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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.

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 (P0.05) 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 (P0.05) 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.

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 (

The objective of the study was to evaluate the effect of trichome (fuzz) removal on the efficacy of ultraviolet-C in inactivating Escherichia coli O157:H7 on peach fruit, and quality of peach [Prunus persica (L.) Batsch, cv. PF25] fruit as affected by fuzz removal and ultraviolet-C. Peach (cultivar PF25) fruit, with and without fuzz removal, were inoculated with a five-strain cocktail of E. coli O157:H7 and treated with ultraviolet-C at doses of 0, 221, and 442 mJ/cm2. Fuzz was rubbed off using damped cloths. Survival of E. coli populations was determined at days 1, 4, and 7 at 20 °C. To study fruit quality, noninoculated fruit with and without fuzz removal were treated with ultraviolet-C at the same doses. Results demonstrated that ultraviolet-C at 442 mJ/cm2 reduced the population of E. coli by 1.2 to 1.4 log colony-forming units (CFU)/fruit on peach with fuzz, and 0.9 to 1.1 log CFU/fruit on fruit without fuzz 1 day after ultraviolet-C treatment. However, E. coli populations of all samples were similar with additional storage time, resulting in no significant difference among the treatments after 7 days of storage at 20 °C. Ultraviolet-C at doses up to 442 mJ/cm2 did not have any significant effect on the surface color of peaches during 7 days of storage, although fruit with fuzz removal increased L*, hue, and chroma values. In addition, fuzz removal promoted the loss of firmness during storage. Furthermore, ultraviolet-C at 442 mJ/cm2 increased antioxidant capacity of peach skin with fuzz. Overall, our results suggested that fuzz removal had marginal effects on the efficacy of ultraviolet-C, and ultraviolet-C did not negatively affect the quality of peaches.

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. HIGHLIGHTS

The interactive effects of mild heat and gaseous chlorine dioxide (ClO2) 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 ClO2 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 ClO2 during treatments were measured, and populations of Salmonella were determined following treatments. Results demonstrated that ClO2 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 ClO2 increased. At 55 and 60°C, >1 mg/L ClO2, and a 4-h treatment time, >4 log CFU/g Salmonella was inactivated. Reductions greater than 4 log of the bacterium by gaseous ClO2 at 55°C were confirmed using a three-strain cocktail of pathogenic Salmonella. Overall, results demonstrated that mild heating is necessary for gaseous ClO2 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 R2 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.

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.

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.

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 pKa 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.

UV-C can inactivate foodborne pathogens on surface of fruit such as apples. However, the practical application of the technology has been limited by lack of methods to measure UV-C doses received by fruit and difficulty of achieving uniform UV-C doses on the same pieces of fruit. In the present study, radiochromic films were evaluated for their suitability to estimate UV-C doses and dose uniformity on apple fruit surface. Parameters investigated included film type, color changes of the films in response to different UV-C doses, color stability of films, UV-C light intensity, and temperature. In addition, apples with films attached to six locations on the surface were exposed to UV-C in a treatment chamber without reflective material, with an aluminum foil and a stainless steel sheet in the bottom of treatment chamber as reflectors, in comparison with the use of a rotating device to study the UV-C dose uniformity of the fruit received. Results showed that the radiochromic films were sensitive to UV-C light as they changed to blue or pink upon exposure to UV. The developed color after UV-C irradiation was stable for at least 15 d at dark and ambient temperature (21 ± 2 °C). The temperature (21 °C vs 4 °C) at which the films were exposed did not affect the changes in color as a result of UV-C exposure. Films exposed to UV-C at an intensity of 60 W m −2 developed a more intense color compared to those at 94 W m −2 . The changes in color of the films as a function of UV-C dose were measured as absorbance at 510 and 600 nm. The relationship between A 510nm and the UV-C dose could be expressed as polynomial equations, and the equations were used to predict UV-C doses on the surface of apples. The rotating device provided higher UV-C uniformity on the apple surface than other UV-C reflective materials as indicated by the coefficient of variations and the ratio of maximum/minimum doses. Therefore, the radiochromic film can be used as a UV dosimeter to determine UV dose distributions on individual fruits. Fruits treated with UV-C on a roller received uniformed UV-C exposure.

