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Strategies to mitigate antibiotic-resistant bacteria (ARB), including human pathogens, on raw vegetables are needed to reduce incidence of resistant infections. The objective of this research was to determine the effectiveness of standard post-harvest interventions, sanitizer washing and cold storage, to reduce ARB, including antibiotic resistant strains of the human pathogen E. coli O15:H7 and a common spoilage bacterium Pseudomonas, on raw carrots. To provide a background inoculum representing potential pre-harvest carryover of ARB, carrots were dip-inoculated in dairy cow manure compost slurry and further inoculated with known ARB. Inoculated carrots were washed with one of three treatments: sodium hypochlorite (50 ppm free chlorine), peroxyacetic acid (40 ppm peroxyacetic acid; 11.2% hydrogen peroxide), tap water (no sanitizer), or no washing (control). Washed carrots were air dried, packaged and stored at 10 °C for 7d or 2 °C for up to 60 d. Enumeration was performed using total heterotrophic plate counts (HPC), HPCs on antibiotic-containing media ("ARBs"), E. coli O157:H7, and Pseudomonas sp. immediately after washing (0 d) and after 7 d of storage. In addition to the cultured bacteria, changes to the surficial carrot microbiota were profiled by sequencing bacterial 16S rRNA gene amplicons to determine the effect of sanitizer wash, storage temperature, and time of storage (0, 1, 7, 14 and 60 d). Storage temperature, addition of a sanitizer during wash, and duration of storage significantly affected the bacterial microbiota (Wilcoxon, p 0.05). Inclusion of either sanitizer in the wash water significantly reduced the log CFU/g of E. coli O157:H7 and Pseudomonas sp., as well as HPCs enumerated on cefotaxime- (10 μg/ml), sulfamethoxazole- (100 μg/ml), or tetracycline (3 μg/ml) supplemented media compared to the unwashed control (ANOVA, p 0.05). However, no significant reductions to bacteria resistant to vancomycin or clindamycin occurred after washing and storage. Members of the Proteobactetria, Firmicutes, Actinobacteria, Planctomycetes, and Acidobacteria comprised the bacterial carrot microbiota. The diversity of the carrot microbiota was significantly affected by the temperature of storage and by extended storage (60 d), when spoilage began to occur. There were no significant differences to the relative abundance of bacterial groups associated with the type of sanitizer used for washing. Results of this study indicate that inclusion of a sanitizer in wash water, followed by storage at 2 °C, might be an effective strategy to prevent re-growth of pathogenic E. coli O157:H7 and reduce levels of bacteria resistant to certain antibiotics on carrots.

Fresh herbs are popular commodities that generally undergo minimal processing. Salmonella and Escherichia coli O157:H7 survival has been observed on herbs; however, little data exists on the survival of Listeria monocytogenes on herbs. The purpose of this study was to evaluate the survival of L. monocytogenes on the surface of basil, cilantro, dill, and parsley plants grown in a greenhouse. Greenhouse production continues to increase worldwide due to a year-round growing season and controllable conditions. Herb plants were grown in a greenhouse with average temperature of 21 ± 5 °C and relative humidity of 62 ± 10%. Each herb plant was inoculated with a five-strain nalidixic acid-resistant cocktail of L. monocytogenes. Samples were enumerated using standard methods at time-points: 0, 0.21, 1, 2, 3, 7, 14, 21, and 28 d. Population levels in log CFU/g of each herb plant were calculated. The initial inoculum was approximately 4–5 log CFU/g for each herb plant. Overall, L. monocytogenes populations did not grow on the studied herb plants; however, L. monocytogenes was able to survive on the surface of the studied herb plants for up to 28 d, except parsley plants, which fell below the limit of detection 7 d post-inoculation. L. monocytogenes populations demonstrated a similar biphasic survival curve on each of the four different herb plants. L. monocytogenes population decline was most evident within the day post-inoculation for all herb plants. Between 0 and 1 d, L. monocytogenes populations significantly decreased by 3.3, 2.4, 2.6, and 3.2 log CFU/g for basil, cilantro, dill, and parsley plants, respectively. Additionally, within that first 24 h, L. monocytogenes populations significantly decreased on the surface of all herb plants at 5 h post-inoculation (approximately 2.4, 1.6, 2.7, and 2.0 log CFU/g for basil, cilantro, dill, and parsley plants, respectively). No significant difference was observed in L. monocytogenes populations between each of the four herb plants after 7 d post-inoculation. Since L. monocytogenes exhibited survival on the herb plants studied, the adoption of pre-harvest best practices (e.g., sanitation, Good Agricultural Practices) is fundamental to limiting the introduction of contamination, especially in greenhouse environments.

Study objective was to determine efficacy of a bacteriophage suspension against Listeria spp. when applied to three common types of materials used in food manufacturing facilities. Materials included two food contact materials (stainless steel and polyurethane thermoplastic belting) and one noncontact material (epoxy flooring). Coupons of each material were inoculated with a cocktail containing L. monocytogenes and L. innocua (4 to 5‐log₁₀ CFU/cm²). Two phage concentrations and a control, 0, 2 × 10⁷ and 1 × 10⁸ PFU/cm² were evaluated. Treated samples were held at 4 or 20°C for 1 and 3 hr to determine the effect of temperature and treatment time. Reductions in Listeria populations ranged from 1.27 to 3.33 log₁₀ CFU/cm² on stainless steel, from 1.17 to 2.76 log₁₀ CFU/cm² on polyurethane thermoplastic belting, and from 1.19 to 1.76 log₁₀ CFU/cm² on epoxy resin flooring. Higher phage concentration (1 × 10⁸ PFU/cm²), longer treatment time (3 hr), and processing area temperature of 20°C showed a greater (p ≤ .05) reduction of Listeria on the stainless‐steel and polyurethane thermoplastic belting coupons. Overall, Listeria reduction by phage treatment occurred on all three materials tested, under all conditions.

