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This study evaluated the efficacy of using sequential forced air ozone followed by an advanced oxidative process (AOP) treatment to inactivate Listeria monocytogenes on and within Empire apples. The forced air ozone treatment consisted of a reactor that introduced ozone (6 g/h) into an airstream that flowed through an apple bed (ca. 30 cm in depth). Before treatment, the apples were conditioned at 4°C to ensure that condensate had formed before the apples were transferred to the reactor. The condensate ensured sufficient relative humidity to enhance the antimicrobial action of ozone. Air was passed through the apple bed at 9.3 m/s, and the ozone was introduced after 10 min. The ozone concentration measured after exiting the apple bed reached a steady state of 23 ppm. A 20-min ozone treatment supported a 2.12- to 3.07-log CFU reduction of L. monocytogenes, with no significant effect of apple position within the bed. The AOP-based method was a continuous process whereby hydrogen peroxide was introduced as a vapor into a reactor illuminated by UV-C and ozone-emitting lamps that collectively generated hydroxyl radicals. Operating the AOP reactor with UV-C light (54-mJ cm² dose), 6% (v/v) hydrogen peroxide, 2 g/h ozone, and a chamber temperature of 488C resulted in a 3-log CFU reduction of L. monocytogenes on the surface of the apples and internally within the scar tissue. Applying a caramel coating, from a molten solution (at 80°C), resulted in a 0.5-log CFU reduction of L. monocytogenes on the apple surface. In apples treated with the sequential process, L. monocytogenes could only be recovered sporadically by enrichment and did not undergo outgrowth when the caramel apples were stored at 22°C for 19 days. However, growth of L. monocytogenes within the core, but not the surface, was observed from caramel apples prepared from nontreated control fruit. [ABSTRACT FROM AUTHOR]

The development of a culture-free method for Salmonella screening of spent irrigation water derived from sprouting mung bean beds is described. The system used tangential flow filtration (TFF) to nonspecifically concentrate cells from large (2- to 10-liter) sample volumes. The retentate (100 ml) from the TFF was then flowed over an anti-Salmonella antibody-modified cellulose acetate membrane. The captured Salmonella was detected by reacting with a secondary anti-Salmonella and goat anti-rabbit biotin labeled antibody, followed by avidin-tagged glucose oxidase. The hydrogen peroxide generated from the enzymic oxidation of glucose was amperometrically detected at an underlying platinum electrode. It was found that 10 liters of Salmonella suspensions of 2 log CFU/ml could be concentrated to 4 log CFU/ml with 60% recovery regardless of the flow rate (112 to 511 ml/min) or transmembrane pressure (0 to 20 lb/in²) applied. The solids content of spent irrigation water negatively affected the filtration rate of TFE This was most evident in spent irrigation water collected in the initial 24 h of the sprouting period, where the solids content was high (4,170 mg/liter) compared with samples collected at 96 h (560 mg/ liter). Trials were performed using mung bean beds inoculated with different Salmonella levels (1.3 to 3.3 log CFU/g). By using the optimized TFF and flowthrough immunoassay it was possible to detect Salmonella in spent irrigation water at levels of 2.43 log CFU/ml within 4 h. The integrated concentration and detection system will provide a useful tool for sprout producers to perform in-house pathogen screening of spent irrigation water. [ABSTRACT FROM AUTHOR]

Raw milk is a well-established vehicle for the carriage of human pathogens, and many regulatory bodies have consequently mandated compulsory pasteurization as a food safety intervention. The residual activity of alkaline phosphatase (ALP) has historically been used to verify the adequacy of pasteurization of cow's milk. However, there is uncertainty on how the current ALP standards and methods of analysis can be applied to sheep's and goat's milk, which naturally contain different levels of the enzyme than that found in cow's milk. The official ALP methods applied in Canada (colorimetric assay; MFO-3) and in the United States (Fluorophos) were assessed for their ability to detect enzyme activity in raw and pasteurized milk derived from cows, sheep, and goats. The detection limit and the limit of quantitation were 0.8 and 2.02 µg/ml phenol, respectively, for the MFO-3 method and 43 and 85 mU/liter, respectively, for the Fluorophos method. The average ALP levels in raw goat's, cow's, and sheep's milk were 165, 1,562, and 3,512 µg/ml phenol, respectively. Raw milk detection limits, which correspond to raw milk phosphatase levels, were 0.051, 0.485, and 0.023% in cow's, goat's, and sheep's milk, respectively, for the MFO-3 method and 0.007, 0.070, and 0.004%, respectively, for the Fluorophos method. Although both methods can be used for ALP determination in cow's, goat's, and sheep's milk, the Fluorophos assay was superior to the colorimetric MFO-3 method based on sensitivity and time required to complete the analysis. [ABSTRACT FROM AUTHOR]

The efficacy of a produce decontamination method based on a combination of UV light (254 nm) and hydrogen peroxide (H2O2) to inactivate the MS2 F(+) coliphage inoculated onto iceberg lettuce was evaluated. Lettuce inoculated with 6.57 log PFU of MS2 was reduced by 0.5 to 1.0 log unit when illuminated with UV light alone for 20 to 60 s (12.64 to 18.96 mJ/ cm²). In contrast, a 3-log reduction in MS2 was achieved with 2% (vol/vol) H2O2 spray delivered at 50°C. No significant increase in log count reduction (LCR) was observed when H2O2 and UV light were applied simultaneously. However, H2O2 sprayed onto lettuce samples for 10 s, followed by a further 20-s UV illumination, resulted in an LCR of 4.12 that compares with the 1.67 obtained with 200 ppm of calcium hypochlorite wash. No further increase in MS2 inactivation was achieved by the use of either longer H2O2 spray or UV illumination times. The extent of MS2 reduction was significantly (P < 0.05) decreased when the H2O2 spray was delivered at 10 or 25°C compared with 50°C. In the course of aerobic storage at 4°C, lettuce treated with UV light and H2O2 (10 or 25°C) developed discoloration (polyphenol accumulation) within 6 days. In contrast, lettuce treated with UV light and H2O2 at 50°C developed less discoloration within this time period and was comparable to untreated controls. This study demonstrated that the combination of UV light and H2O2 represents an alternative to hypochlorite-based washes to reduce the carriage of viruses on fresh produce. [ABSTRACT FROM AUTHOR]

