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Agrarian movements around the world use agroecology to build sovereignty and steward dynamic ecosystems. Research has shown that agroecological farmers steward more resilient crops, more resilient soil biomes, and greater biodiversity than conventional agriculture. GIS and remote sensing offer many tools to detect the impacts of these farmers on the environment, but it is less clear how such technologies fit into agroecological goals. This study asks: what values, experiences and knowledge do smallholder producers in Bernalillo County embody in their soil stewardship practices? Also, what experience or knowledge do smallholder producers in Bernalillo County have about remote sensing, and would they use remotely sensed data to understand the impact of their soil stewardship practices? To answer these questions, I look to political ecology and feminist theory for understanding the embodied knowledge and practices that are fundamental to agroecology. I find that the practice of farmers in this study embody the same values and methods of agroecology farmers worldwide. Additionally, the remote sensing process could be useful to detect the impacts of farmer practices on soil dynamics in Bernalillo County.

Ice recrystallization is a key problem leading to quality deterioration of frozen foods and biomedical materials. Molecules possessing ice recrystallization inhibition (IRI) activity can control ice crystal size after freezing, reducing the degree of freezing-induced damage. While these IRI active molecules have shown promise in maintaining the quality of frozen foods, their slow discovery, limited creation strategies, and lack of mechanistic understanding limit their hypothesis-driven design. In this work, we address these above-stated needs required for the streamlined implementation of these materials into frozen systems by the creation of a machine-learning-based IRI screening method, the development of a new IRI agent creation strategy, and through the study of a small molecule IRI agents’ mode of action. Cellpose, a segmentation algorithm, was used to develop an automated ice measurement protocol to better screen IRI agents using images from the splat and sucrose sandwiching assays. Next, Ni2+ immobilized metal affinity chromatography (IMAC) was used to fractionate pulse peptide hydrolysates by their histidine, cysteine, and tryptophan residues into peptide fractions enriched or lacking these amino acids. The fractionation of these hydrolysates induced IRI independent of their evaluated molecular characteristics, demonstrating for the first time that IMAC fractionation of pulse peptides may act as a viable creation strategy for bio-based IRI agents. Lastly, we present the discovery of a new IRI agent from biomass, sodium copper chlorophyllin (SCC), that exhibits potent IRI in 10 mM NaCl without inducing IRI in 180 mM NaCl and PBS. SCC’s anti-freeze activities only included IRI, lacking dynamic ice shaping and thermal hysteresis. SCC’s IRI was not mediated by ice binding, self-assembly, or governed by its surface orientation within 10 mM NaCl. Hence, this Thesis advances the field of cryopreservation by the novel identification, creation, and characterization of bio-based IRI agents.

The goals of this study were twofold: to further examine sustainability within the brewing industry through a literature analysis across four academic journals, and a benchmarking analysis of a Louisville, Kentucky brewery’s resource consumption and emissions production. Publications related to the brewing industry were examined for the presence of sustainability keywords: “efficiency, sustainability, carbon footprint, climate change and greenhouse gas”. Benchmarking examined the local brewery’s primary production facility to determine their performance compared to other surveyed breweries pursuing sustainability across the country. Literature examination found noticeable discrepancies between the appearance of sustainability topics within article abstracts and their discussion within publications. Benchmarking for the local brewery found deviation in resource use and greenhouse gas production compared to surveyed benchmarks from the larger industry. As brewing proliferates across the nation, further collaboration between industry leaders and researchers will serve as a vital catalyst for applications of sustainability within the brewing industry.

As food prices continue to rise, many consumers are looking at ways to cut back on spending and many look to how much they spend on food. Food waste in an average American household of four leads to about $1,500 worth of unused/wasted food annually, This study was conducted to see how the use of Chlorine Dioxide (ClO2) could be used to treat and extend the shelf life of freshly sliced tomatoes for food service applications. Prior research in this regard has looked at whole tomatoes as opposed to sliced varieties, and many have not utilized a sensory panel to comment on a consumer’s perspective of how the treatments could affect their decision making. Through the use of texture analysis, color readings, as well as sensory were used with statistical analysis performed to determine if any showed significance based on the treatment that was performed on the tomatoes, with treatments ranging from control (no ClO2 treatment), bulk treated by ClO2, and finally bulk treated by ClO2 with a slow-release media sachet in package during storage. It was found that there was no significance with regards to treatment when it came to textural analysis by either TPA or shear blade tests throughout the refrigerated storage period of 20 days. There was a significant difference related to sensory testing. Panelists significantly preferred sliced tomatoes treated in bulk and then stored with in-tray slow-release sachets compared to tomatoes treated in bulk only or the control (untreated) tomatoes. There were no significant differences for tomatoes based on sensory assessment of color, aroma, or taste, indicating chlorine dioxide treatment had no negative effects on the sensory properties of the fresh sliced tomatoes. This is important because the tomatoes can be treated with a quality shelf life extending step while still maintaining sensory acceptability. The results of this study can be used to help determine if use of chlorine dioxide for shelf-life extension of fresh sliced tomatoes is beneficial for use with foodservice operations.

Maintaining sausage analogue quality during a thawing process and storage is important for increasing consumer satisfaction. Following new trends and prevalence of plant-based meats and a flexitarian diet, two experiments were conducted to test how thawing methods (experiment 1) and storage temperatures (experiment 2) affect meat analogue quality. For Experiment 1, Jimmy Dean Original Sausage Patties Made with Pork & Turkey, Morningstar Veggie Original Patties and Beyond Sausage Original Patties were purchased pre-frozen and subsequently thawed in one of three ways: 1. thawed at 4°C for 24 hours, 2. thawed at room temperature (22°C±2°C) for 4 hours and 3. thawed in the microwave for 30-35 seconds. Sausage and sausage analog quality was evaluated by measuring thaw loss percentage, colorimetry, water activity, pH, sensory scoring, GC-headspace volatiles, TBARS values, Texture profile analysis (TPA) and cooking loss percentage. For experiment 2, the same three types of sausage patties were purchased and placed under refrigeration at either 1°C, 4°C or 7°C. Temperature fluctuations and relative humidity data were monitored using sensors. Sausage and sausage analogue quality was evaluated by measuring total aerobic bacteria, drip loss percentage, colorimetry, water activity, pH, GC-headspace volatiles, proximate analysis (moisture, fat, protein content), water holding capacity, sensory, TPA, and cooking yield. The most striking difference in thawing effects was a higher thaw loss in microwaved samples compared to the other methods. Additionally, refrigeration temperatures had an effect on shelf life and quality of samples. The samples that were subjected to a storage temperature of 7°C were associated with lower quality and shortened shelf life as seen by higher microbial counts and less desirable color and texture found through colorimetry and texture profile analysis, respectively. Conversely, a refrigeration temperature of 1°C was associated with higher quality patties as there was lower microbial counts and higher 3 cooking yield. Better textural properties were seen in samples that were refrigerated at 4°C through the use of texture profile analysis. Despite the impact of methodology on samples, patty types were the largest indicator of quality with Morningstar patties seen as the highest quality reflected by low microbial counts, low moisture loss and high scores in sensory trials.

