Search

We present the magnetic properties of a new family of S = 1 molecule-based magnets, NiF2(3,5-lut)4·2H2O and NiX2(3,5-lut)4, where X = HF2, Cl, Br, or I (lut = lutidine C7H9N). Upon creation of isolated Ni–X···X–Ni and Ni–F–H–F···F–H–F–Ni chains separated by bulky and nonbridging lutidine ligands, the effect that halogen substitution has on the magnetic properties of transition-metal-ion complexes can be investigated directly and in isolation from competing processes such as Jahn–Teller distortions. We find that substitution of the larger halide ions turns on increasingly strong antiferromagnetic interactions between adjacent Ni2+ ions via a novel through-space two-halide exchange. In this process, the X···X bond lengths in the Br and I materials are more than double the van der Waals radius of X yet can still mediate significant magnetic interactions. We also find that a simple model based on elongation/compression of the Ni2+ octahedra cannot explain the observed single-ion anisotropy in mixed-ligand compounds. We offer an alternative that takes into account the difference in the electronegativity of axial and equatorial ligands.\ud \ud

Knowledge of construction duration is pertinent to a number of project planning functions prior to detailed design development. Funding, financing, and resource allocation decisions take place early in project design development and are significantly influenced by the construction duration. Currently, there is not an understanding of the project factors having a statistically significant relationship with highway construction duration. Other industry sectors have successfully used statistical regression analysis to identify and model the project parameters related to construction duration. While the need is seen for such work in highway construction, there are very few studies which attempt to identify duration-influential parameters and their relationship with the highway construction duration. This research identifies the project factors, known early in design development, which influence highway construction duration. The factors identified are specific to their respective project types and are those factors which demonstrate a statistically-significant relationship with construction duration. This work also quantifies the relationship between the duration-influential factors and highway construction duration. The quantity, magnitude, and sign of the factor coefficient yields evidence regarding the importance of the project factor to highway construction duration. Finally, the research incorporates the duration-influential project factors and their relationship with highway construction duration into mathematical models which assist in the prediction of construction duration. Full and condensed models are presented for Full-Depth Section and Highway Improvement project types. This research uses statistical regression analysis to identify, quantify, and model these early-known, duration-influential project factors. The results of this research contribute to the body of knowledge of the sponsoring organization (Virginia Department of Transportation), the highway construction industry, and the general construction industry at large. Ph. D.

Farmers markets (FM) have become increasingly popular almost tripling over the past two decades due to the rising interest in local and/or organic foods. Within this same time period, notable farmers market foodborne illness outbreaks; such as E. coli O157:H7 and strawberries and Salmonella and peas; have occurred, and emphasize the importance of food safety practices at farmers markets. Some farmers may be encouraged to follow Good Agricultural Practices (GAPs) to limit the contamination of fresh produce; however, GAPs programs are complex and also very driven by wholesale buyers. Furthermore, many FM growers are unfamiliar with GAPs or do not believe it is applicable to them. The Food Safety Modernization Act (FSMA) was passed in 2011, and shifted the food safety paradigm from a reactive to proactive approach introducing seven different rules including the Preventive Controls for Human Foods (PCHFR) and the Produce Safety Rule (PSR). Each FSMA rule does contain exemptions for smaller-scale operations (e.g., farms, facilities) that allows them to be excluded from the rule, or excluded from certain aspects of the rule. Generally, most FM vendors may satisfy exemptions from the FSMA regulations (e.g., be exempt from the FSMA Produce Safety Rule because of commodities); however, to our knowledge, no studies have assessed the regulatory compliance requirements of FM vendors to the FSMA regulations. This dissertation explored the regulatory requirements of FM vendors, and the accessibility of FSMA materials for these audiences. Additionally, due to the sudden and unprecedented global COVID-19 pandemic, an objective was added that explored how COVID-19 influenced behavior changes among this audience. Interviews were conducted and found that all vendors (100%) were exempt from the PCHFR and that most vendors (67%) were exempt from the PSR, some were qualified exempt (28%), and a few (5%) were covered. This study also found that farmers market vendors received information mainly from University Cooperative Extension sources. The COVID-19 pandemic greatly impacted hygiene and health practices at FMs. Market personnel (managers and vendors) implemented many different hygiene and health practices at their markets. The major source of COVID-19 food safety information for market personnel was local and federal government; as well as University Cooperative Extension. Accessibility analyses showed that many FM or COVID-19 resources were not accessible due to populations that rely on produce safety resources generated by Cooperative Extension due to (i) navigation or web accessibility errors, (ii) high literacy level requirements, and (iii) lack of keep (i.e., resources were not up-to-date, or continuously managed or monitored). These findings will inform the development of FM targeted resources, that are also, accessible to a more diverse and inclusive audience. One example is a produce safety resource on the updated agricultural water requirements that is developed to an 8th grade reading level, with no broken links or additional navigations errors, and if a PDF version is available, proper headers and titles. Doctor of Philosophy Farmers markets (FMs) have increased in popularity over the two decades with because of interest by consumers to support local agriculture, buy organic products, or because they believe these foods to be safer from bacteria and viruses. Shopping at a FMs does not necessarily make the products any safer compared to buying it from a grocery store. It is important that farmers markets follow proper food safety practices during the growing, harvesting, and preparation of produce for sale. One example is the prevention of animal feces from getting on the produce or in a water source that would be used. Another would be to follow proper hygiene practices such as handwashing after using the bathroom. These two examples and others are detailed in a law passed by President Obama in 2011 called the Food Safety Modernization Act (FSMA), which allows the Food and Drug Administration to begin its proactive instead of reactive food safety approach. FSMA does not have to be followed by all food entities. Depending on how much a farm makes in revenue (

Solar radiation influences virtually all biological process on earth. Yeasts, the microbial driver of ethanol fermentation, evolved on the surface of vegetation and had to adapt to survive photonic assault. Past research has demonstrated that white light affects yeast metabolism along with the ability to entrain circadian rhythms, although no known genetic mechanism accounts for this. High intensity narrow wavelength light-emitting diodes were employed to illuminate synthetic cultures under fermentation. Multiple colors along the visible spectrum were used, corresponding to the peak absorbance wavelengths of Saccharomyces sp. yeast. Impacts in primary metabolite evolution were found, dependent on wavelength. Longer wavelengths produced higher amounts of acetic acid and glycerol; shorter wavelengths produced more ethanol. Because past research showed light timing had pronounced effects, illumination schemes on the scale of milliseconds to hours were tested for ethanol production. Light schemes on the scale of enzymatic reactions, yeast generation times, and circadian rhythms produced the most ethanol. Discrete blocks and duration of illumination were used to elucidate where light had the most influence over yeast metabolism and fermentation. Late lag phase and mid log phase illumination impacted ethanol fermentation more than any other period of time. Light effects were tested on apple juice to see if they extended from synthetic media to natural products. Significant impacts on ethanol production were discovered and flavor/aroma impacts were noted. Light, color, intensity, and timing have all been shown to control and affect fermentation with both positive and negative effects established. Doctor of Philosophy Sun light influences virtually all biological process on earth. Yeasts, the microbial drivers of ethanol fermentation, evolved on the surface of vegetation and had to adapt to survive destructive effects of the sun. Past research has demonstrated that white light affects yeast metabolism along with the ability to develop growth cycles similar to day / night patterns, although it is currently not believed this possible due to the biology of yeast. High intensity single color light-emitting diodes were employed to illuminate laboratory formulated cultures under fermentation. Multiple colors along the visible spectrum were used, corresponding to the peak absorbance wavelengths of Saccharomyces sp. yeast. Green/yellow/red wavelengths produced higher amounts of acetic acid (vinegar) and glycerol; blue and ultraviolet wavelengths produced more ethanol. Because past research showed light timing could change how yeast grow and consumed carbohydrates, light timing on the scale of milliseconds to hours were tested for ethanol production. Light timing on the scale of milliseconds, hours, and daylight cycles produces the most ethanol. Discrete blocks and duration of illumination were used to find where during fermentation light had the most impact. It was found that from immediately after the beginning of fermentation to the middle of fermentation is where yeast responded the most strongly. Light effects were tested on apple juice to see if they extended laboratory cultures to natural products. Significant changes in the amount of ethanol produced were discovered and changes in the taste and smell of fermented apple juice were noted. Light, color, intensity, and timing have all been shown to control and affect fermentation with both positive and negative effects established.

Public health costs of foodborne campylobacteriosis are estimated to cost more than a billion dollars in the United States annually. This pathogen has been primarily associated with chicken production and processing which is a ~$33 billion industry. To further identify practices that could reduce Campylobacter presence, concentration, and persistence in chicken prior to consumption a quantitative microbial risk assessment (QMRA) was conducted for boneless, skinless chicken breast meals prepared and consumed domestically in the contiguous United States. This QMRA model was developed with @RISK software (Palisade Corp., Ithaca, NY) and data inputs can be easily modified and updated. QMRA is a powerful analytic method that can be utilized to model the dynamics between the food pathogen, food commodity, and ingestion. It provides insight into the impacts of the process in its interaction and its surrounding environment. The baseline model determined that consumption of this product resulted in annual means of infections: 328,257, illnesses: 108,174, hospitalizations: 27,754, deaths: 37, cases of Campylobacter-associated Guillain-Barré Syndrome (GBS): 1,373, and cases of Campylobacter-associated Irritable Bowel Syndrome (IBS): 9,501. The associated annual economic burdens were ~$192 million for acute campylobacteriosis, ~$77 million for GBS, and ~$96 million for IBS. The effects of targeting and modifying the baseline model's inputs within the farm-to-fork process were studied as follows: Post grow-out (1) prevalence and (2) concentration of Campylobacter on chickens at the farm prior to transport, (3) transport crate cleaning frequency prior to loading, (4) temperature storage conditions at post processing/ pre-retail, (5) the frequency of handwashing in the preparation and handling of a chicken meal, and (6) the combination Campylobacter mitigation models using the inputs from (2) and (4). Mean yearly illnesses are estimated to decrease by approximately half if on-farm Campylobacter prevalence was lowered to 35-50% from the baseline level of 76%. An additional ~50,000 illnesses can be expected if the proportion of home preparers who do not wash their hands increases from 8.3% (baseline) to 20%. The combined effects of reducing on-farm Campylobacter concentrations and increasing the proportion of product frozen in-plant have greater impacts on reducing yearly campylobacteriosis and associated costs than either intervention alone. Doctor of Philosophy Campylobacter is a major foodborne bacterial pathogen that is not well known by the general public, although public health costs are estimated to cost more than a billion dollars in the United States annually. Chicken production is a ~$33 billion industry that is affected by the contamination of this pathogen. In this study, a quantitative microbial risk assessment (QMRA) was performed to study the impact of Campylobacter on chicken meals prepared in the home. Societal and economic burdens were evaluated as well. The models developed provide comprehensive simulations that describe the spread, persistence, and the concentration of Campylobacter throughout the farm to fork process in the annual consumption of boneless, skinless chicken breast meals prepared and consumed domestically in the contiguous United States. Additional simulation models were created to compare methods for reducing Campylobacter along the food chain that could lead to fewer cases of campylobacteriosis in the United States, or what could happen if there was a breakdown in a hygiene step in the preparation of the chicken meal.

