by conventional methods and underestimation of the pathogen. Despite advances in research done so far, studies comparing conditions that lead Salmonella into the VBNC state are scarce. The main objective of this study was to evaluate different stresses to induce Salmonella to the VNBC state. Osmotic (1.2 M NaCl), acid (peracetic acid, 5.66 mg/mL) and oxidative (hydrogen peroxide, 1.20 mg/mL) stress were used at 4 °C to induce Salmonella enterica serovars Enteritidis and Typhimurium to the VBNC state. The culturability loss was monitored in the brain heart infusion (BHI) broth and agar, and the viability was determined by fluorescence microscopy, using the Live/Dead® kit, and by flow cytometry. Besides, the morphological characterization by atomic force microscopy (AFM) was performed. Storage in 1.2 M NaCl at 4 °C induced the VBNC state in Salmonella cells for periods longer than 121 days, and the percentage of viable cells has reached above 80.9%. More aggressive stress conditions promoted by peracetic acid and hydrogen peroxide induced the VBNC state in periods of, at most 0.14 day, and resulted in percentages of 8.5% to 45.5% viable cells, respectively. The counts of viable cells in the flow cytometer corroborate the results obtained by microscopic counts. The VBNC cells obtained in 1.2 M NaCl at 4 °C showed morphological changes, reducing the size and changing the morphology from bacillary to coccoid. No morphological change was observed on the cells stressed by acid or oxidant compounds.

The present study evaluates the antimicrobial susceptibility of persister cells of Bacillus cereus and Pseudomonas fluorescens after their regrowth in suspension and as biofilms. Two conventional (benzalkonium chloride—BAC and peracetic acid—PAA) and two emerging biocides (glycolic acid—GA and glyoxal—GO) were selected for this study. Persister cells resulted from biofilms subjected to a critical treatment using the selected biocides. All biocide treatments developed B. cereus persister cells, except PAA that effectively reduced the levels of vegetative cells and endospores. P. fluorescens persister cells comprise viable and viable but non-culturable cells. Afterwards, persister cells were regrown in suspension and in biofilms and were subjected to a second biocide treatment. In general, planktonic cultures of regrown persister cells in suspension lost their antimicrobial tolerance, for both bacteria. Regrown biofilms of persister cells had antimicrobial susceptibility close to those regrown biofilms of biocide-untreated cells, except for regrown biofilms of persister P. fluorescens after BAC treatment, which demonstrated increased antimicrobial tolerance. The most active biocide against persister cells was PAA, which did not promote changes in susceptibility after their regrowth. In conclusion, persister cells are ubiquitous within biofilms and survive after critical biocide treatment. The descendant planktonic and biofilms populations showed similar properties as the original ones.

Chlorine and peroxyacetic acid (PAA) are commonly applied in dump tanks and flume systems in commercial fresh apple packing lines; however, little is known about their practical efficacies in dump tank water systems. This study evaluated the efficacies of chlorine and PAA to control Listeria monocytogenes on fresh apples and cross-contamination in simulated dump tank water (SDTW). Efficacies of chlorinated water with initial free chlorine (FC) of 25-100 ppm against L. monocytogenes on apples were significantly impacted by the presence of organic matter, especially for chlorine with 25 ppm initial FC. Chlorine with initial FCs of 50-100 ppm and 2 min contact reduced L. monocytogenes on apples by ∼0.9 log

Fresh fruits, especially strawberries, are usually consumed raw without any step to ensure their food safety. Salmonella enterica is one of the most important etiologic agents for foodborne diseases throughout the world and its ability to respond to some stress responses makes it even more dangerous. In the present investigation, we study the survival of S. Enteritidis (CECT-4300) on strawberries after 2-min of various disinfection steps (NaClO (200 ppm), peracetic acid (PAA; 40 ppm), water-assisted UV-C (WUV-C), and the combination WUV-C and 40 ppm of PAA (WUV-C + PAA)) and after 5 days of cold storage (4 ◦C). Moreover, the pathogenic potential of the surviving bacteria, such as the ability to survive throughout the gastrointestinal tract (GI) and subsequently the capability to adhere to and invade Caco-2 cells, was tested at each sampling point. After 2-min of washing procedures, reductions of S. Enteritidis on strawberries were ≥1.2 log, with no significant differences among treatments. However, the use of WUV-C + PAA treatment achieved the highest reductions in washing water, in which S. Enteritidis was not detected (