Roasted and ground coffee is a shelf stable product yet quite sensitive to oxidative staling. A consumer acceptance-based shelf-life modeling system was proposed with intent for the rapid determination of suitable coffee packages. This model requires as input the oxygen consumption rate (OCR) of the coffee, barrier values of packages, and the size of the packaging. Within the time period tested, it was shown that this model accurately predicted the oxygen uptake of coffee over time. Four bio-based packaging systems with barrier layers including mPLA, mPE, mcellophane, and paper were compared against a control (mPET). These materials displayed a range of effectiveness in containing moisture and oxygen. It was determined that no materials, including the control, were able deliver a 6-month shelf life of roasted and ground coffee in a non-modified atmosphere at high sensorial rigor. However, the mPET and mcellophane materials could sustain a 6-month shelf life at medium sensorial rigor, and that all materials could sustain a 6-month shelf life at low sensorial rigor. High, medium, and low sensorial rigor were defined as an oxygen uptake of 150, 225, and 300 μg per gram of coffee, respectively. Additional research is needed to measure consumer acceptance more precisely over time with this model.

Sous vide cooking has the potential to increase the value of lower quality cuts of beef. Understanding the textural properties following a cook-chill sous vide process is crucial to maximizing this potential. The use of enzymes may further increase the value of these products if able to positively alter the texture of the final product. The first research objective was to use instrumental texture analysis to evaluate potential relationships between enzymatic treatments and sous vide processing time of lower value cuts of beef. Three cuts were evaluated: infraspinatus (top blade), semitendinosus (eye of round), and beef tongue. Infraspinatus and semitendinosus samples were sous vide cooked at 60°C for 4, 8, or 12 hours. The beef tongue was sous vide cooked at 60°C for 36, 48, 60, and 72 hours. Samples were treated with papain (EC 3.4.22.2) or bromelain (EC 3.4.22.33). Warner Bratzler Shear Force (WBSF) and Texture Profile Analysis (TPA) were performed on each treatment. Significant differences (P ≤ 0.05) were observed in the WBSF values of toughness and firmness between sous vide processing times and enzymatic treatments. TPA results indicated significant differences (P ≤ 0.05) between sous vide processing time and enzymatic treatment for hardness, adhesiveness, springiness, cohesiveness, and chewiness. A qualitative test was conducted to estimate the remaining enzymatic activity following the different sous vide processing times. The objective of the sensory study was to evaluate potential consumer opinions and preferences of semitendinosus (eye of round) sous vide and treated with papain and bromelain. The untreated samples were cooked with sous vide at 60°C for either 4 or 8 hours. Samples were also treated with 0.075% w/w papain or bromelain and cooked via sous vide for 4 hours. A non-sous vide cooked sample was prepared by searing to an internal temperature of 60°C to match the doneness of the sous vide cook. A hedonic sensory panel was conducted with 51 consumers, scoring the samples on textural properties including juiciness, tenderness, cohesiveness, mouthfeel, flavor, and overall texture. Significant decreases (P ≤ 0.05) in the panelist scores of enzymatically treated samples for parameters of cohesiveness, mouthfeel, and overall texture were observed. A ranked preference test with 54 untrained panelists was conducted using the same treatments. The samples were ranked for preference (1=most preferred, 5=least preferred) to gain insight into consumer acceptance. The non-enzymatically treated samples were significantly ranked higher (P ≤ 0.01) than the enzymatically treated samples. Future investigation into doneness level, final preparation method, and enzyme treatment levels needs to be conducted to find an enzymatic treatment acceptable for use in a sous vide cook-chill process. Currently, the enzymatically treated samples were found unacceptable by consumers at the dosage of enzyme used in this research. It is evident that the application of cysteine proteases is an effective method of altering the texture of sous vide beef, however, more research needs to be conducted to identify optimal enzymatic treatments.

Excess sodium consumption has been linked to various health risks. The use of taste modulators may be a promising strategy to reduce the salt content in low-sodium foods yet maintain salty perception. Previous research has confirmed the subthreshold synergistic effect among pyroglutamyl dipeptides on saltiness enhancement; however, no studies have been published on the subthreshold synergistic effect of mixture of pyroglutamyl peptides and organic acids. This study aimed to explore the subthreshold synergistic effect of pyroglutamyl peptides and organic acids on saltiness perception. Specifically, pyroglutamyl dipeptides (pgluE and pgluV), pyroglutamyl tripeptides (pgluVL and pgluVC), and organic acids (malic acid and succinic acid) were explored in model system and subsequently in a commercial brown onion sauce. First, the detection threshold of pyroglutamyl peptides (pgluE, pgluV, pgluVL, pgluVC) were determined to be 646 μmol/L (167 ppm), 77 μmol/L (18 ppm), 273 μmol/L (93 ppm), and 221 μmol/L (73.2 ppm) respectively. Next, the subthreshold synergistic effect of the pyroglutamyl tripeptides and organic acids was determined using the isobologram method. One of the eight combinations of pyroglutamyl tripeptides with pyroglutamyl dipeptide (pgluV) showed subthreshold synergistic effect, whereas four combinations of pyroglutamyl tripeptides with malic acid and one combination of pyroglutamyl tripeptides with succinic acid exhibited subthreshold synergistic effect. Later, we conducted a triangle test to determine the maximum achievable salt reduction in a commercial brown onion sauce, accomplishing a notable 20% reduction in salt without the introduction of any supplementary ingredients. Subsequently, we achieved an additional salt reduction in the brown onion sauce by 5% and 10%, through the synergistic combinations of pyroglutamyl tripeptides with malic acid and succinic acid, respectively, resulting in an overall salt reduction of 25% and 30% respectively. This research lays the foundation for future investigations into the potential of combinations of pyroglutamyl peptides and organic acids for saltiness enhancement in low-sodium foods.