Salmonella serotypes linked to tomato-associated outbreaks were evaluated for survival in soil and water over a 40-day period. Salmonella enterica serotypes Anatum, Baildon, Braenderup, Montevideo, Newport, and Javiana were inoculated separately into sterile soil and water, followed by plating onto TSAYE and XLT4 at 10-day intervals. Biofilm production by Salmonella serotypes was measured on both quartz particles (soil surrogate) and glass coverslips, and was evaluated using a crystal violet dye assay. Salmonella populations in soil and water over 40 days indicated no significant differences between Salmonella serotypes tested (p > 0.05). Over a 40-day period, there was a 1.84 ± 0.22 log CFU/g and 1.56 ± 0.54 CFU/mL decrease in populations of Salmonella in soil and water, respectively. Enumeration indicated that Salmonella population fluctuated in water but decreased linearly in soil. All serotypes tested produced the "red dry and rough" morphotype on Congo Red agar. Biofilm produced by all the Salmonella serotypes tested was significantly different on quartz particles than on glass coverslips (p

A multiyear survey of 31 ready-to-eat (RTE) food processing plants in the United States was conducted to determine the incidence of Listeria spp. in various RTE production environments. Samples were collected from 22 RTE plants regulated by the U.S. Department of Agriculture's Food Safety and Inspection Service (FSIS) and from 9 RTE food plants regulated by the U.S. Department of Health and Human Services' Food and Drug Administration (FDA). Only nonfood contact surfaces in the RTE manufacturing areas with exposed RTE product were sampled. Each sample was individually analyzed for the presence of Listeria spp. by using a PCR-based rapid assay. In total, 4,829 samples were collected from various locations, including freezers, equipment framework, floors, walls, wall-floor junctures, drains, floor mats, doors, and cleaning tools. Nine (29%) of the facilities had zero samples positive for Listeria spp. in the production environment, whereas 22 (71%) had one or more samples positive for Listeria spp. The total incidence of Listeria spp. in all RTE food plants was 4.5%. The positive rate in plants regulated by the FSIS ranged from 0 to 9.7%, whereas the positive rate in plants regulated by the FDA ranged from 1.2 to 36%.

Effective disinfection in healthcare facilities prevents healthcare-associated infections. This study evaluated the ability of Environmental Protection Agency-approved disinfectants (quaternary ammonium compound, QAC; sodium hypochlorite, and hydrogen peroxide) applied with 3 wiping substrates (microfiber, nonwoven, and cotton) to remove Staphylococcus aureus from Formica surfaces. All treatments reduced S aureus on Formica squares with the exception of QAC applied with cotton and QAC, nondisinfectant, and control applied with a nonwoven cloth. Sodium hypochlorite or hydrogen peroxide applied with cotton or microfiber, respectively, may be the best choice for disinfection of Formica surfaces in healthcare settings.

A bacterial contamination in processed foods has been a continuing challenge for the food industry. Many chemical preservatives are showing signs of inefficiencies as microorganisms build resistance. Therefore, research into novel antimicrobial compounds is critical in improving food safety. The ethanolic extracts of medicinal mushrooms (MM; Agaricus blazei Murrill, Ganoderma lucidum, Grifola frondosa, Inonotus obliquus, Lentinula edodes, and Phellinus linteus) were tested for antimicrobial activities against the common foodborne pathogens (Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes) and food spoilage yeast (Saccharomyces cerevisiae, Zygosaccharomyces bailii, and Zygosaccharomyces bailii/bisporus). Inhibitory activities were measured using a disk diffusion assay (DD). Additional analysis for L. monocytogenes and Z. bailii were carried out using Optical density measurements (OD). HPLC was used to fractionate MM extracts (6–8 fractions per MM). The results from DD and OD showed that most MMs displayed statistically significant inhibition compared to the ethanol control. Stronger inhibition was observed for yeasts than bacteria. Also, the antimicrobial activities were fraction dependent. Further work is required to identify the bioactive compounds.

The presence of dust is ubiquitous in the produce growing environment and its deposition on edible crops could occur. The potential of wind-distributed soil particulate to serve as a vehicle for S. Newport transfer to tomato blossoms and consequently, to fruits, was explored. Blossoms were challenged with previously autoclaved soil containing S. Newport (9.39log CFU/g) by brushing and airborne transfer. One hundred percent of blossoms brushed with S. Newport-contaminated soil tested positive for presence of the pathogen one week after contact (P

Understanding a food's ability to support the growth and/or survival of a pathogen throughout the supply chain is essential to minimizing large-scale contamination events. The purpose of this study was to examine the behavior (growth and/or survival) of Listeria monocytogenes on broccoli and cauliflower florets stored at different postharvest temperatures utilized along the supply chain. Broccoli and cauliflower samples were inoculated with L. monocytogenes at approximately 3 log CFU/g and stored at 23 ± 2, 12 ± 2, 4 ± 2, and -18 ± 2°C. Samples were evaluated for L. monocytogenes levels after 0, 0.167 (4 h), 1, 2, 3, and 4 days at 23 ± 2°C; 0, 0.167, 1, 2, 3, 4, 7, 10, and 14 days at 12 ± 2°C; 0, 0.167, 1, 2, 3, 4, 7, 10, 14, 21, and 28 days at 4 ± 2°C; and 0, 1, 7, 28, 56, 84, 112, 140, and 168 days at -18 ± 2°C. L. monocytogenes populations were determined by plating samples onto tryptic soy agar and modified Oxford agar supplemented with nalidixic acid. Broccoli and cauliflower supported the growth of L. monocytogenes at 23, 12, and 4°C, and higher growth rates were observed at higher temperatures. Populations of L. monocytogenes on broccoli and cauliflower samples significantly increased within 1 day at 23°C (by 1.6 and 2.0 log CFU/g, respectively) (P ≤ 0.05). At 12°C, populations of L. monocytogenes on broccoli and cauliflower samples significantly increased over 14 days by 1.4 and 1.9 log CFU/g, respectively (P ≤ 0.05). No significant difference over time was observed in L. monocytogenes populations on broccoli and cauliflower samples held under refrigeration until populations began to grow by day 10 in both commodities (P > 0.05). Under frozen storage (-18°C), populations of L. monocytogenes survived on broccoli and cauliflower at least up to 168 days. Storage of broccoli and cauliflower at lower temperatures can minimize L. monocytogenes growth potential; growth rates were lower at 4°C than at 12 and 23°C. HIGHLIGHTS

Describes the correct temperature and thermometer use for meat, poultry, and egg dishes.

