Survival of Listeria Strains and Shelf Life Determination of Fresh Blueberries (Vaccinium corymbosum) Treated with Cold Atmospheric Plasma.

Fresh blueberries are delicate, hand-picked, packaged, and refrigerated fruits vulnerable to spoilage and contamination. Cold atmospheric plasma (CAP) is a promising antimicrobial technology; therefore, this study evaluated the CAP treatment effect on acid-tolerant Listeria innocua and Listeria monocytogenes and evaluated changes in the quality of the treated fruit. Samples were spot-inoculated with pH 5.5 and 6.0 acid-adapted Listeria species. Samples were treated with gliding arc CAP for 15, 30, 45, and 60 s and evaluated after 0, 1, 4, 7, and 11 days of storage at 4 °C and 90% humidity for the following quality parameters: total aerobic counts, yeast and molds, texture, color, soluble solids, pH, and titratable acidity. CAP treatments of 30 s and over demonstrated significant reductions in pathogens under both the resistant strain and pH conditions. Sixty-second CAP achieved a 0.54 Log CFU g-1 reduction in L. monocytogenes (pH 5.5) and 0.28 Log CFU g-1 for L. monocytogenes (pH 6.0). Yeast and mold counts on day 0 showed statistically significant reductions after 30, 45, and 60 s CAP with an average 2.34 Log CFU g-1 reduction when compared to non-CAP treated samples. Quality parameters did not show major significant differences among CAP treatments during shelf life. CAP is an effective antimicrobial treatment that does not significantly affect fruit quality.

Combined treatment of pulsed light and nisin-organic acid based antimicrobial wash for inactivation of Escherichia coli O157:H7 in Romaine lettuce, reduction of microbial loads, and retention of quality.

Microbial safety of fresh produce continues to be a major concern. Novel antimicrobial methods are needed to minimize the risk of contamination. This study investigated the antimicrobial efficacy of pulsed light (PL), a novel nisin-organic acid based antimicrobial wash (AW) and the synergy thereof in inactivating E. coli O157:H7 on Romaine lettuce. Treatment effects on background microbiota and produce quality during storage at 4 °C for 7 days was also investigated. A bacterial cocktail containing three outbreak strains of E. coli O157:H7 was used as inoculum. Lettuce leaves were spot inoculated on the surface before treating with PL (1-60 s), AW (2 min) or combinations of PL with AW. PL treatment for 10 s, equivalent to fluence dose of 10.5 J/cm2, was optimal and resulted in 2.3 log CFU/g reduction of E. coli O157:H7, while a 2 min AW treatment, provided a comparable pathogen reduction of 2.2 log CFU/g. Two possible treatment sequences of PL and AW combinations were investigated. For PL-AW combination, inoculated lettuce leaves were initially exposed to optimum PL dose followed by 2 min AW treatment, whereas for AW-PL combination, inoculated lettuce were subjected to 2 min AW treatment prior to 10 s PL treatment. Both combination treatments (PL-AW and AW-PL) resulted in synergistic inactivation as E. coli cells were not detectable after treatment, indicating >5 log pathogen reductions. Combination treatments significantly (P < 0.05) reduced spoilage microbial populations on Romaine lettuce and also hindered their growth in storage for 7 days. The firmness and visual quality appearance of lettuce were not significantly (P > 0.05) influenced due to combination treatments. Overall, the results reveal that PL and AW combination treatments can be implemented as a novel approach to enhance microbial safety, quality and shelf life of Romaine lettuce.

Transfer of generic Escherichia coli and attenuated Salmonella enterica Typhimurium from the soil to the surface of in-shell pecans during harvest.