The authentication of products with claims regarding protein sources or compositions is a challenge for traditional analytical methods, which generally lack the required specificity whole protein analysis can provide. For example, the establishment of milk as “A2” is achieved through genetic testing of cows before milk production, with no methods to authenticate milk products themselves. Establishment of A2 milk is completed through genetic testing of the cows before milk production, but with no methods to authenticate the milk products themselves. Intact protein mass spectrometry (MS) has the potential to directly authenticate protein products, including specific proteoform claims. The development of an intact MS method to detect and differentiate major bovine milk proteins (αS1-, αS2-, β-, K-caseins, β-lactoglobulins, and α-lactalbumins) and their proteoforms is needed for protein profile claims and can be an effective tool to analyze milk products for protein authentication. This was attained through three major phases: generation of a predicted mass database, optimization of sample preparation and instrument parameters conducive with intact bovine milk proteins, and the selection of deconvolution software for protein identification with a mass error tolerance (10 ppm). Fifteen powdered and HTST liquid milk products with an equal distribution of marked A2 and normal commercial products were selected. Each sample was diluted to 1 mg protein/mL in 50 mM ammonium bicarbonate and then defatted through centrifugation of 15 minutes at 3,000 x g. The samples were then cleaned up, desalted through 3 kDa spin column filters, and then separated and analyzed by liquid chromatography mass spectrometry (LC-MS). Data was deconvoluted using BioPharma Finder sliding windows algorithm that were compared to the predicted database and mass were identified. A mean of 85.27% (± 6.68%, n = 57) of the total signal of powdered and liquid HTST milk could be assigned to the predicted database proteoforms using the finalized method. The average ratio of selected normal commercial products was 25.86% A1 and 0.74.14% A2. Advisors: Melanie Downs and Phillip Johnson

Environmental sustainability can be positively impacted by the inclusion of feed additives to reduce enteric methane production from cattle. Methane production can be affected by feed additives that either alter the rumen environment or act as methanogenesis inhibitors. A reduction in methane from cattle can contribute to meeting carbon neutrality. A metabolism study was conducted to evaluate Alga 1.0, a product containing bromoform, fed to cattle to evaluate the effects on gas emissions. Treatments were (0, 69, or 103 g/d Alga 1.0) fed in a corn-based diet. Headbox-style indirect calorimeters were used to measure gas emissions. Feeding Alga 1.0 linearly decreased dry matter intake (P < 0.01). There was a linear decrease in CH4 when expressed as g/d (P < 0.01) and g/kg DMI (P < 0.01). There was no effect on CO2 expressed as g/kg DMI (P > 0.20). Diet digestibility (P ≥ 0.28) and digestible energy were not affected (P ≥ 0.38) when feeding Alga 1.0. Feeding Alga 1.0 reduced CH4 emissions from cattle. A finishing study was conducted to evaluate black seed oil plus acetic acid on beef cattle performance as well as enteric methane production. The diet consisted of Sweet Bran, high moisture corn, dry rolled corn, alfalfa hay, and supplement with the black seed oil plus acetic acid replacing 2.67% of diet DM of the corn blend. Cattle were rotated through a two-chamber emissions barn equipped with open-circuit indirect calorimeters to measure CH4 & CO2. There was no effect on CH4, or CO2 emissions (P ≥ 0.26) compared to the control diet. Body weight, dry matter intake, and ADG were not affected (P ≥ 0.34) by the inclusion of black seed oil plus acetic acid. There was a tendency for improved feed efficiency (P = 0.08) in the cattle consuming black seed oil plus acetic acid compared to control fed cattle. Feeding black seed oil plus acetic acid did not affect gas emissions. Advisor: Andrea K. Watson

Areas of char or overcooking commonly appear in foods people consume. It has been reported that overcooked food is harmful to human health. However, little research exists on the effect of overcooking on in vitro protein and starch digestibility and gut microbial fermentation. This study aimed to reveal the connection between overcooking and in vitro protein and starch digestibility and gut microbial fermentation. In vitro protein digestibility of an overcooked ground beef patty was almost half that of a standard cooked sample (27 ± 2% versus 48 ± 6%, respectively; p = 0.02). Whole wheat bread protein digestibility was also significantly diminished by overcooking (standard: 56 ± 3% versus overcooked: 43 ± 4%; p = 0.02), while starch digestibility was not influenced by overcooking (standard: 80 ± 1% versus overcooked: 76 ± 2%; p = 0.12). Overcooking decreased acetate, propionate, iso-butyrate, iso-valerate and ammonia production by the gut microbiota during fermentation of the beef sample, and decreased propionate and ammonia production during fermentation of the bread sample. Interestingly, overcooking enhanced butyrate production by the microbiota after 48h of fermentation of the bread sample (4.31 ± 1.26 versus 2.05 ± 0.69 respectively, p = 0.035), while no significant difference was found between overcooked and standard cooked beef samples (7.95 ± 1.34 versus 7.79 ± 1.13 respectively, p = 0.14). Overcooking resulted in reductions in many Pseudomonadota and favored several Bacillota, especially Ruminococcaceae, and Oscillospiraceae, which contain butyrate producers. Overall, overcooking had a strongly reduce in the digestibility and fermentation of proteins. Unexpectedly, overcooking induced several purportedly favorable effects on the gut microbiota due to the decreased protein fermentation, which, in future studies, should be weighed against the previous reports that overcooking is deleterious to human health. Advisor: Devin J. Rose

According to a survey by ACNielsen International Research, over half of Americans want to start their own business to build wealth and achieve independence (Bygrave & Zacharakis, 2011). In the research community, most entrepreneurship education research happens in business schools (Matlay, 2006). As a result of continually researching one form of entrepreneurship, less attention has been paid to other forms contributing to the “definitional obscurity with important consequences for the direction of the field” of entrepreneurship (Baker & Welter, 2017). This has created a need for research on entrepreneurship at the college level in fields outside of business. Clemson University and the University of Georgia (UGA) are two universities located in the southern United States. Both universities offer entrepreneurship programs and opportunities to students–Clemson with the Spiro Institute and UGA with the Terry College of Business. The decision to compare the universities was made due to UGA’s entrepreneurship program being focused primarily at the university level, whereas Clemson has a focus on entrepreneurship from colleges other than business. The impact of entrepreneurship programs and other factors on student interest in entrepreneurship were investigated. Clemson University had a greater variety of majors involved in entrepreneurship programs and competitions. Most students answered that participating in competitions increased their interest in entrepreneurship but considered a fear of public speaking, coming up with innovative ideas, and a lack of time to devote as challenges to participation. A flyer on campus, an email from the college and/or university, social media, and a representative of the competition coming to speak to a class were the most impactful forms of communication about entrepreneurship competitions. For both universities, faculty mentor support, business mentor support, and coming up with innovative ideas were the most statistically significant influences for those that had not done entrepreneurship competitions The author participated in a semester-long course that required participation in an entrepreneurship competition. The course and development of the sustainable, nutrient-dense meal bar provided valuable insights into what is required to create a business from instructor-led ideation to pitching a final product. The author found that the work required to develop a business and pitch to judges is not a deterrent and participation in one competition can increase interest in entrepreneurship and future pitch competitions; however, there is value in initially informing students of the time commitment. Educators can use this information to consider tying entrepreneurship competitions into their course curriculum, increasing participation and potentially exposing a greater variety of students to entrepreneurship. This study was limited by sample size (Clemson n= 200, University of Georgia n= 170), classes visited, and period, therefore additional research with a larger sample, a greater variety of classes, and a longer time frame may yield more conclusive results to help guide future research and applications.

