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Chlorine is commonly used by the fresh produce industry to sanitize water and minimize pathogen cross-contamination during handling. The pH of chlorinated water is often reduced to values of pH 6-7, most commonly with citric acid to stabilize the active antimicrobial, hypochlorous acid (a form of free chlorine). Previous studies have demonstrated that citric acid reacts with chlorine to form trichloromethane, a major chlorine by-product in water and a potential human carcinogen. However, it is unclear if other pH control agents could be used in the place of citric acid to minimize the formation of trichloromethane. The objective of the present study was to determine the reactivity of organic and inorganic pH control agents, with chlorine, to generate trichloromethane. Free chlorine (∼100 mg/L) was mixed with 10 mM of each of twelve organic acids and two inorganic pH control agents (i.e., sodium acid sulfate and phosphoric acid) to effect a pH level of 6.5. Free chlorine and trichloromethane levels were measured over 3 h at 3 and 22°C. Results demonstrated that ascorbic acid, dehydroascorbic acid, citric acid, and malic acid rapidly depleted free chlorine concentrations at both 22°C and 3°C, while tartaric acid and lactic acid decreased chlorine concentrations more slowly. Other pH control agents did not significantly reduce free chlorine either at 22 or 3°C. Citric acid led to the generation of significantly higher concentrations of trichloromethane than did other acids. Chloroacetone was also found in chlorinated water in the presence of citric acid and ascorbic acid. Taking buffering capacity and pKa values into account, phosphoric acid and some organic acids may be used to replace citric acid as pH control agents in chlorinated water for washing fresh produce, to stabilize free chlorine level and reduce the generation of trichloromethane., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier Inc.)

Salmonella is capable of surviving dehydration within various foods, such as dried fruit. Dried fruit, including apple slices, have been the subject of product recalls due to contamination with Salmonella. A study was conducted to determine the fate of Salmonella on apple slices, following immersion in three antimicrobial solutions (viz., ε-polylysine [epsilon-polylysine or EP], sodium bisulfate [SBS], or peracetic acid [PAA]), and subsequent hot air dehydration. Gala apples were aseptically cored and sliced into 0.4 cm thick rings, bisected, and inoculated with a five-strain composite of desiccation-resistant Salmonella, to a population of 8.28 log CFU/slice. Slices were then immersed for 2 min in various concentrations of antimicrobial solutions, including EP (0.005, 0.02, 0.05, and 0.1%), SBS (0.05, 0.1, 0.2, and 0.3%), PAA (18 or 42 ppm), or varying concentrations of PAA + EP, and then dehydrated at 60°C for 5 h. Salmonella populations in positive control samples (inoculated apple slices washed in sterile water) declined by 2.64 log after drying. In the present study, the inactivation of Salmonella, following EP and SBS treatments, increased with increasing concentrations, with maximum reductions of 3.87 and 6.20 log (with 0.1 and 0.3% of the two compounds, respectively). Based on preliminary studies, EP concentrations greater than 0.1% did not result in lower populations of Salmonella. Pretreatment washes with either 18 or 42 ppm of PAA inactivated Salmonella populations by 4.62 and 5.63 log, respectively, following desiccation. Combining PAA with up to 0.1% EP induced no greater population reductions of Salmonella than washing with PAA alone. The addition of EP to PAA solutions appeared to destabilize PAA concentrations, reducing its biocidal efficacy. These results may provide antimicrobial predrying treatment alternatives to promote the reduction of Salmonella during commercial or consumer hot air drying of apple slices., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier Inc.)

Antimicrobial properties of biochar have been attributed to its ability to inactivate foodborne pathogens in soil, to varying degrees. High concentrations of biochar have reduced E. coli O157:H7 in soil and dairy manure compost, based on alkaline pH. Preliminary studies evaluating 31 different biochars determined that two slow pyrolysis biochars (paper biochar and walnut hull cyclone biochar) were the most effective at inactivating E. coli in soil. A study was conducted to determine the lowest percentages of paper and walnut hull cyclone biochars needed to reduce E. coli O157:H7 in soil. A model soil was adjusted to 17.75% moisture, and the two types of biochar were added at concentrations of 1.0, 1.5, 2.0, 2.5, 3.5, 4.5, 5.5, and 6.5%. Nontoxigenic E. coli O157:H7 were inoculated into soil at 6.84 log CFU/g and stored for up to 6 weeks at 21°C. Mean E. coli O157:H7 counts were 6.01-6.86 log CFU/g at all weeks between 1 and 6 in soil-only positive control samples. Populations in all soil amended with 1.0 and 1.5% of either type of biochar (as well as 2.0% of the walnut hull biochar) resulted in ≤0.68 log reductions at week 6, when compared with positive controls. All other concentrations (i.e., ≥2.0% paper and ≥2.5% walnut hull) inactivated ≥2.7 log at all weeks between 1 and 6 (p < 0.05). At the end of 6 weeks, E. coli O157:H7 declined by 2.84 log in 2.0% paper biochar samples, while concentrations of between 2.5 and 6.5% paper biochar completely inactivated E. coli O157:H7, as determined by spiral plating, at weeks 5 and 6. In contrast, 2.0% walnut hull biochar lowered populations by only 0.38 log at week 6, although 2.5-6.5% concentrations of walnut hull biochar resulted in complete inactivation at all weeks between 3 and 6, as assessed by spiral plating. In summary, ≥2.5% paper or walnut hull biochar reduced ≥5.0 log of E. coli O157:H7 during the 6-week storage period, which we attribute to high soil alkalinity. Amended at a 2.5% concentration, the pH of soil with paper or walnut hull biochar was 10.67 and 10.06, respectively. Results from this study may assist growers in the use of alkaline biochar for inactivating E. coli O157:H7 in soil., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier Inc.)

Fresh-cut apples, being rich in antioxidants and other nutrients, have emerged as popular snacks in restaurants, at home, and in school lunch programs, partially due to freshness, convenience, and portion size availability. Two major challenges in processing fresh-cut apples are the browning of cut surfaces and contamination with human pathogens. Regarding human pathogens, contamination by Listeria monocytogenes is a major concern, as evidenced by two outbreaks of whole apples and numerous recalls of fresh-cut apples. Antibrowning agents currently used by the industry have little to no antimicrobial properties. The present review discusses the possible origins of L. monocytogenes in fresh-cut apples, including contaminated whole apples, and contamination via the processing environment and the equipment in fresh-cut facilities. Treatment with antibrowning solutions could possibly be an opportunity for Listeria contamination and represents the last chance to inactivate pathogens. The discussion is focused on the antibrowning treatments where formulations and coatings with antibrowning and antimicrobial properties have been developed and evaluated against Listeria and other microorganisms. In addition, several research needs and considerations are discussed to further reduce the chance of pathogen contamination on fresh-cut apples., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier Inc.)

Abstract: Consumer safety concerns over established fresh produce washing methods and the demand for organic and clean-label food has led to the exploration of novel methods of produce sanitization. Essential oils (EOs), which are extracted from plants, have potential as clean-label sanitizers because they are naturally derived and act as antimicrobials and antioxidants. In this review, the antimicrobial effects of EOs are explored individually and in combination, as emulsions, combined with existing chemical and physical preservation methods, incorporated into films and coatings, and in vapor phase. We examined combinations of EOs with one another, with EO components, with surfactants, and with other preservatives or preservation methods to increase sanitizing efficacy. Components of major EOs were identified, and the chemical mechanisms, potential for antibacterial resistance, and effects on organoleptic properties were examined. Studies have revealed that EOs can be equivalent or better sanitizing agents than chlorine; nevertheless, concentrations must be kept low to avoid adverse sensory effects. For this reason, future studies should address the maximum permissible EO concentrations that do not negatively affect organoleptic properties. This review should be beneficial to food scientists or industry personnel interested in the use of EOs for sanitization and preservation of foods, including fresh produce., (Published 2022 by the International Association for Food Protection.)

A laboratory-acquired E. coli O157:H7 infection with associated severe sequelae including hemolytic uremic syndrome occurred in an individual working in the laboratory with a mixture of nalidixic acid-resistant (Nal R ) O157:H7 mutant strains in a soil-biochar blend. The patient was hospitalized and treated with an intravenous combination of metronidazole and levofloxacin. The present study investigated the source of this severe laboratory acquired infection and further examined the influence of the antibiotics used during treatment on the expression and production of Shiga toxin. Genomes of two Stx 2a -and eae -positive O157:H7 strains isolated from the patient's stool were sequenced along with two pairs of the wt strains and their derived Nal R mutants used in the laboratory experiments. High-resolution SNP typing determined the strains' individual genetic relatedness and unambiguously identified the two laboratory-derived Nal R mutant strains as the source of the researcher's life-threatening disease, rather than a conceivable ingestion of unrelated O157:H7 isolates circulating at the same time. It was further confirmed that in sublethal doses, the antibiotics increased toxin expression and production. Our results support a simultaneous co-infection with clinical strains in the laboratory, which were the causative agents of previous O157:H7 outbreaks, and further that the administration of antibiotics may have impacted the outcome of the infection., 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 © 2022 Eppinger, Almería, Allué-Guardia, Bagi, Kalalah, Gurtler and Fratamico.)