The interaction of a range of Salmonella serovars with pre- and postharvest tomatoes was evaluated. Serovars were selected on the basis of previous association in tomato-linked outbreaks of salmonellosis (Salmonella Javiana, Salmonella Montevideo, and Salmonella Newport) or those typically isolated from animal or clinical infections (Salmonella Dublin, Salmonella Enteritidis, Salmonella Hadar, Salmonella Infantis, Salmonella Typhimurium, and Salmonella Senftenberg). Salmonella serovars introduced onto the flowers of growing plants were recovered on and within the developing tomato fruit. Of ail the Salmonella serovars tested, Montevideo appeared to be more adapted to survival within tomatoes and was recovered from 90% of the fruit screened. Ail of the Salmonella serovars could persist and grow when introduced onto unripened (green) tomato fruit. In general, growth (internal and external) was promoted at the high incubation temperature (25°C) and high relative humidity (95%), although this was serovar dependent. The growth and persistence of Salmonella introduced on and into ripened (red) tomatoes was serovar dependent. Salmonella serovars Enteritidis, Typhimurium, and Dublin were less adapted to grow in or on intact red tomatoes than were serovars Hadar, Montevideo, or Newport. The results illustrated that a diverse range of Salmonella serovars can become established within and/or on preharvest tomatoes. The majority of Salmonella can grow and become established both on and within unripened tomatoes, but growth on ripened fruit was serovar dependent. The results provide a possible explanation why only a narrow range of Salmonella serovars are associated with foodborne illness outbreaks linked to tomatoes. [ABSTRACT FROM AUTHOR]

Aim: To determine the mechanisms by which a stabilized oxychloro (SOC)-based sanitizer, applied to decontaminate seeds destined for sprout production, inactivates Escherichia coli O157:H7 ph1 and Salmonella serotype Meleagridis. Materials and Results: The action of SOC on the metabolism, membrane and DNA integrity of Salmonella and E. coli O157:H7 was studied. In both pathogens, there was an oxidative burst and depletion of intracellular glutathione (GSH) upon initial exposure to 200 ppm SOC. Metabolic activity, measured via bioluminescence, decreased over a 4-h period in E. coli O157:H7 ph1 cells exposed to SOC. Membrane integrity, assessed through viability staining, decreased progressively over 23 h when exposed to SOC. The appearance of auxotrophic mutants suggested that DNA damage had also occurred. Enzymes rich in disulfide bonds (alkaline phosphatase and protease) were sensitive to the chlorite-based sanitizer. Through challenging other microbial types, it was found that Gram positive had higher tolerance to SOC than Gram negatives with the exception of Salmonella. MS2 bacteriophage was highly sensitive; however, Bacillus endospores were not inactivated by SOC. Conclusions: SOC inactivates E. coli O157:H7 and Salmonella through GSH oxidation and disruption of disulfide bonds. Ultimately, membrane damage resulting from prolonged exposure to SOC leads to the loss of cell viability. Significance and Impact of the Study: The results provide a basis for understanding why extended treatment times are required to inactivate bacteria using SOC. [ABSTRACT FROM AUTHOR]

The efficacy of a stabilized oxychloro-based food grade sanitizer to decontaminate seeds destined for sprout production has been evaluated. By using mung bean seeds as a model system, it was demonstrated that the sanitizer could be used to inactivate a five-strain cocktail of Escherichia coli O157:H7 or Salmonella introduced onto beans at 10³ to 104 CFU/g. Salmonella was more tolerant to stabilized oxychloro than was E. coli O157:H7, with sanitizer levels of >150 and >50 ppm, respectively, being required to ensure pathogen-free sprouts. The decontamination efficacy was also found to be dependent on treatment time (>8 h optimal) and the seed-to-sanitizer ratio (>1:4 optimal). Stabilized oxychloro treatment did not exhibit phytotoxic effects, as germination and sprout yields were not significantly (P > 0.05) different as compared with untreated controls. Although human pathogens could be effectively eliminated from mung beans, the aerobic plate count of native microflora on sprouts grown from treated seed was not significantly (P > 0.05) different from the controls. The diversity of microbial populations (determined through 16S rRNA denaturing gradient gel electrophoresis analysis) associated with bean sprouts was not significantly affected by the sanitizer treatment. However, it was noted that Klebsiella and Herbasprillum (both common plant endophytes) were absent in sprouts derived from decontaminated seed but were present in control sprouts. When a further range of seed types was evaluated, it was found that alfalfa, cress, flax, and soybean could be decontaminated with the stabilized oxychloro sanitizer. However, the decontamination efficacy with other seed types was less consistent. It appears that the rate of seed germination and putative activity of sanitizer sequestering system(s), in addition to other factors, may limit the efficacy of the decontamination method. [ABSTRACT FROM AUTHOR]