In May 2016, labeling of certain nonintact mechanically tenderized beef (MTB) products was mandated in the United States. MTB products should be handled differently by the consumer because pathogens can be transferred from the exterior to the interior of the meat during the tenderization process. Without labeling, it is difficult to visually distinguish between some intact beef and MTB products, which is a concern because MTB products require higher internal cooking temperatures for safety. An exploratory study was conducted to understand consumer understanding of MTB products and consumer responses to the new label. Thirteen focus groups were convened in rural and urban settings across Virginia and North Carolina between December 2015 and May 2016. Sessions were audiorecorded, transcribed verbatim, and analyzed through constant-comparison thematic analysis. Although MTB products were commonly bought, prepared, and consumed, consumer awareness of MTB products and the MTB process was limited. Generally, the label confused participants, and they did not understand the message. Specifically, terminology such as "blade tenderized" and "mechanically tenderized" were preferred over the term "needle tenderized" on labels. Once explained, many individuals wanted more information and better messaging. Through a multiprong approach, other messaging methods (e.g., in stores, through technology, and with certifications) were highly valued by consumers and may result in increased message clarity. Ultimately, the intrinsic and extrinsic properties of the beef rather than the MTB product continued to be the primary guide for purchasing and preparation. This study is the first to be conducted regarding American perceptions of MTB products. An understanding of consumer awareness of MTB products and labels is needed to develop targeted risk messaging communication tools.

The use of prewetted disinfectant towelettes in health care settings proves challenging because they may dry quickly, reducing disinfection. This study examined the drying time of various commercial disinfectant towelettes and the efficacy of these towelettes over time to eliminate Staphylococcus aureus from glass surfaces. This study confirms that these disinfectants dry quickly. Further disinfection after drying time on glass is minimal, but dependent on the type of disinfectant.

This document is a checklist form with key points about food safety, and more.

Discusses the development of a food safety plan to prevent farm produce from becoming contaminated, and thus reducing chances of foodborne illnesses.

Some land uses may not be acceptable for growing edible fruits and vegetables. The land uses listed in this brochure may need to undergo soil testing or a waiting period before crops are planted.

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, b-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, b-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 b-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 4C are effective strategies to reduce antibiotic-resistant E. coli O157:H7 and P. aeruginosa and relative abundance of various ARG classes. Funding was supported in part by USDA NIFA-AFRI #2014-05280 and #2017-68003-26498 and the Virginia Agricultural Experiment Station and the Hatch Program of the National Institute of Food and Agriculture, U.S. Department of Agriculture.

Mechanically tenderized beef (MTB) products are non-intact beef products that have been tenderized using blades, needles, or other mechanical processes to improve consumer palatability. MTBs have been associated with outbreaks of Shiga toxin-producing Escherichia coli. MTB labeling was mandated in May 2016; however, consumer awareness of product and label is unclear. A national survey quantified consumer understanding and attitudes surrounding MTBs and its preparation. Many (64%) respondents were unaware that beef is mechanically tenderized; MTB label awareness was also low. Thirty-eight percent of respondents familiar with MTBs held positive attitudes towards MTBs due to price and convenience. Negative attitudes (24%) towards MTBs stemmed from concerns with the process, its “natural”-ness, and safety. Respondents regularly preparing MTBs reported using inside and/or outside color of the beef product (63%), texture (43%), juice/blood appearance (47%), smell (22%), and/or time (51%) to indicate doneness. While 20% of respondents used a thermometer, 62% reported “always” or “most of the time” preparing MTBs to 160℉. Contradictions may result from social desirability bias and optimism bias to portray selves in a positive light while believing personal preparation methods achieve safe results. Timely and targeted messaging with clear and standardized definitions, developed with consumers, are needed to bring awareness of MTBs and other food safety information to all populations.

Signs can provide repetitive training on specific food safety practices for multicultural food processing employees. Posted signs for workers in many food processing facilities tend to be text-heavy and focus specifically on occupational hazard safety. The purpose of this study was to evaluate the effectiveness of newly-developed hand washing pictograms on employees' hand washing behavior using video observation. Five employee hand washing behaviors (soap use, wash completeness, wash time, complete rinsing, and towel use) were evaluated with (a) no intervention, company signs posted and considered the baseline; and compared to (b) hand washing behavior the next day (short term) and two weeks (long term) after experimental hand washing signs were displayed at a raw poultry slaughter facility (Facility A) and a poultry further processing facility (Facility B). Both facilities showed a significant increase (p

Cetylpyridinium chloride (CPC) solutions (0, 0.5, or 1.0%) were applied to cantaloupe (Athena and Hale's Best Jumbo cultivars) rind plugs, either before or after inoculation with a broth culture of Salmonella Michigan (10(9) CFU/mL) and held at 37 degrees C for 1 or 24 hr. Rind plugs were diluted, shaken, and sonicated, and solutions were enumerated. Texture quality and color were evaluated over 14 days storage at 4 degrees C after 0 and 1% CPC spray applications. A 0.5 or 1.0% (vol/vol) application of CPC after Salmonella reduced the pathogen levels between 2.34 log CFU/mL and 5.16 log CFU/mL in comparison to the control (p

Fresh fruits and vegetables are the primary commodity sold at farmers markets in the U.S. The growing popularity of farmers markets and number of foodborne outbreaks linked to produce highlights the need for farmers markets to have a proactive approach to food safety practices to protect farmers, patrons and local economies. Many produce farmers selling at these markets are exempt from the food safety regulations within the Food Safety Modernization Act (FSMA). The lack of regulation for these outlets can lead to gaps in training and understanding of how to manage food safety risks. There may also be increased opportunities for food to become contaminated and/or pathogen growth in these temporary establishment settings because they are outdoor settings that often lack infrastructure such as water and electricity. This chapter provides an overview of fresh produce food safety risks and the current landscape of research surrounding farmers market produce. Recommendations for growers to reduce the risk of selling fresh produce in farmers markets include close attention to growing conditions, food handling, sanitation and prevention of potential cross-contamination.