Whole fresh blueberries were treated using a parallel pulsed electric field (PEF) treatment chamber and a sanitizer solution (60 ppm peracetic acid [PAA]) as PEF treatment medium with square wave bipolar pulses at 2 kV/cm electric field strength, 1μs pulse width, and 100 pulses per second for 2, 4, and 6 min. The effects of PEF on native microbiota and artificially-inoculated Escherichia coli K12 and Listeria innocua populations on blueberries were determined. Color, texture, anthocyanins and total phenolic compound concentrations were also evaluated. The combination of PEF and PAA was able to achieve up to 3 log reduction of E. coli and Listeria as well as 2 log/g reduction of native microbiota. PEF treatments did not cause any changes in color and appearance of the blueberries. The treatments did, however, cause the blueberries to soften in texture. Anthocyanins and phenolic compounds in blueberries increased by 10 and 25%, respectively, after PEF treatments. The results demonstrate the potential of PEF applications to enhance the safety and improve the quality and nutritional value of fruits and their derived products.

The purpose of the present study was to investigate the efficacy of in-package aerosolized aqueous sanitizers in reducing populations of Salmonella enterica serovar Typhimurium on tomato fruit and in maintaining fruit quality. Cherry tomatoes were inoculated with a cocktail of attenuated S. Typhimurium ATCC 53647 and 53648 strains on the smooth skin surface and stem scar area. Next, 200 ppm free chlorine, and peroxyacetic acid (PAA) and aqueous ClO 2 at different concentrations, 2% lactic acid + 2% acetic acid + 2% levulinic acid, and 3% acetic acid + 3% lactic acid were aerosolized into a clamshell container containing cherry tomatoes. Results showed that S. Typhimurium populations on smooth tomato surfaces were reduced by more than 5 log CFU/fruit with 400 ppm PAA, 2% lactic acid + 2% acetic acid + 2% levulinic acid, 3% acetic acid + 3% lactic acid, and aqueous ClO 2 (100 and 400 ppm). On the stem scar area, 400 ppm aqueous ClO 2 was more effective in reducing S . Typhimurium populations than other treatments, achieving 4.89 log CFU/fruit reduction, followed by 400 ppm PAA (2.62 log CFU/fruit). The efficacy of ClO 2 and acid combination treatments increased during 3-week storage at 10 °C, achieving >3 log CFU/fruit inactivation with the acid combination and ca. 6 log with for 400 ppm with ClO 2 . None of the treatments significantly (p > 0.05) affected color, appearance, firmness, vitamin C, lycopene or antioxidant values of tomatoes during 3 weeks of storage; although, an acidic odor was detected for samples treated with the organic acids in the earlier period of the storage. These results suggest that in-package aerosolized sanitizers can be used as a novel method for the inactivation of Salmonella on tomato fruit.

Toxoplasma gondii is a common protozoan parasite, whose environmentally-resistant stage, the oocyst, can contaminate irrigation water and fresh edible produce. Current washing steps in produce processing may not be effective for eliminating T. gondii from at-risk varieties of produce. The objective of this study was to evaluate low-dose irradiation as a means to inactivate T. gondii oocysts on blueberries. Blueberries (10 g) inoculated with T. gondii (5 log oocysts/g) were exposed to an absorbed dose of 0 (control), 0.2, 0.4 or 0.6 kGy gamma radiation at 4°C. After treatment, oocysts were recovered from berries by washing, and excysted sporozoites were enumerated using a plaque assay. Vero cells were maintained as monolayers in a 6-well microplate, exposed to sporozoites recovered from berries, and incubated for 7 days to determine plaque-forming units (PFU). Immediately after treatment, blueberries without inoculated T. gondii were analyzed for compression firmness, surface color, and total anthocyanins. The results from this experiment demonstrate that the viability of T. gondii oocysts was significantly reduced after an irradiation treatment of 0.2 kGy. All treatments produced reductions of 4 log PFU/g beyond the detection limit of 1 log PFU/g. Quality analysis showed that there was no significant change in compression firmness, anthocyanins, or color in berries after the irradiation treatment. Results of this study indicated that low-dose irradiation is a potential intervention measure for controlling T. gondii contamination on blueberries without affecting product quality.

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. HIGHLIGHTS

The interactive effects of mild heat and gaseous chlorine dioxide (ClO

The heat resistance of three heat-resistant strains of

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

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

There are numerous obstacles to the detection of foodborne pathogens in foods that exhibit a low water activity (aw). 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-aw 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-aw foods. A review of relevant rapid and conventional testing methodologies for Salmonella in low-aw foods will also be discussed.

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.

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.

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.

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.

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.

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.

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 ft3/min (1.89 ×10−3 m3/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, respecti...

