The growing popularity of farmers' markets coupled with a high percentage of produce-related foodborne outbreaks highlights the need for an emphasis on food safety within these markets to protect farmers, patrons and local economies. The number of farmers' markets registered in the United States has almost tripled in the last 15 years. Fresh produce constitutes the majority of food sold at farmers'markets. Between 1998 and 2008, raw produce accounted for almost half of the 4,589 foodborne illness outbreaks linked to a specific commodity. This research was conducted to identify practices at farmers' markets which may contribute to an increased risk of contamination, assess the microbial quality of produce sold at farmers' markets, as well as to develop a food safety management plan template for market managers to utilize to build their own food safety plan. Using an observational data collection method, risky food safety practices were identified at Southwest Virginia farmers' markets. While market managers and vendors in three of the five markets observed had formal food safety training, numerous risky food safety behaviors were still observed including temperature abuse, cross contamination opportunities, and poor personal hygiene and sanitation. Additionally, the microbial quality of produce from Southwest Virginia farmers' markets was compared to produce sold at retail using culture based microbiological plating and molecular methods. Total aerobic bacteria and coliforms were enumerated, and the presence of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp., Staphylococcus aureus and generic E. coli were determined. A significantly greater quantity of total aerobic bacteria was isolated from farmers' market leafy greens, onions and tomatoes when compared to a retail grocery store (P=0.0011, P=0.0395, and P<0.0001, respectively). Additionally, one or more target pathogen was isolated from 28 farmers' market samples and 16 retail grocery store samples. The observed risky food safety behaviors along with the bacterial data collected emphasize the need for a pathogen reduction focus on fresh produce not only at farmers' markets, but also with growers and other retail outlets. To help promote proper food safety practices at farmers' markets, a farmers' market food safety management plan (FSMP) template was developed to address the top five risk factors contributing to foodborne illness as identified by the Centers for Disease Control and Prevention (CDC). The FSMP was evaluated for practicality and feasibility through interviews with market mangers in North Carolina and Virginia. Most market managers (66.7%) agreed that the FSMP was practical for their market while only 33.3% agreed that they could implement the plan immediately. Revisions suggested to the FSMP will be made and it will be made available in Virginia and North Carolina in spring 2016. / Ph. D.

Recent demands for minimally-processed foods, has led to the exploration of plant-derived essential oil (EO) compounds as an alternative means of preservation. While some of these compounds are effective against foodborne pathogens, their strong aroma and "spicy" flavor are not compatible with the flavor of juice. The purpose of this research was to evaluate the antimicrobial activity of three EO compounds (thymol, eugenol, and trans-cinnamaldehyde) alone and in combination with three naturally-occurring apple aroma compounds (hexanal, trans-2-hexenal and 1-hexanol) in order to identify combinations that lower the concentrations needed to destroy foodborne pathogens in apple juice. The standard agar dilution method (SAD) and the Spiral Gradient Endpoint method (SGE) were compared for their abilities to determine minimum inhibitory concentrations (MIC) of the EO compounds. Both methods produced similar patterns of inhibition; however, the MICs produced by the SGE system were significantly higher than those produced by the SAD method of analysis (P<0.05). Since the results produced by the SAD method were more comparable with those published in literature, this method was selected for further testing. In general, the EO compounds were significantly more effective against the test pathogens (Listeria monocytogenes, Salmonella Typhimurium and Staphylococcus aurues) than were the apple aroma compounds (P<0.05). Cinnamaldehye exhibited the highest degree of activity, followed by thymol and eugenol. Eugenol was the only compound that acted synergistically with the apple aroma compounds. The most effective compounds (cinnamaldehyde, eugenol and trans-2-hexenal) were then used to inactivate L. monocytogens and S. Typhimurium in preservative-free apple juice. In most cases, treatment with 0.05% of each compound resulted in a 5 log CFU/ml reduction in bacterial numbers following one day of storage at 4°C or 25°C. Likewise, treatment with antimicrobial combinations (containing 0.025% of trans-2-hexenal in combination with 0.025% trans-cinnamaldehyde or eugenol) also resulted in a 5 log CFU/ml reduction in bacterial numbers, following one day of storage at 4°C or 25°C. Since these combinations contained half the effective concentration of the essential oil compounds, they may be used to preserve the microbial quality of apple juice, while reducing the likelihood of off flavors in the final juice product. / Ph. D.

Hot peppers are gaining popularity in the United States as both a vegetable and a spice. In 2008, jalapeño peppers were involved in a multistate outbreak of Salmonella Saintpaul. This is the first outbreak implicating jalapeño as a vehicle for foodborne illness. Hot peppers contain many compounds thought to possess antimicrobial characteristics. This research was conducted to provide more information on the interactions of pathogenic bacteria and jalapeño peppers, as well as to identify properties of Capsicum annuum that affect bacterial survival, growth, and inhibition. Behavior of pathogens associated with jalapeños was investigated by inoculating jalapeño fruits with a cocktail of Listeria monocytogenes, Salmonella enterica, or Escherichia coli O157:H7 on the intact external surface, injured external surface, or intact internal cavity and storing the jalapeños at 7°C or 12°C. Intact external jalapeñosurfaces did not support the growth of the bacteria tested under storage conditions of 7°C. However, L. monocytogenes populations remained detectable throughout the 2 week study. At 7°C, pathogenic bacteria were able to survive but not grow on injured and internally inoculated jalapeño, but populations increased at 12°C (p=0.05). The most supportive growth environment for the pathogenic bacteria was the internal cavity of jalapeño held at 12°C. This study demonstrated the importance of intact uninjured produce and proper storage temperatures for food microbial safety. Inhibitory properties of jalapeños were studied by making extracts from fresh jalapeño peppers to test for antimicrobial activity. A disk diffusion assay determined that the extracts were capable of inhibiting the growth of the pathogenic bacteria tested. Listeria monocytogenes was especially sensitive to the extracts. jalapeño extracts were fractionated using HPLC and used for inhibition assays using disk diffusion and growth curve generation. Two fractions stimulated bacterial growth (p=0.05), while two other fractions inhibited bacterial growth. The inhibitory fractions were separated further using HPLC and tested for antimicrobial activity. Fraction E1 suppressed the growth of L. monocytogenes. HPLC-MS analysis revealed that Fraction E1 contained compounds known as capsianosides. To prove that inhibition is caused by capsianoside(s) and determine minimum inhibitory concentrations, a method to isolate the pure compound should be developed. / Ph. D.