Abstract: A study was conducted to evaluate a new organic mixed peroxyacid solution produce wash composed of a combination of organic acids (lactic acid and one or more fruit acids) and hydrogen peroxide for activity against foodborne pathogens. The mixed peroxyacid was challenged against Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes in suspension or on the surface of dip-inoculated cherry tomatoes. Cherry tomatoes were also treated with 8 ppm of free chlorine in the form of sodium hypochlorite. When tested against planktonic cells of Salmonella and E. coli O157:H7 in pure culture for 120 s, these pathogens were reduced by 7.5 and 7.1 log CFU/mL, respectively, by the 0.40% peroxyacid solution, and L. monocytogenes was decreased by 5.0 log CFU/mL by the 0.80% solution. When cherry tomatoes were dip inoculated and treated with 8 ppm of free chlorine, Salmonella and E. coli O157:H7 populations decreased by 2.5 and 2.6 log CFU/g, respectively; these reductions were not significantly different from those obtained after sterile water rinses. However, the 1.0% peroxyacid solution reduced the same microorganisms by 3.8 and 3.4 log CFU/g, respectively, which was significantly greater (P < 0.05) than the reductions achieved by the 2-min sterile water rinse. For tomatoes dip inoculated with L. monocytogenes, populations were reduced by 3.5 log CFU/g by the 1.0% peroxyacid solution, which was significantly greater (P < 0.05) than reductions achieved by 8 ppm of free chlorine (2.6 log CFU/g) or sterile water (1.7 log CFU/g). These results indicate that this peroxyacid combination is an effective organic antimicrobial agent for preventing cross-contamination during the washing of cherry tomatoes and can inactivate S. enterica, E. coli O157:H7, and L. monocytogenes by up to 3.8, 3.4, and 3.5 log CFU/g, respectively., (Published 2022 by the International Association for Food Protection. Not subject to U.S. Copyright.)

Abstract: Mesquite flour with endogenous high sugar content is often contaminated with Bacillus cereus. The purpose of the present study was to evaluate the thermal resistance of Bacillus spp. in naturally contaminated mesquite flour. Flours with and without adjusted water activity (aw) were treated at various temperatures (100 to 140°C) and times (up to 2 h). Total mesophilic bacteria and Bacillus spp. were enumerated using tryptic soy agar and Brilliance Bacillus cereus Agar, respectively. Results revealed that naturally contaminated Bacillus spp. and other mesophilic bacteria in mesquite flour (aw = 0.34) were highly resistant to heat. To reduce the initial populations (4.75 log CFU/g) of Bacillus spp. to nondetectable levels (<1.18 log CFU/g), thermal treatments of 120°C for 2 h were required. D100°C-values for total mesophilic bacteria were 5.6-fold higher than those of Bacillus spp. With increasing treatment temperature, the difference in D-value between total mesophilic bacteria and Bacillus spp. became smaller. When the aw of flour was adjusted from 0.34 to 0.71, the D-values for Bacillus decreased significantly. Treatment at 100°C for 1 h reduced Bacillus spp. populations to nondetectable levels. Our results demonstrate that naturally present Bacillus spp. in flour are highly resistant to heat, whereas increasing the aw increased their heat sensitivity. The high thermal resistance of microbes in mesquite flour warrants further investigations., (Published 2021 by the International Association for Food Protection. Not subject to U.S. Copyright.)

Abstract: Soils in which fresh produce is grown can become contaminated with foodborne pathogens and are sometimes then abandoned or removed from production. The application of biochar has been proposed as a method of bioremediating such pathogen-contaminated soils. The objectives of the present study were to evaluate three fast-pyrolysis-generated biochars (FPBC; pyrolyzed in house at 450, 500, and 600°C in a newly designed pyrolysis reactor) and 10 United Kingdom Biochar Research Center (UKBRC) standard slow-pyrolysis biochars to determine their effects on the viability of four surrogate strains of Escherichia coli O157:H7 in soil. A previously validated biocidal FPBC that was aged for 2 years was also tested with E. coli to determine changes in antibacterial efficacy over time. Although neither the UKBRC slow-pyrolysis biochars or the 450 and 500°C FPBC from the new reactor were antimicrobial, the 600°C biochar was biocidal (P < 0.05); E. coli populations were significantly reduced at 3 and 3.5% biochar concentrations (reductions of 5.34 and 5.84 log CFU/g, respectively) compared with 0.0 to 2.0% biochar concentrations. The aged 500°C FPBC from the older reactor, which was previously validated as antimicrobial, lost efficacy after aging for 2 years. These results indicate that the biocidal activity of FPBC varies based on production temperature and/or age., (Published 2020 by the International Association for Food Protection. Not subject to U.S. Copyright.)

Abstract: Dehydrated fruits, including dried coconut (Cocos nucifera) and dried apple (Malus sp.) slices, have been the subject of manufacturer recalls due to contamination with Salmonella. A study was conducted to determine the survival of Salmonella on apple slices of six apple cultivars after dehydration and also following treatment with antimicrobial solutions (0.5%, w/w) and dehydration. Samples of six apple cultivars (Envy, Gala, Red Delicious, Fuji, Pink Lady, Granny Smith) were cored and sliced into 0.4-cm rings, halved, inoculated with a five-strain composite of desiccation-resistant Salmonella, and dehydrated at 60°C for 5 h. Subsequently, Gala apple slices were treated in 0.5% solutions of one of eight antimicrobial rinses for 2 min and then dehydrated at 60°C for 5 h. Antimicrobial solutions used were potassium sorbate, sodium benzoate, ascorbic acid, propionic acid, lactic acid, citric acid, fumaric acid, and sodium bisulfate. Reduction of Salmonella populations varied according to apple cultivar. Salmonella survival on Envy, Gala, Red Delicious, Fuji, Pink Lady, and Granny Smith was 5.92, 5.58, 4.83, 4.68, 4.45, and 3.84 log CFU, respectively. There was significantly greater (P < 0.05) Salmonella inactivation on Granny Smith, Pink Lady, and Fuji apples than on Gala and Envy. Survival of Salmonella on Gala apple slices following dehydration was 5.58 log CFU for the untreated control and 4.76, 3.90, 3.29, 3.13, 2.89, 2.83, 2.64, and 0.0 log CFU for those treated with potassium sorbate, sodium benzoate, ascorbic acid, propionic acid, lactic acid, citric acid, fumaric acid, and sodium bisulfate, respectively. Pretreatment of apple slices with either fumaric acid or sodium bisulfate before dehydration led to lower Salmonella survival than pretreatment with all other antimicrobial treatments. Lower apple pH was statistically correlated (P < 0.05) with decreasing survival of Salmonella following dehydration. These results may provide methodology applicable to the food industry for increasing the inactivation of Salmonella during the dehydration of apple slices., (Published 2020 by the International Association for Food Protection. Not subject to U.S. Copyright.)

Abstract: Contamination of fresh produce with the foodborne pathogens Salmonella enterica, Listeria monocytogenes, and Escherichia coli O157:H7 continues to be problematic, resulting in outbreaks of foodborne illness and costly corporate recalls. Various individual concentrations of citric or lactic acids (0.35 to 0.61%) or isopropyl citrate (0.16 to 0.54%) combined with two generally recognized as safe surfactants, 0.025% sodium-2-ethyl-hexyl sulfate and 0.025% sodium dodecylbenzene-sulfonate, were tested against these three pathogens in suspension and when inoculated and dried on the surface of grape tomatoes. The efficacy of sodium hypochlorite (NaClO; at 46 ppm) was also evaluated under dirty and clean conditions in suspension after addition of 0.3 or 0.03% bovine serum albumin, respectively, as an organic load. NaClO (46 ppm) inactivated the three pathogens in suspension by <0.76 log CFU/mL after 5 min in the presence of 0.3% bovine serum albumin, whereas 9 and 15 ppm of free chlorine inactivated the pathogens by 0.64 and 2.77 log CFU/mL, respectively, after 5 min under clean conditions. Isopropyl citrate (0.16% acidulant) plus 0.05% total concentration of the two surfactants inactivated the pathogens in suspension by up to 7.0 log CFU/mL within 2 min. When applied to grape tomatoes for 2 min, 0.54% isopropyl citrate plus 0.025% concentrations of each of the two surfactants reduced Salmonella, E. coli O157:H7, and L. monocytogenes by as much as ca. 5.47, 4.89, and 4.19 log CFU/g, respectively. These reductions were significantly greater than those achieved with 49 ppm of free chlorine. Citric acid and lactic acid plus surfactant washes achieved greater inactivation than water-only washes, reducing Salmonella, E. coli O157:H7, and L. monocytogenes on tomatoes by up to 4.90, 4.37, and 3.98 log CFU/g, respectively. These results suggest that these combinations of acidulants and surfactants may be an effective tool for preventing cross-contamination during the washing of grape tomatoes, for reducing pathogens on the fruit itself, and as an alternative to chlorine for washing fresh produce., (Published 2020 by the International Association for Food Protection. Not subject to U.S. Copyright.)