Black seed oil concentrations of 0%, 0.1%, 1% and 5% were added to milk inoculated with kefir grains and incubated at 25°C for 22 h. The pH and microbial count indicated 1% black seed oil caused low inhibition (P > 0.05) of fermentation, but 5% black seed oil caused significant inhibition of the kefir microorganisms (P < 0.05). Cobb 500 male chicks (n = 256) were distributed in a randomized block design and received one of four treatments: CTRL1 (Non-medicated, no kefir, no Clostridium perfringens), CTRL2 (Non-medicated, no kefir, C. perfringens inoculated), CTRL3 (BMD medicated, no kefir, C. perfringens inoculated), KTRT (Non-medicated, kefir treated, C. perfringens inoculated). Kefir was supplied via a 1:10 kefir to water ratio in the chicken water supply. Birds were orally inoculated with Eimeria maxima and C. perfringens on Day 11 and Days 16 through 18, respectively. KTRT (kefir) weight gain was comparable to the medicated CTRL3. CTRL3 and KTRT feed conversion ratios were significantly lower than CTRL2 (untreated). There was no significant difference (P > 0.05) between CTRL3 and KTRT lesion scores. CTRL3 (antibiotic) mortality rate was significantly lower than other treatments. KTRL (kefir) reduced mortality but not as effectively as CTRL3. Cobb 500 male chicks (n = 320) were placed in randomized block design and received one of five treatments: CTRL1 (nonmedicated, uninoculated, no kefir, no black seed), CTRL2 (nonmedicated, inoculated, no kefir, no black seed), CTRL3 (medicated, inoculated, no kefir, no black seed), KTRT1 (nonmedicated, inoculated, 2:10 kefir to water supplied in drinking water, no black seed), KTRT2 (nonmedicated, inoculated, kefir, black seed), KH2O (sample of 2:10 kefir-water mixture from water trough). Birds were orally inoculated with E. maxima and C. perfringens on Day 14 and on Days 19 and 20, respectively. Weight gain of CTRL3 (antibiotic) and KTRT2 (kefir + black seed) increased during D14-D28. Feed intake of KTRT2 was significantly (P < 0.05) higher than CTRL3 and KTRT1 (kefir). CTRL3 and KTRT2 lesion scores were the lowest of all treatments. KTRT1 was as effective as CTRL3 in the prevention of mortalities. KTRT2 mortality rates were the lowest of all treatments. Results suggest KTRT2 was effective against necrotic enteritis in broiler chickens.

Salmonella contamination poses a significant risk to food safety, particularly in poultry products and public health. This study aims to investigate the effectiveness of peracetic acid (PAA) treatment in mitigating Salmonella cross-contamination in chicken thighs within simulated post-chill tanks, considering different inoculum concentrations and organic matter (OM) levels. The experiment involved the use of five distinct Salmonella serovars to inoculate chicken thighs, which were categorized into two groups: higher inoculation level (ca. 7 log CFU/mL) and lower inoculation level (ca, 4 log CFU/mL). These groups underwent treatment with four different solutions, including 540 ppm PAA with 1.5% OM, 540 ppm PAA with 3% OM, pre-chilled distilled water with 1.5% OM, and pre-chilled distilled water with 3% OM. Each treatment included immersion of one inoculated thigh and five non-inoculated thighs in separate tubs for 30 seconds, allowing for the assessment of cross-contamination potential. Microbiological analysis was conducted, and non-inoculated chicken thigh samples that tested negative for Salmonella were further subjected to incubation at 37 °C for 24 h for the identification of low levels of the bacteria that may have been initially undetectable. Simulated post-chill water quality parameters, including pH, chemical oxygen demand (COD), and total nitrogen (TN), were characterized before and after each treatment cycle. The results demonstrated the significant (P < 0.05) reduction in bacterial contamination achieved through PAA treatment compared to distilled chilled water for both highly and lowly inoculated chicken thigh samples. The efficacy of PAA against Salmonella was unaffected by the presence of OM at varying concentrations. Moreover, the study confirms the occurrence of cross-contamination during the immersion process in post-chill tanks, underscoring the necessity of implementing preventive measures throughout poultry processing. Additionally, the treatment led to notable changes in the chemical composition of the samples, evident through increased levels of chemical oxygen demand (COD), total nitrogen (TN), and pH. These findings underscore the importance of comprehensive preventive measures within the industry and provide valuable insights into the efficacy of PAA treatment for Salmonella cross-contamination. Advisor: Byron D. Chaves

Soil nutrient management is of the utmost importance to ensure crop yields are maximized to feed the world’s growing population. Micro and macronutrient deficiencies can be detrimental to plant growth and cause a drastic reduction in yield. Optimization of nutrient management needs extensive research. Nutrients such as nitrogen increase crop yield, even in legumes capable of using a large amount of nitrogen (N) via biological N fixation. Iron (Fe) fertilizers are also crucial for legumes as they are susceptible to Fe deficiency induced chlorosis. Dry edible beans (Phaseolus vulgaris) are leguminous plants and excellent dietary protein sources. Great Northern (GN) beans are a market class of dry edible beans and a major agricultural commodity in Nebraska. Soil nutrient management with N fertilizer can enhance bean production by increasing N uptake, improving protein quantity and quality, and result in a potential economic benefit to bean farmers. Soil nutrient management with Fe fertilizers can combat iron chlorosis and increase bean yield. Thus, this study aims to evaluate and optimize the effects of N treatment on GN bean yield, total protein, soluble protein, digestibility, and intestinal absorption. This study also aims to evaluate the effects of Fe fertilizers on GN bean yield. Results concluded that N treatment at a rate of 140 kg/ha was successful at increasing GN bean yield, total protein, soluble protein, digestibility, and intestinal absorption compared to the control, which had no added N. Results also concluded that Fe fertilizer, regardless of source and application type, had no significant effect on GN bean yield. As the demand for plant-based protein grows, it brings a large market for legume proteins that can be optimized with N management. N management to enhance dry bean yield, protein content, digestibility, and intestinal absorption will benefit the growing population and ensure nutritious plant-based protein sources are available. Advisors: Kaustav Majumder and Bijesh Maharjan