During harvest pecan nuts are at risk of contamination with foodborne pathogens from extended contact with the ground. The objective of this study was to determine the potential transfer of Escherichia coli and Salmonella from the ground to in-shell pecans during the harvesting process. Plots (2 m2) were sprayed with 1 L of a rifampicin (rif) resistant strain of either E. coli TVS 353 or an attenuated Salmonella Typhimurium inoculum at a low (∼4 log CFU/ml), mid (∼6 log CFU/ml) or high (∼8 log CFU/ml) concentrations. The following day, nuts were mechanically harvested and samples from each plot were collected at 1 min, 4 h, and 24 h. Samples were enumerated for Salmonella and E. coli on tryptic soy agar supplemented with rif. The Salmonella levels in the soil from the inoculated plots were 2.0 ± 0.3, 4.1 ± 0.1, and 6.4 ± 0.2 log CFU/g for the low, mid, and high inocula, respectively. The E. coli levels in the soil from the inoculated plots were 1.5 ± 0.4, 3.7 ± 0.3, and 5.8 ± 0.1 log CFU/g for the low, mid, and high inocula, respectively. There was a significant difference in the average daily rainfall among the three trials. Trial 3 received 23.8 ± 9.2 cm, while trials 1 and 2 received much less (0.1 ± 0.1 0.0 ± 0.0 cm, respectively). Inoculation concentration and trial were significant (P<0.05) factors that influenced the transfer of E. coli and Salmonella to pecans. For the high inoculum treatment, bacterial transfer to pecans ranged from 0.7 ± 0.3 to 4.1 ± 0.2 for E. coli and 1.3 ± 0.7 to 4.3 ± 0.4 log CFU/g for Salmonella. For the medium inoculum treatment, transfer ranged from <0.3 to 1.5 ± 0.1 for E. coli and <0.3 to 1.9 ± 0.2 log CFU/g for Salmonella. For the low treatment, transfer ranged from <0.3 to 0.4 ± 0.2 and <0.3 to 0.5 ± 0.1 log CFU/g for E. coli and Salmonella, respectively. These results show the need for implementing agricultural practices that prevent potential transfer of foodborne pathogens onto the surface of in-shell pecans during harvest.

A review on plasma-activated water and its application in the meat industry.

Meat is a nutritious food with a short shelf life, making it challenging to ensure safety, quality, and nutritional value. Foodborne pathogens and oxidation are the main concerns that lead to health risks and economic losses. Conventional approaches like hot water, steam pasteurization, and chemical washes for meat decontamination improve safety but cause nutritional and quality issues. Plasma-activated water (PAW) is a potential alternative to thermal treatment that can reduce oxidation and microbial growth, an essential factor in ensuring safety, quality, and nutritional value. This review explores the different types of PAW and their physiochemical properties. It also outlines the reaction pathways involved in the generation of short-lived and long-lived reactive nitrogen and oxygen species (RONS) in PAW, which contribute to its antimicrobial abilities. The review also highlights current studies on PAW inactivation against various planktonic bacteria, as well as critical processing parameters that can improve PAW inactivation efficacy. Promising applications of PAW for meat curing, thawing, and decontamination are discussed, with emphasis on the need to understand how RONS in PAW affect meat quality. Recent reports on combining PAW with ultrasound, mild heating, and non-thermal plasma to improve inactivation efficacy are also presented. Finally, the need to develop energy-efficient systems for the production and scalability of PAW is discussed for its use as a potential meat disinfectant without compromising meat quality.

Radio Frequency Treatment of Food: A Review on Pasteurization and Disinfestation.

Radio frequency (RF) is a novel technology with several food processing and preservation applications. It is based on the volumetric heating generated from the product's dielectric properties. The dielectric properties of each material are unique and a function of several factors (i.e., temperature, moisture content). This review presents a list of dielectric properties of several foods and describes the use of RF as an innovative technology for the food industry. This paper includes several examples of pasteurization, fungi inactivation, and disinfestation in selected food products. The aim of this review is to present the potential applications of RF in pasteurization and disinfestation and research needs that should be addressed. RF has been successfully applied in the inactivation of pathogens such as Salmonella spp., Listeria monocytogenes, and Escherichia coli in low- and high-moisture food. The disinfestation of crops is possible using RF because of selective heating. This process inactivates the insects first because of the different dielectric properties between the pests and the food. The products' final quality can be considerably better than conventional thermal processes. The processing time is reduced compared to traditional heating, and thermal damage to the food is minimized. The main drawback of the technology is the lack of uniform heating, mainly when the product is surrounded by a packaging material with different dielectric properties from the food.