Fresh produce-associated outbreaks of foodborne illnesses continue to occur every year in the U.S., suggesting limitations of current practices and the need for effective intervention technologies. Advanced oxidation process involves production of hydrogen radicals, which are the strongest oxidant. The objective of the present study was to evaluate the effectiveness of advanced oxidation process by combining gaseous ozone and aerosolized hydrogen peroxide. Grape tomatoes were inoculated with a 2-strain cocktail of Salmonella typhimurium on both stem scar and smooth surface. Gaseous ozone (800 and 1600 ppm) and aerosolized hydrogen peroxide (2.5, 5 and 10%) were separately or simultaneously introduced into a treatment chamber where the inoculated tomatoes were placed. During the 30 min treatments, hydrogen peroxide was aerosolized using an atomizer operated in two modes: continuously or 15 s on/50 s off. After the treatments, surviving Salmonella on the smooth surface and stem scar were enumerated. Results showed that ozone alone reduced Salmonella populations by <0.6 log CFU/fruit on both the smooth surface and the stem scar area, and aerosolized hydrogen peroxide alone reduced the populations by up to 2.1 log CFU/fruit on the smooth surface and 0.8 log CFU/fruit on stem scar area. However, the combination treatments reduced the populations by up to 5.2 log CFU/fruit on smooth surface and 4.2 log CFU/fruit on the stem scar. Overall, our results demonstrate that gaseous ozone and aerosolized hydrogen peroxide have synergistic effects on the reduction of Salmonella populations on tomatoes., (Published by Elsevier B.V.)

The interactive effects of mild heat and gaseous chlorine dioxide (ClO 2 ) on populations of Salmonella on almonds were studied. Almonds, dip inoculated with a two-strain cocktail of attenuated Salmonella Typhimurium, were treated with three concentrations of ClO 2 at ambient temperature (ca. 22°C), and at 45, 50, 55, and 60°C for 4 h and with more than 90% relative humidity. Concentrations of ClO 2 during treatments were measured, and populations of Salmonella were determined following treatments. Results demonstrated that ClO 2 at concentrations of more than 4 mg/L and ambient temperature only reduced populations of Salmonella by 1.46 log CFU/g. With increasing treatment temperature, the efficacy of gaseous ClO 2 increased. At 55 and 60°C, >1 mg/L ClO 2 , and a 4-h treatment time, >4 log CFU/g Salmonella was inactivated. Reductions greater than 4 log of the bacterium by gaseous ClO 2 at 55°C were confirmed using a three-strain cocktail of pathogenic Salmonella . Overall, results demonstrated that mild heating is necessary for gaseous ClO 2 to achieve more than 4 log CFU/g inactivation of Salmonella on almonds.

The heat resistance of three heat-resistant strains of Salmonella was determined in whole liquid egg (WLE). Inoculated samples in glass capillary tubes were completely immersed in a circulating water bath and held at 56, 58, 60, 62, and 64°C for predetermined lengths of time. The recovery medium was tryptic soy agar with 0.1% sodium pyruvate and 50 ppm of nalidixic acid. Survival data were fitted using log-linear, log-linear with shoulder, and Weibull models using GInaFiT version 1.7. Based on the R 2 and mean square error, the log-linear with shoulder and Weibull models consistently produced a better fit to Salmonella survival curves obtained at these temperatures. Contaminated WLE must be heated at 56, 60, and 64°C for at least 33.2, 2.7, and 0.31 min, respectively, to achieve a 4-log reduction of Salmonella ; 39.0, 3.1, and 0.34 min, respectively, for a 5-log reduction; and 45.0, 3.5, and 0.39 min, respectively, for a 6-log reduction. The z -values calculated from the D -values were 3.67 and 4.18°C for the log-linear with shoulder and Weibull models, respectively. Thermal death times presented in this study will be beneficial for WLE distributors and regulatory agencies when designing pasteurization processes to effectively eliminate Salmonella in WLE, thereby ensuring the microbiological safety of the product.

There are numerous obstacles to the detection of foodborne pathogens in foods that exhibit a low water activity (a w ). These obstacles include the presence of antimicrobial compounds, particulates, PCR inhibitors, and fatty matrices. New approaches should be sought to increase the sensitivity of pathogen testing in low-a w foods and to overcome the effects of various inhibitors and antimicrobials. The U.S. Food and Drug Administration and other laboratories are working toward this goal. This review will address these issues while delineating specific inhibitors and antimicrobials that impede testing of low-a w foods. A review of relevant rapid and conventional testing methodologies for Salmonella in low-a w foods will also be discussed.

Highlights: Consumers are demanding more natural and organic foods and ingredients. Many additives alone are insufficient to inactivate pathogens or prevent growth. Mild heat combined with antimicrobial agents synergistically inactivate pathogens. Here we review studies using thermally assisted antimicrobial inactivation. Many opportunities exist for the application of this hurdle technology in foods.

Spices in the desiccated state provide an environment that allows the survival of many foodborne pathogens. Currently, the incidence of pathogen-positive spices imported into the United States is 1.9 times higher than for any other imported food. Correspondingly, imported spices have been associated with numerous foodborne outbreaks and multiple product recalls. Despite the association with recalls and outbreaks, the actual pathogen populations in spices, when found, are frequently extremely small. In addition to pathogenic bacterial species, toxigenic molds have been frequently recovered from spices, and aflatoxins have been found in as many as 58% of the spices sampled. The presence of toxigenic molds is especially problematic to the immunocompromised or those on immunosuppressive therapy and has been linked to gut aspergillosis. Numerous detection methods, including both traditional and advanced DNA regimes, are being tested to optimize recovery of pathogens from spices. Further, a number of new inactivation intervention methods to decontaminate spices are examined and discussed.

Compost is organic material that has been degraded into a nutrient-stabilized humus-like substance through intense microbial activity, which can provide essential plant nutrients (nitrogen, phosphorus) to aid in the growth of fruits and vegetables. Compost can be generated from animal waste feedstocks; these can contain human pathogens, which can be inactivated through the heat and microbial competition promoted during the composting process. Outbreaks of infections caused by bacterial pathogens such as Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes on fruit and vegetable commodities consumed raw emphasize the importance of minimizing the risk of pathogenic contamination on produce commodities. This review article investigates factors that affect the reduction and survival of bacterial foodborne pathogens during the composting process. Interactions with indigenous microorganisms, carbon:nitrogen ratios, and temperature changes influence pathogen survival, growth, and persistence in finished compost. Understanding the mechanisms of pathogen survival during the composting process and mechanisms that reduce pathogen populations can minimize the risk of pathogen contamination in the cultivation of fruits and vegetables.

A study was conducted to evaluate the effects of post-treatment rinsing with water on the inactivation efficacy of acid treatments against Salmonella inoculated onto stem scar areas of two types of tomatoes. In addition, impact on fruit quality was investigated during 21 days post-treatment storage at 10 °C. A four-strain cocktail of Salmonella enterica (S. Montevideo, S. Newport, S. Saintpaul, and S. Typhimurium) was inoculated onto stem scar areas of grape and large round tomatoes. The inoculated fruits were then treated for 2 min with the following solutions: water, 2% lactic acid +2% acetic acid +2% levulinic acid, 1.7% lactic acid +1.7% acetic acid +1.7% levulinic acid, and 3% lactic acid +3% acetic acid. After treatments, half of the fruits were rinsed with water while another half were not rinsed. Non-inoculated grape tomatoes for quality analysis were treated with the same solutions with and without subsequent water rinse. Results demonstrated that the acid combinations reduced populations of Salmonella enterica on the stem scar area of grape tomatoes by 1.52-1.90 log CFU/fruit, compared with the non-treated control while water wash and rinse removed the bacterium by only 0.23-0.30 log CFU/fruit. On the stem scar of large round tomatoes, the same acid treatments achieved 3.54 log CFU/fruit reduction of the pathogen. The varying response to the acid washes between grape and large round tomatoes seems to be related to the differences in surface characteristics of stem scar areas observed with SEM. Rinsing with water after acid combination treatments did not significantly affect the efficacy of the treatments in either grape or large round tomatoes. Acidic off-odor was detected on fruits treated with acid combination without water rinse 1 day after treatment while water rinse eliminated the off-odor. The acid treatments with and without water rinse did not consistently affect appearance, color, firmness, or lycopene or ascorbic acid contents of tomatoes during 21-days storage at 10 °C. Considering the similarity in antimicrobial efficacy between the fruits with and without water rinse following acid treatments, and the elimination of acidic odor by water rinse, fruits should be rinsed with water after acid treatments. Overall, our results demonstrated that the acids were more effective in inactivating Salmonella on large round tomatoes than on grape tomatoes, and water rinses following acid treatments eliminated the acidic odor without affecting the efficacy of the acids against Salmonella., (Published by Elsevier B.V.)