Listeria monocytogenes biofilms are ubiquitous in the food-processing environment, where they frequently show resistance against treatment with disinfectants such as peracetic acid (PAA) due to sub-lethal damage resulting in biofilm persistence or the formation of secondary biofilms. L. monocytogenes serovar ½a EGD-e biofilms were cultivated under continuous flow conditions at 10 °C, 22 °C, and 37 °C and exposed to industrially relevant PAA concentrations. The effect of PAA on biofilm metabolic activity and biomass was monitored in real-time using the CEMS-BioSpec system, in addition to daily measurement of biofilm-derived planktonic cell production. Biofilm-derived planktonic cell yields proved to be consistent with high yields during biofilm establishment (≥106 CFU.mL−1). The exposure of biofilms to the minimum inhibitory PAA concentration (0.16%) resulted in only a brief disruption in whole-biofilm metabolic activity and biofilm biomass accumulation. The recovered biofilm accumulated more biomass and greater activity, but cell yields remained similar. Increasing concentrations of PAA (0.50%, 1.5%, and 4.0%) had a longer-lasting inhibitory effect. Only the maximum dose resulted in a lasting inhibition of biofilm activity and biomass–a factor that needs due consideration in view of dilution in industrial settings. Better disinfection monitoring tools and protocols are required to adequately address the problem of Listeria biofilms in the food-processing environment, and more emphasis should be placed on biofilms serving as a “factory” for cell proliferation rather than only a survival mechanism.

Peracetic acid (PAA) applied to whole poultry carcasses can reduce the number of Campylobacter, a leading cause of human gastroenteritis. However, previous modelling experiments indicated that Campylobacter survived in greater numbers when pre-treated with a thermal stress equivalent to poultry processing scalding prior to chilling with PAA than when subject to chilling with PAA only. To better understand how Campylobacter responds to PAA, proteomes of C. jejuni poultry strain 2704 were measured after exposure to PAA (60 ppm, pH 4.0) for 45 min under laboratory ambient conditions (approximately 23 °C) to establish a foundational map of survival mechanism before combining with other stresses. Analysis of 580 quantified proteins did not indicate a triggered "peroxide shock" response, nor were common heat shock responses detected. Thioredoxin, iron homeostatic, peroxiredoxins and cytochrome c peroxidases became more abundant suggesting that PAA disturbed cytoplasmic redox homeostasis resulting in antioxidant activation and increased prioritisation of iron homeostasis. The PAA treatment led to responses that included an increased priority for oxidative phosphorylation and a simultaneous decrease in central metabolism associated protein abundances. Lon protease was induced suggesting it has a role in maintaining homeostasis during non-thermal stress. Proteins in flagella and chemotaxis became more abundant though whether PAA has a chemorepellent effect requires further investigation. Overall, the proteome data suggests there was a rapid cellular response to applied PAA stress in the first 15 min with the adaptation to the stress completing between 30 and 45 min. The findings will help guide PAA implementation in commercial poultry processing in terms of processing location and length of application.

Chicken meat is frequently contaminated with zoonotic bacterial pathogens such as Campylobacter spp and Salmonella spp. These two bacterial genera are commonly linked with cases of human gastrointestinal disease, thus mitigating their presence in the poultry meat supply chain is paramount. Here, the efficacy of two sanitizers, peroxyacetic acid (PAA) and acidified sodium chlorite (ASC), was tested using whole chicken carcasses obtained either prior to the inside/outside wash or the post-immersion spin chill steps of processing. Two concentrations of PAA (100 and 200 ppm) and ASC (450 and 900 ppm) were tested, and both significantly reduced total viable bacteria and Campylobacter counts per carcass. Both sanitizers also reduced the prevalence of Salmonella on whole carcasses from both processing steps. Log reduction of both the total viable and Campylobacter counts were, however, temperature and processing stage dependent. The efficacy of both PAA and ASC were also compared with sodium hypochlorite. No significant difference between the three sanitizers was observed for the reduction of TVC, Campylobacter or Salmonella using carcasses obtained at either processing step. These results demonstrate that PAA or ASC could be implemented as a replacement or used in addition to sodium hypochlorite to effectively reduce bacteria on whole carcasses.