Abstract: The dissemination of enteric contaminants (generic Escherichia coli and Salmonella) associated with pork carcasses and contact surfaces within a high capacity (6000 carcasses per day) pork slaughter line was evaluated. Sponge samples were taken periodically from the holding area floor and carcasses at different points in the line throughout an 8.75 h production period. E. coli levels within the holding area were high (ca. 6 log cfu 100 cm−2) during the initial phase of processing and did not significantly increase throughout the activity period. In the course of dehairing carcasses, the levels of E. coli were significantly (p <0.05) reduced by scalding but increased during the scraping process. A combination of polishing and triple singeing reduced E. coli populations and the bacterium was only recovered sporadically on eviscerated carcasses. The E. coli populations associated with the slaughter line had a low diversity considering the large number of carcasses processed. In Visit I, the 665 E. coli isolates typed using ERIC–PCR could be grouped into 41 genotypes. In Visit II, 141 genotypes were identified among the 855 E. coli isolates tested. This would suggest that contamination on incoming pigs was of only minor significance compared to that present within the slaughterhouse environment. The holding area was shown to act as a reservoir for endemic E. coli genotypes that could be systematically transferred throughout the dressing line on carcasses. Indeed, the majority of genotypes could be re-isolated throughout the 8.75-h processing period. E. coli isolated from carcasses within the evisceration area could be traced to up-stream operations. The holding area and scraper operation were found to be the most important sites of cross-contamination. Fourteen genotypes recovered (primarily within the holding area) on Visit I were re-isolated on Visit II. Despite the presence of endemic E. coli populations, Salmonella was recovered from only two sites (holding area floor and a carcass within the cooler) on a single occasion. The two Salmonella recovered were genetically distinct (similarity index=22%) suggesting that they originated from different sources and were not part of an endemic population. The study has further illustrated the utility of molecular typing of generic E. coli isolates to establish the dynamics of enteric contamination within pork slaughter lines. However, the extent to which the distribution of E. coli can be extrapolated to that of Salmonella remains uncertain. [Copyright &y& Elsevier]

The reliability of testing spent irrigation water to assess the microbiological status of sprouting mung bean beds has been investigated. In commercial trials, the distribution of opportunistic contaminants within 32 bean sprout beds (25 kg of mung beans per bin) was assessed 48 h after germination. The prevalence of generic Escherichia coli, thermotolerant coliforms, and Aeromonas in sprouts (n = 288) was 5, 11, and 39%, respectively, and 57, 70, and 79% in the corresponding spent irrigation water samples (n = 96). Contamination was heterogeneously distributed within the seedbed. In laboratory trials, beans inoculated with a five-strain cocktail of either Salmonella or E. coli O157:H7 (10³ to 104 CFU/g) were introduced (1 g/500 g of noninoculated seeds) at defined locations (top, middle, or base), and the beans were then sprouted for 48 h. When seeds inoculated with pathogens were introduced at the base or top of the seedbed, the pathogens were typically restricted to these sites and resulted in 44% of the spent irrigation water samples returning false-negative results. Introducing inoculated beans into the middle or at the presoak stage enhanced the distribution of both pathogens within the subsequent sprout bed and resulted in comparable levels recovered in spent irrigation water. The study demonstrated that even though screening a single sample of spent irrigation water is more reliable than testing sprouts directly, it does not provide an accurate assessment of the microbiological status of sprouting mung bean beds. Such limitations may be addressed by ensuring that bean batches are mixed prior to use and by taking spent irrigation water samples from multiple sites at the latter stages of the sprouting process. [ABSTRACT FROM AUTHOR]

Abstract Background Clostridium difficile infection (CDI) is an important cause of morbidity and mortality internationally, yet there are important regional differences in the epidemiology and microbiology of disease. Most reports have come from North America and Europe, with limited information from other regions, including the Middle East. Given the changes in the epidemiology of CDI in developed countries, particularly associated with the dissemination of hypervirulent epidemic clones, an understanding of the epidemiology and microbiology of CDI in diverse regions is warranted. This study involved collection of stool samples from individuals with diarrhea at the Isfahan University of Medical Sciences Teaching Hospital, Isfahan, Iran, between October 2010 and March 2011. Selective enrichment culture for C. difficile was performed and isolates were characterised using ribotyping, PCR for the detection of tcdA, tcdB and cdtB genes, and tcdC sequence analysis. Findings Clostridium difficile was isolated from 19/89 (21%) stool samples of 17/86 (20%) patients. 13/17 (77%) cases of CDI were hospital-associated. Patients with CDI were significantly older (43 ± 28y) than those with non-CDI diarrhea (24, ± 26y)(P = 0.018). All isolates were toxigenic, and possessed genes encoding for toxins A and B. Six (32%) of 19 isolates also possessed cdtB. Twelve ribotypes were identified. Ribotype 078/toxinotype V was most common, accounting for 4 (21%) of isolates. A single isolate of a different toxinotype V ribotype was identified, as was a toxinotype XXIV isolate. The remaining isolates consisted of 9 different toxinotype 0 ribotypes. Conclusions CDI is an important cause of diarrhea in patients in this hospital. The diversity of ribotypes was striking, and the number of different types suggests the presence of a broad range of strains in the community, the hospital or both. The predominance of toxinotype V strains, which have been associated with community-associated disease and food animals, was unexpected and possible sources of this type require further investigation.