Antimicrobial washing (AW), antimicrobial coating (AC), and a combination of washing followed by coating (AW+AC) were evaluated for their ability to inactivate artificially inoculated foodborne pathogens and native microbiota on strawberries stored at 4°C. Strawberries were inoculated with a six-strain composite of Escherichia coli O157:H7 and Salmonella; treated by AW, AC, or AW+AC; and stored at 4°C for 3 weeks. The washing solution contained 90 ppm of peracetic acid, and the coating solution consisted of chitosan (1%, w/v), allyl isothiocyanate (1%, v/v), and corn-bio fiber gum (5%, w/v). The effectiveness of the antimicrobial treatments against E. coli O157:H7 and Salmonella pathogens and native microflora on strawberries and their impact on fruit quality (appearance, weight loss, color, and firmness) were determined. By the end of storage, pathogen populations on strawberries were 2.5 (AW+AC), 2.9 (AC), 3.8 (AW), and 4.2 log CFU for the positive (untreated) control. AW+AC treatments also inactivated the greatest population of native microflora, followed by the AC treatment alone. AW+AC treatments showed additional antimicrobial effectiveness against these two pathogens and native microflora. Both AW+AC and AC treatments preserved the color, texture, and appearance of strawberries throughout storage. The coating treatments (AW+AC and AC alone) further reduced the loss of moisture throughout storage. The AW treatment was the least effective in reducing populations of pathogens and native microflora and in maintaining the quality of strawberries throughout storage. This study demonstrates a method to improve the microbiological safety, shelf life, and quality of strawberries.

Salmonella contamination associated with market fresh tomatoes has been problematic for the industry and consumers. A number of outbreaks have occurred, and dollar losses for the industry, including indirect collateral impact to agriculturally connected communities, have run into the hundreds of millions of dollars. This review covers these issues and an array of problems and potential solutions surrounding Salmonella contamination in tomatoes. Some other areas discussed include (i) the use of case-control studies and DNA fingerprinting to identify sources of contamination, (ii) the predilection for contamination based on Salmonella serovar and tomato cultivar, (iii) internalization, survival, and growth of Salmonella in or on tomatoes and the tomato plant, in biofilms, and in niches ancillary to tomato production and processing, (iv) the prevalence of Salmonella in tomatoes, especially in endogenous regions, and potential sources of contamination, and (v) effective and experimental means of decontaminating Salmonella from the surface and stem scar regions of the tomato. Future research should be directed in many of the areas discussed in this review, including determining and eliminating sources of contamination and targeting regions of the country where Salmonella is endemic and contamination is most likely to occur. Agriculturalists, horticulturalists, microbiologists, and epidemiologists may make the largest impact by working together to solve other unanswered questions regarding tomatoes and Salmonella contamination.

Combination treatments of surfactants and phenolic or short-chain organic acids (SCOA) may act synergistically or additively as sanitizers to inactive foodborne viruses and prevent outbreaks. The purpose of this study was to investigate the effect of gallic acid (GA), tannic acid, p-coumaric acid, lactic acid (LA), or acetic acid (AA), in combination with sodium dodecyl sulfate (SDS), against Tulane virus (TV), a surrogate for human norovirus. An aqueous stock solution of phenolic acids or SCOA with or without SDS was prepared and diluted in a twofold dilution series to 2× the desired concentration with cell growth media (M119 plus 10% fetal bovine serum). The solution was inoculated with an equal proportion of 6 log PFU/mL TV with a treatment time of 5 min. The survival of TV was quantified using a plaque assay with LLC-MK2 cells. The minimum virucidal concentration was 0.5:0.7% (v/v) for LA-SDS at pH 3.5 (4.5-PFU/mL reduction) and 0.5:0.7% (v/v) AA-SDS at pH 4.0 (2.6-log PFU/mL reduction). GA and SDS demonstrated a minimum virucidal concentration of 12.5 mM GA-SDS at pH 7.0 (0.2:0.3% GA-SDS) with an 0.8-log PFU/mL reduction and 50 mM GA-SDS (0.8:1.4% GA-SDS at pH 7.0) increased log reduction to 1.6 log PFU/mL. The combination treatments of AA or LA with SDS at pH 7.0 did not produce significant log reduction, nor did individual treatments of tannic acid, GA, p-coumaric acid, AA, LA, or SDS. This study demonstrates that a surfactant, such as SDS, aids in the phenolic acid and SCOA toxicities against viruses. However, inactivation of TV by combination treatments is contingent upon the pH of the sanitizing solution being lower than the pK a value of the organic acid being used. This information can be used to develop sanitizing washes to disinfect food contact surfaces, thereby aiding in the prevention of foodborne outbreaks.

The purpose of this review is to delineate means of decontaminating soil. This information might be used to mitigate soil-associated risks of foodborne pathogens. The majority of the research in the published literature involves inactivation of plant pathogens in soil, i.e., those pathogens harmful to fruit and vegetable production and ornamental plants. Very little has been published regarding the inactivation of foodborne human pathogens in crop soil. Nevertheless, because decontamination techniques for plant pathogens might also be useful methods for eliminating foodborne pathogens, this review also includes inactivation of plant pathogens, with appropriate discussion and comparisons, in the hopes that these methods may one day be validated against foodborne pathogens. Some of the major soil decontamination methods that have been investigated and are covered include chemical decontamination (chemigation), solarization, steaming, biofumigation, bacterial competitive exclusion, torch flaming, microwave treatment, and amendment with biochar. Other innovative means of inactivating foodborne pathogens in soils may be discovered and explored in the future, provided that these techniques are economically feasible in terms of chemicals, equipment, and labor. Food microbiology and food safety researchers should reach out to soil scientists and plant pathologists to create links where they do not currently exist and strengthen relationships where they do exist to take advantage of multidisciplinary skills. In time, agricultural output and the demand for fresh produce will increase. With advances in the sensitivity of pathogen testing and epidemiological tracebacks, the need to mitigate preharvest bacterial contamination of fresh produce will become paramount. Hence, soil decontamination technologies may become more economically feasible and practical in light of increasing the microbial safety of fresh produce.

Nonpathogenic surrogate bacteria are prevalently used in a variety of food challenge studies in place of foodborne pathogens such as Listeria monocytogenes, Salmonella, Escherichia coli O157:H7, and Clostridium botulinum because of safety and sanitary concerns. Surrogate bacteria should have growth characteristics and/or inactivation kinetics similar to those of target pathogens under given conditions in challenge studies. It is of great importance to carefully select and validate potential surrogate bacteria when verifying microbial inactivation processes. A validated surrogate responds similar to the targeted pathogen when tested for inactivation kinetics, growth parameters, or survivability under given conditions in agreement with appropriate statistical analyses. However, a considerable number of food studies involving putative surrogate bacteria lack convincing validation sources or adequate validation processes. Most of the validation information for surrogates in these studies is anecdotal and has been collected from previous publications but may not be sufficient for given conditions in the study at hand. This review is limited to an overview of select studies and discussion of the general criteria and approaches for selecting potential surrogate bacteria under given conditions. The review also includes a list of documented bacterial pathogen surrogates and their corresponding food products and treatments to provide guidance for future studies.