While shrimp is the most-consumed seafood product in the United States, there is a lack of research into the extent of short-weighting and mislabeling of shrimp in the marketplace. The objective of this study was to investigate frozen shrimp for Country of Origin Labeling (COOL) compliance, species authentication, acceptable market names, net weights, and percent glaze. A total of 106 frozen shrimp packages were purchased from grocery stores in Southern California. Samples were determined to be COOL compliant if both the procurement method and country of origin were reported at the point of sale. Species authentication and acceptable market names were determined by comparing the species identification based on DNA barcoding to the acceptable market names on the FDA Seafood List. Net weights and percent glaze were determined by taking the weight of each sample before and after deglazing according to AOAC methods. The determined net weight of each product was compared to the declared net weight to determine if samples had been short-weighted, taking into account the maximum allowed variance (MAV) by the National Institute of Standards and Technology (NIST) standards. Overall, 94% of samples were compliant with COOL; two samples did not indicate the production method and three had conflicting country of origin information on the label and placard. The average percent glaze was 16.6%, with 26% of samples having >20% glaze. Short-weighting was detected in 37% of samples, with the greatest proportion of incidents recorded for the super/extra colossal shrimp category (57.1%). Species mislabeling was observed in 37% of samples, with conflicting market names, species substitution, and/or use of unacceptable market names. The results of this study indicate a high level of COOL compliance but suggest a need for increased scrutiny of species mislabeling and short-weighting of frozen shrimp.

Ice cream’s main shelf-life defect is ice recrystallization. This refers to the growth of larger ice crystals at the expense of smaller ones, which are currently stabilized mainly by polysaccharide gums. These gums work decently but have become increasingly expensive over the years. Using the sucrose sandwich assay, corn cob hemicelluloses’ and holocellulose nanocrystals’ ice recrystallization inhibition (IRI) activity was investigated using the sucrose sandwich assay. Although holocellulose nanocrystals (holoCNCs) did not exhibit activity, 0.5% w/w hemicelluloses (hemiCs) were able to inhibit ice recrystallization by 67.70 ± 1.09% in a model ice cream system after 7 days and were active at concentrations as low as 0.1% w/w. In a commercial ice cream mix 0.5% w/w hemiCs inhibited ice recrystallization by 51.75± 0.40% after 7 days, performing better than two common commercial stabilizers—guar gum and locust bean gum (LBG). Some preliminary physicochemical experiments were conducted to ensure that hemiCs were a practical stabilizer that could actually be added to ice cream without any major defects.