Low water activity foods (LWAF), specifically nuts and dried fruits, have been generally considered safe because they do not support the growth of foodborne pathogens. However, many pathogens have been noted to survive in LWAF for considerable periods of time, and a number of recent outbreaks and recalls have implicated various types of nuts and dried fruits. The Food Safety Modernization Act requires food processors to develop preventive control plans that make ready-to-eat LWAF safer for consumers. The presented research was designed to investigate several aspects of LWAF safety by evaluating a steam process as a strategy to remove pathogen contamination from LWAF, modeling the inactivation of such treatments, and studying the thermal resistances of two E. coli strains in low-water activity solutions. Low-temperature, vacuum-assisted steam (vacuum-steam) was evaluated as a potential intervention and preventive control to remove pathogens from the surface of LWAF without using high-heat treatments that could damage product quality. The presented work examined the efficacy of vacuum-steam (4 log CFU/g from the surfaces of the foods. Additionally, Pediococcus acidilactici, proved to be a surrogate organism for these pathogens and could be used to challenge and validate similar treatments within processing plants. The data were fit to models, which showed that food type significantly impacted the fit, with the Weibull model best describing bacterial inactivation kinetics on raisins and macadamia nuts, and the Gompertz model best describing reductions on the apricot halves. The models were challenged for validation of their abilities to predict times required for 3-log reductions using internal and external datasets, determining the usefulness to industry members who wish to design similar thermal treatments for LWAF. Comparing predicted values from internally constructed models to observed values generated from external data, models were shown to be limited in scope and application and could only be applied to pathogen inactivation on different LWAF or thermal processes in certain circumstances. First-order and Weibull model predictions of bacterial reductions on dried apricots had varied success in predicting times for 3-log reductions on other thermally treated LWAF. However, the models of bacterial reductions on thermally treated macadamia nuts frequently overestimated the times required for 3-log bacterial reductions for other LWAF. In an effort to understand the effect that reduced water activity has specifically on STEC, two strains were investigated for induced thermal resistance due to osmotic stress. Thermal resistance of STEC strains (O121:H19 and O157:H7) were evaluated on the basis of strain variation, culture preparation, and water activity (D- and z-values). At the lowest treatment temperature (56°C), O121 displayed greater heat resistance than O157, and the broth-grown samples exhibited greater heat resistance than the lawn-grown cells, but significant differences were not observed at higher temperatures. Samples in reduced-water activity solutions displayed reduced thermal resistance at 56°C, but the z-values were 29-43% higher than those of high-water activity samples. While water activity has been shown to impact thermal resistance of pathogens, comparisons of STEC thermal resistance according to the D- and z-values revealed that other factors also play roles in pathogen thermal resistance on LWAF. Results from the collection of experiments conclude that efficacy of thermal treatments is impacted by the physiological state of the cells, stress experienced in the food matrix, and characteristics of the food, including water activity and composition. Doctor of Philosophy Consumers expect foods they purchase to be safe to consume by themselves and family members, particularly those that are ready-to-eat with no additional cooking requirements. Many of these foods are low-water activity foods (LWAF), like nuts and dried fruits, with very little water content that could be used by bacteria. These foods may be preferred snack foods due to their affordability, long shelf lives, and health benefits over other types of snack foods. Until recently, LWAF were generally considered safe because they do not support the growth of foodborne pathogens due to the lack of moisture or water within the food. However, a number of recent outbreaks related to various types of nuts and dried fruits have proven that many pathogens can survive in dried foods, even if not actively growing, for considerable amounts of time. Designed to address these types of food safety issues, the Food Safety Modernization Act recognizes risks associated with foods and responded with regulations requiring food processors to take steps to make ready-to-eat LWAF, like nuts and dried fruits, safer for consumers. A popular strategy is to treat foods with heat to destroy pathogens, however the quality attributes of some nuts and dried fruits could be damaged by high-heat treatments like roasting. An alternative process uses a vacuum to form steam at lower temperatures, allowing for efficient heat transfer through water droplets to the surface of the foods, thus causing less damage to the foods without introducing too much moisture. This research evaluated how this process could be used by food processors to remove harmful bacteria from the surfaces of whole macadamia nuts, dried apricot halves, and raisins. Results indicated that the low-temperature steam treatments successfully reduced Salmonella, Listeria monocytogenes, and Shiga toxin-producing Escherichia coli (STEC) by >4 log CFU/g (>99.99%) from the surfaces of the foods. Additionally, a nonpathogenic lactic acid bacterium, Pediococcus acidilactici, exhibited similar or greater heat tolerance, which would allow food processors to use it as a substitute, or surrogate, for in-plant studies without introducing harmful bacteria into the food processing environment. Mathematical models were used to describe the trends of bacterial death due to the steam treatments, and the results indicated that the type of food significantly impacted the reduction of bacteria. The models were tested using additional data collected within our own laboratory, as well as others. Results indicated that some of the models could be used as predictors of bacterial death for similar LWAF but can only be applied with caution and consideration for the type of food and process. Additionally, two different E. coli strains associated with outbreaks (O121:H19 and O157:H7) were investigated to understand impacts of strain variation, growth method, and water activity on thermal resistance. Some differences in heat resistance were observed between the strains and between the growth methods. Additionally, the reduced water activity seemed to decrease the bacteria's ability to withstand some heat treatments. Overall, thermal resistance studies indicated that several factors, in addition to water activity, impact pathogens' development of resistance to heat treatments. The experiments' results show that there are complex relationships between bacteria and the food they inhabit. Food processors must consider these relationships in order to design the best thermal processes to make LWAF safe for consumers.

Solanaceous crops such as peppers and more commonly tomatoes have been linked to Salmonella outbreaks that have occurred in the United States. The source of contamination for some of these outbreaks was traced back to the preharvest environment. Sources of contamination at the preharvest level of production often include irrigation water, soil, and the use of biological soil amendments of animal origin (BSAAO). This dissertation aims to (i) determine how factors such as the use of BSAAO's, Salmonella strain, soil type, and irrigation influence the survival of Salmonella in agricultural soils and (ii) determine the risk of Salmonella internalization in to transplanted solanaceous crops (tomatoes and bell peppers) and identify factors that influence internalization through the roots such as mode of contamination, soil contamination level, and root damage. Sandy loam (SL) and clay loam (CL) soils both unamended and amended with poultry litter (PL), irrigated either daily or weekly were inoculated with one of twelve strains of Salmonella enterica and monitored for growth and survival until no longer detected. Tomato plants were transplanted into one of three contamination treatments: contaminated soil (A) or irrigation water administered either once (B, single exposure event) or for 7 d following transplantation (C, repeated exposure event). Plants were sampled 1, 2, 3, 7 and 90 d post-transplantation for Salmonella internalization. Pepper plant roots were left intact, damaged, or removed and placed in inoculated water (6.2 log CFU/ml). Pepper plants in the soil contamination study, were transplanted into soil inoculated with Salmonella at different contamination levels (High, Mid, and Low). Samples were collected 1, 2, 3, and 7d post-transplant. Plants were surface sanitized and enumerated for Salmonella internalization by plant section. Strains in soils treated with PL survived significantly (P

Water used in fresh produce washing is a potential vehicle of foodborne pathogen contamination. This work focused on assessing the sanitizing efficacy of ethanol mist to reduce Salmonella populations on the surfaces of tomatoes and cantaloupes. Ethanol (70%) mist was applied to whole tomatoes and cantaloupe rind plugs using a Biomist sanitation system, which uses CO2 as a carrier gas to spray vapors through a fine droplet spray nozzle. Fresh red tomatoes (Solanum lycopersicum) and cantaloupe (Cucumis melo) plugs were inoculated with Salmonella enterica Newport to a concentration of log 7 CFU/tomato and log 7 CFU/cm2 respectively. Application time (5, 10, and 15 sec), dry vs. wet surface, and stem scar contamination were evaluated on tomatoes, while only application time was evaluated on cantaloupe. Application of ethanol mist for 10 seconds was the most effective treatment time, reducing Salmonella by 3.3 log CFU/tomato. Application of ethanol mist was more effective on dry opposed to wet tomato surfaces by approximately 0.7 log CFU/tomato. Ethanol mist application to inoculated tomato stem scars reduced Salmonella 1.2 log CFU/tomato. Additionally, Salmonella decreased by 1.3 log CFU/cm2 on cantaloupe rind plugs at 10 seconds, again the most effective treatment time. Ethanol mist resulted in sufficient reductions of Salmonella populations on dry tomato surfaces, but was limited in effectiveness if the surface was wet, or if Salmonella adhered to the stem scar. Furthermore, this technique was overall not an efficient method to sanitize cantaloupe surfaces. Master of Science in Life Sciences Water is often used in washing and moving fresh produce during harvesting. However, this water is often found to be a source of contamination that can cause the fruits or vegetables to become unsafe to eat. In order to resolve this problem, a mist-type sanitizing system is being explored to wash fresh produce in packinghouses. In this experiment, a mist of ethanol (70%) was applied to whole tomato and cantaloupe plugs using a Biomist sanitation system, which turns liquids into a mist. Whole fresh tomatoes and cantaloupe plugs were infected with Salmonella enterica bacteria. Tomatoes and cantaloupe plugs were treated with ethanol mist for various times of 5, 10, or 15 seconds. Additionally, tomatoes were treated with ethanol to compare a wet surfaced tomato, as well as the tomatoes stem scar (where the vine of the tomato is attached during growing). Ethanol mist application was most effective at 10 seconds, but there was very little difference in bacterial elimination when all the times were compared to the untreated tomatoes. In addition, the mists effectiveness decreased if the surface of the tomato was wet compared to dry, and was even less effective if the bacteria were located where the vine attaches. Also, the ethanol mist has very little effect if bacterial contamination is present on a cantaloupes surface. If a harvested tomato remains dry post harvest, ethanol mist may make for a good washing system. However, it is probably not the best for rough surfaced produce like cantaloupes.

Listeria monocytogenes has emerged as a food safety concern for a number of produce commodities. While L. monocytogenes contamination can occur throughout the supply chain, contamination from the packinghouse environment represents a particular challenge and has been linked to recalls. This study aimed to investigate the prevalence, persistence, and diversity of Listeria monocytogenes (LM) and other Listeria species (LS) in produce packinghouses. A longitudinal study was performed in 11 packinghouses (commodities included micro-green, peach, apple, tomato, broccoli, cauliflower, and cucumber) in three US states. In each packinghouse, 34 to 46 sites representing zones 2-4 were selected and swabbed. Packinghouses were visited 4 times and samples were processed for Listeria by US Food and Drug Administration's Bacteriological Analytical Manual methods. Presumptive Listeria-positive isolates were confirmed by PCR. Species and allelic type (AT) were identified by sigB sequencing. Among the 1,584 samples tested, 3.2%, 2.7%, and 0.6% of the samples were positive for LM, LS, and both LM and LS, respectively. Five different species of Listeria were identified with L. monocytogenes being the most prevalent species. A high AT diversity (0.95 Simpson's Diversity Index) was observed amongst Listeria isolates. There were 15 instances of Listeria repeated isolation (site testing positive ≥2 times). Upon analysis of subtype data, only 3 sites tested positive for the same Listeria AT >2 times. Data showed in this longitudinal study that Listeria prevalence and persistence in packinghouses was low (e.g.