A review on egg pasteurization and disinfection: Traditional and novel processing technologies.

Salmonella Enteritidis is a pathogen related to many foodborne outbreaks involving eggs and egg products. Regulations about whether eggs should be pasteurized are very different and inconsistent worldwide. In the United States, eggs are not required to be pasteurized. Hence, less than 3% of the eggs in the country are pasteurized. The standard pasteurization method (57°C, 57.5 min) uses a long thermal process that increases the cost of the product and affects its quality. Foodborne outbreaks can be reduced if eggs are properly pasteurized to inactivate Salmonella spp. However, the technology to pasteurize eggs needs to offer a faster and more reliable method that can be scaled up to industry settings at a low cost and without affecting product quality. Several novel technologies have been tested for eggshell disinfection and egg pasteurization. Some thermal technologies have been evaluated for the pasteurization of eggs. Microwave has limited penetration depth and is a technical challenge for egg pasteurization. However, radio frequency can penetrate eggshells effectively to inactivate Salmonella, considerably reduce processing time, and maintain the quality of the product. Nonthermal technologies such as ultraviolet, pulsed light, cold plasma, ozone, pressure carbon dioxide, electrolyzed water, and natural antimicrobials have been explored for surface cleaning of the intact egg as alternatives without affecting the internal quality. This review presents some of these novel technologies and the current challenges. It discusses the possible combination of factors to achieve the egg's internal pasteurization and the eggshell's disinfection without affecting the quality at a low cost for the consumer.

Effects of intense pulsed light and gamma irradiation on Bacillus cereus spores in mesquite pod flour.

This study was conducted to evaluate the inactivation of Bacillus cereus spore in mesquite flour with intense pulsed light (IPL) and gamma radiation. The physical, chemical, and toxicity of treated mesquite flour were also investigated. The results showed that up to 3.51 log10CFU/g B. cereus spore inactivation was achieved with 8 kGy of gamma radiation, and up to 1.69 log10CFU/g reductions could be achieved after 28s of catalytic IPL exposure. Although chemometric analysis showed 9-hydroxy-10,12-octadecadienoic acid was slightly increased after a 28s-catalytic IPL treatment, the concentration is within the acceptable range. No significant increase in acetic or propionic acids (typical off-flavor volatile compounds) was observed after either treatment. For cytotoxicity, the Caco-2 cell viability analysis revealed that these two technologies did not induce significant cytotoxicity to the treated mesquite flour. Overall, these two technologies exhibit strong potential for the decontamination of B. cereus in mesquite flour.

Cold Atmospheric Plasma Jet Inactivates Cryptosporidium parvum Oocysts on Cilantro.

ABSTRACT: Cilantro was recently identified as a vehicle for protozoan illness. Current postharvest practices are not sufficient to inactivate protozoa on cilantro. Cold plasma is an emerging nonthermal waterless technology with potential applications in food processing that are currently being investigated to enhance the safety of herbs. The purpose of this study was to determine the impact of cold atmospheric plasma (CP) on the viability of Cryptosporidium parvum oocysts on cilantro. C. parvum oocysts were inoculated onto cilantro and treated with a CP jet for 0, 30, 90, and 180 s at a working distance of 10 cm with a flow of 1.42 × 10-3 m3/s. Oocyst viability was determined using HCT-8 cell culture infectivity assays. Overall, each treatment significantly reduced oocyst infectivity compared with the 0-s treatment control (P ≤ 0.02). Log inactivations of oocysts observed on cilantro were 0.84, 1.23, and 2.03 for the 30-, 90-, and 180-s treatment times, respectively. Drying and darkening of cilantro leaves was observed with treatments longer than 30 s. CP can reduce C. parvum infectivity on cilantro. With further research and optimization, this treatment technology has potential applications in postharvest processing of cilantro.

Nisin-Based Organic Acid Inactivation of Salmonella on Grape Tomatoes: Efficacy of Treatment with Bioluminescence ATP Assay.