Food safety at the market begins before the food reaches the market, continues while at the market, and extends after the market has ended for the day. The entire chain, from farmers to consumers, must be well-maintained in order to reduce the risk of contamination of a food product at the point of consumption. There are specific behaviors and practices that a market should maintain in order to establish a food safety culture and reduce the risk of foodborne illness. The five main contributing factors associated with foodborne illness are: (1) poor personal hygiene; (2) improper holding/time and temperature; (3) contaminated equipment/lack of protection of food from contamination; (4) inadequate cooking; and (5) food from unsafe sources. Controlling these factors can reduce the occurrence of foodborne illness. Attention to preventing these contributing factors is the cornerstone for establishing a market culture that values food safety. This chapter focuses on specific practices that a farmers market manager and vendors can follow to enhance safety.

Over the past decade, the Eastern Shore of Virginia (ESV) has been implicated in at least four outbreaks of salmonellosis associated with tomato, all originating from the same serovar, Salmonella enterica serovar Newport. In addition to Salmonella Newport contamination, the devastating plant disease bacterial wilt, caused by the phytopathogen Ralstonia solanacearum, threatens the sustainability of ESV tomato production. Bacterial wilt is present in most ESV tomato fields and causes devastating yield losses each year. Although the connection between bacterial wilt and tomato-related salmonellosis outbreaks in ESV is of interest, the relationship between the two pathogens has never been investigated. In this study, tomato plants were root dip inoculated with one of four treatments: (i) 8 log CFU of Salmonella Newport per ml, (ii) 5 log CFU of R. solanacearum per ml, (iii) a coinoculation of 8 log CFU of Salmonella Newport per ml plus 5 log CFU of R. solanacearum per ml, and (iv) sterile water as control. Leaf, stem, and fruit samples were collected at the early-green-fruit stage, and S. enterica contamination in the internal tissues was detected. S. enterica was recovered in 1.4 and 2.9% of leaf samples from plants inoculated with Salmonella Newport only and from plants coinoculated with Salmonella Newport plus R. solanacearum, respectively. S. enterica was recovered from 1.7 and 3.5% of fruit samples from plants inoculated with Salmonella Newport only and from plants coinoculated with Salmonella Newport plus R. solanacearum, respectively. There were significantly more stem samples from plants coinoculated with Salmonella Newport plus R. solanacearum that were positive for S. enterica (18.6%) than stem samples collected from plants inoculated with Salmonella Newport only (5.7%). Results suggested that R. solanacearum could influence S. enterica survival and transportation throughout the internal tissues of tomato plants.

Grocery store associates in deli/bakery departments prepare and serve an increasing number of ready-to-eat (RTE) foods and full meals. The shift to more convenience foods and take-home meals highlights the need for effective food safety training programs in retail grocery establishments to prevent foodborne illness. Through qualitative and quantitative methods, food safety knowledge, training preferences, needs, and current practices of grocery stores deli/bakery food handlers in Southwest and Southern Virginia were explored. Deli employees (n = 78) completed a 34 question survey eliciting information on demographics, food safety training needs, preferences and knowledge. In an additional phase of this project, a subset of those originally surveyed (n = 15) were observed for food handling practices (6 h per person). Observational data collection focused on cross-contamination, glove use and hand washing. Most grocery store deli/bakery food handlers reported wanting frequent hands-on, interactive one-on-one training lasting less than 2 h. The target audience's largest knowledge gaps included correct temperatures for cooking, reheating and cooling foods. Observed behaviors did not correlate with food safety knowledge. For example, greater than 95% of participants reported using correct hand washing techniques; however, observational behavior data showed less than 50% of hand washing events observed were correct. Additionally, food handlers were observed not washing hands prior to putting gloves on as well as practicing bare hand contact with RTE foods. The creation of short, hands-on or interactive trainings for retail grocery food handlers that focuses on changing food handling and preparation behaviors may enhance the food handler's safe food handling practices.

Cranberry products have long been used to treat urinary tract infections. It is believed that the A-type proanthocyanidins in cranberries contribute to this function. Peanut is one of the other, few food sources that primarily contain A-type proanthocyanidins. The skin on the outside of the peanut kernels (testa), which is treated as an agriculture waste product, contains high levels of A-type proanthocyanidins. In this study, an HPLC diol column separation method and MALDI-TOF MS were used to characterize and compare the proanthocyanidin compositions of peanut skins and cranberries. MALDI-TOF MS in linear mode was able to detect a group of proanthocyanidins with DP (degree of polymerization) 10 in peanut skin extract, but was only able to detect DP 8 in cranberry extract. The reflectron mode showed clusters of clear narrow peaks at DP 7 in peanut skin extract, while the highest DP resolved for cranberry extract was only 3 in reflectron mode. This might be due to the low response intensity of the cranberry samples with the current cleanup method and the matrix. Based on the resolved peaks in reflectron mode, pPeanut skins and cranberries have similar proanthocyanidins composition; they contain both A-type and B-type proanthocyanidins, with the A-type being predominant. This result may inspire future studies on the comparison of biological functions between peanut skins and cranberries and further comparison of their polymeric proanthocyanidin composition.