Biofilm formation on food-contact surfaces is a matter of major concern causing food safety and spoilage issues to this sector. The aim of this study was to assess the durability of the anti-biofilm capacity of a plasma-polymerized coating composed of a base coating of (3-aminopropyl)triethoxysilane (APTES) and a functional coating of acrylic acid (AcAc). Coated and uncoated AISI 316 stainless steel (SS) plates were subjected to five sanitization cycles with sodium hypochlorite (0.05%) and peracetic acid (0.5%). The effectiveness of the coating for the inhibition of multi-strain Listeria monocytogenes biofilm formation was confirmed using a three-strain cocktail, which was grown on the SS plates at 12 °C for 6 days. Compared to the uncoated SS, relative biofilm productions of 14.6% on the non-sanitized coating, 27.9% on the coating after sanitization with sodium hypochlorite, and 82.3% on the coating after sanitization with peracetic acid were obtained. Morphological and physicochemical characterization of the coatings suggested that the greater anti-biofilm effectiveness after sanitization with sodium hypochlorite was due to the high pH of this solution, which caused a deprotonation of the carboxylic acid groups of the functional coating. This fact conferred it a strong hydrophilicity and negatively charged its surface, which was favorable for preventing bacterial attachment and biofilm formation.

Human papillomaviruses (HPV) are non-enveloped DNA viruses infecting cutaneous and mucosal squamous epithelia. Sexually transmitted HPV-types that are carcinogenic to humans such as HPV16 can induce cervical and other anogenital cancers. Virus transmission through fomites such as inadequately disinfected gynecological equipment is a further potential transmission route. Since HPV cannot be easily grown in cell culture, polyomavirus SV40 has been used as a surrogate virus when testing the virucidal activity of chemical disinfectants. So far, studies that have compared the virucidal activity of different disinfectants against HPV and SV40 are lacking. Here, we evaluated the susceptibility of HPV16 pseudovirus and SV40 to seven active biocidal substances using quantitative suspension tests. Ethanol, glutaraldehyde (GTA), dodecyldipropylentriamin (DPTA), and ortho-phthalaldehydes (OPA) were able to reduce the infectivity of HPV16 pseudovirus &gt, 99.99% after 5 min. In contrast, isopropanol, peracetic acid (PAA), and quaternary ammonium compounds with alkylamines (QAC) only led to a slight or no reduction in infectivity. Concerning SV40, only GTA (60 min contact time), PAA, and OPA had virus-inactivating effects. In conclusion, the virucidal activity of three out of seven disinfectants tested was different for HPV16 pseudovirus and SV40. In this study, SV40 was shown to be a reliable surrogate virus for HPV when testing isopropanol-, GTA-, QAC-, and OPA-based disinfectants.

Antibiotics are commonly used as the first line of defense in the treatment of infectious diseases. However, the rise of antimicrobial resistance (AMR) is rendering many antibiotics less effective. Consequently, effective non-antibiotic antimicrobial strategies are urgently needed to combat AMR. This paper presents a strategy utilizing cold plasma for the "on-demand"activation of acetyl donor molecules. The process generates an aqueous-based antimicrobial formulation comprising a rich mixture of highly oxidizing molecules: peracetic acid, hydrogen peroxide, and other reactive oxygen and nitrogen species. The synergistic potent oxidative action between these molecules is shown to be highly effective at eradicating common wound pathogenic bacteria (Pseudomonas aeruginosa and Staphylococcus aureus) and at inactivating a virus (SARS-CoV-2). Refereed/Peer-reviewed

Apples are naturally coated with a water-repelling hydrophobic wax layer, which may limit the antimicrobial efficacies of surface sanitizer solutions. Lauric arginate (LAE) is a cationic surfactant with antimicrobial efficacy against Listeria monocytogenes. In this study, we investigated the antimicrobial and the wettability effects of LAE in enhancing anti-L. monocytogenes efficacy of peracetic acid (PAA) and further verified the optimized treatment combinations in a pilot spray-bar brush bed system. Apples after 48 h of inoculation were treated with PAA surface sanitation in combination with different concentrations of LAE at 22 or 46°C. The effectiveness of PAA with LAE solutions in decontaminating L. monocytogenes significantly increased with the increased concentration of PAA (60–80 ppm) or LAE (0.01–0.05%) or the treatment temperature (from 22 to 46°C). A 30–120-sec wash by 80 ppm PAA with 0.01 and 0.05% LAE at 22°C reduced L. monocytogenes on apples by 2.10–2.25 and 2.48–2.58 log10 CFU/apple, respectively. Including LAE in the PAA solution decreased contact angles on apple surfaces. However, the increased wettability of the sanitizer solution may not be the main contributor to the enhanced antimicrobial efficacy of the PAA solution, given that the addition of Tween 80 or Tween 85 only slightly boosted the anti-L. monocytogenes efficacy of PAA solutions though both increased the wettability of the PAA solutions. The synergistic effects of PAA and LAE were further validated in a pilot spray-bar brush bed packing system, where a 30-sec spray wash with 80 ppm PAA and 0.05% LAE at 22 and 46°C caused 1.68 and 2.08 log reduction of Listeria on fresh apples, respectively. This study provides an improved PAA process/preventive strategy for ensuring microbial food safety of fresh apples that is applicable to commercial apple packing lines.