Consumers desire tender, juicy, and flavorful cuts of beef. Mechanical tenderization (MT) and enhancement methods applied to lower valued beef cuts can improve tenderness, flavor or juiciness, increasing desirability for the consumer. However, these processes can introduce pathogens that may be present on the exterior of the meat into the sterile interior. This process renders an ‘intact’ product ‘non-intact’ and requires altered cooking methods to ensure safety. The primary pathogens of concern for beef products are Shiga-toxin producing Escherichia coli (STEC). STEC are associated with approximately 265,000 illnesses and 3,600 hospitalizations annually. Since 2006, there have been 6 STEC outbreaks in the United States and 18 cases in Canada attributed to MT beef (MTB). The pathogen has also been implicated in 136 non-intact beef-related recalls. Due to the potential food safety hazards associated with MTB, mandatory labeling of these products was mandated in 2015 to inform consumers on how to safely handle the product. While this is a good step to inform consumers, it is unclear how familiar they are with the terms associated with these processes. Consumer’s knowledge, beliefs, attitudes, awareness, and behaviors related to MTBs is quite limited. This study uses an exploratory sequential mixed-methods design, to assess consumer knowledge of MTB. Qualitative focus groups conducted throughout urban and rural North Carolina and Virginia found that although participants purchased MTBs, they were unaware of the process, did not prepare MTBs properly, wanted to know more about the process, and wanted applicable risk messages. A nationwide survey developed from the focus group findings found that demographic differences were associated with knowledge of; and how participants interact with MTBs. How demographics influence consumer’s beef safety knowledge, practices, and risky behaviors was further explored. Demographic characteristics were highly correlated with consumers’ behaviors surrounding beef storage, refrigerator temperature knowledge, defrosting behaviors, meat washing, and meat preparation behaviors. Collectively, the mixed methods research design provided insight into specific demographic characteristics related to consumer attitudes, beliefs, and behaviors surrounding beef safety. This data will help inform the development of well-crafted, culturally, and socially relevant risk messaging that may promote safe handling behaviors. Ph. D. In the United States, every year, the Center for Disease Control estimates that 48M people are sickened, 128,000 people are hospitalized, and 3,000 people die from foodborne illnesses. The most common illnesses arise from Salmonella, Norovirus, Campylobacter, Escherichia coli [E. coli], Listeria, and Clostridium perfringens; these bacteria and viruses have been on the news as being associated with flour recalls, cantaloupe, eggs, cheeses, berries, and even at restaurants like Chipotle! The bacteria of concern in this study is E. coli. Most E. coli is not pathogenic, and extremely common in the environment; living in mammalian (e.g. humans, cows, sheep, insects, etc.) gut and within the environment (e.g. in the soil). However, there are some pathogenic variants, like Shiga-toxin producing E. coli [STEC] that have been associated with 265,000 annual illnesses and deaths. The main reservoirs of many pathogenic E. coli are within the intestines of ruminant mammals, including cattle. If mishandled, feces can contaminate and cause human illness as a result of improper handling and preparation. The contamination can occur through meat, water, and fecal-oral routes; often, improperly cooking and handling beef products can lead to illness. In the U.S., there have been 27 multi-state STEC associated food outbreaks. While the U.S. beef industry is the largest in the world, five multi-state outbreaks were related to E. coli O157:H7 contamination in ground beef products. Additionally, between 2005 – 2018, 136 of 171 recalls were due to STEC-contaminated beef products. Non-intact beef products (e.g. ground beef) are the most commonly recalled types of beef products. Consumers purchase beef products on extrinsic (i.e. price, weight, cut) and intrinsic (i.e. color, fat, safety) factors with a desire for tenderness, juiciness, and flavor. Tenderization processes (e.g. mechanical tenderization) or other enhancement processes (e.g. marinades) can increase tenderness on lower-value cuts but may introduce pathogens from the exterior to the sterile interior. To prevent illness, it is necessary to prepare beef products to the recommended USDA-specified temperature using a thermometer to check. Up until this point, consumer knowledge of and behaviors towards mechanically tenderized beef products [MTB]s and other enhanced beef products had not been characterized. This study uses an exploratory sequential mixed-methods study design (qualitative study guides quantitative study) to assess consumer knowledge of MTBs, enhanced beef products, and food safety / beef safety handling behaviors. Focus groups conducted throughout urban and rural North Carolina and Virginia found that although participants purchased MTBs, they were unaware of the process, did not prepare them properly, wanted to know more about MTBs, and requested different ways of obtaining information. A nationwide survey developed from the focus group findings found that demographic differences were associated with knowledge of and interaction with MTBs and enhanced beef products. However, people were still generally unaware of MTBs, despite a recent 2016 labeling mandate for all MTBs. The Theory of Planned Behavior was used to further explore reported consumer’s food safety knowledge, practices, and risky behaviors in the kitchen. Demographic characteristics were highly correlated with consumers’ behaviors of beef storage, refrigerator temperature knowledge, defrosting behaviors, meat washing, and meat preparation behaviors. Collectively, the mixed methods research design provided insight into specific demographic characteristics related to consumer attitudes, beliefs, and behaviors surrounding beef safety. Additional research, alternative non-didactic strategies, and collaborations within health and public services is necessary to accommodate for specific demographics, cultures, and social groups. Well-crafted, culturally, and socially relevant targeted risk messaging must be developed to increase awareness and promote ease-of-access.

Aquacultured products assist the human demands for seafood so that foodfish supplies can remain sustainable and consistent. Although the fish-farming industry has seen dramatic growth and intensification in recent years, the latter has led to an increase in bacterial diseases and fish health management problems, resulting in major economic losses around the world. In addition to the lack of understanding of fish physiology, these complications are exacerbated by the inappropriate and controversial use of antibiotics. This work addressed these issues in striped catfish (Pangasius hypophthalmus) and Nile tilapia (Oreochromis niloticus), two economically important foodfish, by investigating alternative, more cost-effective options to promote fish health. The first two studies established reference intervals for immunology, hematology and plasma chemistry analytes in striped catfish in a recirculating aquaculture system (RAS). In a third study, the immunomodulatory effects after directly feeding probiotic strains of Bacillus subtilis NZ86 and O14VRQ in Nile tilapia were ascertained. This last study revealed that supplementation with both of the probiotic strains for 51 days stimulated several local and systemic innate immune responses of tilapia. When these transient probiotic bacteria were present in the gut, a pro-inflammatory environment was developed as evidenced by the localized higher expression of the cytokines tumor necrosis factor (TNF) – α and interleukin (IL) – 1β. Significant increases (p < 0.05) were noted differentially by both probiotic strains throughout the trial in plasma lysozyme content, alternative complement activity, and in the peripheral blood leukocyte profiles. Additionally, there were trends for increased levels of phagocytosis and respiratory burst in leukocytes of the anterior kidney and spleen at the end of the trial, suggesting the potential use of these probiotic strains for improved immune-competence. These findings help to understand and clarify the potential mechanism of action associated with the increased disease resistance recorded in preliminary studies with the same probiotic strains. Implementation of the tools established and validated in this work could be useful in evaluating fish welfare situations involving striped catfish grown in RAS conditions, and also show promise for a healthier foodfish supply where antibiotic applications practices could be minimized. Ph. D.

Airborne pathogenic bacteria can present a significant public health risk. Pathogenic Listeria monocytogenes can colonize numerous surfaces as well, through direct and indirect cross contamination. The physical environment can also affect the transmission and viability of Listeria (distance from the source, temperature, humidity, air flow). The purpose of this work was to explore the ability of Listeria innocua (a surrogate for L. monocytogenes) to contaminate a surface after it has become aerosolized in a bioaerosol chamber and a walk-in cooler. L. innocua was nebulized into a 154 L biosafety chamber (~5 log CFU in 1 mL) at two relative humidity (RH) levels (83% and 65%). Oxford Listeria agar plates, stainless steel coupons and polyethylene (HDPE) coupons in the chamber were exposed to the aerosolized bacteria for 5, 10, 20 or 40 minutes. Also, at these times, air samples (100 L) were collected on to gelatin filters which were transferred to Oxford agar plates. In the second part of the research, L. innocua was nebulized into an 11 m3 walk-in cooler where RH ranged from ~29-37%. Aerosolized bacteria were collected on to Oxford agar plates for 10 min intervals and with 50 or 100 L air samples. Recovery of L. innocua from steel, plastic and agar was significantly higher at 83% RH (2.7 cells/cm2) compared to 65% RH (0.45 cells/cm2). Mean cell recovery from air samples (gelatin filters) was significantly higher (p

In this study, potential of high hydrostatic pressure (HHP) and essential oils (EOs) as natural antimicrobials was evaluated to produce E. coli safe and quality beef product. First, the individual and combined effects of antimicrobial activity (minimum inhibitory concentration) of basil, black cumin, cilantro, cumin, fenugreek, ginger, oregano, black pepper, rosemary, thyme, turmeric oil emulsions on E. coli ATCC 25922 with and without HHP treatment were evaluated. Cumin, oregano and thyme EOs showed highest antimicrobial activity against E. coli ATCC 25922. The synergy of selected EOs against E. coli ATCC 25922 was determined using the checkerboard method to obtain fractional inhibitory concentration index. Although their combinations did not show synergy, they expressed synergy when combined with HHP (400 MPa, 10 min, 20 °C) and the best combination was cumin and oregano EOs with HHP. Effects of HHP and EO combinations on inactivation of E. coli ATCC 25922 in beef were investigated using response surface methodology (RSM). Statistical analysis showed the model was significant for predicting log reduction with high accuracy. The significant model terms were pressure and time. Compared to control, HHP/EO treated samples showed no-post growth when stored up to 120 days at 4°C. Presented results suggests that the combination of HHP and antimicrobials has not only improved the process parameters (lowered pressure, time, and EO concentration) but also prevented recovery of E. coli ATCC 25922 during storage. RSM was employed to analyze the synergistic effects of HHP and EOs on beef quality (color, texture and lipid oxidation). Color indices were significantly affected by pressure, time and their interactions. Above 400 MPa the discoloration was similar to cooked beef and EO addition did not help color improvement. However, EOs showed significant antioxidant activity on both treated and untreated samples during storage. In conclusion, there is a great potential of HHP and EO combinations to enhance pathogen inactivation while keeping the quality of beef. Moreover, presence of EOs can prolong the shelf life of pressure treated beef. Therefore, the combination of HHP and EO is very promising for meat industry. Ph. D.