Abstract: The effect of different processing parameters on the efficacy of commercial post-harvest biocidal washes to decrease the bacterial loading on spinach and lettuce has been evaluated. Sampling was performed at two spinach processors (Facility A & B) and a shredded lettuce producer (Facility C). Aerobic colony counts (ACC) and coliform counts were determined on samples taken at pre- and post-wash. In parallel, the heterotrophic plate count (HPC) and coliform levels in wash water was also determined. Processing parameters measured were the temperature of leafy greens (pre- and post-washing) and wash water. The sanitizer levels (peroxyacetic acid, oxidation–reduction potential), pH, conductivity and turbidity were also measured. The wash process in Facility B had a residence time of 50s for the spinach, maintained a constant hypochlorite concentration and continuously re-charged the tanks with fresh water. In contrast, Facility A had a short residence time (15s) did not maintain a constant sanitizer (peroxyacetic acid) concentration or re-charge tanks with fresh water. Despite the differences in processing operations there was no statistical difference between the log count reductions (LCR) obtained in ACC and coliform counts although counts were only reduced by <0.6logcfu/g. The carriage of Escherichia coli on pre-wash spinach was 19% and 25% in Facility A and B respectively. There was a high prevalence (57% positive) of E. coli in the wash water of Facility A with none being recovered in water samples taken from Facility B. Yet, the carriage of E. coli on post-wash spinach was the same in the two facilities (7%). Lettuce harboured a lower level of both ACC and coliforms with LCR being significantly greater than spinach. In general, the LCR in ACC and coliforms could be positively correlated to bacterial counts of pre-washed leafy greens and conductivity (solids content) of the wash water. A negative correlation was found between LCR and water temperature. Interestingly, within the ranges measured the LCR was independent of the bacterial loading of the water. The results of the study confirmed the limited efficacy of biocidal washes to remove field acquired contamination. Although it is thought maintaining a low microbial loading in the wash water and maintaining sanitizer concentration is key the current study suggests high conductivity and low temperature of the wash water enhances the LCR achieved. [Copyright &y& Elsevier]

Air flow characteristics and decontamination efficacy are investigated within a forced-air ozone reactor designed to control Listeria monocytogenes on apple fruit. The commercial reactor introduced an air-ozone mix through a column of apples (540 kg contained within two bins; dimensions 1.21 m × 1.21 m × 0.72 m) for a defined air velocity and time period. The apple bed had high porosity (ɛ = 0.533) with a corresponding low pressure differential (10 Pa/m). Incoming air flowed through the ozone generator with low air velocities (0.022 m/s) introducing 39 ppm of the antimicrobial gas into the chamber. As the air flow was increased to 0.147 m/s, the corresponding ozone concentration at the entry point into the chamber dropped to 4 ppm through dilution. At the lowest air velocity the log reduction of Lactobacillus (surrogate bacteria for Listeria) inoculated onto applies varied between 0.5 and 2.5 log CFU across the bed, compared to 1.6 to 2.5 log CFU reductions that were observed at air velocities greater than 0.088 m/s. It was found the homogeneity in decontamination efficacy across the bed was primarily dependent on the applied air velocity with ozone concentration (4–39 ppm) and treatment time (5–40 min) being less influential. [ABSTRACT FROM AUTHOR]

Abstract: As part of investigative sampling there is a need to rapidly concentrate the target from large sample volumes to enable downstream detection using suitable diagnostic platforms. To this end the following describes a method based on tangential flow filtration (0.2 μm pore size) for concentrating Bacillus endospores from sample matrices (water, raw whole milk, apple juice concentrate and lettuce rinse samples). Bacillus endospores were selected as a model system as a representative select agent of significance. From optimization studies it was found that the recovery of endospores was in the order of 2.5% and could not be enhanced through increasing the flow rate or transmembrane pressure (TMP). However, recoveries could be increased to >80% (concentration factor 6.74–8.86) by backwashing the system with 10% w/v Tween 80 solution to detach spores adsorbed onto the internal surface of the filtration unit. The recovery of endospores from raw milk and apple juice concentrate was problematic due to the presence of solids or high viscosity. However, by performing an initial dilution (1:10) prior to passing through the Tangential Flow Filtration (TFF) system it was possible to achieve over 80% endospore recovery with no significant decrease in filter performance (Concentration Factor of 6–10). The recovery of endospores from lettuce surfaces was facilitated by rinsing with 4% w/v Tween 80 solution. When used in conjunction with tangential flow filtration system it was possible to recover endospores derived from lettuce rinse solutions at levels in the order of 300 cfu/ml. The results of the study illustrate that TFF is a useful approach for concentrating microbial targets from a diverse range of food matrices. [Copyright &y& Elsevier]

Abstract: There is interest in using bacteriophage as an indicator for the presence of pathogens, such as Salmonella, in health care and food processing environments. However, the current plaque assay technique to detect bacteriophages is time consuming and laboratory based. The following reports on a bacteriophage sensor based on conducting polymer organic electrodes modified with phage host cells (Salmonella Newport). Conducting polymer electrodes were fabricated by chemical deposition of polypyrrole onto the surface of a microporous polycarbonate membrane. The formed films exhibited quasi-reversible redox behaviour which was dominated by anion exchange although cations also contributed to the charge transfer kinetics. Salmonella host cells were absorbed onto the surface of the film and reacted with infecting bacteriophage in LBM broth at 37 °C. Upon bacteriophage mediated host cell lysis the impedance of the supporting polypyrrole electrode increased especially in relation to Z″. It is proposed that the increase in the dielectric properties of the polypyrrole layer was caused by the interaction of cellular constituents derived from the lysed cells. From dose response curves it was found that the sensor could detect 3 log Plaque Forming Units (pfu)/ml within 270 min although no linear correlation between phage concentration and sensor response was observed. [Copyright &y& Elsevier]