Fresh produce and poultry have been associated with several foodborne illness outbreaks. Strategies to mitigate these risks are a much-needed research priority; therefore, five experiments were conducted to analyze pathogen cross-contamination capabilities on produce and poultry. Microbial contamination prevention and reduction strategies were also studied. In Chapter Two, three antimicrobial delivery methods in combination with increasing concentrations of hydrogen peroxide-peroxyacetic acid (H2O2-PAA) (0.05-0.25 ml/dL) were compared to evaluate their ability to reduce and prevent cross-contamination of Listeria monocytogenes (L. monocytogenes) and Listeria innocua (L. innocua) on inoculated apples. Apples have been identified as a source of L. monocytogenes contamination and were responsible for several multistate outbreaks in the United States. Additionally, the surrogate L. innocua is well-established as an acceptable surrogate for L. monocytogenes in thermal inactivation studies, however, few studies have compared the surrogate to the pathogen using antimicrobial parameters. The objectives of this study were to: 1) compare the three antimicrobial delivery methods, conventional garden spray (GS), electrostatic spray (ES), and a single immersion dip using a H2O2-PAA sanitizer to mitigate microbial cross-contamination of L. monocytogenes on apples; 2) determine whether L. innocua is an appropriate surrogate for L. monocytogenes for use in antimicrobial challenge studies. The antimicrobial used in this study was SaniDate® 5.0 (H2O2-PAA), which is recommended by the wholesale distribution company (Appalachian Harvest). Appalachian Harvest is a buyer of produce from several small-quantity produce growers in WV. Based on the results of this study, the dip method caused the highest reductions (P < 0.05) of both the pathogen and surrogate bacteria (> 3 log10MPN/ml); whereas the GS method caused the lowest L. monocytogenes (-0.35 log10MPN/ml) and L. innocua (0.19 log10MPN/ml) (P < 0.05) cross-contamination on to uninoculated apples. The concentration 0.25 ml/dL was the most effective at reducing bacteria and preventing cross-contamination. This study concluded that since L. innocua behaved similarly (P > 0.05) to L. monocytogenes, L. innocua is an acceptable surrogate for L. monocytogenes for H2O2-PAA antimicrobial studies. Results from this challenge study also indicated that the dip and GS methods were more effective at reducing and preventing cross-contamination of bacteria on apples, respectively. In addition to apples, tomatoes were another culprit of several multi-state outbreaks in the United States. Nationally, and locally, tomatoes were associated with Salmonella contamination. Therefore, in Chapter Three, the triple wash methods (TWM) water+antimicrobial+water (WAW) or water+water+antimicrobial (WWA) were analyzed to determine their efficacy in reducing and preventing cross-contamination of Salmonella Typhimurium (S. Typhimurium) and the surrogate Enterococcus faecium (E. faecium) on tomatoes. The TWM was used in this study because it is the washing method recommended by the WV Small Farm Center (WVU-SFC) to control pathogens on locally grown fruits and vegetables. The first objective of this study was to evaluate the efficacy of reducing and preventing cross-contamination of S. Typhimurium and E. faecium on tomatoes using two TWMs with different concentrations of a H2O2-PAA sanitizer. The second objective of this study was to evaluate the feasibility of E. faecium as a surrogate for future antimicrobial challenge studies. The results showed that the application of 0.25 ml/dL of H2O2-PAA in WAW or WWA TWM resulted in the lowest (P < 0.05) cell counts compared to lower concentration treatments for S. Typhimurium (0.16-0.69 log10MPN/g) and E. faecium (-0.41 log10MPN/g). There were no significant differences (P > 0.05) in survival between S. Typhimurium and E. faecium after TWM treatments. It was concluded that E. faecium is an acceptable surrogate for S. Typhimurium in antimicrobial challenge studies. It was also concluded that although 0.25 ml/dL significantly (P < 0.05) reduced bacteria, it did not completely prevent cross-contamination of the pathogen. Therefore, future studies are needed to analyze what concentrations of H2O2-PAA could significantly reduce and prevent cross-contamination of bacteria on tomatoes. (0, 0.0064, 0.1, and 0.25 ml/dL) of Previous concentrations analyzed in Chapter Three (0-0.25 mL/ dL) did not completely inactivate the surrogate or pathogen on tomatoes; therefore, in Chapter Four, a follow-up study to the experiment described in Chapter Three was conducted. Higher, off-label concentrations (0.5-0.7ml/dL) of the sanitizer SaniDate® 5.0 (H2O2-PAA) were tested to determine their efficacy in reducing and preventing the cross-contamination of S. Typhimurium and E. faecium on tomatoes. This study aimed to find the minimal concentrations of H2O2-PAA required to prevent microbial cross-contamination of S. Typhimurium and surrogate E. faecium on tomatoes. The sanitizer was applied to inoculated tomatoes by washing fruits for 45 s using TWM methods (WWA or WWA). Significant reductions (P < 0.05) of Salmonella were only achieved when the off-label concentration of 0.7% H2O2-PAA was applied using the WWA strategy. Results showed cross-contamination of S. Typhimurium was prevented at 0.7% of H2O2-PAA using the WWA strategy. Additionally, E. faecium was prevented at and above 0.50% of H2O2-PAA, regardless of wash strategy. Though 0.7% H2O2-PAA is effective at mitigating cross-contamination and achieving more than a 4-log reduction of S. Typhimurium on tomatoes, future research is needed to determine the safety of these greater antimicrobial concentrations. Additionally, approval by the federal government is required before changing the recommendations for the use of these concentrations. Similar to the previous study, results of this study also suggested that E. faecium could be an acceptable surrogate for S. Typhimurium when validating antimicrobial washing systems on tomatoes. In the previous Chapter, we found that the TWM can be an effective washing strategy to reduce pathogen growth and cross-contamination on produce. However, it is unknown if local growers are aware of the WVU-SFC recommended TWM or how much they know about produce cross-contamination. In Chapter Five, a survey was conducted at Farmers’ Markets (FMs) in WV to analyze small and very small-quantity produce growers’ knowledge of food safety. Their awareness of the WVU-SFC recommended TWM and willingness to implement the washing method was also assessed. The information obtained in these surveys were important because previous studies have found that produce at local FMs have been infected with pathogens. Surveys filled out by consumers revealed that most FM consumers consider it the vendors’ responsibility to wash the produce and they generally view FM produce as safe. However, few surveys have analyzed the FM vendors’ perception of food safety on their products. The survey results revealed knowledge gaps about food safety among FM produce vendors. Additionally, 53.7% of FM vendors did not wash their produce due to increased spoilage (61.4%; P < 0.05). Among participants who washed produce, only 28.2% were familiar with the TWM. Most respondents who washed produce (65.3%; P < 0.05) stated they would be interested in attending good agricultural practices/ Food Safety Modernization Act (GAP/FSMA) and TWM training. Understanding FM vendors’ perceptions and current wash methods allows local state government agencies to make better-informed decisions regarding food safety policies to ensure produce safety. Using the information from the findings of this study, the authors’ team plans to work closely with the WVU-SFC to develop outreach activities on training WV small to very small-quantity produce growers on the application of the TWM. In addition to produce being a cross-contamination risk, chicken is a risk both nationally and locally because it is the number one meat commodity associated with foodborne illness. Poultry is also one of the most harvested meats in WV. The risk of pathogen contamination on chickens in the state has only increased since the rise in popularity of mobile poultry units (MPPUs) in WV. Therefore, the objectives of Chapter Six were to characterize the microbial quality of broilers (total aerobic plate counts and generic E. coli), and the presence/absence of Salmonella and Campylobacter after processing carcasses in a pilot commercial MPPU at the West Virginia University Poultry Farm. Aerobic plate counts (APCs), Escherichia coli (E. coli), and the presence/absence of Salmonella and Campylobacter on 96 broiler carcasses after processing in the MPPU were assessed. Samples were taken after the processing steps, immersion scalding, manual evisceration, and immersion chilling in water or water with 5ppm chlorine (Cl2). Results showed that the highest number of bacteria recovered from carcasses were APCs (4.21 log10CFU/mL) and E. coli (3.77 log10CFU/mL). Overall, results show that E. coli, Salmonella, and Campylobacter were significantly reduced (P < 0.05) from the initial scalding to the chilling step. However, Salmonella frequency doubled (15.63-34.38%) after the evisceration step, indicating evisceration may be a critical control point in preventing cross-contamination of Salmonella. Results also show that proper chilling after evisceration is an important microbial mitigation step in MPPU processing. Campylobacter (18.75% positive) was more resistant (P < 0.05) to water chilling than Salmonella (0% positive). Additionally, Campylobacter was not completely inactivated (0/16) until carcasses were chilled in CW, whereas W chilling was sufficient to completely inactivate Salmonella (0/16) on carcasses. Although the addition of 5ppm Cl2 in chill water was significant in reducing bacteria (P < 0.05) on chilled broiler carcasses, it did not necessarily cause a more significant reduction than water chilling alone. Therefore, more MPPU processing studies are needed to research what post- evisceration wash methods and chill-water sanitizers are most effective at preventing cross-contamination, while preserving the integrity of the final product.