Salmonella enterica has been associated with several outbreaks due to consumption of low water activity foods including spices. Consequently, to improve microbiological quality, spices are commonly treated before ultimately reaching consumers. These processes may result in sub-lethal injury to cells, which can lead to an underestimation of microbial populations when plating on selective media. Sub-lethally injured cells are difficult to enumerate due to reduced ability to grow on selective microbiological media. Poor recovery of sub-lethally-injured cells may obscure process validation results and lead to overestimation of process effect. Therefore, this work was performed to determine the influence of agar overlay and media supplementation methods on the recovery of Salmonella from steam and ethylene oxide treated peppercorns and cumin seeds. Traditional agar overlay allowed recovery of significantly more Salmonella (p < 0.05) from inoculated peppercorns treated with steam (65.5C, 15 sec.) than selective media (XLT4) or solid agar overlay. Additional supplementation of the TSA base to contain 3,3'-thiodipropionic acid further improved Salmonella recovery from steam treated peppercorns (p < 0.05). For peppercorns and cumin seeds subjected to ethylene oxide, neither sodium pyruvate + yeast extract, 3,3'-thiodipropionic acid, glycerophosphate, ATP, nor magnesium enhanced recovery compared to overlay alone but both methods recovered significantly higher numbers than XLT-4 alone (p < 0.05) for both cumin seeds and peppercorns. Master of Science in Life Sciences

Current methods to reduce the native microbiota and potential pathogens on spices include steam treatments and ethylene oxide (EtO) fumigation. The objectives of this research were to identify the effectiveness of a lab-scale steam apparatus and a commercial EtO process on the inactivation of Salmonella enterica or Enterococcus faecium NRRL B-2354 inoculated whole black peppercorns and cumin seeds. Peppercorns and cumin seeds were inoculated with Salmonella or Enterococcus and processed in a lab-scale steam apparatus at 16.9 PSIA and two references temperatures (165°F and 180°F) and in a commercial ethylene oxide fumigation chamber using a standard commercial EtO fumigation process. Cells were enumerated by serial dilution and plating onto TSA with a thin overlay of selective media. Inoculation preparation influenced inactivation of Salmonella on peppercorns with greater reductions reported for TSA-grown cells compared to within a biofilm. To achieve an assured 5-log reduction of TSA-inoculated Salmonella on peppercorns exposure for 125s and 100s at 165°F and 180°F, respectively is required. For cumin seeds temperatures of 165°F for 110s were needed or 65s at 180°F to assure 5 log reduction. EtO fumigation significantly reduced both microorganisms on both spices (p

Salmonella contamination of spices and other low water activity foods is a growing concern for the food industry due to increased frequency of salmonellosis outbreaks and detection-based product recalls. The impact of inoculation preparation on the survival of a Salmonella enterica and its proposed surrogate, Enterococcus faecium NRRL B-2385, on the whole black peppercorns and cumin seeds was examined. Three liquid inoculation methods (biofilm-inclusion, agar-grown, broth-grown) for Salmonella enterica and surrogate Enterococcus faecium and one dry transfer method for Salmonella enterica were developed then applied to whole peppercorn and cumin seeds. Spices were returned to original water activity (aw 0.3) and stored for 28 days with periodic sampling (0, 1, 7, 14, 21, 28 days) and surviving bacteria enumerated. Average log reductions (LR) over time were statistically analyzed to determine differences in stability during storage. Inoculation preparation was associated with significant differences in recovered Salmonella and Enterococcus from both peppercorn and cumin over the storage period. At 28 days, the most stable inoculations of Salmonella resulted from the biofilm-inclusion (-0.04 CFU/g LR) and agar grown (-0.75 CFU/g LR) methods on peppercorn and the biofilm inclusion method (-0.28 CFU/g LR) on cumin. Log reductions of Enterococcus faecium (-0.02 CFU/g LR biofilm-inclusion-peppercorn, -0.19 CFU/g LR agar-grown-peppercorn, -0.61 CFU/g LR biofilm-inclusion-cumin) were comparable to Salmonella after 28d desiccated storage. These results will guide the inoculation strategies for validating inactivation processes for reducing Salmonella on whole spices, and for comparisons of inactivation of Salmonella and its proposed surrogate Enterococcus faecium. Master of Science in Life Sciences

Salmonella embedded biofilms were formed on whole black peppercorns and treated with ethanol vapor under atmospheric pressure conditions and under vacuum assisted pressure conditions. The effect of ethanol vapor, heat and vacuum pressure on the survival of multiple Salmonella enterica serovars within a biofilm contained on low moisture food products (Aw 0.30-0.40) was observed in this study. Samples were exposed to zero, one, five and ten minute ethanol vapor treatments at atmospheric boiling temperature (Atmospheric Pressure Boiling Method, AB), and at reduced temperature and pressure conditions, -20 inHg (Vacuum-Assisted Boiling Method, VB). The AB treatments showed 4.0 log CFU/g reductions on nonselective media that included native microbiota, and 6.0 log CFU/g reductions on Salmonella selective media. The VB treatments showed 2.69 log reductions on Tryptic Soy Agar and 4.55 log reductions on Xylose Lysine Tergitol-4 agar. Ethanol vapor treatments should be further investigated as an alternative to ethylene oxide or ionizing radiation processes to treat dry spices to control Salmonella. Master of Science in Life Sciences

Fresh market tomatoes have been associated with 15 multistate Salmonella outbreaks between 1973 and 2010. While, S. enterica survival has been studied in tomato plants, field studies have been limited. To understand pathogen growth and survival, in crop fields, surrogate or attenuated organisms must be developed and validated. The purpose of this study was to compare the growth and survival of seven attenuated S. enterica Typhimurium and Newport strains against virulent strains S. Typhimurium ATCC14028 and S. Newport J1892 in optimum (TSB and TSB+kan) and minimal M9 growth media, and in commercial, red ripe tomatoes. Bacterial growth in media was assessed via BioScreen. Tomatoes were separately inoculated with 7 Log CFU/g of each isolate via vacuum infiltration, surface spot inoculation, or diced inoculation. Populations of each strain were determined on Days 0, 1, 3, and 5. In media, there were few differences in overall growth and growth rates between mutant isolates and wild-type (P0.05); however, Salmonella strains in diced tomatoes had a higher growth rate than that in spot treated tomatoes (P>0.05). The growths of all the isolates in tomatoes indicated that under the tested conditions, isolates acted similarly to their WT counterparts. Thus, these strains may be able to be used as surrogate organisms in field studies. Master of Science in Life Sciences

Sour rot of tomatoes, caused by Geotrichum candidum, occurs in the field and postharvest settings regularly, although postharvest losses are severe only in some years on the Eastern Shore of Virginia (ESV) and other tomato production regions. Fungicide products and cultural control methods are tested for efficacy utilizing a traditional wounding technique that does not properly reflect natural sour rot infections. A new inoculation technique was optimized for G. candidum using negative pressure to infiltrate the tomato stem scar with pathogenic spores. This new method creates consistently high rates of infection and more successfully creates infections in mature green and breaker fruit. The population of G. candidum on the Eastern Shore of VA (ESV) was characterized using multilocus sequencing technique. The resulting phylogenetic tree defines four distinct groups, including two with uncommon loci that distinguish them from the majority of the population. Thirty-seven G. candidum isolates were inoculated to media amended with ten fungicides and antimicrobial compounds commonly used in tomato production and postharvest treatments. Propiconazole and tebuconazole completely inhibited growth of all colonies. Cultivar trials were conducted to determine if resistance or tolerance to G. candidum occurs. Ten commonly grown round and Roma cultivars on the ESV were similarly susceptible to G. candidum, even at low inoculum levels. Field and postharvest surveys of sour rot on tomato fruit attempted to correlate disease incidence with weather conditions in order to better understand the cause of sporadic infection. Few patterns were seen consistently throughout harvest periods and years. Rainfall was positively correlated with disease 2-3 days before surveys and temperature was negatively correlated with disease 5-7 days before surveys. No in-field weather conditions were correlated with postharvest disease incidence. Greenhouse trials were conducted to assess the influence of water congested tomato fruit on susceptibility to sour rot. Tomato plants were exposed to water inundation to mimic rainfall and varying levels of irrigation, both in order to congest tomato fruit. Though water congestion was achieved, tomato fruit were equally susceptible to sour rot infections. Ph. D.

Grape seed extract (GSE) and peanut skin extract (PSE) are waste products in the wine and peanut industries. Both extracts have high concentrations of polyphenols, known to possess antioxidant and antimicrobial properties. A subcategory of polyphenol is procyanidin, which can be divided into two types, type A and type B. Type A (PSE), contains two single bonds connecting the phenolic groups while type B (GSE), contains one single bond connecting the phenolic groups. The minimum inhibitory concentration (MIC) of the two extracts was evaluated for their antimicrobial effect on Listeria monocytogenes, Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella Typhimurium using the pour plate method. GSE was found to have a significantly lower MIC (p ≤ 0.05) than PSE for L. monocytogenes (GSE=60.60ppm, PSE=not found), S. aureus (GSE=38.63ppm, PSE=51.36ppm), and S. Typhimurium (GSE=45.73ppm, PSE=60.60ppm). There was no significant difference in inhibition of E. coli O157:H7 (GSE=47.44ppm, PSE=51.13ppm). Since GSE, contributed to greater pathogen inhibition, its extract was fractionated into monomer and oligomers components. Growth curves of all four pathogens inoculated in the monomer and oligomer fractions were compared using the BioScreen method. Oligomers inhibited growth of L. monocytogenes, S. aureus, and E. coli O157:H7 while monomers inhibited growth of S. Typhimurium. These results indicate that an extract with type B procyanidins that are high in oligomers may be more effective as antimicrobials. Type B procyanidins have also been shown to prevent bacterial adhesion, as is the case with urinary tract infections, and may aid in the prevention of biofilms. Master of Science in Life Sciences

Bacterial biofilms attached to food contact surfaces are an ongoing concern for the food industry due to the resistance of bacteria within biofilms to detergents and sanitizers. Within food manufacturing facilities, stainless steel is a common food-contact surface in which microbial cell attachment and biofilm formation may occur. Identifying methods to prevent and remove biofilms during standard cleaning and sanitation practices could prove useful, as mature biofilms can release planktonic cells into an aqueous environment, causing continual low-level contamination. Dental studies involving delmopinol hydrochloride, a cationic surfactant, have found a preventative and dissociating affect on biofilms, where food applications have scarcely been researched. This study demonstrates the prevention and removal of Listeria monocytogenes 1/2a and S. enterica Agona biofilms on stainless steel with pre- and post-exposures of delmopinol hydrochloride. Stainless steel blanks (#304, 16 gauge, 2cm x 2cm, finish #4) were submerged in a 0.2% or 0.5% delmopinol solution before or after biofilm formation. Treatment times were 1, 5 or 10 minutes, whereas controls were not exposed to the delmopinol solution. Disinfected stainless steel blanks were spot-inoculated with 20µL of a 10⁹ CFU/mL liquid culture, and pre-exposed blanks were additionally submerged in delmopinol and dried prior to inoculation. Biofilms were exclusively formed on the finished and inoculated side by placing the surface face-down on TSA. After cell attachment and biofilm development for 24 hours at 25°C, blanks were rinsed with phosphate buffer. Post-exposed blanks were submerged in 0.2% or 0.5% delmopinol for 1, 5 or 10 minutes before all blanks were individually vortexed for 90 seconds to dislodge films. Bacterial populations were determined by surface plating onto TSA followed by incubation at 32°C for L. monocytogenes and 37°C for S. Agona for 48 hours. Treatments were in-duplicate and repeated three times for each microorganism. Pre-exposure of 0.2% delmopinol resulted in a significant decrease in L. monocytogenes concentration at 1, 5 and 10 minute exposures (P < 0.05). Pre-exposures with the 0.5% solution had no significant effect on L. monocytogenes biofilm populations (P > 0.05), whereas all post-exposures lead to a significant decline in biofilm concentrations (P < 0.0001). Post-exposures of 10 minutes exhibited a mean log₁₀ reduction of 5.59 and 6.40 log₁₀ for 0.2% and 0.5% delmopinol solutions, respectively. For S. Agona, 0.2% pre-exposure resulted in no significant log10 reduction (P > 0.05), while the 10 minute 0.5% pre-exposure exhibited a minimal reduction in bacterial growth (P < 0.05). Post-exposures of 10 minutes exhibited a mean log10 reduction of 7.65 and 7.75 log10 for 0.2% and 0.5% delmopinol solutions, respectively. For L. monocytogenes and S. Agona, post-exposure to delmopinol hydrochloride caused a notable log10 reduction. The removal effect of delmopinol on biofilms is significantly greater the preventative effect. Master of Science in Life Sciences