Chlorpyrifos is a widely used insecticide in apple production but there are concerns that residues ingested by consumers can lead to chronic neurological conditions. In the following, a method based on Advanced Oxidation Process (AOP) was validated for the degradation of chlorpyrifos on apples and inactivating Escherichia coli O157:H7, along with Aspergillus niger spores. AOP is a process that generates free-radicals from the UV-C (at 254 nm) mediated degradation of hydrogen peroxide thereby leading to highly oxidative species. By using Response Surface Methodology (RSM) it was found that the degradation of chlorpyrifos was primarily dependent on UV-C dose and synergistically enhanced by applying the hydrogen peroxide at temperatures up to 66 °C. However, the degradation of chlorpyrifos was independent on the hydrogen peroxide concentration up to 6% v/v. Maximal degradation of chlorpyrifos on apples was achieved through an AOP treatment applying 68.4 kJ/m2 UV dose and 1.22% v/v H 2 O 2 at 66 °C that resulted in 118 μg degradation (representing 59% of the original level) of the pesticide with <20 ppb accumulation of the degradative photoproduct, CPY-O. The same treatment supported a >6.6 log CFU reduction in E. coli O157:H7 and >4.7 log CFU reduction of A. niger spores. Applying UV alone in the case of E. coli O157:H7 or hydrogen peroxide alone against A. niger spores supported a lower level of lethality compared to AOP. In conclusion, the study has demonstrated that an AOP based process can support the degradation of chlorpyrifos and address microbiological hazards associated with apples. • An Advanced Oxidation Process was optimized for degrading chlopyrifos on apples. • UV-C contributed to degradation reaction to a greater extent than hydrogen peroxide. • An AOP process comprised of 69 kJ/m2 and 1,22% hydrogen peroxide supported 59% chlorpyrifos degradation. • Optimized treatment decreased E. coli O157 and Aspergillus niger spores to below the limit of detection. • The process developed can be applied to address chemical and microbial hazards associated with apples. [ABSTRACT FROM AUTHOR]

Wheat kernels harbor a diverse microflora that can negatively affect the suitability of the grains for further processing. To reduce surface microflora, a kernel disinfection method is required that does not affect grain functionality. Three different versions of gas phase hydroxyl-radical processes were compared with the common method for grain disinfection, that is, a bleach treatment. The gas phase hydroxyl-radicals are generated by the UV-C mediated degradation of hydrogen peroxide and/or ozone in a near water-free process. It was found that treating kernels with a bleach solution could reduce total aerobic count (TAC) and fungal count to below the level of enumeration. In comparison, the gas phase hydroxyl-radical treatment, that is, H 2 O 2 -UV-ozone treatment, could support a 1.3 log count reduction (LCR) in TAC and a 1.1 LCR in fungal count. The microbial load reduction for the wholemeal samples was less pronounced as endophytic microorganisms were less affected by all treatments, hinting at a limited penetration depth of the treatments. Despite reducing the microbial load on the kernel surface through the bleach and H 2 O 2 -UV-ozone treatments, none of these treatments resulted in a reduced microbial count on grains that underwent sprouting after the treatments. No negative effect on germination power or development of the seedling was observed for any of the treatments. The gluten aggregation behavior and xylanase activity of the wholemeal also remained unchanged after the gas phase hydroxyl-radical treatments. Our findings suggest that UV-H 2 O 2 -ozone treatment shows promise for dry-kernel disinfection, but further optimization of the processing parameters is required., (© 2023 The Authors. Journal of Food Science published by Wiley Periodicals LLC on behalf of Institute of Food Technologists.)

Eggs represent a significant vehicle for Salmonella Enteritidis with the pathogen being transferred to chicks in the hatchery, or to consumers via table eggs. In the following, the efficacy of a gas-phase hydroxyl-radical process for decontaminating hatchery and table eggs was evaluated. Recovery of Salmonella was maximized through holding eggs in tryptic soy broth containing 20% w/v glycerol for 1 h prior to plating. By using this technique, it was possible to recover 63% of the theoretical Salmonella inoculated onto eggs. The continuous hydroxyl-radical reactor consisted of a bank of UV-C lamps (254 nm) that generated hydroxyl-radicals from the degradation of hydrogen peroxide (H 2 O 2 ) mist and ozone gas. The optimal treatment was defined as that which supports a 5 log CFU/egg reduction of Salmonella without negatively affecting egg quality or leaving H 2 O 2 residues. A process of 2% v/v H 2 O 2 delivered at 30 mL/min with a UV-C dose of 19 mJ/cm 2 and ozone (20 ppm) with a total treatment time of 10s was selected. The egg quality metrics (Haugh value, yolk index, albumin pH, yolk pH) did not negatively differ over a 35-day shelf-life at 4 or 25℃ compared to washed eggs or nontreated controls. The cuticle layer of eggs remained intact following hydroxyl-radical treatment. Fertilized eggs (n = 61) treated with the hydroxyl-radicals exhibited the same hatchery rate (75%) as nontreated controls (71-79%) with no defects (unhealed navels or red hocks) being observed. The same hydroxyl-radical treatment could be applied to table eggs to support >5 log CFU/egg reduction of Salmonella and was compatible with egg washing regimes practiced in industry. In comparison, the egg washing process based on sodium hydroxide and chlorine supported a 2.76 ± 0.38 log CFU/egg reduction of Salmonella. The hydroxyl-radical treatment represents a preventative control step to reduce the carriage of Salmonella on hatchery and table eggs., 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 The Author(s). Published by Elsevier Inc. All rights reserved.)

The emergence of antibiotic-resistant bacteria due to the overuse or inappropriate use of antibiotics has become a significant public health concern. The agri-food chain, which serves as a vital link between the environment, food, and human, contributes to the large-scale dissemination of antibiotic resistance, posing a concern to both food safety and human health. Identification and evaluation of antibiotic resistance of foodborne bacteria is a crucial priority to avoid antibiotic abuse and ensure food safety. However, the conventional approach for detecting antibiotic resistance heavily relies on culture-based methods, which are laborious and time-consuming. Therefore, there is an urgent need to develop accurate and rapid tools for diagnosing antibiotic resistance in foodborne pathogens. This review aims to provide an overview of the mechanisms of antibiotic resistance at both phenotypic and genetic levels, with a focus on identifying potential biomarkers for diagnosing antibiotic resistance in foodborne pathogens. Furthermore, an overview of advances in the strategies based on the potential biomarkers (antibiotic resistance genes, antibiotic resistance-associated mutations, antibiotic resistance phenotypes) for antibiotic resistance analysis of foodborne pathogens is systematically exhibited. This work aims to provide guidance for the advancement of efficient and accurate diagnostic techniques for antibiotic resistance analysis in the food industry., (© 2023 Institute of Food Technologists®.)