Viruses are currently the leading cause of foodborne outbreaks, most of which are associated with foods consumed raw. Cold plasma (CP) is an emerging novel nonthermal technology that can be used to surface decontaminate foods. This study investigated CP technology for the nonthermal inactivation of human norovirus surrogates, Tulane virus (TV) and murine norovirus (MNV), on the surface of blueberries. Blueberries (5 g) were weighed into sterile 4 oz. glass jars and inoculated with TV, 5 log PFU/g. Samples were treated with atmospheric CP for 0, 15, 30, 45, and 60 s at a working distance of 7.5 cm with 4 cubic feet/minute (cfm) of CP jet. Temperature readings were taken with an infrared camera prior to, and immediately following, CP treatments. In order to establish the impact of air flow during CP treatment (4 cfm), an additional 7 cfm jet of room temperature air was introduced from a separate nozzle. The experiment was repeated with 90 and 120 s as additional treatment time points. Viral titers were measured immediately after each treatment with a plaque assay using LLC-MK2 cells (TV) or RAW 264.7 cells (MNV). TV was significantly reduced 1.5 PFU/g compared to the control after treatment time of 45s, which was achieved regardless of temperature conditions. With the addition of 7 cfm of ambient air, the maximum log reduction for TV was 3.5 log PFU/g after 120s of treatment. MNV was significantly reduced by 0.5 log PFU/g compare to the control at 15s, and further treatment of MNV with ambient air brought the log reduction to greater than 5 log PFU/g at 90 s of treatment (Fig. 3). These results demonstrate that CP viral inactivation does not rely on thermal inactivation, and is therefore nonthermal in nature. With further optimization, CP may be used by food processors as a means of nonthermal inactivation of foodborne viruses., (Published by Elsevier Ltd.)

The disinfecting properties of high-intensity monochromatic blue light (MBL) were investigated against Escherichia coli O157:H7, Salmonella , and nonpathogenic bacteria inoculated onto the surface of almonds. MBL was generated from an array of narrow-band 405-nm light-emitting diodes. Almonds were inoculated with higher or lower levels (8 or 5 CFU/g) of pathogenic E. coli O157:H7 and Salmonella , as well as nonpathogenic E. coli K-12 and an avirulent strain of Salmonella Typhimurium, for evaluation as potential surrogates for their respective pathogens. Inoculated almonds were treated with MBL for 0, 1, 2, 4, 6, 8, and 10 min at a working distance of 7 cm. Simultaneous to treatment, cooling air was directed onto the almonds at a rate of 4 ft 3 /min (1.89 ×10 -3 m 3 /s), sourced through a container of dry ice. An infrared camera was used to monitor the temperature readings after each run. For E. coli K-12, reductions of up to 1.85 or 1.63 log CFU/g were seen for higher and lower inoculum levels, respectively; reductions up to 2.44 and 1.44 log CFU/g were seen for E. coli O157:H7 (higher and lower inoculation levels, respectively). Attenuated Salmonella was reduced by up to 0.54 and 0.97 log CFU/g, whereas pathogenic Salmonella was reduced by up to 0.70 and 0.55 log CFU/g (higher and lower inoculation levels, respectively). Inoculation level did not significantly impact minimum effective treatment times, which ranged from 1 to 4 min. Temperatures remained below ambient throughout treatment, indicating that MBL is a nonthermal antimicrobial process. The nonpathogenic strains of E. coli and Salmonella each responded to MBL in a comparable manner to their pathogenic counterparts. These results suggest that these nonpathogenic strains may be useful in experiments with MBL in which a surrogate is required, and that MBL warrants further investigation as a potential antimicrobial treatment for low-moisture foods.

Thermal inactivation kinetics of heat resistant strains of Salmonella Enteritidis in shell eggs processed by hot water immersion were determined and the effects of the processing on egg quality were evaluated. Shell eggs were inoculated with a composite of heat resistant Salmonella Enteritidis (SE) strains PT8 C405, 2 (FSIS #OB030832), and 6 (FSIS #OB040159). Eggs were immersed in a circulating hot water bath for various times and temperatures. Come-up time of the coldest location within the egg was 21 min. SE was reduced by 4.5 log at both hot water immersion treatments of 56.7 C for 60 min and 55.6 °C for 100 min. Decimal reduction times (D-values) at 54.4, 55.6, and 56.7 °C were 51.8, 14.6, and 9.33 min, respectively. The z-value was 3.07 °C. Following treatments that resulted in a 4.5 log reduction (56.7 °C/60 min and 55.6 °C/100 min), the surviving population of SE remained static during 4 wk of refrigerated storage. After processing under conditions resulting in 4.5 log reductions, the Haugh unit and albumen height significantly increased (P < 0.01) and yolk index significantly decreased (P < 0.05). The shell dynamic stiffness significantly increased (P < 0.05), while static compression shell strength showed no significant difference (P < 0.05). Vitelline membrane strength significantly increased (P < 0.05); although, no significant difference (P < 0.05) was observed in vitelline membrane elasticity. In summary, the hot water immersion process inactivated heat resistant SE in shell eggs by 4.5 log, but also significantly affected several egg quality characteristics., (Published 2016. This article is a U.S. Government work and is in the public domain in the USA.)

The Egg Products Inspection Act of 1970 requires that egg products in the U.S. must be pasteurized prior to release into commerce. The USDA Food Safety and Inspection Service (FSIS) is responsible for regulating egg products. Salmonellae are infrequently isolated from pasteurized egg products by food manufacturers or the FSIS and may be present as a result of either pasteurization-resistant bacteria or post-processing contamination. In this study, seventeen strains of Salmonella isolated from pasteurized egg products and three heat-resistant control strains were compared for the following attributes: thermal resistance in liquid whole egg (LWE) at 60 °C, enzymatic profiles, and serotyping and phage typing, antibiotic susceptibility, fatty acid analysis and strain morphological variation evaluated by scanning electron microscopy. Isolates were serotyped as Heidelberg (4 isolates), Widemarsh, Mbandaka, Cerro, Thompson, 4,12:i:-, and Enteritidis (8 isolates). All 20 isolates were sensitive to all 14 antibiotics tested for. The D60 values in LWE ranged from 0.34 to 0.58 min. All 20 strains were recovered from LWE inoculated with 8.5 logCFU/mL of Salmonella and pasteurized at 60 °C for 3.5 min; however, some isolates were not recovered from pasteurized LWE that had been inoculated with only 4.5 logCFU/mL Salmonella and treated at 60 °C for 3.5 min. Although some strains exhibited atypical enzymatic activity (e.g., reduction of adonitol, hydrolysis of proline nitroanilide or p-n-p-beta-glucuronide, and nonreduction of melibiose), differences in biochemical reactions could not be correlated with differences in thermal resistance. Furthermore, fatty acid analysis revealed that differences insaturate/unsaturated profiles may be correlated with differences in heat resistance, in two instances. One heat resistant strain (#13, Enteritidis) had the statistically lowest unsaturated/saturate ratio at 39%. However, one heat sensitive strain (#3, serovar 4,12:i:-) had the highest unsaturated/saturate ratio at 81%, and also the lowest concentration of stearic acid. This data represents the first steps in determining whether Salmonella contamination in pasteurized egg products may be the result of either thermally-resistant isolates or post-processing contamination. Contamination of LWE by Salmonella strains with higher heat resistance, (e.g., isolate #'s 2, 6, 10 and 12) may indicate the ability of Salmonella to survive pasteurization, while contamination of LWE strains with lower heat resistance (e.g., isolate #'s 1, 3, 5, 7, 8, 11, and 15) may indicate post-processing contamination of LWE by this foodborne pathogen., (Copyright © 2015. Published by Elsevier B.V.)

Cold plasma (CP) is a novel nonthermal technology, potentially useful in food processing settings. Berries were treated with atmospheric CP for 0, 15, 30, 45, 60, 90, or 120 s at a working distance of 7.5 cm with a mixture of 4 cubic feet/minute (cfm) of CP jet and 7 cfm of ambient air. Blueberries were sampled for total aerobic plate count (APC) and yeast/molds immediately after treatment and at 1, 2, and 7 days. Blueberries were also analyzed for compression firmness, surface color, and total anthocyanins immediately after each treatment. All treatments with CP significantly (P < 0.05) reduced APC after exposure, with reductions ranging from 0.8 to 1.6 log CFU/g and 1.5 to 2.0 log CFU/g compared to the control after 1 and 7 days, respectively. Treatments longer than 60s resulted in significant reductions in firmness, although it was demonstrated that collisions between the berries and the container contributed significantly to softening. A significant reduction in anthocyanins was observed after 90 s. The surface color measurements were significantly impacted after 120 s for the L* and a* values and 45 s for the b* values. CP can inactivate microorganisms on blueberries and could be optimized to improve the safety and quality of produce., (Published by Elsevier Ltd.)