Juices have been implicated in numerous foodborne outbreaks over the last couple of decades. The FDA requires a 5-log10 reduction in juice products, which is most commonly achieved through pasteurization. However, pasteurization deteriorates some sensorial properties and nutritive value. Coconut water (CW; classified as a juice), is rapidly gaining popularity increasing over 300% since 2005. CW has not been implicated in a microbial outbreak, but is thermally processed to achieve the required 5-log10 CFU/ml reduction, which results in negative organoleptic properties. The objectives of this study are to determine whether E. coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes can grow in CW, and evaluate the use of High Pressure Processing (HPP) to reduce populations of these bacteria in CW. The three pathogens were inoculated separately into CW and bacterial populations were enumerated over 24 hours. All three bacteria reached at least 8-log10 CFU/ml after 24 hours, which was not significantly different from the control (TSB). CW was then inoculated with each pathogen and processed using HPP (400, 5000, or 600 MPa) for 120 seconds. The D-glucose, D-fructose, sucrose, and phenol oxidase levels in the CW were assessed before and after treatments. Following processing, the pathogens were enumerated from the CW. All three pathogens were reduced by more than 6-log10 CFU/ml following treatments of 500 and 600 MPa, enough to achieve the mandatory 5-log CFu/m reduction. There were no significant changes in the D-glucose, D-fructose, sucrose, and phenol oxidase activity after any of the treatments. Master of Science in Life Sciences

Since the surfaces of cantaloupes are highly rough or irregular, Salmonella enteric and other bacteria can easily attach to these surfaces and are difficult to remove. Cetylpyridinium chloride (CPC) is the active ingredient of some antiseptic oral mouth rinses and has a broad antimicrobial spectrum with a rapid bactericidal effect on Gram-positive pathogens. Delmopinol hydrochloride (delmopinol) is a cationic surfactant that is effective for treating and preventing gingivitis and periodontitis. The application of delmopinol or CPC to cantaloupe surfaces may be an alternative post harvest technique to reduce the frequency and level of Salmonella contamination. Cantaloupe (Athena and Hale's Best Jumbo (HBJ) cultivars) rind plugs were inoculated with a broth culture of Salmonella Michigan. After 15 min, plugs were sprayed with 10 ml of a 1% delmopinol solution, or a CPC solution (0.5 or 1.0%) or distilled water (Control), and held at 37 oC for 1 hr or 24 hr. For additional samples, the chemical treatments were applied 15 min before pathogen inoculation. Melon plugs were submerged in Butterfield's Phosphate Buffer, shaken, sonicated and solutions were enumerated on Tryptic Soy Agar. The texture quality and color of additional melon samples were evaluated after delmopinol or CPC spray treatments and storage at 4 oC. A 1.0% application of CPC reduced Salmonella levels up to 2.34 log CFU/ml (Athena) and 4.95 log CFU/ml (HBJ) in comparison to the control (p

Listeria monocytogenes represents a high risk for consumers, because it causes severe illness. This work studied in-vitro growth and survival of L. monocytogenes in media acidified with malic acid, lactic acid or blueberry extract. The growth of L. monocytogenes and shelf life extension of fresh blueberries were evaluated after storage at 4 deg C or 12 deg C under different controlled atmosphere conditions, including air (control); 5% O2 and 15% CO2, 80% N2 (CAS); or ozone gas (O3) 4ppm at 4 deg C or 2.5ppm at 12 deg C, at high relative humidity (90-95%) for a total of 10 days. L. monocytogenes growth in tryptic soy broth with yeast extract (TSB+YE) mixed with different acid solutions (malic acid, lactic acid and blueberry extract) and incubated at 25 deg C for 24h, was calculated measuring optical density. Complete inhibition occurred in the presence of treatments including malic acid pH 2.0 and pH 3.0; lactic acid pH 2.0, pH 3.0 and pH 4.0; and with blueberry extract pH 2.0 in the mixture. After 6h, there were significant differences among growing treatments. At 18h, there were no significant differences in turbidity among media mixed with blueberry extract at pH 3.0, 4.0 and 5.0, and their optical density values were higher than treatments including media mixed with malic acid 5.0 or lactic acid 5.0. Blueberry extract was not an effective acidifying media and acid adapted L. monocytogenes grew in acidified media. Fresh blueberries inoculated with L. monocytogenes were stored at 4 deg C or 12 deg C under different controlled atmosphere conditions (Air, CAS or O3 4ppm at 4 deg C or 2.5ppm at 12 deg C) and sampled on day 0, 1, 4, 7 and 10 for bacterial growth, weight loss, firmness and yeast and molds counts. CAS did not delay or inhibit Listeria monocytogenes, yeast, or molds by day 10. Storage at 4 deg C showed lower weight loss values compared to 12 deg C. Ozone controlled weight loss and firmness loss. Moreover, gaseous ozone achieved 3 and 2 log reductions when compared with air at 4 deg C and 12 deg C, respectively. Ph. D.

Campylobacter spp. are the second leading bacterial cause of food borne illness in the U.S. New antimicrobials that prevent bacterial attachment may be effective for reducing Campylobacter. Delmopinol hydrochloride (delmopinol) is a cationic surfactant that is effective for treating and preventing gingivitis and periodontitis. This study evaluated the effectiveness of delmopinol for reducing attachment of Campylobacter jejuni to chicken, stainless steel and high-density polyethylene. Chicken pieces, steel and HDPE coupons were spot-inoculated with 0.1 mL of a Campylobacter jejuni culture. After 10 min, samples were sprayed with 0.5% or 1.0% delmopinol, 0.01% sodium hypochlorite, or distilled water. Contact times were 1, 10, or 20 min prior to rinsing with buffered peptone water. Rinses were serially diluted onto Campy Cefex Agar for enumeration. For additional samples, solutions were applied first, followed by inoculation with C. jejuni after 10 min. Cultures remained undisturbed for 1, 10, or 20 min. Then samples were rinsed and plated as above. When C. jejuni was inoculated before treatments, 1% delmopinol application led to mean log reductions of 1.26, 3.70, and 3.72 log CFU/mL, greater than distilled water, for chicken, steel and HDPE respectively. When C. jejuni was inoculated after spray treatments, 1% delmopinol reduced C. jejuni by 2.72, 3.20, and 3.99 mean log CFU/mL more than distilled water for chicken, steel and HDPE respectively. Application of 1% delmopinol, either before or after bacteria inoculation, resulted in a significantly (p

In recent years, there have been a number of studies looking at inhibition of microorganisms by spices, herbs or their extracts. Many of these products have been shown to have antimicrobial activity against foodborne pathogens. The purpose of this research was to evaluate the antimicrobial activity of three essential oil (EO) compounds (thymol, eugenol, and trans-cinnamaldehyde) alone and in combination with three naturally-occurring apple aroma (AA) compounds (hexanal, trans-2-hexenal and 1-hexanol) to identify the minimum inhibitory concentrations necessary to inhibit E. coli O157:H7. Three commercial apple juice flavoring mixtures (natural apple cinnamon, natural apple spice and natural red apple) were additionally tested alone for antimicrobial activity against E. coli O157:H7. The standard agar dilution method (SAD) and checkerboard assay were used to evaluate the efficacy of the nine compounds, alone and in combination against E. coli O157:H7. In general, the EO compounds were significantly more effective against E. coli O157:H7 than the AA compounds (P

Contaminated wheat flour has been identified as the probable vehicle of a multi-state outbreak of Escherichia coli O157:H7 associated with ready-to-bake cookie dough. Several cookie dough manufacturers are currently using heat-treated flour for ready-to-bake products, although data on thermal inactivation of foodborne pathogens in wheat flour remains scarce. The objective of this research was to first determine appropriate methods and parameters for bacterial inoculation and thermal treatment of wheat flour, and to subsequently determine the population reductions of E. coli O157:H7 and Salmonella Agona in artificially contaminated wheat flour following thermal treatment for 1, 5, 15 or 30 minutes at 55, 60, 65 or 70°C in a shaking water bath. Flour samples (aw = 0.55) in sterile plastic bags were individually inoculated (~109 CFU/g), pulsified to distribute cultures, and pressed to a uniform thickness (1mm) prior to heat treatment. Following treatment, samples were rapidly cooled and diluted with peptone water; then plated onto Tryptic Soy Agar (TSA) and incubated at 37°C for 24 h prior to enumeration. The minimum heat treatments required for a 5-log reduction in microbial populations (~ 109CFU/g to ~ 104CFU/g) were 5 minutes at 70°C and 30 minutes at 70°C for E. coli O157:H7 and S. Agona, respectively. This research supports the hypothesis that the microbiological safety of ready-to-bake products may be improved by the use of heat-treated flour. Master of Science in Life Sciences

The demand for biodegradable packaging materials as an alternative to synthetic ones to reduce environmental cost has seen an increase in recent years. In addition, functionalizing the packaging film to provide specific advantages like antimicrobial properties has yet to be explored thoroughly. This study considers adding antimicrobial agents to improve the quality and safety of actively packaged fresh produce using an antimicrobial enzyme (lysozyme) immobilized on a biopolymer based packaging film (corn-zein). The developed packaging material is aimed as an active biodegradable packaging to reduce bacterial contamination on the surface of fresh organic produce, specifically tomatoes. The study uses glutaraldehyde and glyoxal as binding agents to immobilize the enzyme on the packaging film. The effect of concentration of glutaraldehyde and glyoxal on the controlled release of the enzyme was studied. Concentrations of 0.1 and 0.2 g/g lysozyme: cross linking agent had controlled release properties. However, concentrations of 0 or 0.05 are about 20-30% more effective at inactivating bacteria. Antimicrobial activity in the constructed zein films are also tested against selected pathogens (Salmonella Newport and Listeria monocytogenes). Developed zein based film is tested against inoculated tomatoes to determine the efficacy of the films in reducing the pathogen population. The inoculated tomatoes are stored at room temperature over a storage period of one week. The film was able to reduce Listeria monocytogenes population by three logs but was unable to reduce the population of Salmonella Newport. Master of Science

Contaminated tomatoes have become a commonly implicated vehicle of Salmonella outbreaks. Exposure of tomatoes to pathogen could occur in the field. Blossom inoculation with Salmonella can result in contaminated fruit but natural routes of blossom contamination are not well known. Salmonellae have been known to survive in agricultural soil. Since dispersal of soil particulate by wind is a common phenomenon, the potential of airborne soil particulate as a vehicle of Salmonella contamination in tomato blossoms was examined. It was determined that Salmonella enterica serotype Anatum, Baildon, Braenderup, Montevideo, Newport, Javiana had similar survival patterns in both soil and water. At the end of 40 days, populations of Salmonella in soil dropped by 2.59 log CFU/g and 5.11 log CFU/g when enumerated on Tryptic Soy Agar Yeast Extract (TSAYE) and xylose lysine Tergitol 4 (XLT4) agar respectively. Salmonella populations in water reduced by 2.55 log CFU/ml (TSAYE, enumeration) and 2.94 log CFU/ml (XLT4, enumeration). Blossom to fruit formation takes 20-30 days in tomatoes hence the introduction or presence of the pathogen in agricultural soil and water could increase risk of blossom contamination. Also, it was determined that all Salmonella serotypes tested were capable of biofilm production on glass coverslips and quartz particles. Biofilm based attachment of Salmonella to sand might aid in its dispersal. To visualize transfer of pathogen from soil particulate to blossom in real-time, bioluminescent S. Baildon, S. Braenderup, S. Newport, S. Javiana and S. Anatum were created.Heat shock procedure was developed to improve electrotransformation efficiency in Salmonella. Transformed strains were compared for bioluminescence production and plasmid stability. S. Newport had the best bioluminescence properties but no difference was observed between strains for plasmid stability. Imaging of soil particulate - S. Newport mixture inoculated blossoms, indicated that the event led to pathogen transfer to blossom. It was also determined that S. Newport â soil particulate contaminated blossoms developed into fruits that were positive for S. Newport. S. Newport presence in blossom, fruit surface and internal tissue indicates that contaminated soil particulate could serve as a vehicle of tomato contamination. Ph. D.