Antarctic krill (Euphausiacea superba) is an abundant and sustainable but underutilized natural resource with well-studied protein isolation methodology using Isoelectric solubilization-precipitation (ISP). The resulting krill protein isolate (KPI) has high yield and sufficient impurity removal in addition to desirable nutritional and functional properties but has yet to be used in food systems. Extrusion is a versatile and scalable food processing method with many benefits; thus, the aim of this study was to determine the impact of KPI as an additive in extrusion processing with the eventual goal of introducing it to human and animal food systems. KPI was isolated with standard ISP methodology and incorporated into four recipe formulations with moisture balanced control formulations. The moisture levels and dry ingredients for each formulation were 15% and 20% moisture 100% rice flour, 25% moisture pet food mix, and 33% moisture aquatic feed mix. Specific mechanical energy (SME), expansion, density, crushing force, and toughness were measured and the effects of altering isolate pH on KPI functionality was investigated for foaming, emulsion, and water retention properties. KPI elevated SME in rice flour blends but reduced SME in pet and aquatic feeds. In rice flour blends expansion and texture were largely unaffected. Pet food saw a decrease in radial expansion and an increase in density, and an increase in specific length with KPI addition. These changes did not significantly affect texture. Aquatic feed was significantly less dense and had slightly larger radial expansion with KPI addition and had reduced average crushing force, maximum crushing force, and toughness. KPI displayed better functionality when pH was altered compared to the isoelectric pH. Overall, KPI addition succeeded in this pilot study displaying strong potential for further research and use in food systems.

Oat (Avena sativa L.), a specialty cereal grain belonging to the Poaceae family, is relatively abundant in nutritious protein (12–20%) and dietary fiber (β-glucan). The balanced hydrophilic/hydrophobic amino acid profile in oat protein relative to other plant sources provides numerous possibilities for formulating oat protein isolate (OPI) into a wide range of food products. Yet, there has been limited research on OPI and the application of this valuable protein source is hampered by its low solubility at neutral and mildly acidic conditions. This dissertation research aimed to elucidate the efficacy of high-intensity ultrasound (HIU) for structural modification of OPI and investigate the complementary roles of high pressure, disulfide-breaking agents (mainly cysteine), and flavonoids for the ultimate improvement of OPI functionalities, including solubility, emulsification, and gelation. OPI (mainly 12S globulins) was isolated from oat groats by alkaline extraction–isoelectric precipitation. Because protein functionality is influenced by environmental factors, such as pH, ionic strength, and temperature, it is essential to understand the surface properties of native protein (charge, polarity, and amphiphilicity). Therefore, Experiment 1 was conducted to examine the solubilization behavior of OPI in relation to surface properties when subjected to varying pH and ionic strength conditions. A characteristic U-shaped solubility curve was observed within pH 2.0–8.0 at low ionic strengths (I < 0.01) and a minimum solubility was found at pH 4.0–5.0 and I of 0.03–0.2. Particle size, ξ-potential, and SDS–PAGE patterns supported the solubility profile. In seeking effective modification strategies, Experiment 2 was carried out to determine the effect of internal disulfide cleavage on OPI solubility and emulsifying activity. Differential scanning calorimetry revealed that disulfide bonds contributed to the remarkable high stability (Tm of 111.1 °C) of OPI but impeded its molecular flexibility and functionality. Hence, cysteine was applied to disrupt inter-subunit S–S bonds of OPI. Cysteine at concentrations higher than 1.0 mM/mg protein induced disulfide cleavages, and the 12S globulin subunit dissociation reached a maximum (80%) at 3.3 to 6.7 mM/mg protein. Correspondingly, emulsions prepared with cysteine-treated OPI showed superior interfacial protein coverage (0.170 m2/g compared to 0.092 m2/g protein for control) and reduced emulsion particle size from 4722 to 2238 nm. While cysteine was able to modulate OPI emulsifying activity, more robust techniques were necessary to modify protein structure for functionality improvement. Hence, Experiment 3 sought to apply high-intensity ultrasound (HIU) to dissociate 12S globulins and disrupt tertiary structure to overcome the low-solubility constraints. Through cavitation-induced microscopic shearing and pressure, HIU treatment (5 minutes at 70% amplitude) significantly reduced OPI particle size (up to 37%) and increased solubility (up to 48%) at pH 5.5–8. Fluorescence spectrometry and hydrophobicity measurements revealed protein particle dissociation and major conformational changes, with the exposure of previously occluded hydrophilic/hydrophobic groups predisposing OPI to increased interfacial activity. To enhance the effectiveness of HIU, high pressure (HP) at 30 MPa was applied to complement the effect on modulating OPI functionality (Experiment 4). With further increased emulsifying activity of OPI by the HUI+HP combination treatment at 20 and 60 °C, the emulsions prepared exhibited an additional particle size reduction (from 1653 to 955 nm). Confocal microscopy revealed increased distribution uniformity of oil droplets and emulsion stability. Although in theory both HP and the moderate temperature (60 °C) could aid HIU in dissociating protein aggregates, the improvement on emulsifying activity remained predominated by the HIU process. Based on the above findings, cysteine was reintroduced into Experiment 5 to assist in the formation of oat protein-based gel networks with the pretreatment of HIU. Native OPI samples treated with cysteine alone up to 400 mg/g protein had a significantly increased gel strength (from 0.0049 to 0.23 N) and decreased cooking loss (up to 44%). For HIU treated OPI, the efficacy of cysteine increased 3-fold (gel strength improved to 0.68 N). The combination of cysteine and HIU also enhanced the rheological properties (storage modulus), structural, and water-immobilizing capacity of the heat-induced gel. Polyphenols are naturally present in oat groats and are increasingly utilized to modulate plant protein functionality, Experiment 6 was conducted to investigate the potential collaborative effect of three di-phenol flavonoids with varying hydroxyl groups (1-kaempferol, 2-quercetin, and 3-myricetin) with HIU for enhancing OPI emulsifying activity. Binding with flavonoids (fluorescence quenching) subjected HIU treated OPI to further increased hydrophobicity with 0.5 mmol/L myricetin producing the maximum effect. The HIU+flavonoid treatment improved the emulsion stability. Additionally, all flavonoid-treated samples exhibited a higher oxidative stability against free radicals. Overall, this dissertation research demonstrated that natural chemical compounds (cysteine and flavonoids) and physical treatment (high pressure) provided synergistic effects for HIU on increasing structural flexibility and improving amphiphilicity of oat globulins. The physicochemical changes led to enhanced emulsifying and gelling properties accentuating the potential of these structure-modifying techniques for oat protein.