The antimicrobial activity of a new nisin-based organic acid sanitizer (NOAS), developed in our laboratory, was tested against viable aerobic mesophilic bacteria and Salmonella populations inoculated on produce surfaces. The activity of NOAS was compared with 200 ppm of chlorinated wash water and a bioluminescence ATP technique to determine the efficacy of treatments compared with plate count methods. The activity of the 10% final concentration of NOAS against viable populations of 109 CFU/mL Salmonella in phosphate-buffered saline (PBS), sterile deionized distilled water, and buffered peptone water was tested in vitro and on grape tomatoes inoculated with Salmonella at 2.5 log CFU/g. A similar batch of inoculated tomatoes were treated with 200 ppm of total available chlorinated water. All treatments for inactivation of viable Salmonella in vitro was tested up to 30 min and 5 min for the attached populations on tomatoes. Inactivation of viable Salmonella at 109 log CFU/mL by 10% the NOAS solution averaged >107 log CFU/mL in PBS, sterile deionized distilled water, and buffered peptone water. Similarly, Salmonella bacteria inactivated on tomato surfaces by the NOAS solution was significantly (P < 0.05) greater than numbers on chlorinated washed tomatoes, and surviving bacterial populations on NOAS and chlorine-treated tomatoes were <1 and 4 CFU/g, respectively. A significant linear correlation coefficient (r2 = 0.99) between the bioluminescence ATP assay and aerobic plate counts of inoculated and untreated grape tomatoes were recorded but not with NOAS and chlorine-treated tomatoes, as bacterial populations were less than the minimum baseline for determination. Also, the results indicated that the NOAS solution is a better alternative antimicrobial wash solution than 200 ppm of chlorinated water.

Gamma Irradiation Influences the Survival and Regrowth of Antibiotic-Resistant Bacteria and Antibiotic-Resistance Genes on Romaine Lettuce.

Contamination of romaine lettuce with human pathogens, antibiotic-resistant bacteria (ARB), and antibiotic resistance genes (ARGs) occurs during production. Post-harvest interventions are emplaced to mitigate pathogens, but could also mitigate ARB and ARGs on vegetables. The objective of this research was to determine changes to lettuce phyllosphere microbiota, inoculated ARB, and the resistome (profile of ARGs) following washing with a sanitizer, gamma irradiation, and cold storage. To simulate potential sources of pre-harvest contamination, romaine lettuce leaves were inoculated with compost slurry containing antibiotic-resistant strains of pathogenic (Escherichia coli O157:H7) and representative of spoilage bacteria (Pseudomonas aeruginosa). Various combinations of washing with sodium hypochlorite (50 ppm free chlorine), packaging under modified atmosphere (98% nitrogen), irradiating (1.0 kGy) and storing at 4°C for 1 day versus 14 days were compared. Effects of post-harvest treatments on the resistome were profiled by shotgun metagenomic sequencing. Bacterial 16S rRNA gene amplicon sequencing was performed to determine changes to the phyllosphere microbiota. Survival and regrowth of inoculated ARB were evaluated by enumeration on selective media. Washing lettuce in water containing sanitizer was associated with reduced abundance of ARG classes that confer resistance to glycopeptides, ß-lactams, phenicols, and sulfonamides (Wilcoxon, p < 0.05). Washing followed by irradiation resulted in a different resistome chiefly due to reductions in multidrug, triclosan, polymyxin, ß-lactam, and quinolone ARG classes (Wilcoxon, p < 0.05). Irradiation followed by storage at 4°C for 14 days led to distinct changes to the ß-diversity of the host bacteria of ARGs compared to 1 day after treatment (ANOSIM, R = 0.331; p = 0.003). Storage of washed and irradiated lettuce at 4°C for 14 days increased the relative abundance of Pseudomonadaceae and Carnobacteriaceae (Wilcoxon, p < 0.05), two groups whose presence correlated with detection of 10 ARG classes on the lettuce phyllosphere (p < 0.05). Irradiation resulted in a significant reduction (∼3.5 log CFU/g) of inoculated strains of E. coli O157:H7 and P. aeruginosa (ANOVA, p < 0.05). Results indicate that washing, irradiation and storage of modified atmosphere packaged lettuce at 4°C are effective strategies to reduce antibiotic-resistant E. coli O157:H7 and P. aeruginosa and relative abundance of various ARG classes.