A continuous Photo-Fenton Advanced-Oxidation-Process (AOP) for reducing the chlorine-demand of spent lettuce wash water was developed based on the generation of hydroxyl-radicals from the UV-C degradation of hydrogen peroxide in the presence of ferric-catalyst. It was found that an interaction between UV-C and hydrogen peroxide or ferric-catalyst concentration was associated with high hydroxyl-radical generation as determined from the oxidation of methylene blue. The optimal AOP treatment was identified as 320 mJ/cm 2 UV-C dose, 9.6 mg/L H 2 O 2, and 9 mg/L ferric-catalyst. When the treatment was applied to simulated lettuce spent wash water (6.6 g romaine lettuce per liter of distilled water containing 100 mg bentonite; pH 6.9) the chlorine demand was reduced from 150 ppm to 130 ppm. The chlorination of AOP treated water did not result in a greater log reduction of pathogens (Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella) on lettuce but did reduce cross-contamination between batches during washing. The chlorinated byproducts formed in AOP treated water exhibited higher antimicrobial activity compared to untreated controls. Although the treatment was successful in reducing cross-contamination of lettuce batches the cytotoxicity of disinfection byproducts requires to be assessed., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The following reports on the generation of hydroxyl-radical activated water prepared by passing a hydrogen peroxide solution containing Fe(III) catalyst through a UV-C reactor. The activated water was subsequently evaluated for antimicrobial activity against Escherichia coli O157:H7 in suspension or when inoculated onto mung beans. Hydroxyl-radical generation was assessed through the oxidation of methylene blue when reacted with activated water prepared from solutions of different pH (4-10), UV-C dose (32-128 mJ/cm 2 ), hydrogen peroxide (0-1000 mg/L) and Fe(III) concentration (0-100 mg/L). Methylene blue oxidation was associated with high concentrations of each reactant with a positive correlation with Fe(III) concentration. Inactivation curves of E. coli O157:H7 in activated water were diphasic with an initial slow rate that increased after 15 min contact time. In contrast to the methylene blue assay, the antimicrobial action of activated water was associated with high hydrogen peroxide (500 mg/mL) and low Fe(III) catalyst (1 mg/L) with no significant interaction with UV-C dose. Evidence would suggest that the mode-of-inactivation was through a radical propagation reaction that is rate-limited by the reduction of Fe (III) to Fe (II). Here, the initial activation process via UV-C illumination results in photo-reduction of Fe(III) and propagates the formation of hydroxyl-radicals. Fe(III) to Fe(II) cycling continues with oxidation of cell structures that ultimately leads to loss of viability due to accumulation of cellular damage. When activated water was used to soak mung beans inoculated with E. coli O157:H7 a 1 log reduction was obtained with a 19% increase in germinated beans and 8.5% higher sprout yield relative to controls. The oxidation reduction potential decreased from 477 mV to 288 mV and pH increased from 3.97 to 5.47, over the 24 h mung bean soak period. The reduction of Salmonella and Listeria monocytogenes on mung beans soaked in activated water was <1 log CFU/g with all three pathogens growing back over the sprouting period. From the results it can be concluded that activated water can enhance the germination of mung beans along with sprout yield but has limited capacity when applied alone as a seed disinfection method., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Abstract: Processes based on generating vapor-phase hydroxyl radicals or chlorine radicals were developed for inactivating Listeria monocytogenes on mushrooms without negatively affecting quality. Antimicrobial radicals were generated from the UV-C degradation of hydrogen peroxide or hypochlorite and ozone gas. Response surface modeling was used to identify the interaction among the operating parameters for the hydroxyl radical process: UV-C254nm intensity, hydrogen peroxide concentration, and ozone delivered. There was an inverse relationship between hydrogen peroxide concentration and UV-C intensity in terms of the log reduction of L. monocytogenes. The independent parameters for the chlorine radical process were hypochlorite concentration, pH, and UV-C intensity. From predictive models, the optimal hydroxyl radical treatment was found to be 5% (v/v) H2O2, 2.86 mW/cm2 UV-C intensity (total UV-C dose 144 mJ/cm2), and 16.5 mg of ozone. The optimal parameters for the chlorine radical process were 10 ppm of hypochlorite (pH 3.0), 11.0 mg of ozone, and 4.60 mW/cm2 UV-C intensity. When inoculated mushrooms were treated with the optimal hydroxyl radical and chlorine radical processes, the reduction of L. monocytogenes was found to be 2.42 ± 0.42 and 2.61 ± 0.30 log CFU, respectively, without any negative effects on mushroom quality (weight loss and Browning index during 14 days of storage at 4°C). These reductions were significantly greater than those from application of the individual elements of the radical processes and those in the control process, which used a 90-s dip in 1% (v/v) hydrogen peroxide. The study has demonstrated that hydroxyl radical and chlorine radical vapor-phase treatments are equally effective at inactivating L. monocytogenes on mushrooms and can be considered as a preventative control step., (Copyright ©, International Association for Food Protection.)