Proanthocyanidins were extracted from peanut skins and investigated for their antimicrobial activity against Saccharomyces cerevisiae, Zygosaccharomyces bailii, and Zygosaccharomyces bisporus in traditional growth media (Sabouraud Dextrose and Maltose broth) and a simulated apple juice beverage. Peanut skins extracts (PSE) were prepared through a multisolvent extraction procedure. The PSE extended the lag phase growth of the 3 yeasts studied at a concentration of 1 mg/mL and at 10 mg/mL yeast growth was totally inhibited for 120 h. PSE was fractionated by normal phase high performance liquid chromatography and the active components/fractions were determined. Compounds present in the fractions were identified by liquid chromatography-mass spectrometry to determine the compounds responsible for inhibition. Fractions consisting mostly of A-type proanthocyanidin dimers, trimers, and tetramers showed the highest percent inhibition toward the yeasts tested in this study. Both optical density (OD) and standard enumeration plating methods were performed in this study. The OD method led to an overestimation of the inhibitory effects of PSE, the 2 methods agreed in respect to treatment effects but not the severity of the inhibition. Practical Application: There is a growing consumer demand for “fresh like” products containing reduced amounts of chemical preservatives without compromising food safety and quality. Therefore, the goal of this study was to determine if an extract of peanut skins containing flavonoid rich compounds could function as a natural antimicrobial in a model beverage system. Proteins were removed through the process of producing the peanut skin extract, thus it is unlikely to contain peanut allergens. The antimicrobial compounds mentioned in this study were successfully integrated into a model beverage system, and were found to have antimicrobial effect. However, the incorporation of these compounds would likely lead to negative sensory attributes at the concentration needed to achieve an appreciable antimicrobial effect alone.

There is a lack of general knowledge regarding the behavior of foodborne pathogenic bacteria associated with jalapeño peppers. The survival and growth behaviors of Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella enterica on the interior and exterior of jalapeño peppers were determined under different storage conditions. Jalapeños were inoculated with a five-strain cocktail of L. monocytogenes, E. coli O157:H7, or S. enterica on the intact external surface, injured external surface, or intact internal cavity of jalapeño peppers and held at 7 or 12°C for a period of 14 days. Populations of each pathogen were determined at 0, 1, 2, 5, 7 10, and 14 days throughout storage. The uninjured, intact external surface of jalapeño peppers did not support growth of the pathogens tested under both storage conditions, with the exception of L. monocytogenes at 12°C. Populations of E. coli and S. enterica declined on the external injured surface of peppers at 7°C, but populations of L. monocytogenes remained consistent throughout the length of storage. At 12°C, L. monocytogenes and S. enterica populations increased throughout storage, and E. coli populations remained unchanged on injured surfaces. The uninjured internal cavity of the jalapeño supported growth of all pathogens at 12°C. Overall, L. monocytogenes was the microorganism most capable of growth and survival in association with jalapeño peppers for the scenarios tested. Results emphasize the importance of jalapeño pepper quality and proper storage conditions in preventing or reducing pathogen survival and growth.

This study characterized the types of interactions between Escherichia coli O157:H7 and spinach phylloepiphytic bacteria and identified those that influence persistence of E. coli O157:H7 on edible plants. A total of 1512 phylloepiphytic bacterial isolates were screened for their ability to inhibit or to enhance the growth of E. coli O157:H7 in vitro and on spinach leaf surfaces. Fifteen different genera, the majority belonging to Firmicutes and Enterobacteriaceae, reduced growth rates of E. coli O157:H7 in vitro by either nutrient competition or acid production. Reduced numbers of E. coli O157:H7 were recovered from detached spinach leaves that were co-inoculated with epiphytic isolates belonging to five genera. A 1.8 log reduction in E. coli O157:H7 was achieved when co-inoculated with Erwinina perscinia and 20% cellobiose, a carbon source used by the phylloepiphytes but not E. coli O157:H7. The reduction on leaves was significantly less than reduction measured in vitro. Phylloepiphytic bacteria belonging to eight different genera, increased numbers of E. coli O157:H7 when co-cultured in vitro on spent medium and when co-cultured on detached spinach leaves. The results, showing reduction of E. coli O157:H7 numbers by natural epiphytic bacteria, support the hypothesis that native plant microbiota can be used for bio-control of foodborne pathogens, however, other epiphytes may promote the persistence of enteric pathogens on the phyllosphere.

Fresh and fresh-cut tomatoes have been associated with numerous outbreaks of salmonellosis in recent years. One effective post harvest treatment to reduce Salmonella enterica in tomatoes may be high pressure processing (HPP). The objectives of the study were to determine the potential for HPP to reduce S. enterica serovars Newport, Javiana, Braenderup and Anatum in tryptic soy broth (TSB) and to determine the effect of HPP to reduce the most pressure resistant of the four serovars from fresh diced and whole tomatoes. To evaluate pressure resistance, TSB containing 8 log CFU/ml of one of the four serovars was packaged in sterile stomacher bags and subjected to one of three different pressures (350, 450 or 550MPa) for 120s. The most pressure resistant S. enterica serovar evaluated was Braenderup. Subjecting the broth culture to 350, 450 and 550MPa resulted in a 4.53, 5.74 and 7.09 log reduction in S. Braenderup, respectively. Diced tomatoes (150g) and whole red round tomatoes (approximately 150g) were inoculated with 0.1ml of 9.1 log CFU/ml S. Braenderup, and subjected to the same pressure treatments (350, 450 or 550MPa). Significant reductions of S. Braenderup concentrations in diced tomatoes (P0.05) were seen after processing at 350 (0.46CFU/g), 450 (1.44 log CFU/g), and 550MPa (3.67 log CFU/g). In whole tomatoes, significant reductions (P0.05) were also seen at 350 (1.41 log CFU/g), 450 (2.25 log CFU/g) and 550MPa (3.35 log CFU/g). HPP may be an effective post harvest strategy to reduce low levels of S. enterica contamination in whole and diced tomatoes.