Influence of Antimicrobial Agents on the Thermal Sensitivity of Foodborne Pathogens: A Review.

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.

In-package atmospheric cold plasma treatment of bulk grape tomatoes for microbiological safety and preservation.

Effects of dielectric barrier discharge atmospheric cold plasma (DACP) treatment on the inactivation of Salmonella and the storability of grape tomato were investigated. Grape tomatoes, with or without inoculation with Salmonella, were packaged in a polyethylene terephthalate (PET) commercial clamshell container and cold plasma-treated at 35 kV at 1.1 A for 3 min using a DACP system equipped with a pin-type high-voltage electrode. DACP treatment inactivated Salmonella (p < 0.05) without altering the color or firmness of the grape tomatoes (p > 0.05). DACP treatment inactivated Salmonella uniformly in both layers of the double-layer configuration of the grape tomatoes regardless of the position of the tomatoes in each layer. Salmonella was most efficiently inactivated when the headspace to tomato volume ratio of the container was highest. Integration of rolling of tomatoes during treatment significantly increased the Salmonella reduction rates from 0.9 ±â€¯0.2 log CFU/tomato to 3.3 ±â€¯0.5 log CFU/tomato in the double-layer configuration of the tomato samples. Rolling-integrated DACP also initially reduced the number of total mesophilic aerobic bacteria and yeast and molds in the double-layer configuration of tomato samples by 1.3 ±â€¯0.3 and 1.5 ±â€¯0.2 log CFU/tomato, respectively. DACP treatment effectively reduced the growth of Salmonella and indigenous microorganisms at 10 and 25 °C, and did not influence the surface color, firmness, weight loss, lycopene concentration and residual ascorbic acid of grape tomatoes during storage at 10 and 25 °C. DACP treatment holds promise as a post-packaging process for improving microbial safety against Salmonella and storability of fresh grape tomatoes.

Evaluation of gaseous chlorine dioxide for the inactivation of Tulane virus on blueberries.

To determine the effectiveness of gaseous chlorine dioxide (gClO2) against a human norovirus surrogate on produce, gClO2 was generated and applied to Tulane virus-coated blueberries in a 240 ml-treatment chamber. gClO2 was produced by an acidifying sodium chlorite solution. Initial assessments indicated that blueberries treated with gClO2 generated from ≤1 mg acidified sodium chlorite in the small chamber appeared unaffected while gClO2 generated from ≥10 mg of acidified sodium chlorite solution altered the appearance and quality of the blueberries. Treatments of inoculated blueberries with gClO2 generated from 0.1 mg sodium chlorite reduced the virus populations by >1 log after exposure for 30 to 330 min. For the 1 mg sodium chlorite treatments, the virus populations were reduced by >2.2 log after 15 min exposure and to non-detectable levels (>3.3 logs reductions) after 180 min exposure. Measured concentrations of gClO2 peaked in the treatment chamber at 0.9 µg/l after 10 min for 0.1 mg treatments and 600 µg/l after around 20 min for 1 mg treatment. Overall results indicate that gClO2 could be a feasible waterless intervention for blueberries and other produce.

Surfactant-Enhanced Organic Acid Inactivation of Tulane Virus, a Human Norovirus Surrogate.

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.

Cold Plasma Inactivation of Salmonella in Prepackaged, Mixed Salads Is Influenced by Cross-Contamination Sequence.