The objectives of this study were to determine the influence of a symbiotic arbuscular mycorrhizal (AM) fungus on persistence of Salmonella and enterohemorrhagic Escherichia coli O157:H7 (EHEC) within soil, and survival within Romaine lettuce. Romaine seedlings were grown with or without AM fungi. Soil surrounding plants was inoculated with ca. 8 log CFU/plant of either Salmonella enterica or E. coli EHEC composites. Samples (soil, root, and shoot) were analyzed on days 1, 8, 15 and 22 for Salmonella and EHEC by direct plating and selective enrichment. Twenty-four hours after inoculation, populations of Salmonella and EHEC, respectively, were 4.20 and 3.24 log CFU/root, 2.52 and 1.17 log CFU/shoot, and 5.46 and 5.17 log CFU/g soil. By selective enrichment, samples tested positive for Salmonella or EHEC at day 22 at rates of 94 and 68% (shoot), 97 and 56% (root), and 100 and 75% (soil), respectively, suggesting that Salmonella has a greater propensity for survival than EHEC. Salmonella populations in soil remained as high as 4.35 log CFU/g by day 22, while EHEC populations dropped to 1.12 log CFU/g in the same amount of time. Ninety-two percent of all Romaine leaves in our study were positive for internalized Salmonella from days 8 to 22 and remained as high as 1.26 log CFU/shoot on day 22 in AM fungi+Romaine plants. There were no differences (P>0.05) between the survival of either pathogen based on the presence or absence of mycorrhizal fungi. Results of this study suggest that AM fungi do not affect the internalization and/or survival of either S. enterica or E. coli O157:H7 in Romaine lettuce seedlings. Our results should provide Romaine lettuce farmers confidence that the presence and/or application of AM fungi to crop soil is not a contributing factor to the internalization and survival of Salmonella or E. coli O157:H7 within Romaine lettuce plants., (Published by Elsevier B.V.)

A 2011 outbreak of hemorrhagic colitis, which resulted in the death of two individuals, was associated with contaminated strawberries. A study was conducted to identify antimicrobial washes effective at reducing E. coli O157:H7 and Salmonella enterica from the surface of fresh whole strawberries during two-minute immersion washes. Twenty-seven antimicrobial treatments were tested. Vacuum perfusion was applied to strawberries during chlorine and peracetic acid treatments to promote infiltration of sanitizer into porous strawberry tissue. Strawberries were inoculated to 7.1logCFU/strawberry with a seven-strain bacterial composite, consisting of three strains of E. coli O157:H7 and four serovars of Salmonella enterica. Berries were air-dried for 2h and immersed in circulating antimicrobial solutions for 120s at 22°C. Four treatments reduced ≥3.0logCFU/strawberry, including (a) 1% acetic acid+1% H2O2, (b) 30% ethanol+1% H2O2, (c) 90ppm peracetic acid, and (d) 1% lactic acid+1% H2O2. Two additional treatments that reduced 2.8logCFU/strawberry were (a) 40% ethanol, and (b) 1% each of phosphoric+fumaric acids. Eight treatments reduced 2.0-2.6logCFU/strawberry. Five treatments reduced <1.45CFU/strawberry, including (a) 1% citric acid, (b) 1% lactic acid, (c) 1% acetic acid, (d) 0.5% each of acetic+citric acids and (e) 0.5% each of acetic+lactic acids. The use of vacuum perfusion with 200ppm chlorine or 90ppm peracetic acid did not reduce greater populations of pathogens than did the same treatments without vacuum perfusion. Fourteen treatments reduced no more pathogens (p<0.05) than did sterile deionized water. Results from this study provide some options for end-point decontamination of strawberries for retail operations just prior to serving to customers., (Published by Elsevier B.V.)

An exploratory study was performed to determine the influence of fast pyrolysis (FP) and slow pyrolysis (SP) biochars on enterohemorrhagic Escherichia coli O157:H7 (EHEC) in soil. Soil + EHEC (inoculated at 7 log colony-forming units [CFU]/g of soil) + 1 of 12 types of biochar (10% total weight:weight in soil) was stored at 22°C and sampled for 8 weeks. FP switchgrass and FP horse litter biochars inactivated 2.8 and 2.1 log CFU/g more EHEC than no-biochar soils by day 14. EHEC was undetectable by surface plating at weeks 4 and 5 in standard FP switchgrass, FP oak, and FP switchgrass pellet biochars. Conversely, EHEC populations in no-biochar control samples remained as high as 5.8 and 4.0 log CFU/g at weeks 4 and 5, respectively. Additionally, three more SP hardwood pellet biochars (generated at 500°C for 1 h, or 2 h, or generated at 700°C for 30 min) inactivated greater numbers of EHEC than did the no-biochar control samples during weeks 4 and 5. These results suggest that biochar can inactivate E. coli O157:H7 in cultivable soil, which might mitigate risks associated with EHEC contamination on fresh produce.

The goal of this study was to determine the inactivation kinetics of Salmonella in commercial 10% salted liquid whole egg (LWE) to assist the U.S. Department of Agriculture in writing new liquid egg pasteurization guidelines. Current data are not sufficient for predicting thermal inactivation kinetics of Salmonella spp. for use in updating pasteurization guidelines for many types of liquid egg products, including salted LWE (SLWE). This is, in part, due to variations in Salmonella strains and changes in the processing of liquid egg products that have arisen in the past 40 years. Pasteurization guidelines are currently being reevaluated in light of recent risk assessments. Heat-resistant Salmonella serovars Enteritidis and Oranienburg were composited and mixed into 10% SLWE, resulting in final populations of approximately 5.7-7.8 log colony-forming units (CFU)/mL. Inoculated egg was injected into glass capillary tubes, flame-sealed, and heated in a water bath at 60, 62.2, 63.3, 64.3, or 66°C. Contents were surface-plated and incubated at 37°C for 24 h. Survival curves were not log-linear (log levels versus time), but decreased rapidly, and after initial periods became linear. Asymptotic decimal reduction values at each temperature were calculated from survivor curves with a minimum inactivation of 5.0 log CFU/mL. The asymptotic thermal D-values for SLWE were 3.47, 2.23, 1.79, 1.46, and 1.04 min at 60, 62.2, 63.3, 64.3, or 66°C, respectively. The calculated thermal z-value was 11.5°C. A model that predicts lethality for given times and temperatures that was developed predicted that the current pasteurization requirements for 10% SLWE (i.e., 63.3°C for 3.5 min, or 62.2°C for 6.2 min) are not sufficient to inactivate 7 log CFU/mL of Salmonella and only achieve approximately 4 log CFU/mL inactivation. This model will assist egg-products manufacturers and regulatory agencies in designing pasteurization processes to ensure product safety.

The effects of storage conditions on subsequent high-hydrostatic pressure (HHP) inactivation of Vibrio parahaemolyticus and Vibrio vulnificus in oysters were investigated. Live oysters were inoculated with V. parahaemolyticus or V. vulnificus to ca. 7-8 log MPN/g by feeding and stored at varying conditions (i.e., 21 or 35 °C for 5h, 4 or 10 °C for 1 and 2 days and -18 °C for 2 weeks). Oyster meats were then treated at 225-300 MPa for 2 min at 4, 21 or 35 °C. HHP at 300 MPa for 2 min achieved a >5-log MPN/g reduction of V. parahaemolyticus, completely inactivating V. vulnificus (negative by enrichment) in oysters. Treatment temperatures of 4, 21 and 35 °C did not significantly affect pressure inactivation of V. parahaemolyticus or V. vulnificus (P>0.05). Cold storage at -18, 4 and 10 °C, prior to HHP, decreased V. parahaemolyticus or V. vulnificus populations by 1.5-3.0 log MPN/g, but did not increase their sensitivity to subsequent HHP treatments. The effects of cold storage after HHP on inactivation of V. parahaemolyticus in oysters were also determined. Oysters were inoculated with V. parahaemolyticus and stored at 21 °C for 5h or 4 °C for 1 day. Oyster meats were then treated at 250-300 MPa for 2 min at 21 or 35 °C and stored for 15 days in ice or in a freezer. V. parahaemolyticus populations in HHP-treated oysters gradually decreased during post-HHP ice or frozen storage. A validation study using whole-shell oysters was conducted to determine whether the presence of oyster shells influenced HHP inactivation of V. parahaemolyticus. No appreciable differences in inactivation between shucked oyster meat and whole-shell oysters were observed. HPP at 300 MPa for 2 min at 21 °C, followed by 5-day ice storage or 7-day frozen storage, and HPP at 250 MPa for 2 min at 21 °C, followed by 10-day ice or 7-day frozen storage, completely inactivated V. parahaemolyticus in whole-shell oysters (>7 log reductions). The combination of HHP at a relatively low pressure (e.g., 250 MPa) followed by short-term frozen storage (7 days) could potentially be applied by the shellfish industry as a post-harvest process to eliminate V. parahaemolyticus in oysters., (Copyright © 2013 Elsevier B.V. All rights reserved.)