Large amounts of water used in the apple packing process, the presence of organic matter, and difficult-to-clean equipment are vectors for contaminating apples with foodborne pathogens, such as Listeria monocytogenes. There is a need to develop new cleaning methods for fresh apples and evaluate their antimicrobial efficacy. A series of surfactants of different chemical properties (cationic lauric arginate [LAE], anionic sodium dodecyl sulfate [SDS], and nonionic Tween 20 [T20]) alone and combined with peracetic acid (PAA) were evaluated for their efficiency in the removal of L. innocua, a surrogate of L. monocytogenes, from fresh apples. Whole fresh apples were inoculated with L. innocua (7 log CFU/mL) by a dipping method, dried, and subjected to wash treatments with selected cleaning solutions (water, PAA, PAA-LAE, PAA-SDS, and PAA-T20). The contact angle between the cleaning solutions and the surface of the apples was measured. The antimicrobial activity of the cleaning solutions and the efficacy of the cleaning treatment were evaluated by enumeration of L. innocua from treated apples and visualized by scanning electron microscopy. Stem bowl and calyx cavities of the apple harbored higher bacteria concentrations (∼4.82 log CFU per apple), compared with the equatorial section (∼2.66 log CFU per apple). Addition of 0.1% of selected surfactants led to a significant decrease in surface tension of cleaning solutions and allowed better spreading on the apple surface. Surfactants combined with PAA solution resulted in higher L. innocua reduction compared with PAA alone; however, their efficacy was limited. The most effective cleaning solution was PAA-T20, with reduction of approximately 2.2 log. Scanning electron microscopy imaging confirmed that most bacteria were harbored inside the surface irregularities. PAA alone and with surfactants caused damage and deformation of bacteria cells. Cleaning apples with PAA combined with surfactants may improve microbial safety in whole apples; however, the efficiency of the decontamination treatment can be reduced because of variations in apple morphology.

Vibrio parahaemolyticus is a prevalent seafoodborne enteropathogen that has become a global concern since the spread of its pandemic strain in 1996. This study investigates the responses of this pathogen to the oxidative disinfectants hydrogen peroxide, chlorine dioxide, and peracetic acid. Expression of the regulator genes oxyR and rpoS, determined by reverse transcription PCR, in V. parahaemolyticus wild-type, oxyR mutant, and rpoS mutant strains exhibited similar patterns in response to the tested oxidative disinfectants. The transcription of the rpoS gene was markedly enhanced in the oxyR mutant strain in the exponential phase. The expression of catalase KatE1 was tracked by using a LacZ fusion reporter in these strains. The experimental results revealed that KatE1 was a significant scavenger of hydrogen peroxide and peracetic acid in V. parahaemolyticus, and RpoS may partially compensate for the regulatory role of OxyR in the oxyR mutant strain. In contrast to its responses to hydrogen peroxide and paracetic acid, KatE1 was not the primary scavenger of chlorine dioxide in these V. parahaemolyticus strains. This study shows that these disinfectants activated a basic oxidative response in this pathogen with different features.

Peracetic acid (PAA) is an emerging disinfectant with a low disinfection by-product formation potential, but how PAA destroys gene function after killing bacteria remains to be studied. Bacterial plasmid DNA is a mobile genetic element that often harbors undesirable genes encoding antibiotic resistance and virulence factors. Even though PAA efficiently kills bacteria, bacterial plasmids and other mobile genetic elements might still be intact and functional after PAA disinfection, posing potential public health and environmental risks. This study evaluated the impact of PAA disinfection on the functionality of plasmid DNA in vivo and compared the results with those from chlorination. We delivered a plasmid DNA harboring two antibiotic resistance genes to Escherichia coli TOP10 to form an antibiotic-resistant bacterium (ARB). The planktonic ARB was treated with PAA and chlorine to find the minimum doses inhibiting the regrowth of the strain. PAA and chlorine stopped the regrowth at 8 ± 1 mg PAA·L−1 and 20 ± 9 mg Cl2·L−1, respectively. The functionality of the plasmid DNA after PAA and chlorine disinfection was then determined at higher doses in vivo. Neither PAA nor chlorine completely destroyed the plasmid DNA. However, chlorine was more efficient than PAA in eliminating the plasmid DNA. PAA at 25 mg PAA·L−1 reduced the transforming activity of the plasmid DNA by less than 0.3 log10 units, whereas chlorine at 25 mg Cl2·L−1 reduced the transforming activity by approximately 1.7 log10 units. Chlorine had a more pronounced impact on the functionality of the plasmid DNA because it oxidizes or destroys bacterial components including plasmid DNA faster than PAA. In addition, environmental scanning electron microscopy shows that chlorination desiccated the cells resulting in the flat cellular structure and possibly more complete loss of plasmid DNA, whereas PAA disinfection had a less impact on cell structure and morphology. This study demonstrates that more plasmid DNA remains functional in water after PAA disinfection than after chlorination. These functional genetic elements could be acquired by other microorganisms via horizontal gene transfer to pose potential public health and environmental risks.