Customer demand for convenient food products has led to an increased production of prepackaged and ready-to-eat food products. Most of these products rely mainly on surface disinfection and other traditional approaches to ensure shelf life and safety. Novel processing techniques, such as cold plasma, are currently being investigated to enhance the safety and shelf life of prepacked foods. The purpose of this study was to determine the effects of cold plasma corona discharge on the inactivation of Salmonella on prepackaged, tomato-and-lettuce mixed salads. Two different inoculation methods were evaluated to address cross-contamination of Salmonella from cherry tomatoes to lettuce and vice versa. In separate studies, a sample of either cherry tomatoes (55 g) or romaine lettuce (10 g) was inoculated with a Salmonella cocktail (6.93 ± 0.99 log CFU/mL), placed into a commercial polyethylene terephthalate plastic container, and thoroughly mixed together with its noninoculated counterpart. Mixed salads were allowed to dry in a biosafety cabinet for 1 h. Samples were treated with 35 kV cold plasma corona discharge inside plastic containers for 3 min. Samples were stomached and serially diluted in buffered peptone water and then were plated onto aerobic plate count Petrifilm and incubated for 18 h at 37°C. When lettuce was the inoculated counterpart, log kill of Salmonella was significantly greater on tomatoes (0.75 log CFU/g) compared with lettuce (0.34 log CFU/g) (P = 0.0001). Salmonella was reduced on mixed salad only when lettuce was the inoculated counterpart (0.29 log CFU/g) (P = 0.002). Cold plasma can kill Salmonella in a prepackaged mixed salad, with efficacy dependent on the nature of contamination, direction of transfer, and the surface topography of the contaminated commodity.

Evaluation of Chlorine Treatment Levels for Inactivation of Human Norovirus and MS2 Bacteriophage during Sewage Treatment.

This study examined the inactivation of human norovirus (HuNoV) GI.1 and GII.4 by chlorine under conditions mimicking sewage treatment. Using a porcine gastric mucin-magnetic bead (PGM-MB) assay, no statistically significant loss in HuNoV binding (inactivation) was observed for secondary effluent treatments of ≤25 ppm total chlorine; for both strains, 50 and 100 ppm treatments resulted in ≤0.8-log10 unit and ≥3.9-log10 unit reductions, respectively. Treatments of 10, 25, 50, and 100 ppm chlorine inactivated 0.31, 1.35, >5, and >5 log10 units, respectively, of the norovirus indicator MS2 bacteriophage. Evaluation of treatment time indicated that the vast majority of MS2 and HuNoV inactivation occurred in the first 5 min for 0.2-µm-filtered, prechlorinated secondary effluent. Free chlorine measurements of secondary effluent seeded with MS2 and HuNoV demonstrated substantial oxidative burdens. With 25, 50, and 100 ppm treatments, free chlorine levels after 5 min of exposure ranged from 0.21 to 0.58 ppm, from 0.28 to 16.7 ppm, and from 11.6 to 53 ppm, respectively. At chlorine treatment levels of >50 ppm, statistically significant differences were observed between reductions for PGM-MB-bound HuNoV (potentially infectious) particles and those for unbound (noninfectious) HuNoV particles or total norovirus particles. While results suggested that MS2 and HuNoV (measured as PGM-MB binding) behave similarly, although not identically, both have limited susceptibility to chlorine treatments of ≤25 ppm total chlorine. Since sewage treatment is performed at ≤25 ppm total chlorine, targeting free chlorine levels of 0.5 to 1.0 ppm, these results suggest that traditional chlorine-based sewage treatment does not inactivate HuNoV efficiently.IMPORTANCE HuNoV is ubiquitous in sewage. A receptor binding assay was used to assess inactivation of HuNoV by chlorine-based sewage treatment, given that the virus cannot be routinely propagated in vitro Results reported here indicate that chlorine treatment of sewage is not effective for inactivating HuNoV unless chlorine levels are above those routinely used for sewage treatment.

In-package inhibition of E. coli O157:H7 on bulk Romaine lettuce using cold plasma.