The objective of this work was to determine the effect of high pressure processing (HPP) on fresh tomato-associated outbreak isolates of Salmonella enterica in broth and on green mature tomato fruits. Nalidixic acid resistant (to 50 ppm) cultures of Salmonella enterica ser. Newport and Salmonella enterica ser. Braenderup were suspended in tryptic soy broth to a concentration of approximately 8 log CFU/ml and subjected to 350, 450, and 550 MPa for 120 s. Samples were serially diluted in peptone water, and surface plated onto tryptic soy agar supplemented with nalidixic acid (50 ppm; TSAN) and incubated at 35°C for 48 h. Reductions of 5.64, 6.30, and 6.61 log CFU/ml in S. Newport, and reductions of 4.10, 5.22, and 6.35 log CFU/ml in S. Braenderup at 350, 450, and 550 MPa, respectively, were observed. Green tomato fruits inoculated with S. Newport or S. Braenderup to an initial concentration of approximately 6 log CFU/g were sealed in a bag containing 350 ml of 1% CaCl2 and subjected to the same pressure treatments described above. The whole tomato fruits were pummeled in a stomacher and samples were surface plated onto TSAN supplemented with 1% pyruvic acid. Reductions of 1.55, 2.89, and 4.26 log CFU/g for S. Newport and 1.22, 2.26, and 3.77 log CFU/g for S. Braenderup at 350, 450, and 550 MPa, respectively, were observed. Bagged (350 ml 1% CaCl₂) samples of non-inoculated green tomato fruits were subjected to the same conditions described above. HPP treated tomatoes were then subjected to an ethylene gas (125 ppm; 0.7 cc/min) for 5 to 6 days. Pressured tomato fruits did not ripen. Even though HPP effectively reduced populations of S. enterica, it adversely affects the ripening characteristics of green mature tomato fruits. Master of Science in Life Sciences

Leafy greens like lettuce and spinach are a common vehicle for foodborne illness in United States. It is unknown if native plant epiphytic bacteria may play a role in the establishment of enteric pathogens on leaf surfaces. The objective of this study was to characterize the bacterial communities of fresh and packaged spinach leaves and to explore interactions with E. coli O157:H7. We assessed the bacterial diversity present on the spinach leaf surfaces and how parameters such as spinach cultivar, field conditions, post-harvest operations and the presence of E. coli O157:H7 affected its diversity. Differences in bacterial population size and species richness were associated with differences in plant topography; flat leaves had smaller bacterial populations than savoy leaves, which correlated with larger number of stomata and trichomes in savoy leaves. During spinach growing season shifts in environmental conditions affected richness and population size of the spinach bacterial community. Decreases in the overall soil and ambient temperature and increased rainfall decreased richness and bacterial population size. Fresh spinach richness and composition assessed by parallel pyrosequencing of 16S rRNA elucidated 600 operational taxonomic units, with 11 different bacterial phyla. During postharvest operations diversity indexes and evenness tended to decrease, likely attributed to storage at low temperature and time of storage (4°C and 10°C), that promoted the dominance of g-Proteobacteria. Bacteria isolated from fresh spinach elicited growth inhibition of E. coli O157:H7 in vitro, which was associated with nutrient competition. In contrast growth enhancement produced by epiphytes was associated to low correlations in carbon source utilization and the ability of E. coli O157:H7 to rapidly utilize carbon resources. In packaged spinach, E. coli O157:H7 altered the composition of the bacterial community and its growth was promoted on packaged spinach when a disinfection and temperature abuse occurred, removal of the epiphytic bacteria resulted in significant increases in numbers of E. coli O157:H7 at 10°C and was associated with increased expression of E. coli O157:H7 virulence and stress response genes. The large diversity present on the surface of spinach leaves significantly impacted the ecology of enteric pathogens like E. coli O157:H7 on the phyllosphere. Ph. D.

Gastrointestinal tract (GIT) microorganisms play important roles in animal health, including providing energy and vitamins, improving the host immune response and preventing pathogenic microorganisms from colonizing. Prebiotic feed supplementation offers an alternative to antimicrobial growth promoters by stimulating key populations of the GIT bacteria that can ferment these non-digestible compounds, producing various short chain fatty acids used by the animal. The objective of this study was to quantify the effects of a proprietary Saccharomyces cerevisiae fermentation product (XPC, Diamond V Mills, Inc., Cedar Rapids, IA) inclusion in nursery diets on the microbial diversity and growth performance of pigs before, during and after an oral challenge with Salmonella. Pigs (n= 40) were weaned at 21 d of age, blocked by body weight (BW) and assigned in a 2Ã 2 factorial arrangement consisting of diet (control or 0.2% XPC) and inoculation (broth or Salmonella). Diet had no effect on pig growth performance prior to inoculation; however, consumption of XPC altered the composition of the gastrointestinal microbial community resulting in increased growth performance prior to inoculation. After Salmonella infection, XPC altered the composition of the gastrointestinal microbial community resulting in increased (P < 0.05) populations of Bacteroidetes and Lactobacillus. Infection with Salmonella and treatment of the piglets with ceftiofur-HCl resulted in alterations to the species richness and abundance of key members of the GIT community. The addition of XPC to the diets of weaning pigs results in greater compensatory gains after infection with Salmonella and an increase in beneficial bacteria within the GIT. Master of Science

Current trends indicate that American consumers are increasingly selecting products that they believe to be environmentally friendly or "natural." In the kitchen, this trend has been expressed through greater desire for using alternative or "green" sanitizers instead of bleach or other common chemical sanitizers. The purpose of this work was to evaluate the effectiveness of one suggested alternative, tea tree oil, as a food contact surface sanitizer. Three foodborne bacterial pathogens (Listeria monocytogenes N3-031 serotype 1/2a, Escherichia coli O157:H7 strain E009, and Salmonella Typhimurium ATCC 14028) were applied separately onto three different food contact surfaces (high density polyethylene, glass, and Formica® laminate). Tea tree oil (TTO), borax, and vinegar (5% acetic acid) were applied individually as well as in combination for a total of seven treatment solutions. In addition, household bleach (6.15% sodium hypochlorite), sterile reverse osmosis (RO) water, and no applied treatment were used as controls. Treatments were tested using an adaptation of the Environmental Protection Agency DIS/TSS-10 test method, whereby each contaminated surface was treated with 100 µl of test solution and held for 1 min followed by submersion in neutralizing buffer and microbiological plating. Samples (0.1 ml) were plated onto TSA and incubated at 35°C for 48 h prior to colony counting. Bleach reduced microbial populations significantly with greater than 5-log reduction reported for all surfaces (Formica® laminate, glass, and HDPE), against E. coli O157:H7, L. monocytogenes, and S. Typhimurium. TTO produced reductions between four and five logs for E. coli O157:H7, L. monocytogenes, and S. Typhimurium and was not statistically different from the vinegar treatment (P>0.05). All combination recipes, including the borax treatment, failed to produce reductions in microbial populations at levels considered to be appropriate for food contact surface sanitizers. Surface type did not play a significant role in the effectiveness of the treatment (P>0.05). Although TTO and vinegar did reduce pathogen populations on surfaces, reductions were not sufficient enough to be considered an equally effective alternative to household bleach. Master of Science in Life Sciences

Fresh and fresh-cut tomatoes have been associated with numerous outbreaks of salmonellosis in recent years. While the exact routes of contamination are unknown, high pressure processing (HPP) is being evaluated as a post harvest treatment to eliminate Salmonella enterica from tomatoes. The objectives of the study were to determine the potential for of HPP to reduce S. enterica serovars Newport, Javiana, Braenderup and Anatum (clinical isolates from tomato outbreaks) in tryptic soy broth (TSB) and to determine the effect of HPP to reduce the most pressure resistant S. enterica serovar from fresh diced and whole tomatoes. Five ml portions of broth containing 8 log CFU/ml of one of the four serovars (nalidixic acid resistant) were packaged in sterile stomacher bags and subjected to one of three different pressures (350, 450, or 550 MPa) for 120s. Samples were enumerated by surface plating onto tryptic soy agar supplemented with 50 ppm nalidixic acid (TSAN) and incubated at 35°C for 48 hours. The most pressure resistant S. enterica serovar evaluated was Braenderup. Subjecting the broth culture to 350, 450 and 550 MPa resulted in a 4.53, 5.74 and 7.09 log reduction in S. Braenderup, respectively. Diced tomatoes (150g) and whole red round tomatoes (150g; packaged in 350ml of 1% CaCl2) were inoculated with S. Braenderup, to obtain 6 log CFU/g throughout the sample and subjected to the same pressure treatments as described above. After HPP, diced tomatoes were homogenized for 1 minute and then plated on TSAN. Whole tomatoes were surface sampled, and then homogenized for 1 minute. Surface and homogenate samples were plated on TSAN supplemented with 1% pyruvic acid (TSANP). Significant reductions of S. Braenderup concentrations in diced tomatoes (P < 0.05) were seen after processing at 350 (0.46 CFU/g), 450 (1.44 log CFU/g), and 550 MPa (3.67 log CFU/g). In whole tomatoes, significant reductions (P < 0.05) were also seen at 350 (1.41 log CFU/g), 450 (2.25 log CFU/g) and 550 MPa (3.35 log CFU/g). There were no differences in visual appearance between fresh and HPP diced and whole tomatoes. HPP may be an effective post harvest strategy to reduce low levels of S. enterica contamination in diced tomatoes. Master of Science in Life Sciences

Today students confront more than writing, reading, and arithmetic in school. Students witness and participate in various forms of bullying at an alarming rate. As educators we must help create an environment that is conducive for all students to learn. This study examines college students' definitions and perceptions of incidents of bullying that they witnessed, or where they were the victim or perpetrator. Through 41 in-depth interviews and utilizing the constant comparative method of analysis, themes were identified including reasons students bully and are bullied, such as: weight, size, clothing, being perceived as different, sexual preferences, and placement in special education. Clothing as a reason for bullying emerged as a theme that was echoed by many of those interviewed. Whether it was the brand name of the clothes, where they were purchased, or the style of the clothes, several participants were bullied and bullied others because of clothing. Participants' definitions of bullying were from the perspective of those who are bullied, bully, and who have witnessed bullying, and included defining emotional bullying. Other students noted in their definitions the role of groups and the role of power in incidents of bullying. Unique to this study were participants' recollections of regrets. These regrets were from those participants who had participated as a bully or as a witness. In addition, themes that emerged in the data included: how students perceive teachers' involvement in incidents of bullying, ways to reduce bullying in schools, where bullying occurs, and why some people bully. Ph. D.