Persisters are a form of dormancy in bacteria that provide temporary resistance to antibiotics. The following reports on the formation of Escherichia coli O157:H7 E318 type II persisters from a protracted (8 days) challenge with ampicillin. Escherichia  coli O157:H7 followed a multiphasic die-off pattern with an initial rapid decline (Phase I) of susceptible cells that transitioned to a slower rate representing tolerant cells (Phase II). After 24 h post-antibiotic challenge, the E. coli O157:H7 levels remained relatively constant at 2 log CFU/mL (Phase III), but became non-culturable within 8-days (Phase IV). The revival of persisters in Phase III could be achieved by the removal of antibiotic stress, although those in Phase IV required an extended incubation period or application of acid-shock. The carbon utilization profile of persister cells was less diverse compared with non-persisters, with only methyl pyruvate being utilized from the range tested. Inclusion of methyl pyruvate in tryptic soy agar revived non-cultural persisters, presumably by stimulating metabolism. The results suggest that persisters could be subdivided into culturable or non-culturable cells, with the former representing a transition state to the latter. The study provided insights into how to revive cells from dormancy to aid enumeration and control.

Feeding practices have been found to influence gut microbiota which play a major role in immunity of poultry. In the present study, changes in cecal microbiota and humoral responses resulting in the 55 ppm bacitracin (BACI), 1% each of cranberry (CP1) and wild blueberry (BP1) pomace alone or in combination (CP+BP) feeding in broiler Cobb 500 vaccinated or not against coccidiosis were investigated. In the non-vaccinated group, no significant treatment effects were observed on performance parameters. Vaccination significantly affected bird's performance parameters particularly during the growing phase from 10 to 20 days of age. In general, the prevalence of coccidiosis and necrotic enteritis (NE) was reduced by vaccination ( P < 0.05). BACI-treated birds showed low intestinal lesion scores, and both CP1 and BP1 feed supplementations reduced Eimeria acervulina and Clostridium perfringens incidences similar to BACI. Vaccination induced change in serum enzymes, minerals, and lipid levels in 21-day old birds while, levels of triglyceride (TRIG) and non-esterified fatty acids (NEFA) were higher ( P < 0.05) in CP1 treated non-vaccinated group than in the control. The levels of NEFA were lower in BACI- and CP1-fed birds than in the control in non-vaccinated day 28 old birds. The highest levels of all estimated three immunoglobulins (IgY, IgM, and IgA) were found in the vaccinated birds. Metagenomics analysis of the cecal bacterial community in 21-day old birds showed the presence of Firmicutes (90%), Proteobacteria (5%), Actinobacteria (2%), and Bacteroidetes (2%). In the vaccinated group, an effect of BACI was noted on Proteobacteria ( P = 0.03). Vaccination and/or dietary treatments influenced the population of Lactobacillaceae , Enterobacteriaceae, Clostridiaceae , and Streptococcaceae which were among the most abundant families. Overall, this study revealed that besides their beneficial effects on performance, alike bacitracin, berry pomaces in poultry feed have profound impacts on the chicken cecal microbiota and blood metabolites that could be influenced by vaccination against coccidiosis., Competing Interests: 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 © 2021 Her Majesty the Queen in Right of Canada, as represented by the Minister of Agriculture and Agri-Food Canada.)

A gas-phase Advanced Oxidation Process (gAOP) was evaluated for decontaminating N95 and surgical masks. The continuous process was based on the generation of hydroxyl-radicals via the UV-C (254 nm) photo-degradation of hydrogen peroxide and ozone. The decontamination efficacy of the gAOP was dependent on the orientation of the N95 mask passing through the gAOP unit with those positioned horizontally enabling greater exposure to hydroxyl-radicals compared to when arranged vertically. The lethality of gAOP was independent of the applied hydrogen peroxide concentration (2-6% v/v) but was significantly (P<0.05) higher when H2O2 was introduced into the unit at 40 ml/min compared to 20 ml/min. A suitable treatment for N95 masks was identified as 3% v/v hydrogen peroxide delivered into the gAOP reactor at 40 ml/min with continuous introduction of ozone gas and a UV-C dose of 113 mJ/cm2 (30 s processing time). The treatment supported >6 log CFU decrease in Geobacillus stearothermophilus endospores, > 8 log reduction of human coronavirus 229E, and no detection of Escherichia coli K12 on the interior and exterior of masks. There was no negative effect on the N95 mask fitting or particulate efficacy after 20 passes through the gAOP system. No visual changes or hydrogen peroxide residues were detected (<1 ppm) in gAOP treated masks. The optimized gAOP treatment could also support >6 log CFU reduction of endospores inoculated on the interior or exterior of surgical masks. G. stearothermophilus Apex spore strips could be applied as a biological indicator to verify the performance of gAOP treatment. Also, a chemical indicator based on the oxidative polymerization of pyrrole was found suitable for reporting the generation of hydroxyl-radicals. In conclusion, gAOP is a verifiable treatment that can be applied to decontaminate N95 and surgical masks without any negative effects on functionality., Competing Interests: The authors have declared that no competing interests exist.

Agricultural pesticide use is ongoing and consumer concern regarding the safety of pesticide residues on produce has generated interest in techniques that can safely reduce residues post-harvest. Recently an advanced oxidative process has shown promise in substantial residue reduction on the surface of produce. Given the potential for oxidative transformation of pesticides to generate transformation products with greater toxicity than the parent residue, take for example the oxon products of the organophosphorus insecticides, it is important to consider what transformation products are generated by pesticide exposure to an oxidative process and their potential toxicity. In this study, previously published transformation products of boscalid, pyraclostrobin, fenbuconazole and glyphosate were identified after exposure to 3% hydrogen peroxide, UV-C irradiation or their combination in an advanced oxidative process on glass, their oral toxicity, carcinogenicity and developmental toxicity were identified in-silico and an initial tier hazard assessment was conducted. Of the 87 total structures that were searched for, 53 were detected by UPLC-QTOF-MS and identified by mass spectra: 15, 13, 22 and 3 structures for boscalid, pyraclostrobin, fenbuconazole and glyphosate respectively, including the parent residues. Oral toxicity of the transformation products of pyraclostrobin and glyphosate was similar to or lower than the parent residue. Several transformation products of boscalid and fenbuconazole were estimated to be significantly more orally toxic than their parent residues. While the majority of the transformation products of boscalid, pyraclostrobin and fenbuconazole were predicted to be carcinogenic there were 11 that were consistently identified to have carcinogenic potential by several assessments. 29 of the 53 molecules were predicted to be probable developmental toxicants. An initial tier hazard assessment was conducted for Cramer rules classification and mutagenicity using the threshold of toxicological concern approach and predicted rat oral LD 50 . Two exposure scenarios were considered, one highly protective considering each transformation product to be at the highest maximum residue limit (MRL) for the pesticide and whole produce consumption at the highest consumption rate from the USEPA Exposures Handbook, the other considering only apple consumption with the relevant MRL. As indicated by the hazard assessment, several transformation products of boscalid, pyraclostrobin and fenbuconazole should be strongly considered for further testing, either by quantifying their production or in-vivo and in-vitro toxicity tests due to their predicted toxicity and associated hazard., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