Dielectric barrier discharge atmospheric cold plasma (DACP) treatment was evaluated for the inactivation of Escherichia coli O157:H7, surface morphology, color, carbon dioxide generation, and weight loss of bulk Romaine lettuce in a commercial plastic clamshell container. The lettuce samples were packed in a model bulk packaging configuration (three rows with either 1, 3, 5, or 7 layers) in the container and treated by DACP (42.6 kV, 10 min). DACP treatment reduced the number of E. coli O157:H7 in the leaf samples in the 1-, 3-, and 5-layer configurations by 0.4-0.8 log CFU/g lettuce, with no significant correlation to the sample location (P > 0.05). In the largest bulk stacking with 7 layers, a greater degree of reduction (1.1 log CFU/g lettuce) was observed at the top layer, but shaking the container increased the uniformity of the inhibition. DACP did not significantly change the surface morphology, color, respiration rate, or weight loss of the samples, nor did these properties differ significantly according to their location in the bulk stack. DACP treatment inhibited E. coli O157:H7 on bulk lettuce in clamshell containers in a uniform manner, without affecting the physical and biological properties and thus holds promise as a post-packaging process for fresh and fresh-cut fruits and vegetables.

Inactivation of Escherichia coli O157:H7 and Aerobic Microorganisms in Romaine Lettuce Packaged in a Commercial Polyethylene Terephthalate Container Using Atmospheric Cold Plasma.

The effects of dielectric barrier discharge atmospheric cold plasma (DACP) treatment on the inactivation of Escherichia coli O157:H7 and aerobic microorganisms in romaine lettuce packaged in a conventional commercial plastic container were evaluated during storage at 4°C for 7 days. Effects investigated included the color, carbon dioxide (CO2) generation, weight loss, and surface morphology of the lettuce during storage. Romaine lettuce pieces, with or without inoculation with a cocktail of three strains of E. coli O157:H7 (~6 log CFU/g of lettuce), were packaged in a polyethylene terephthalate commercial clamshell container and treated at 34.8 kV at 1.1 kHz for 5 min by using a DACP treatment system equipped with a pin-type high-voltage electrode. Romaine lettuce samples were analyzed for inactivation of E. coli O157:H7, total mesophilic aerobes, and yeasts and molds, color, CO2 generation, weight loss, and surface morphology during storage at 4°C for 7 days. The DACP treatment reduced the initial counts of E. coli O157:H7 and total aerobic microorganisms by ~1 log CFU/g, with negligible temperature change from 24.5 ± 1.4°C to 26.6 ± 1.7°C. The reductions in the numbers of E. coli O157:H7, total mesophilic aerobes, and yeasts and molds during storage were 0.8 to 1.5, 0.7 to 1.9, and 0.9 to 1.7 log CFU/g, respectively. DACP treatment, however, did not significantly affect the color, CO2 generation, weight, and surface morphology of lettuce during storage (P > 0.05). Some mesophilic aerobic bacteria were sublethally injured by DACP treatment. The results from this study demonstrate the potential of applying DACP as a postpackaging treatment to decontaminate lettuce contained in conventional plastic packages without altering color and leaf respiration during posttreatment cold storage.

Nonthermal inactivation of norovirus surrogates on blueberries using atmospheric cold plasma.

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.

Survival of Salmonella Typhimurium on soybean sprouts following treatments with gaseous chlorine dioxide and biocontrol Pseudomonas bacteria.

Control of Salmonella Typhimurium on sprouts is crucial for food and consumer safety. In this study, natural microflora on soybean seed was assessed and effects of gaseous chlorine dioxide (ClO2) and biocontrol Pseudomonas on the survival of S. Typhimurium on soybean sprouts were evaluated. Sprouts were dip-inoculated with S. Typhimurium prior to the application of the biocontrol (P. chlororaphis and P. fluorescens). After inoculation with S. Typhimurium, the sprouts were treated with ClO2 at 0.4 mg/L for 1 h (90% R.H., 13°C). Pseudomonas strains and Salmonella were recovered on Pseudomonas Agar F (PAF) and xylose lysine tergitol-4 (XLT-4) media, respectively. Pseudomonas strains reduced Salmonella by <1 log colony forming units (CFU)/g of sprouts, whereas S. Typhimurium on soybean sprouts was reduced from 2.55 to 5.35 logs CFU/g by ClO2. Gaseous ClO2 treatment reduced S. Typhimurium by 3.90 (0 h), 4.47 (24 h), and 3.61 log CFU/g (168 h). It was concluded that ClO2 and biocontrol treatment can enhance sprout safety.