In response to a need for a natural antimicrobial to replace sodium benzoate, cinnamic acid was chosen. Due to cinnamic acid's solubility issues, α-cyclodextrin was used as a host molecule to form an inclusion complex with the cinnamic acid molecule. The cinnamic acid: α-cyclodextrin inclusion complex was then characterized using phase solubility analysis, proton nuclear magnetic resonance (H-NMR), and solid inclusion. Phase solubility analysis verified the maximum amount of cinnamic acid that α-cyclodextrin was able to host. H-NMR was used to determine the complex association constant, determine the chemical shifts of available protons, and yield a stoichiometry for the complex. The solid inclusion complex allowed for a physical formation of the complex, yielding further information in support of the complex stoichiometry. Microbiological tests were also performed to quantify the antimicrobial abilities of the complex, the guest, and the host against the yeast Saccharomyces cerevisiae and mold Paecilomyces variotii. Results indicated that approximately 990.29 ppm in aqueous solution was the maximum amount of cinnamic acid in the complex. The 2:1 stoichiometry yields an association constant of 21.7 M-1. Results also indicated that the cinnamic acid readily conformed to fit within the α-cyclodextrin host molecule, which remained a rigid structure. An 8.9% weight to weight of cinnamic acid was calculated for the solid inclusion again reinforcing a 2:1 stoichiometry. Microbiological studies showed little to no inhibition power by the complex at varying concentrations against S. cerevisiae and P. variotii. Free cinnamic acid showed greater antimicrobial activity compared with free α-cyclodextrin and the complex. Master of Science in Life Sciences

Chill brines used in ready-to-eat meat processing may be an important source of post-processing contamination by Listeria monocytogenes. The purpose of this study was to determine the efficacy of ultraviolet light (UV) in combination with antimicrobials to reduce L. monocytogenes in fresh and used chill brines. Three different antimicrobials were used in combination with UV; citric acid (CA, 0.2 and 0.5%), dimethyl dicarbonate (DMDC, 250 and 500 ppm), and hydrogen peroxide (HP, 2000 and 4000 ppm). For fresh brine studies, brine (8.0% w/v NaCl) was prepared and inoculated with a cocktail of three L. monocytogenes strains (approximately 6 log CFU/mL). Brine was treated with UV alone, antimicrobials alone, and combination of UV and antimicrobials. Moreover, to observe the effect of treatment temperature and brine circulation through the UV system on survival of listeriae cells, inoculated brine was circulated through the system without any treatment that served as control for all the treatments. For UV treatment, inoculated brine solution was exposed to UV in an Ultraviolet Water Treatment Unit (Model: AMD 150B/1/2T D; Aquionics Inc., Peak output: 254 nm) fitted with an inline chiller to maintain brine temperature of -1°C. Samples were withdrawn at regular intervals for 120 minutes. When L. monocytogenes population was no longer detectable via direct plating on MOX, enrichment was performed and suspect colonies were confirmed using API-Listeria. For antimicrobial-only (i.e., no UV) treatments, a specific concentration of antimicrobial was added in inoculated brine and samples were taken for 120 minutes. For the brine that received combination of UV and antimicrobial treatments, UV was turned on once a specific concentration of antimicrobial was added in inoculated brine and samples were withdrawn at regular intervals for 120 minutes. When treated with UV alone, L. monocytogenes population decreased from approximately 6 log CFU/mL to below the detection limit (i.e., 1 log CFU/mL) in 15 minutes with the reduction rate of 0.87 log CFU/mL per minute. However, cells were detectable by enrichment through 120 minutes. The highest rate of decline (0.90 log CFU/mL per minute) was achieved by the combination of UV and 500 ppm DMDC (UV+500 ppm DMDC), which was not significantly different from the reduction rates of UV and UV+0.5% CA. UV+500 ppm DMDC reduced L. monocytogenes to the detection limit in 15 minutes and the organism was not detected by enrichment after 60 minutes. Though the reduction rate of UV+0.5% CA was not significantly lower than the rate of UV+500 ppm DMDC (P>0.05), the former treatment resulted in non-detectable levels more quickly (45 minutes) than the latter (60 minutes). Thus, based on enrichment studies UV+0.5% CA was the most effective treatment in reducing the population of L. monocytogenes in fresh brine. Moreover, when brine was treated with 0.5% CA alone the population decreased to below detection limit in 15 minutes with the rate significantly lower than UV+500 ppm DMDC and UV+0.5% CA (P

Cinnamic acid (CA) is a naturally occurring organic acid that is found in some fruits and a number of spices. CA has antimicrobial activity against certain spoilage microorganisms and pathogenic bacteria. However, the acid is poorly soluble in water. Cyclodextrin molecules have a hydrophobic cavity that allows them to serve as a host for insoluble molecules in aqueous matrices. This study was conducted to determine if the aqueous solubility of cinnamic acid could be improved via complexation with α- or β-cyclodextrins, and if these complexes could be used to control bacterial pathogens in juices. Based upon phase solubility analysis, α-cyclodextrin was chosen as the host molecule for the remainder of this study. In complex with α-cyclodextrin, the solubility of cinnamic acid increased from approximately 400 mg/L to 3800 mg/L. Prepared cinnamic acid complexed with α-cyclodextrin was aseptically added (400 mg/L and 1000 mg/L) to orange juice inoculated with a Salmonella enterica (7 log CFU/mL) and apple cider inoculated with Escherichia coli O157:H7 (7 log CFU/mL). Cider and orange juice samples were extracted on day 0 and at 24 h intervals for seven days and spread plated onto Tryptic Soy Agar. Cinnamic acid was effective for reducing populations of both bacterial pathogens in juice. Populations of E. coli O157:H7 in the apple cider were significantly reduced after 7 days at 25.6 ± 0.42°C at concentrations of 400 mg/L (5-log CFU/mL reduction) and 1000 mg/L (6-log CFU/mL reduction) cyclodextrin-cinnamic acid. S. enterica counts were also reduced in orange juice at 4° C treated with 400 mg/L (2.7-log CFU/mL reduction) and 1000 mg/L (3.2-log CFU/mL reduction) complexed cinnamic acid. The much improved solubility of this compound provides food processors with greater flexibility in using cinnamic acid in their product formulations. Master of Science

Batches of fromage blanc, a soft white cheese were prepared from whole pasteurized cow's milk. Processing and formulation methods were used in cheese making to reduce Listeria monocytogenes in artificially contaminated cheese. Treatments implemented included use of additional starter culture in formulation (25% more starter culture than original formulation), use of a higher temperature draining process (at 45oC instead of 22oC), addition of the anti-listerial bacteriocin nisin (Danisco Nisaplin) in formulation at different levels (125 ppm, 250 ppm, 400 ppm), and combinations of these treatments. Characteristics including pH, fat content, protein content, and color were evaluated for each treatment cheese. Statistically significant differences (p0.05) differences found between colorimetric values for different cheese treatments. Master of Science

Amylopectin granules in Apicomplexan protozoa are hypothesized to be used as an energy source to aid the parasites in surviving in the environment allow latent stages to excyst and release infective stages, allow them to be mobile, invade host cells, and to continue their life cycle. The objective of this project was to determine if parasite glycolytic enzymes: alpha-amylase, amyloglucosidase, enolase, lactate dehydrogenase, and phosphorylase could be used to decrease amylopectin stores and subsequently infectivity of Cryptosporidium parvum oocysts/sporozoites in both fresh oocysts and stored oocysts. In addition, glycolytic enzymes and substrates: glucose, glucose-1-phosphate, and glycogen synthase were investigated to determine if they can be used to increase amylopectin stores and thus increase infectivity to aid in detection/storage of oocysts. Oocysts of Cryptosporidium parvum were suspended in 1mg/ml glycolytic enzymes or substrates (except glucose - 0.05M and glycogen synthase - 1U/ml) and electroporated. Oocysts were incubated at 37°C for one hour to allow treatments to react with amylopectin followed by incubation on HCT-8 cells for 24 hours for infection. Real-time PCR and immunohistochemistry were performed to determine the effect of the enzymes on infectivity. An amylopectin assay and excystation assay was performed to determine if the enzymes degraded amylopectin and if decreased amylopectin reduced excystation. Alpha amylase and amyloglucosidase had the greatest impact on reducing both amylopectin and infectivity of fresh oocysts with reductions of 99.5% and 99.1% in infective oocysts, respectively (P

Greenhouse grown tomato fruits and tissues were tested for the presence of Salmonella after the plants had been treated with Salmonella contaminated irrigation water or grown from contaminated seeds. Greenhouse grown tomato plants were placed into eight different groups. Groups one through six consisted of five plants each and were treated with 350 ml of 10^6 Salmonella contaminated irrigation water over a course of 70 days; group one received one 350 ml 10^7 Salmonella treatment, group two received two treatments, and so on, the treatments were scheduled every 14 days. Group seven was the control that consisted of five plants and received no Salmonella treatment. Group eight was grown from seeds that had been contaminated with Salmonella by soaking the seeds in a 10^8 Salmonella suspension for 24 hours at room temperature, and received no Salmonella watering treatment. A total of 128 tomatoes were sampled from the tomato plants of all three groups and none tested positive for Salmonella. Tissue samples consisting of roots, leaves, and stems, and were collected from one plant per each of three replications. No leaves or stems contained Salmonella, however, five of the twenty-four root samples were positive for Salmonella. In a second study, Salmonella was tested for its ability to survive in three concentrated fertilizer stock solutions and 1.6% diluted solutions of the fertilizer. Fertilizer sample CF-S was a stock solution of commercial 20N-4.4P-16.6K fertilizer, US-S was a mix of 11.3 kg UltraSol, 4.5 kg Epsom Salts, and 2.3 kg 0N-0P-43.2K in 114 L water, Fertilizer CN-S is a mix of 11.3 kg Calcium Nitrate and 56.7 g Iron chelate (10%) to 30 L water; Fertilizers CF-1.6, US-1.6, and CN-1.6 were the 1.6% fertilizer dilutions respectively. There was no significant difference (p

The ability of Listeria monocytogenes, to attach to various food contact surfaces such as stainless steel, polypropylene, and rubber compounds is well documented. The retention of these or other pathogenic bacteria on food contact surfaces increases the risk of transmission to food products. The objective of this study was to compare several methods for quantitative recovery of Listeria monocytogenes from stainless steel surfaces. A cocktail of four serotypes of Listeria monocytogenes (Scott A (4b)), 1/2b, 3b, and 4b) were mixed in equivalent concentrations and inoculated onto type 304 stainless steel coupons in a 2cm x 2cm area. After a one hour exposure, coupons were sampled by one of the following methods: 1) swabbing using a pre-moistened Dacron swab, 2) rinsing with phosphate buffered saline, 3) direct contact onto a Tryptic Soy Agar containing 0.6% yeast extract (TSA+YE) plate surface for 10 seconds, 4) sonication in an ultrasonic water bath (40 kHz), 5) contact with the bristles of a sonicating brush head for 1 min, and 6) indirect contact (2-4 mm) with the bristles of a sonicating brush head for 1 min. Coupon rinses were plated onto TSA containing 0.6% yeast extract and incubated for 24 hours at 35°C. The three sonication methods yielded higher recovery than the other three methods (p < 0.05). Brushing the coupons with the sonicating brush head yielded a recovery level of 58% and indirect exposure to the sonicating brush head permitted a recovery level of 65% from the initial microbial load. The lowest cell recovery (~20%) was observed with the swab and direct agar contact methods. Master of Science