Environmental factors encountered during growing and harvesting may contribute to Escherichia coli O157:H7 contamination of lettuce. Limited nutrients and extended exposure to water may cause E. coli O157:H7 to shed its O antigen. Absence of the O157-polysaccharide antigen could affect the cell's physicochemical properties (hydrophobicity and cell charge) and ultimately influence its attachment to surfaces. The objectives of this study were to evaluate the effect of the E. coli O157:H7 O-antigen on the cell's overall hydrophobicity, charge and ability to attach to cut edge and whole leaf iceberg lettuce surfaces. Three strains of E. coli O157:H7 (86-24 wild type; F-12, mutant lacking the O-antigen and pRFBE, plasmid for O157 gene reintroduced) were examined for their hydrophobicity, overall charge and ability to attach to lettuce. Overall, E. coli O157:H7 attached at higher levels to cut surfaces over whole leaf surfaces (P=0.008) for all strains and treatments. Additionally, the strain lacking the O-antigen (F12)-attached significantly less to lettuce (P=0.015) than the strains expressing the antigen (WT and pRFBE). Cells lacking the O antigen (strain F-12) were also significantly more hydrophobic than strains 86-24 or pRFBE (P≤0.05). Surface charge differed among the strains tested (P≤0.05); however, it did not appear to influence bacterial attachment to lettuce surfaces. The charge was not fully restored in the pRFBE strain (expression of O-antigen reintroduced), therefore, no conclusions can be made pertaining to the effect of charge on attachment in this study. Results indicate that E. coli O157:H7 cells which lack the O-antigen have greater hydrophobicity and attach at lower concentrations than cells expressing the O-antigen, to iceberg lettuce surfaces.

Aims: To investigate the changes in bacterial diversity on fresh spinach phyllosphere associated with storage at refrigeration temperatures. Methods and Results: Community structure and population dynamics of spinach phylloepiphytic bacteria associated with packaging and refrigeration of ready-to-eat fresh produce were evaluated using pyrosequencing of 16S rRNA gene amplicons. A diverse community responsive to storage at refrigerated temperatures was detected belonging to over 1000 operational taxonomic units, including many diverse members not previously described on the phyllosphere. Of the approx. 8800 unique sequences examined from fresh spinach leaves, 75% were from previously undescribed taxa. The classified sequences from the fresh spinach phyllosphere were assigned to 11 different phyla with the largest number of reads belonging to Proteobacteria and Firmicutes. Packaging and storage of spinach under refrigerated conditions decreased the richness, diversity and evenness of the bacterial community. Refrigeration at 4 and 10°C and storage resulted in a decrease in number of taxa represented from 11 phyla in fresh spinach to only 5 phyla after 1 day of storage. Sequences belonging to γ-Proteobacteria, particularly Pseudomonas spp. and members of the Enterobacteriaceae, were the most numerous after 15 days of storage at both temperatures. Growth inhibition of the genera Escherichia was achieved at 4°C but not at 10°C storage, thus highlighting the importance of temperature in fresh packaged spinach. Conclusions: The application of pyrosequencing to describe composition and diversity of the phyllosphere on spinach leaves provided a broader outlook of the bacterial composition of this community complementing other phyllosphere studies that have used culture- and nonculture-dependent approaches. Significance and Impact of the Study: Pyrosequencing allowed a broader description of the bacterial composition and diversity of the spinach leaf surface than previously obtained using culture-based detection and will be a powerful tool to help ensure the future safety and quality of packaged spinach.

Sodium benzoate has been shown to produce benzene in combination with ascorbic acid. This has led to research for safe alternatives from plant essential oils and parabens that have shown some antimicrobial activity, but many of these compounds exhibit poor solubility in aqueous solutions. Cyclodextrins can increase the solubility of many compounds. This work aimed to investigate the solubility of 23 plant essential oils and 4 parabens in water and an apple juice medium. Four of these compounds were chosen for their low aqueous solubility to determine if complexing the compound with α- and β-cyclodextrin would increase solubility. Three of the complexes were dissolved in an acidified aqueous solution and then studied in glass and polyethylene terephthalate (PET) to determine if storage material would affect the stability. Solubility of the 27 compounds in distilled water ranged from 1.6 mg/L to 2460.6 mg/L and the solubility of 18 of the compounds decreased from 2.5 to 84.7% in apple juice medium (pH = 3.4, 12-13 °Brix). Complexation with cyclodextrin dramatically increased the solubility of the compounds, up to 10-fold. Packaging material had no effect on concentration of compounds present over 7 days. Cyclodextrins were able to increase solubility of these compounds to more suitable concentrations, and may lead to viable natural alternatives to sodium benzoate.

This study investigated the effects of packaging and storage temperature on the spinach phylloepiphytic bacterial community and fate of Escherichia coli O157:H7. Freshly harvested spinach was rinsed and/or disinfected, packaged and stored under typical retail conditions (4 degrees C) or under temperature abuse conditions (10 degrees C) for a period of 15 days. The final population size of culturable epiphytic bacteria after 15 days of storage was not affected by the temperature of storage or the presence of E. coli O157:H7. However, analysis of the bacterial community using denaturing gradient gel electrophoresis of 16s rDNA revealed changes with time of storage and the presence of E. coli O157:H7. Excision and sequencing of prominent DGGE bands identified that the majority of sequences belonged to the phyla Actinobacteria, Bacteroidetes, Firmicutes and Alphaprotebacteria. After 10 days of storage at 4 degrees C or 10 degrees C the population became more dominated by psychrotrophic bacteria. Removal of the epiphytic bacteria resulted in significant increases in numbers of E coli O157:H7 at 10 degrees C and was associated with decreased expression of E. coli O157:H7 virulence (stxA, curli, eaeA) and stress response (rpoS, sodB) genes. In conclusion, storage temperature and time of storage of packaged spinach affected the diversity of the epiphytic spinach microbiota which influenced the growth, establishment, physiology and potentially virulence of E. coli O157:H7.