With the pressure to reduce antibiotics use in poultry production, cost-effective alternative products need to be developed to enhance the bird's immunity. The present study evaluated the efficacy of cranberry fruit by-products to modulate immunity in broiler chickens. Broiler Cobb 500 chicks were fed a control basal diet, basal diet supplemented with bacitracin (BACI, 55 ppm), cranberry pomace at 1% and 2% (CP2), or cranberry pomace ethanolic extract at 150 and 300 ppm (COH300) for 30 d. Blood sera were analyzed at days 21 and 28 of age for Ig levels by ELISA. The innate and adaptive immune-related gene expression levels in the liver and bursa of Fabricius were investigated at 21 d of age by quantitative polymerase chain reaction arrays. At day 21, the highest IgY level was found in the blood serum of the CP2-fed birds. In the liver, 13 of the 22 differentially expressed genes were downregulated across all treatments compared with the control. Expression of genes belonging to innate immunity such as caspase 1 apoptosis-related cysteine peptidase, chemokine receptor 5, interferon gamma, myeloid differentiation primary response gene 88, and Toll-like receptor 3 were significantly downregulated mainly in BACI- and COH300-fed birds. In the bursa, 5 of 9 genes associated with the innate immunity were differentially expressed. The expression of anti-inflammatory IL-10 gene was upregulated in all treatment groups in bursa compared with the control. The expression of transferrin gene was significantly upregulated in livers of birds fed COH300 and in bursa of birds fed BACI, indicating feeding practices and organ-dependant modulation of this gene in broiler. Overall results of this study showed that cranberry product feed supplementation modulated the innate immune and suppressed proinflammatory cytokines in broilers, providing a platform for future investigations to develop berry products in poultry feeding., (Crown Copyright © 2020. Published by Elsevier Inc. 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.)

A method based on vapor-phase advanced oxidation process (AOP) for decontaminating red or green grapes was validated for inactivating Listeria monocytogenes and spoilage molds. A Central Composite Design (CCD) and Response Surface Methodology (RSM) were applied to determine the contribution of UV-C (254 nm) dose, hydrogen peroxide, and ozone concentration on the lethality toward Aspergillus niger spores (biodensiometer) and changes to the grape quality (firmness and color over 14-day post-treatment storage at 4 °C). A high UV-C dose (>129 mJ/cm 2 ) or >4.0 % v/v hydrogen peroxide induced-blistering and darkening of grapes at the end of the storage period. Yet, an optimized AOP treatment (with regards to preserving grape quality) was derived to be 1.3% v/v hydrogen peroxide (5 mL/10 berries) with 9-mg ozone gas and a UV-C dose of 123 mJ/cm 2 (10 s at UV-C intensity of 12 mW/cm 2 ). A predictive model was constructed and verified based on the log reduction of A. niger spores and changes in quality characteristics of red grapes. The optimal AOP treatment supported a 1.6-log CFU/g reduction of Aspergillus spores and decreased L. monocytogenes counts by 3.92 ± 0.17 and 4.77 ± 0.30 log CFU/g on green and red grapes, respectively, that were not significantly different to the surrogate, Lactobacillus fructivorans. There was no significant difference in the reduction of L. monocytogenes with grapes arranged in a single or double layer. Botrytis cinerea counts were reduced by 1.08 to 1.35 log CFU/g using the optimized AOP treatment with no change in grape color or firmness during storage. A sensory panel could not differentiate AOP-treated grapes from nontreated controls although 3 of 15 panelists did note subtle flavor notes. PRACTICAL APPLICATION: Postharvest washing of fresh produce has limited efficacy in removing foodborne pathogens and spoilage microbes. This is especially relevant to berries, such as grapes, that are susceptible to spoilage following washing. The vapor-phase AOP treatment provides a supplemental or alternative approach for produce decontamination. However, the operating parameters need to be optimized to ensure that decontamination of grapes is not at the expense of quality. In the current study, this was achieved by ensuring a balance between hydrogen peroxide, ozone, and UV-C dose that form the elements of an AOP treatment., (© 2020 Institute of Food Technologists®.)

Evidence supporting the effectiveness of the PEERS® social skills intervention for adolescents with autism spectrum disorder is relatively strong. Less is known about whether the duration of the program impacts participant outcomes. The current study compared outcomes of participants randomly assigned to participate in an accelerated version of PEERS that met twice weekly for 7 weeks (n = 11) or the traditional PEERS program that met once weekly for 14 weeks (n = 10). The accelerated PEERS group demonstrated improvements consistent with previous research on the program, and treatment response did not differ significantly between the accelerated PEERS and traditional PEERS groups. Together, findings provide preliminary evidence that PEERS is effective when administered as a 7-week program.