This study was conducted to develop antimicrobial coatings to decontaminate and prevent cross-contamination of shell eggs. Egg shells were inoculated with nalidixic acid-resistant Salmonella enterica Enteritidis strains OB030832, OB040159, and C405 and treated with antimicrobial coatings. Polylactic acid served as a nonedible polymer, and chitosan served as an edible polymer carrier of natural antimicrobials, including nisin, allyl isothiocyanate (AIT), lauric arginate ester (LAE), and organic acids. Increases of AIT concentrations or addition of nisin to AIT in either the polylactic acid or chitosan coating solutions resulted in greater reductions of Salmonella. Chitosan coatings with 0.1, 0.5, and 1.0% LAE reduced Salmonella by 1.7, 2.5, and 5.2 log CFU/cm(2), respectively. Shell eggs treated with 1.0 and 0.5% LAE in chitosan coatings had nondetectable Salmonella cells (< 0.5 log CFU/cm(2)) after 3 and 7 days of storage at 7°C, respectively, and no outgrowth was observed up to 28 days. Coating treatments significantly reduced weight loss of shell eggs during 12 weeks of storage at 7 or 4°C. This study demonstrates an alternative and effective intervention technology for decontaminating shell eggs and provides an alternative approach to reduce possible recalls and outbreaks associated with pathogen contamination on shell eggs and in egg products.

A study was conducted to determine the influence of arbuscular mycorrhizal (AM) fungi on Salmonella and enterohemorrhagic Escherichia coli O157:H7 (EHEC) in autoclaved soil and translocation into leek plants. Six-week-old leek plants (with [Myc+] or without [Myc-] AM fungi) were inoculated with composite suspensions of Salmonella or EHEC at ca. 8.2 log CFU/plant into soil. Soil, root, and shoot samples were analyzed for pathogens on days 1, 8, 15, and 22 postinoculation. Initial populations (day 1) were ca. 3.1 and 2.1 log CFU/root, ca. 2.0 and 1.5 log CFU/shoot, and ca. 5.5 and 5.1 CFU/g of soil for Salmonella and EHEC, respectively. Enrichments indicated that at days 8 and 22, only 31% of root samples were positive for EHEC, versus 73% positive for Salmonella. The mean Salmonella level in soil was 3.4 log CFU/g at day 22, while EHEC populations dropped to ≤ 0.75 log CFU/g by day 15. Overall, Salmonella survived in a greater number of shoot, root, and soil samples, compared with the survival of EHEC. EHEC was not present in Myc- shoots after day 8 (0/16 samples positive); however, EHEC persisted in higher numbers (P = 0.05) in Myc+ shoots (4/16 positive) at days 15 and 22. Salmonella, likewise, survived in statistically higher numbers of Myc+ shoot samples (8/8) at day 8, compared with survival in Myc- shoots (i.e., only 4/8). These results suggest that AM fungi may potentially enhance the survival of E. coli O157:H7 and Salmonella in the stems of growing leek plants.

Background: Toxin-antitoxin (TA) systems are commonly found in bacteria and Archaea, and it is the most common mechanism involved in bacterial programmed cell death or apoptosis. Recently, MazF, the toxin component of the toxin-antitoxin module, has been categorized as an endoribonuclease, or it may have a function similar to that of a RNA interference enzyme., Results: In this paper, with comparative data and phylogenetic analyses, we are able to identify several potential MazF-conserved motifs in limited subsets of foodborne pathogens and probiotic strains and further provide a molecular basis for the development of engineered/synthetic probiotic strains for the mitigation of foodborne illnesses. Our findings also show that some probiotic strains, as fit as many bacterial foodborne pathogens, can be genetically categorized into three major groups based on phylogenetic analysis of MazF. In each group, potential functional motifs are conserved in phylogenetically distant species, including foodborne pathogens and probiotic strains., Conclusion: These data provide important knowledge for the identification and computational prediction of functional motifs related to programmed cell death. Potential implications of these findings include the use of engineered probiotic interventions in food or use of a natural probiotic cocktail with specificity for controlling targeted foodborne pathogens.

A study was conducted to identify sanitizing solutions effective at inactivating ca. 5logCFU of Salmonella enterica inoculated onto the stem scar of red round tomatoes during two-minute immersion treatments. Sixty-three antimicrobial combinations were tested. Vacuum perfusion was applied to tomatoes during selected treatments to promote infiltration of sanitizer into porous tomato stem scar tissue. Red round tomatoes were inoculated to ca. 6.9logCFU/stem scar with a four-serovar composite of Salmonella enterica, air dried, and tomatoes were immersed in circulating sanitizing solutions for 120s at ca. 22°C. Stem scars were aseptically excised, macerated in DE neutralizing broth, and the homogenate was spiral plated. Twenty-four washes inactivated ≥3.0logCFU/stem scar. Seven treatments reduced ≥4.8 log (viz., 40% EtOH, sulfuric acid, and organic acid combinations). LogCFU/stem scar reductions for various sanitizers are listed in parenthesis, as follows: 90ppm peroxyacetic acid (1.31), 200ppm chlorine (1.53), 190ppm chlorine+15″ Hg vacuum perfusion (2.23), 0.2N sodium hydroxide (NaOH) (3.78), 2% total of lactic+acetic acid (4.35), 3% total of phosphoric+lactic acids (4.51), and 40% ethanol (4.81). Solutions that achieved ≥4.95 log reductions were 5.1% total of lactic+acetic+levulinic acids, 49% ethanol, 6% total of lactic+acetic acids, and a 0.2M H(2)SO(4) (sulfuric acid) solution. The use of vacuum perfusion with 200ppm chlorine increased inactivation by 0.7logCFU over chlorine alone, however, P>0.05. Results from this study provide tomato processers with some sanitization options effective at inactivating Salmonella from the stem scars of tomatoes. These results may also help processors and scientists design future decontamination studies by incorporating combinations of these chemical treatments., (Published by Elsevier B.V.)

Unlabelled: Cronobacter sakazakii is a life-threatening bacterium, infrequently implicated in illnesses associated with the consumption of powdered infant formula (PIF). It can cause rare but invasive infections in neonatal infants who consume contaminated PIF. The objective of this research was to investigate the growth kinetics and develop mathematical models to predict the growth of heat-injured C. sakazakii in reconstituted PIF (RPIF). RPIF, inoculated with a 6-strain cocktail of non-heat-treated (uninjured) or heat-injured C. sakazakii, was incubated at different temperatures to develop growth models. Except for storage at 6 °C, C. sakazakii grew well at all test temperatures (10 to 48 °C). Uninjured C. sakazakii exhibited no observable lag phase, while a lag phase was apparent in heat-treated cells. A simple 3-parameter logistic equation was used to fit growth curves for non-heat-treated cells, while both Baranyi and Huang models were suitable for heat-treated C. sakazakii. Calculated minimum and maximum growth temperatures were 6.5 and 51.4 °C for non-heat-treated cells, and 6.9 and 50.1 °C for heat-treated cells of C. sakazakii in RPIF, respectively. There was no significant difference between growth rates of non-heat-treated and heat-injured cells in RPIF. For heat-treated cells of C. sakazakii, the lag phase was temperature-dependent and very short (between 25 °C and 48 °C). These results suggest that both non-heat-treated and heat-injured C. sakazakii cells may present a risk to infants if the pathogens are not completely destroyed by heat in RPIF and then exposed to subsequent temperature abuse., Practical Application: C. sakazakii is a life-threatening bacterium found in powdered infant formula (PIF). This study shows that the uninjured bacterium exhibits very short or no lag phase if not refrigerated and can grow well in reconstituted PIF (RPIF), while the heat-injured cells can multiply at an equivalent rate following metabolic recovery. Temperature abuse may allow C. sakazakii to grow and endanger infants fed with RPIF. Predictive models developed in this study can be used to estimate the growth and conduct risk assessments of this pathogen., (Journal of Food Science copy; 2012 Institute of Food Technologists® No claim to original US government works.)

A novel in-package ozonation device was evaluated for its efficacy in inactivating three microorganisms (viz., Listeria innocua, attenuated Salmonella Typhimurium, and Escherichia coli O157:H7) on tomatoes and for its effect on fruit quality. The device produced ozone inside sealed film bags, reaching a concentration of 1,000 ppm within 1 min of activation. The three bacterial cultures were inoculated onto either the smooth surface or the stem scar areas of the tomatoes, which were then sealed in plastic film bags and subjected to in-package ozonation. L. innocua on tomatoes was reduced to nondetectable levels within 40 s of treatment on the tomato surface, with inactivation of ca. 4 log CFU per fruit on the stem scar area. An increase in treatment time did not result in a proportional increase in bacterial reduction. For E. coli O157:H7 and Salmonella, there was little difference (<1 log) in the effectiveness of the system when comparing surface and scar-inoculated bacteria. Both bacteria were typically reduced by 2 to 3 log CFU per fruit after 2- to 3-min treatments. No negative effects on fruit color or texture were observed during a 22-day posttreatment storage study of ozone-treated tomatoes. These results suggest that the three bacteria responded differently to ozonation and that in-package ozonation may provide an alternative to chemical sanitizers commonly used by the industry.