Tomatoes have been associated with numerous outbreaks of salmonellosis in recent years. Trace-backs suggest tomato fruits may become contaminated during preharvest. The objective of this study was to determine the potential for Salmonella enterica serotype Newport to be internalized into the roots, stems, leaves, and fruit of red round tomato plants through contaminated irrigation water at various stages of plant development. Tomato plants were irrigated with 250 or 350 mL (depending on growth stage) of 7 log CFU·mL−1 S. Newport-contaminated irrigation water every 7 days. Roots, stems, leaves, and two tomato fruit from plants irrigated with S. Newport or water (negative control) were sampled for contamination at five stages of growth. Twenty-five of the 92 total samples taken from plants irrigated with S. Newport were confirmed positive (serovar specificity was not evaluated). Sixty-five percent of confirmed samples were roots, 40% were stems, 10% were leaves, and 6% were fruit. There was a significant difference in the presence of S. enterica according to tissue sampled (roots > stems > leaves, and fruit) (P < 0.05) and no association between growth stage and contamination (P > 0.05). Contamination of tomato fruit with S. Newport introduced through irrigation water is low because a high level of persistent contamination of a plant in the agricultural setting is unlikely.

Cinnamic acid (CA), a naturally occurring organic acid found in fruits and spices, has antimicrobial activity against spoilage and pathogenic bacteria, but low aqueous solubility limits its use. The purpose of this study was to determine the effectiveness of solubility-enhancing alpha-cyclodextrin-CA inclusion complexes against Escherichia coli O157:H7 and Salmonella enterica serovars suspended in apple cider or orange juice at two different incubation temperatures (4 and 26 degrees Celsius). Two concentrations (400 and 1,000 mg/liter) of alpha-cyclodextrin-CA inclusion complex were aseptically added to apple cider inoculated with E. coli O157:H7 (7 log CFU/ml) and orange juice inoculated with a cocktail of six Salmonella enterica serovars (7 log CFU/ml). Samples were extracted at 0 min, at 2 min, and at 24-h intervals for 7 days, serially diluted in 0.1 % peptone, spread plated in duplicate onto tryptic soy agar, and incubated at 35 degrees Celsius for 24 h. Populations of E. coli O157:H7 in apple cider were significantly reduced (Por = 0.05) during the 7-day sampling period in all solutions regardless of temperature. Compared with the controls, populations were significantly reduced by the addition of 400 and 1,000 mg/liter inclusion complex, but reductions were not significantly different (Por = 0.05) between the two treatment groups (400 and 1,000 mg/liter). Salmonella was significantly reduced in all solutions regardless of temperature. There were significant differences between the control and each inclusion complex concentration at 4 and 26 degrees Celsius. Coupled with additional processing steps, alpha-cyclodextrin-CA inclusion complexes may provide an alternative to traditional heat processes.

Tomatoes have been linked to outbreaks of salmonellosis, demonstrating the need to identify sources of contamination. Objectives of this study included determining the ability for Salmonella enterica serovar Montevideo to be internalized into tomatoes from contaminated irrigation water and seed stock, and establishing whether Salmonella Montevideo can survive in fertilizer solutions. Six treatment groups (five plants per group) were irrigated with 350 ml of 7 log CFU/ml of Salmonella Montevideo every 14 days for 70 days, each group receiving an increased number of contaminated water events progressively: group 1 received one contaminated watering at day 0, and group 6 received a total of six contaminated waterings. Group 7 was a control, and group 8 was grown from seeds soaked in 8 log CFU/ml of Salmonella Montevideo for 24 h. All plants were watered daily with uncontaminated water. Three replications were completed. Fruit from every plant, and roots, stems, and leaves of one plant per treatment were sampled. All tomatoes were negative for Salmonella Montevideo; five root samples tested positive. For fertilizer studies, a commercially available fertilizer, two custom mixed and 1.0% dilutions of each (total of six solutions), and sterile water were inoculated with 8 log CFU/ml of Salmonella Montevideo and stored at 25 degrees C. Solutions were sampled at 24, 48, and 72 h. There were no differences (Por = 0.05) between survival of Salmonella Montevideo in diluted fertilizers and the control. Results indicate Salmonella Montevideo is unable to contaminate tomato fruit via irrigation water and seed stock but can survive in fertilizer solutions.

Ready-to-eat meat products have been implicated in several foodborne listeriosis outbreaks. Microbial contamination of these products can occur after thermal processing when products are chilled in salt brines. The objective of this study was to evaluate UV radiation on the inactivation of Listeria monocytogenes and lactic acid bacteria in a model brine chiller system. Two concentrations of brine (7.9% [wt/wt] or 13.2% [wt/wt]) were inoculated with a approximately 6.0 log CFU/ml cocktail of L. monocytogenes or lactic acid bacteria and passed through a UV treatment system for 60 min. Three replications of each bacteria-and-brine combination were performed and resulted in at least a 4.5-log reduction in microbial numbers in all treated brines after exposure to UV light. Bacterial populations were significantly reduced after 5 min of exposure to UV light in the model brine chiller compared with the control, which received no UV light exposure (P0.05). The maximum rate of inactivation for both microorganisms in treated brines occurred between minutes 1 and 15 of UV exposure. Results indicate that in-line treatment of chill brines with UV light reduces the number of L. monocytogenes and lactic acid bacteria.