This chapter provides an overview regarding the advantages of virulotyping over historic serology-based, PCR, based on genes that identify an organism, or enzymatic and biochemical-based analyses of foodborne pathogens in clinical diagnostics and food industry microbiology testing. Traditional identification and characterization methods are designed to detect a given genus, species, serovar or genetic variant of bacteria via enzymatic, genetic or biochemical characterization. Virulotyping is a technique that identifies specific virulence genes within the pathogen’s genome; thus having the discriminatory power to differentiate between a virulent and an avirulent (nonpathogenic) strain of bacterium within a species. As an example, the United States Department of Agriculture, Food Safety and Inspection Service (USDA-FSIS) methods for detection, isolation and identification of Shiga toxin producing E. coli (STEC) use the presence of the eae and stx genes as an initial screen after selective culture enrichment to determine if an adulterant STEC may be present in a raw beef verification sample and as a confirmatory screen of STEC isolates. While virulence testing for STEC is just one example of potential advantages of virulotyping, further benefits may also be realized from virulotyping other common foodborne pathogens (e.g., Campylobacter spp., Salmonella enterica, and Listeria monocytogenes), should avirulent subgroups one day be identified. While PCR and microarray analyses are currently used to perform virulotyping, genome sequencing-based methods may be a more robust and discriminatory technique, when they become practicable for food safety testing and clinical diagnostics. A potential future virulotyping-based medical diagnostics and food testing system might be envisioned in the United States, which could prevent illnesses.

A UV-C treatment system (two treatment chambers connected by an inclined belt to rotate apricots between chambers) was tested in a commercial setting. Escherichia coli ATCC 25922, used as a surrogate for E. coli O157:H7 to determine the system's antimicrobial efficacy, was inoculated onto fruit surfaces at a population of 6.8 log CFU/fruit. UV-C dosage was evaluated by attaching film dosimeters to six fixed locations on each apricot. Results suggested that reduction of inoculated E. coli ATCC 25922 populations on the apricot fruit by UV-C treatment was small (only 0.5–0.7 logs). There were large variations in UV-C doses among varying apricot surface locations. Approximately 1/3 of apricots had individual surfaces receiving less than 0.2 kJ m −2 UV-C exposure, even though fruit received, on average, more than 1 kJ m −2 . Low reductions of E. coli may be attributed, in part, to non-uniform UV-C exposure. This study demonstrates the need to use a fruit rotation device more capable of delivering uniform UV-C dosage to the surface of apricots for inactivating bacteria in a commercial setting.

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.

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 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 (P0.05) 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.

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 (p0.05) 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.

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.

Current FDA regulations require that juice processors achieve a 5 log CFU/ml reduction of a target pathogen prior to distributing products. Whereas thermal pasteurization reduces the sensory characteristics of juice, pulsed electric field (PEF) treatments can be conducted at lower temperatures and may preserve sensory characteristics. Escherichia coli O157:H7 (ATCC 43895) and a non-pathogenic E. coli (ATCC 35218), respectively, were inoculated into single-strength strawberry juice with or without 750 ppm sodium benzoate (SB), 350 ppm potassium sorbate (PS), and 2.7% citric acid (CA). Juice was treated at outlet temperatures of 45, 50 and 55 °C at a field strength of 18.6 kV/cm for 150 μs with a laboratory-scale PEF unit. Inactivation of surrogate E. coli at 45, 50, and 55 °C were 2.86, 3.12, and 3.79 log CFU/ml, respectively, in plain juice (pH 3.4), and 2.75, 3.52, and 5.11 with the addition of benzoic and sorbic acids (pH 3.5). Inactivation of E. coli O157:H7 under the same conditions were 3.09, 4.08, and 4.71 log CFU/ml, respectively, and 2.27, 3.29, and 5.40 with antimicrobials. E. coli O157:H7 in juice with antimicrobials and 2.7% CA (pH 2.7) treated with PEF was reduced by 2.60, 4.32 and 6.95 log CFU/ml at 45, 50 and 55 °C while the surrogate E. coli decreased by 3.54, 5.69, and 7.13 log under the same conditions. When juice (pH 2.7) was held for 6 h without PEF treatment, higher numbers of E. coli 35218 (7.17 log CFU/ml) were inactivated than of acid-resistant E. coli O157:H7 (3.89 log). Slightly greater PEF inactivation of E. coli O157:H7 than of the surrogate bacterium indicates that E. coli ATCC 35218 provides a margin of safety when used as a surrogate for O157:H7 in plain strawberry juice or in juice + SB + PS at 45–50 °C, or with SB + PS and CA at 55 °C.