Pale, soft, and exudative (PSE) poultry originates during rigor mortis when the muscle pH drops rapidly in high temperature carcasses. This condition results from antemortem stress and/or genetic material in the live animal. PSE poultry is pale in color, has low water-holding capacity, and forms products that are unappealing, dry, and unacceptable to consumers. Since value added products processed with PSE turkey meat display poor protein bind, color, and water retention, enhanced usability could add value to this low value raw material through locating a niche for PSE meat currently utilized in further processed products. Experiment 1 consisted of four broiler breast treatments: 100% PSE, 100% PSE + 1.5% chicken collagen, 100% normal, and 100% normal + 1.5% chicken collagen to test the effects of raw material and chicken collagen. Incorporation of collagen improved (p0.05) CIE L* and CIE a* values to that of normal treatments. No differences (p>0.05) in consumer acceptability existed among the treatments. Master of Science

The effect of pressure on the log reduction of six strains of E. coli O157:H7 and five serovars of Salmonella enterica were investigated in tryptic soy broth, sterile distilled water and commercially sterile orange and apple juice. Samples were subjected to high hydrostatic pressure (HHP) at 300 and 550 MPa for 2 minutes at 6°C, and then held for 24 hours at 4°C following treatment. E. coli O157:H7 strain E009 was the most pressure resistant, having a decrease of only 0.77 log10 CFU/ml directly after pressurization in TSB. S. Agona was the most pressure resistant Salmonella serovar tested with a decrease of 3.79 log¬10 CFU/ml in TSB at 550 MPa. The two most pressure resistant cultures were then used in a subsequent study using HHP in conjunction with antimicrobials (dimethyl dicarbonate [DMDC] at 62.5 and 125 ppm, hydrogen peroxide at 150 and 300 ppm, cinnamic acid, potassium salt at 125 and 250 ppm, potassium sorbate [KS] at 500 and 1000 ppm and sodium benzoate [NaB] at 500 and 1000). For both E. coli O157:H7 and Salmonella, the most effective antimicrobial was DMDC having a 5.79 and 5.96 log10 CFU/ml decrease directly following pressurization, respectively. Other treatments that were significantly different from the samples treated with no antimicrobial were hydrogen peroxide, and NaB at 500 ppm for E. coli O157:H7 and a treatment of NaB at 1000 ppm for S. Agona. After 24 hours at 4°C, S. Agona samples with added antimicrobials had a close to or above 5-log10 CFU/ml reduction. DMDC should be further investigated as an antimicrobial agent that can work in conjunction with HHP. Master of Science

Hazard Analysis Critical Control Point (HACCP) regulations for juice manufacture require the application of a process that will result in a 5-log reduction (99.999%) of the pertinent pathogen in the juice being processed. The use of ultraviolet (UV) light, as an alternative to traditional thermal processing, has been adopted by some juice processors as a means of meeting the HACCP 5-log performance standard. However, little research had been performed to determine the effect of UV when used in combination with antimicrobial agents that are commonly added to juice products. Therefore, the objectives of this work were (1) to determine if chemical preservatives and ultraviolet light have a combined effect on the reduction of Escherichia coli in apple cider, and (2) to determine the influence of adding chemical preservatives at different points in the processing of juice (i.e., either prior to or after ultraviolet light processing) on the reduction of Escherichia coli in apple cider. In this study, refrigerated (4°C) pasteurized apple cider that contained no added preservatives was inoculated with E. coli ATCC 25922, a surrogate strain for E. coli O157:H7, and exposed to UV (peak output: 254 nm). The following chemical preservatives were added to apple cider either prior to or after UV exposure: dimethyl dicarbonate (75 and 150 ppm), hydrogen peroxide (75 and 150 ppm), potassium sorbate (1000 and 2000 ppm), and sodium benzoate (1000 and 2000 ppm). Following UV exposure and chemical preservative application, inoculated juices were stored at 4°C for 72 hours. Samples were collected prior to and immediately after UV exposure and at 24, 48, and 72 hours of storage. At each sampling point, juice portions (0.1 ml) were serially diluted in peptone diluent (0.1%) and surface plated onto Tryptic Soy Agar (TSA). Counts of the bacterial colonies were made 48 hours after incubating plates at 35°C. Overall, reductions of E. coli were greater in cider treated with preservatives after UV processing than when preservatives were added prior to UV processing (P < 0.05). Furthermore, dimethyl dicarbonate and hydrogen peroxide were more effective than potassium sorbate and sodium benzoate in reducing E. coli populations in conjunction with UV (P < 0.05). When added prior to UV exposure, potassium sorbate was the least effective, allowing for the greatest survival (P < 0.05). This study describes the use of UV in conjunction with hydrogen peroxide and dimethyl dicarbonate as an effective method for producing a 5-log or greater reduction of E. coli O157:H7 in apple cider. Master of Science

Environmental sampling is a pathogen monitoring technique that has become important in the food industry. Many food processing companies have adopted environmental sampling as a way to verify good manufacturing practices and sanitation plans in their facilities. Environmental sampling is helpful because it gives better information on the source of product contamination than end product sampling. Two specific pathogens of concern to the food industry are Listeria monocytogenes and Salmonella enterica. Environmental samples are rarely analyzed immediately, but instead may be batched for later analysis or shipped to an off site testing facility. Multiple media on the market today is used for storage and transport of environmental samples. These various media types, differences in holding temperatures and time create variability in test sample conditions. Select time, temperature and media combinations were tested to determine their effect on Listeria monocytogenes and Salmonella enterica populations during transport and storage of samples. Cocktails of Listeria monocytogenes and Salmonella enterica were added separately to sample tubes containing D/E Neutralizing Broth, Neutralizing Buffer or Copan SRK Solution. Bacterial counts at 0, 12, 24 and 48 hours post inoculation were compared. Neutralizing Buffer and Copan SRK Solution maintained consistent bacterial populations at all temperatures. At 10° and 15°C, D/E Broth supported bacterial growth. This study helps validate the use of D/E Neutralizing Broth, Neutralizing Buffer and Copan SRK Solution for environmental sample transport and storage at proper holding temperatures. At temperatures >10°C Neutralizing Buffer or Copan SRK solution should be used if quantifying microbial recovery. Master of Science

Preventing flavor fade requires an understanding of the relationship between carbonyl amine and lipid oxidation reactions. The polyunsaturated fatty acid content of lipids in peanuts makes them more susceptible to lipid oxidation. The major by-products of the oxidation reaction are nonanal, hexanal, octanal, and decanal. These chemicals are associated with cardboardy, painty, and oxidized flavors associated with flavor fade. The carbonyl-amine reaction yields a variety of pyrazines with positive flavor attributes. Initial flavor notes were explored through sensory work, Gas Chromatography-Olfactometry, and chemical analysis. The fresh roasted volatiles produced from roasted peanuts and the aldehydes resulting from oxidation were also evaluated using GC-FID to quantify and identify the pyrazines and hexanal over a 21 day storage period. Electronic Nose was used to determine differences between storage periods. Gas chromatography-Olfactometry identified potent pyrazines contributing to fresh roasted peanutty aroma in fresh peanuts. Using GC-FID a significant decrease (p0.05) was noted in the other pyrazines evaluated. A significant increase (p

Fish products under vacuum (VAC) and/or modified atmosphere packaging (MAP) conditions can have a significantly extended shelf life. Prevention of toxin production by Clostridium botulinum is essential for processors of VAC and MAP refrigerated fishery products. The objective of this study was to determine if C. botulinum toxin development precedes microbiological spoilage and sensory rejection in fully cooked breaded and battered Alaskan Pollock or raw aquacultured flounder fillets. Aquacultured summer flounder (Paralichthys dentatus) fillets and fully cooked breaded and battered Alaskan pollock (Theragra chalcogramma) were either aerobically packed (Oxygen Transmission Rate (OTR) of 3,000 cc/m2/24h@70°F for flounder and 6,000 cc/m2/24h@70°F for Pollock), vacuum packed or MAP packaged in a 100% CO₂ atmosphere (OTR of 7.3 cc/m2/24h@70°F). Flounder fillets were stored at either 4 or 10°C while pollock portions were stored at 8 and 12°C. Based on the time to spoilage (counts >107 CFU/g), additional samples were inoculated with five strains of nonproteolytic C. botulinum and analyzed qualitatively for botulinum toxin using a mouse bioassay. For flounder at 4°C, toxin formation did not occur after 35 days in aerobically packed fillets. Vacuum packed and 100% CO2 fillets produced toxin before spoilage at days 20 and 25, respectively. In the aerobic packages at 10°C, toxin production occurred after spoilage at day 8, but before spoilage in the vacuum and 100% CO₂ packages at day 9. Sensory evaluation of toxic vacuum and 100% CO₂ packages at 4°C revealed toxin production proceeded spoilage and absolute sensory rejection. However, at 10°C toxin production was evident only after absolute sensory rejection and microbiological spoilage for aerobically packed fillets. Vacuum packages and 100% CO₂ packages were toxic during spoilage but before absolute sensory rejection. Toxin was not present in the aerobically and 100% CO₂ packed pollock samples at 8°C and the 100% CO2 packed samples at 12°C after 35 days. Aerobically packed portions stored at 12°C first produced toxin at day 25; toxicity occurred after absolute sensory rejection and before spoilage. The vacuum packed portions first formed toxin at day 25 for 8 and 12°C storage before spoilage and absolute sensory rejection. Ph. D.

Listeria monocytogenes is the major pathogen in ready-to-eat meat products such as deli meats and frankfurters. Contamination can occur via the salt brines that are used to cool thermally processed meats. Both L. monocytogenes and lactic acid bacteria can grow and thrive under these brine conditions, and may become competitive with each other for available nutrients. The objective of this study was to determine the effect of a three strain cocktail of lactic acid bacteria Enterococcus faecalis, Carnobacterium gallinarum, and Lactobacillus plantarum on the survival of Listeria monocytogenes and Listeria innocua in brines stored under low temperatures up to 10 days. Three brine concentrations (0%, 7.9%, and 13.2% NaCl) were inoculated with ~7.0 log₁₀ cfu/ml of one of five cocktails (L. monocytogenes, L. innocua, lactic acid bacteria (LAB), L. monocytogenes + LAB, or L. innocua + LAB) and stored for 10 days at either 4°C or 12°C. Three replications of each brine/cocktail/temperature combination were performed. No reductions of L. monocytogenes were seen in 7.9 or 13.2% NaCl with LAB; however, reductions of L. monocytogenes were seen in the 0% NaCl with LAB (1.43 log at 4°C and 3.02 log at 12°C). Listeria innocua was significantly less resilient to environmental stresses than L. monocytogenes, both with and without LAB present (p

The effects of packaging treatments, post harvest cooling delay and storage duration on color, texture, ascorbic acid content, weight loss and glucosinolate retention in crown-cut heads of broccoli were studied. Oxygen and CO2 levels inside shrink wrap packages were also monitored. Shrink wrap packaging had a significant positive effect on hue angle (p