The utilization of non-conventional chicken parts as human food varies widely across countries. The high prevalence of Salmonella, especially in the giblets, poses a high risk to public health. Poultry processors have implemented multiple hurdle technology to reduce this gram-negative pathogen in chicken parts. This study was conducted to evaluate the antimicrobial efficacy of a commercial blend of citric acid/ hydrochloric acid (CP), peroxyacetic acid (PAA), and sulfuric acid (SA) in reducing Salmonella inoculated on two chicken giblets: livers and hearts. Chicken hearts and livers were inoculated (6 log CFU/ml of rinsate) by individually immersing them in a cocktail of five poultry-borne strains of Salmonella enterica subsp. Enterica each for 30 s achieving initial mean Salmonella recovery of 4.75 ± 0.10 and 4.69 ± 0.10 log CFU/g for chicken hearts and livers respectively. Inoculated chicken hearts and livers were treated by immersing into solutions (4 °C) containing PAA (500 ppm, 90 s), 5% v/v CP (pH 0.66; 30 s), 2% v/v SA (pH 0.93; 30 s), or distilled water (control; 90 s) and analyzed for survivors immediately after treatment (0 h), after one (24 h), two (48 h) and three (72 h) days of aerobic storage at 4 °C. The effect of these treatments on the growth of aerobic mesophilic bacteria and their effect on the color of the chicken hearts and livers were also investigated at the same time points. Results for Salmonella log reductions and aerobic plate count (APC) showed that there was no interaction between the type of antimicrobial treatment and storage time. Salmonella survivors recovered in chicken hearts following treatment (0 h) with PAA, SA, or CP were not significantly different (p < 0.05) from the control. However, SA-treated chicken hearts had significantly lower Salmonella counts than distilled water immediately after treatment. Unlike distilled water, all antimicrobials achieved greater than one-log reductions of Salmonella on both chicken hearts and livers, which indicated that immersing in antimicrobial solutions was more effective in reducing Salmonella. All treatments were effective in minimizing the growth of aerobic mesophilic microflora throughout the three days of storage with no significant differences (p < 0.05) in L*(lightness), a* (redness), b*(yellowness) values on the third day of storage in both chicken parts. Hence the results of the study showed PAA, SA, and CP may be used in the poultry industry as part of a multiple hurdle system to reduce Salmonella in non-conventional chicken parts. Advisor: Byron D. Chaves

Recently, there has been a profound increase in demand of plant-based proteins, especially pulse proteins. However, pulses contain high concentrations of antinutritional factors that hinder the digestibility of proteins. Processing techniques such as enzymatic hydrolysis, microbial fermentation, and physical processing modulate nutritional quality and functionality of pulses. Processing also releases peptides from parent proteins that exhibit health-beneficial bioactivity against various disease risk factors. In this study, germination, gastrointestinal digestion, and microbial fermentation were evaluated to determine the nutritional quality and release of bioactive peptides during and after processing. Germination of chickpeas (Cicer arietinum L.) modulated the protein profile, and the addition of simulated gastrointestinal digestion improved digestibility of chickpea proteins. Germinated and digested chickpeas exhibited high antioxidant activity in an in vitro cell model, but the peptide profile obtained through mass spectrometry revealed no significant differences in di- and tripeptides identified in chickpeas. Microbial fermentation was performed on Great Northern beans (Phaseolus vulgaris L.) with Limosilactobacillus reuteri LTH5448, which is a strain known for producing bioactive gamma-glutamyl peptides. Microbial fermentation effectively modulated the total peptide concentration of Great Northern beans. Mass spectrometry analysis revealed changes in concentration of several gamma-glutamyl peptides and the fermented extract exhibited in vitro anti-inflammatory activity. This study empowers the plant protein industry on how processing methods can modulate the nutritional profile and produce health beneficial compounds from whole pulses. Advisor: Kaustav Majumder

ClO2 as an antimicrobial gas in the headspace of produce package is a relatively novel approach. Gaseous ClO2 is more effective than aqueous ClO2 and can be used in the headspace of fresh food packages. ClO2 gas can diffuse into product surfaces and films. As an oxidizer, it can react with and change polymeric package components, possibly affecting the product’s shelflife. This research studied effects of ClO2 gas treatments on produce packaging materials (APET, two PE types, Nylon). The treatment group (with ClO2) and the control group (without ClO2) of packaging materials were stored at room temperature and at three different relative humidities (49%, 84%, 99%) in sealed chambers. Dart drop, tensile tests, Tg, Tm, Tc, and water vapor transmission rate of materials were performed at six times over a 21-day period. A low dose (2 was a suitable application in these produce packaging materials. This research allows further experimenting with the literature on varying RH’s for the general application (ppm) of ClO2 on each polymer used for common fresh produce applications. This research also studied the use of ClO2 gas sachets to determine the effects on the sensory properties of strawberries. Conditions included typical strawberry storage systems (open pallet system under cold conditions (0-2 0C) and a controlled atmosphere storage system (99% RH, 0-2 0C). Both conditions were tested for 21 days with and without ClO2. ClO2 gas preserved strawberry quality with or without maximum RH. In the absence of 99% RH, ClO2 treated strawberries exhibited better quality than the untreated berries. High RH (99%) alone has significant quality retention of strawberries in a chamber system with or without ClO2. Either ClO2 or high (99%) RH plays a significant role in strawberry preservation in each other’s absence.

Raw, ready-to-eat (RTE) seafood products have become increasingly popular globally, but they are vulnerable to species mislabeling and substitution. DNA barcoding allows for fish species identification by extracting, amplifying, and sequencing a standardized gene target. A wide variety of fish products have been studied with DNA barcoding, but notably missing are ceviche and poke, particularly in the United States. Sushi is known to be a target of mislabeling but has not been extensively studied in Orange County, CA. The objective of this study was to investigate species labeling and use of acceptable market names for sushi, poke, and ceviche sold at restaurants in Orange County, CA. A total of 105 raw, RTE seafood products were collected, including sushi (n = 35), poke (n = 35), and ceviche (n = 35). All samples were sequenced utilizing DNA barcoding or mini-barcoding. The identified species were compared against the menu names, verbal declarations by staff, and the Food and Drug Administration (FDA) Seafood List to verify whether acceptable market names were used for each product. Of the 103 samples identified with DNA barcoding, species substitution was detected at a rate of 23.3% and unacceptable market names were found in 45.6% of samples. Overall, 63.1% of samples had some form of mislabeling. Six samples had an unacceptable market name based on the menu declaration and were also determined to be targets of species substitution based on verbal declarations by restaurant staff. When the mislabeling rates were divided based on product category, ceviche had the highest overall mislabeling rate (85.3%), followed by poke (61.8%), and sushi (42.9%). Mislabeling of ceviche and poke was primarily driven by the use of unacceptable market names, while species substitution was more common in sushi dishes. These rates reveal widespread mislabeling among raw, RTE seafood products and suggest the need for outreach efforts to ensure proper labeling of fish using acceptable market names.