Curli fibers are produced by some Escherichia coli cells in response to environmental stimuli. These extracellular proteins enhance the cell's ability to form biofilms on various abiotic surfaces. E. coli 0157:H7 cells readily attach to a variety of fruit and vegetable surfaces. It is not known whether the expression of curli influences the cell's ability to attach to produce surfaces. In this study, the effect of curli expression on the cell's overall hydrophobicity, charge, and ability to attach to cut and whole iceberg lettuce surfaces was examined. All strains, regardless of curli expression, attached preferentially to the cut edges of lettuce (P < 0.05). The curli-producing cells of E. coli 0157:H7 strain E0018 attached in significantly greater numbers to both cut and whole lettuce pieces than did the non-curli-producing E0018 cells (P < 0.05); however, no significant attachment differences were observed between the curli-producing and non-curli-producing cells of E. coli 0157:H7 strains 43894 and 43895. All curli-producing E. coli 0157:H7 strains were significantly more hydrophobic (P < 0.01); however, no association between the cells' hydrophobic characteristics and lettuce attachment was observed. Overall surface charge of the cells did not differ among strains or curli phenotypes. Results indicate that overall hydrophobicity and cell charge in E. coli 0157:H7 strains do not influence attachment to iceberg lettuce surfaces. The presence of curli may not have any influence on attachment of E. coli 0157:H7 cells to produce items. Additional factorsmay influence the attachment of E. coli 0157:H7 to plant surfaces and should be further examined.

As produce consumption has increased, so have foodborne disease outbreaks associated with fresh produce. Little research has addressed food safety practices used on small to medium-sized farms selling locally or in farmers markets. This study evaluated current food safety practices used by farmers on small to medium-sized farms and managers of farmers markets in Georgia, Virginia, and South Carolina based on responses to surveys. Surveys were developed, pretested, and revised before implementation with target audiences and were implemented via mail and the Web to maximize participation, with reminders sent to nonrespondents. Data were collected from 226 farmers and 45 market managers. Frequencies and percentages were calculated for all response variables. Responses from farmers indicated that more than 56% of them use manures. Of those who use manures, 34% use raw or mixtures of raw and composted manure, and over 26% wait fewer than 90 days between application of raw manure and harvest. Over 27% use water sources that have not been tested for safety for irrigation, and 16% use such water sources for washing produce. Over 43% do not sanitize surfaces that touch produce at the farm. Only 33% of farmers always clean transport containers between uses. Responses from market managers indicated that over 42% have no food safety standards in place for the market. Only 2 to 11% ask farmers specific questions about conditions on the farm that could affect product safety. Less than 25% of managers sanitize market surfaces. Only 11% always clean market containers between uses. Over 75% of markets offer no sanitation training to workers or vendors. While farmers and market managers are using many good practices, the results indicate that some practices being used may put consumers at risk of foodborne illness. Consequently, there is a need for training for both farmers and market managers.

Ready-to-eat (RTE) deli meats are considered a food at high risk for causing foodborne illness. Deli meats are listed as the highest risk RTE food vehicle for Listeria monocytogenes. Cross-contamination in the retail deli market may contribute to spread of pathogens to deli meats. Understanding potential cross-contamination pathways is essential for reducing the risk of contaminating various products. The objective of this study was to track cross-contamination pathways through a mock retail deli market using an abiotic surrogate, GloGerm, to visually represent how pathogens may spread through the deli environment via direct contact with food surfaces. Six contamination origination sites (slicer blade, meat chub, floor drain, preparation table, employee's glove, and employee's hands) were evaluated separately. Each site was inoculated with 20 ml of GloGerm, and a series of standard deli operations were completed (approximately 10 min of work). Photographs were then taken under UV illumination to visualize spread of GloGerm throughout the deli. A sensory panel evaluated the levels of contamination on the resulting contaminated surfaces. Five of the six contamination origination sites were associated with transfer of GloGerm to the deli case door handle, slicer blade, meat chub, preparation table, and the employee's gloves. Additional locations became contaminated (i.e., deli case shelf, prep table sink, and glove box), but this contamination was not consistent across all trials. Contamination did not spread from the floor drain to any food contact surfaces. The findings of this study reinforce the need for consistent equipment cleaning and food safety practices among deli workers to minimize cross-contamination.

Listeria monocytogenes is the pathogen of concern in ready-to-eat (RTE) meat products. Salt brines are used to chill processed meats. L. monocytogenes and lactic acid bacteria (LAB) can grow under saline conditions, and may compete with each other for nutrients. The objective of this study was to determine the effect of lactic acid bacteria (Enterococcus faecalis, Carnobacterium gallinarum, and Lactobacillus plantarum) on the survival of L. monocytogenes and Listeria innocua in brines stored under low temperatures for 10 d. Sterile tap water (STW) and 2 brine solutions (7.9% and 13.2% NaCl) were inoculated with 1 of 5 cocktails (L. monocytogenes, L. innocua, LAB, L. monocytogenes+ LAB, or L. innocua+ LAB) at initial concentrations of 7 log CFU/mL. Brines were stored for 10 d at 4 or 12 degrees C. Three replications of each brine concentration/cocktail/temperature combination were completed. No significant reductions of L. monocytogenes occurred in 7.9%[w/v] or 13.2%[w/v] brines when LAB were present; however, there were significant reductions after 10 d of L. monocytogenes in the STW solution when LAB were present (1.43 log CFU/mL at 4 degrees C and 3.02 log CFU/mL at 12 degrees C). L. innocua was significantly less resilient to environmental stresses of the brines than L. monocytogenes, both with and without LAB present (P< or = 0.05). These strains of lactic acid bacteria are not effective at reducing L. monocytogenes in brines at low temperatures. Furthermore, use of L. innocua as a model for L. monocytogenes is not appropriate under these environmental conditions.

A key step in promoting good hygiene is ensuring that there are handwashing stations and toilet facilities available on the farm in close proximity (not more than ï__ mile) to the workers.

Ensuring that you have clean water on the farm is an important part of minimizing contamination by disease-causing microorganisms called pathogens. Topics covered include irrigation and surface water, water testing and produce rinsing.

Training and certification programs establish some standard procedures for production, worker hygiene, harvesting, post-harvest handling and even transporting produce to market.