This study was conducted to investigate the efficacy of antimicrobial coatings for inactivation of Salmonella on the surface of tomato stem scars. Scars were inoculated with a four-strain cocktail of Salmonella (serovars Montevideo, Newport, Saintpaul, and Typhimurium) and coated with acid-chitosan solutions. The chitosan coating with three acids (3A plus chitosan), the chitosan coating with one acid, and the three-acid solution without chitosan reduced the populations of Salmonella by 6.0, 3.6, and 5.3 log CFU per stem scar, respectively. Addition of allyl isothiocyanate (10 μl/ml) to the 3A plus chitosan coating did not significantly increase (P > 0.05) the antimicrobial efficacy. Although the populations of Salmonella in the controls (ca. 7.5 log CFU per stem scar) did not change significantly throughout the 14-day storage period at 10° C, Salmonella cells were reduced to undetectable levels (< 0.7 log CFU per stem scar) in the samples treated with 3A plus chitosan coating after two days of storage, and no growth was observed for the remaining storage period. Results from this study demonstrate that coatings of acid plus chitosan provide an alternative antimicrobial intervention for decontamination of tomatoes.

Human norovirus (NoV) is the leading cause of nonbacterial acute gastroenteritis epidemics worldwide. High-pressure processing (HPP) has been considered a promising nonthermal processing technology to inactivate food- and waterborne viral pathogens. Due to the lack of an effective cell culture method for human NoV, the effectiveness of HPP in inactivating human NoV remains poorly understood. In this study, we evaluated the effectiveness of HPP in disrupting the capsid of human NoV based on the structural and functional integrity of virus-like particles (VLPs) and histo-blood group antigen (HBGA) receptor binding assays. We found that pressurization at 500 to 600 MPa for 2 min, a pressure level that completely inactivates murine norovirus and feline calicivirus, was not sufficient to disrupt the structure and function of human NoV VLPs, even with a holding time of 60 min. Degradation of VLPs increased commensurate with increasing pressure levels more than increasing time. The times required for complete disruption of human NoV VLPs at 700, 800, and 900 MPa were 45, 15, and 2 min, respectively. Human NoV VLPs were more resistant to HPP in their ability to bind type A than type B and O HBGAs. Additionally, the 23-nm VLPs appeared to be much more stable than the 38-nm VLPs. Taken together, our results demonstrated that the human NoV capsid is highly resistant to HPP. While human NoV VLPs may not be fully representative of viable human NoV, destruction of the VLP capsid is highly suggestive of a typical response for viable human NoV.

This study investigated the antimicrobial effect of a chitosan coating+allyl isothiocyanate (AIT) and nisin against Salmonella on whole fresh cantaloupes. Cantaloupes were inoculated with a cocktail of three Salmonella strains and treated with chitosan, chitosan+AIT, chitosan+nisin, and chitosan+AIT+nisin coatings. With AIT concentrations increasing from 10 to 60 μl/ml, the antibacterial effects of coating treatments against Salmonella increased. Chitosan coatings with 60 μl/ml AIT (chitosan+60AIT) reduced more than 5 log₁₀ CFU/cm² of Salmonella. The addition of nisin to the chitosan-AIT coating synergistically increased the antibacterial effect; coatings with nisin (25 mg/ml or 25,000 IU/ml)+30 μl/ml AIT resulted in a 4.8 log₁₀ reduction of Salmonella. The chitosan+60AIT coating significantly (p<0.05) reduced populations of native bacteria on cantaloupes to ca. 2 log₁₀ CFU/cm² during the first 6 days and populations remained unchanged through day 14 at 10 °C. The same coating treatment completely inactivated mold and yeast on cantaloupe at day 1 and no regrowth occurred even up to 14 days of storage. Scanning electron microscopy revealed that cell membrane damage and leakage of intercellular components occurred as a result of the chitosan-AIT coating treatments. No visual changes in overall appearance and color of cantaloupe rind and flesh due to coating treatments were observed. These results indicate that the application of an antimicrobial coating may be an effective method for decontamination of cantaloupes., (Published by Elsevier B.V.)

Propyl p-hydroxybenzoic acid (propylparaben [PRPA]) is a phenolic antioxidant, known to occur in nature and used as a microbiostat in foods, feeds, pharmaceuticals, cosmetics, and medications. The U.S. Department of Agriculture, Food Safety and Inspection Service (FSIS) requires that liquid egg white (LEW) be pasteurized at 56.7°C for 3.5 min. This study evaluated the effects of PRPA on the pasteurization sensitivity of Salmonella in LEW. When LEW (pH 7.8) was pasteurized under FSIS conditions, salmonellae declined by 0.5, 4.6, 4.5, > 7.0, and > 7.0 log CFU/ml, with 0, 125, 250, 500, or 1,000 ppm of PRPA, respectively, and D(56.7°C)-values were 2.99, 1.05, 0.68, 0.26 and ≤0.16 min. Albumen (pH 8.9) pasteurized under FSIS standards incurred salmonellae reductions of 3.3, 2.8, 5.2, > 7.0, and > 7.0 log CFU/ml, with 0, 125, 250, 500, or 1,000 ppm of PRPA, respectively, while D(56.7°C)-values were 0.87, 0.99, 0.66, 0.22, and 0.09 min. Adding 500 ppm of PRPA to albumen (pH 7.8) reduced D(56.7°C)-values more than 11-fold, and reduced the time to achieve a 5-log reduction from 15.0 to only 1.3 min. A 7-log reduction in plain LEW (pH 7.8) at 56.7°C required 20.9 min, versus only 1.8 and 1.1 min with 500 and 1,000 ppm of PRPA, respectively. Furthermore, a 7-log reduction in plain LEW (pH 8.9) required 6.1 min, versus only 1.5 and 0.6 min with 500 and 1,000 ppm of PRPA, respectively. This study is the first to report the efficacy of PRPA (pK(a) = 8.4) in sensitizing Salmonella in LEW to thermal pasteurization, while documenting that PRPA retains its antibacterial efficacy at pH levels as high as 8.9.

There is no suitable model for predicting thermal inactivation kinetics of Salmonella spp. for many types of liquid egg products, including salted liquid egg yolk, for use in updating U.S. Department of Agriculture (USDA) pasteurization guidelines. This is because, in part, of the variations in Salmonella strains and the changes in the processing of liquid egg products over the past 40 years. The objectives of the present study were to determine the thermal inactivation kinetics and to create a general thermal inactivation kinetics model that can be used for estimating log reductions of salmonellae in 10% salted liquid egg yolk for temperatures between 62.2 and 69°C. This model can be used by processors to help ensure adequate pasteurization. This was accomplished by studying the inactivation kinetics of a three-strain composite of heat-resistant Salmonella serovars Enteritidis and Oranienburg, inoculated into commercially processed 10% salted liquid egg yolk. The survival curves were convex, with asymptotic D-values. From these curves, a general model was developed to predict log reductions for given times at specified temperatures. For example, at a temperature of 67.3°C (153.1°F) for 3.5 min, our model predicts a 5-log reduction would be obtained, whereas with the current USDA minimum required pasteurization regimen (63.33°C [146°F] for 3.5 min), our model predicts that a reduction of only 2.7 log would be obtained. The results of this study provide information that can be used by processors to aid in producing safe, pasteurized egg yolk products, and for satisfying USDA pasteurization performance standards and developing industry guidance.

Aims: To develop an antimicrobial bottle coating effective at inhibiting the growth of Salmonella in liquid egg albumen (egg white) and reduce the risk of human Salmonellosis., Methods and Results: Four-ounce glass jars were coated with a mixture of polylactic acid (PLA) polymer and antimicrobial compounds containing 100-500 μl allyl isothiocyanate (AIT), 250 mg nisin, 250 mg zinc oxide nanoparticles per jar or their combinations. The coated jars contained 100 ml of liquid egg white (LEW) inoculated with a three-strain Salmonella enterica ssp. enterica cocktail at populations of 10(3) or 10(7) CFU ml(-1) and stored at 10°C for 28 days. The PLA coating with 500 μl AIT completely inactivated 3 and 7 log CFU ml(-1) of Salmonella after 7 and 21 days of storage, respectively. The PLA coating with 200 μl AIT in combination with 250 mg nisin reduced Salmonella populations to an undetectable level (<10 CFU ml(-1) ) after 21 days of storage., Conclusions: PLA coatings containing AIT alone or in combination with nisin effectively inactivated salmonellae in LEW., Significance and Impact of the Study: This study demonstrated the commercial potential of applying the antimicrobial bottle coating method to liquid eggs and possibly other fluid food products., (Journal of Applied Microbiology © 2011 The Society for Applied Microbiology. No claim to US Government works.)