Information on the sensitivity of spoilage fungi of bakery products to sanitizing agents is scarce in the literature. Thus, the aim of this study was to evaluate the antifungal activity of different classes of commercial sanitizers, which have permitted use in the food industry, on the main fungi involved in spoiling bakery products. The tests were carried out according to the protocol for testing the antifungal effect of chemical sanitizers of the European Committee for Standardization (CEN), with adaptations. Different strains of six isolated fungal species responsible for spoiling bakery products (Penicillium roqueforti, Penicillium paneum, Hyphopichia burtonii, and Aspergillus pseudoglaucus) were tested against five sanitizers at three concentrations: benzalkonium chloride (0.3%, 2.5%, 5%), biguanide (2%, 3.5%, 5%), peracetic acid (0.15%, 1.5%, 3%), quaternary ammonium (0.3%, 2.5%, 5%), and sodium hypochlorite (0.01%, 0.1%, 0.2%). Peracetic acid was the most effective sanitizes considering the genera, species, and concentrations evaluated, generally being capable of reductions between 2 and 4 logs of initial control tested. Biguanide should not be the compound of choice when the main goal of the bakery industry is fungal control.

The main goal of this work was to approach food industry conditions in the comparison of the susceptibility of biofilms of Listeria monocytogenes to the biocides benzalkonium chloride (BAC) and peracetic acid (PAA). Twelve isolates of L. monocytogenes, including nine well characterized BAC resistant strains were used. Biofilms were produced on stainless steel coupons (SSC), at 11 °C (refrigeration temperature) or at 25 °C (room temperature), in culture media simulating clean (nutrient limiting) or soiled (nutrient rich) growth conditions. Neither different nutrient availability nor growth temperature showed significant effect (p > .05) on biofilm formation. PAA confirmed to be more effective than BAC in biofilm elimination. Biofilms formed under nutritional stress tended to differentiate more the response to BAC of the resistant or sensitive strains, but the resistant or sensitive phenotype of the planktonic cells did not dictate biofilm susceptibility.

In Canada, Salmonella-related foodborne illness accounts for more than 88,000 cases annually. Poultry products represent one of the major vectors for the zoonotic transmission of Salmonell...

Raw chia and flax seeds are increasingly associated with Salmonella contamination. However, intervention technologies for these seeds that maintain them in a raw state, without causing clumping because of mucilage production upon moisture exposure, are limited. In this study, a commercial ethanol and paracetic acid sanitizing solution meeting these criteria was evaluated for efficacy against Salmonella and Enterococcus faecium NRRL B-2354, a known Salmonella surrogate for thermal intervention technologies. Samples (100 g each) of chia and flax seeds ( n = 5) were inoculated with either a cocktail of Salmonella Newport, Senftenberg, Oranienburg, Saintpaul, Typhimurium DT104, and Cubana or E. faecium NRRL B-2354. After overnight acclimatization, samples were treated with 4 mL of sanitizing solution per sample and then held at ambient temperature (20 to 25°C) for 1 h before bacterial enumeration. Separate 1-kg-treated batches were evaluated for germination ability (4 replicates of 100-g samples), as well as nutrient content and rancidity ( n = 3), compared with untreated control. Following the posttreatment holding time, these batches were dried back to original moisture content at 70°C to evaporate residual sanitizing solution, thereby stopping treatment. The sanitizing solution was found to be an effective intervention method for chia and flax seeds, reducing Salmonella to below the level of detection by more than 4 and more than 5 average log CFU/g, respectively. Germination was not significantly affected ( P ≥ 0.05) for chia seed. For both seeds, nutrition and rancidity were not significantly affected ( P ≥ 0.05). Furthermore, E. faecium NRRL B-2354 was found to be an appropriate Salmonella surrogate for treatment of chia and flax seeds with this sanitizing solution, showing comparable but higher resistance to treatment with the sanitizing solution than the Salmonella cocktail.

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.