Browsing by Author "Bunning, Marisa, advisor"
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Item Open Access Assessment of airborne microorganisms in a craft brewery(Colorado State University. Libraries, 2011) Miller, Amanda Leah, author; Bunning, Marisa, advisor; Stone, Martha, committee member; Hyatt, Doreene, committee memberAlthough pathogenic bacteria have little chance of surviving in beer due to its intrinsic antimicrobial hurdles, there are other microorganisms capable of surviving and spoiling beer. The quality of all food products including beer are not only affected by the integrity of the raw materials, and cleanliness of the equipment and packaging materials, but also by the purity of the environmental air surrounding the processing area. The purpose of this project was to examine the environmental microbial air quality within various areas of a craft brewery with special emphasis on potential beer spoiling bacteria. First, samples inside the brewery and samples outside the brewery were collected to establish a baseline of data, identify areas of concern, and to examine the effect of seasonality. Those areas of concern then were sampled more often and also were sampled based on the risk of product contamination. The canning line within the brewery was identified as a specific area of concern. Bottling and canning lines in breweries often are considered non-closed production equipment and have the ability to become contaminated from outside sources including the environment. The air was sampled 307 times over a period of 22 months using an automated impaction sieve sampler pulling 80 liters of air. Samples were plated both aerobically and anaerobically. The aerobic plates were used for a general cleanliness of the area while the anaerobic plates were included to examine for beer spoiling organisms. The standard (specification limit) used for the indication of a contaminated area was a plate with 40 colony forming units (CFU) or more per 80 liters of air sampled. The results of this study revealed that testing for airborne microorganisms is highly recommended in the craft brewing industry due to the potential for the impurity of the environmental air surrounding the processing area. Seasonality had an effect on total number of aerobic airborne microorganisms with the spring months being approximately five times higher than other months. The canning line in the brewery was found to be contaminated with beer spoiling bacteria on average 75% of the time. Critical areas in the brewery, such as the bottling and canning lines, should be routinely tested for airborne microorganisms as they could lead to final product contamination. Routine microbial environmental air testing is a good indicator of overall brewery cleanliness.Item Open Access Brewing analysis and quality control: development of an undergraduate course and integration within a university fermentation science and technology curriculum(Colorado State University. Libraries, 2020) Strevey, Daniel, author; Avens, John S., advisor; Bunning, Marisa, advisor; Laybourn, Paul, committee memberThe development of the course FTEC 422 Brewing Analysis and Quality Control (BAQC) originated at Colorado State University to alleviate the lack of university level course offerings available to future brewing professionals in the United States; particularly a course that addressed the analytical and quality control aspects of beer production. At the time of development only one course was available in Fermentation Science and Technology (FTEC) at CSU, with one more course in the planning stages. This indicated a lack of educational opportunities to support a quickly growing brewing industry of 2,051 breweries in the U.S. as of April 2012 (Gatza, 2012). In 2018 there where 7,346 breweries and more than 500,000 industry jobs (Watson, 2018). BAQC was designed to introduce and educate students in quality analysis and control topics related to the brewing industry through weekly lecture, lab exercises, and industry related field trips. BAQC was first offered as a pilot course in the spring semester of 2012 as FTEC 480 (Year 1 n=8), then again in spring 2014 as FTEC 422 (Year 2 n=12). During both years, the course had a set enrollment limit of 20 students. Instructor approval was required for student admittance in both years. As a result, registered students represented a diverse demographic of science disciplines. In Year 2, the course included students enrolled in the newly formed Fermentation Science and Technology Bachelors of Science degree. BAQC was designed to be a foundational course within the new major. Statistical analysis was used to determine if students' education backgrounds would have an impact on their success in the class. Results did not indicate that a student enrolled in a differing degree program would do better or worse in BAQC (P = 0.80). This result was somewhat expected since instructor approval was required for admittance. As a result some students were excluded from the course who may have performed poorly. An open admittance policy could have produced different results. Course evaluation forms were completed by students at the end of the semester with overwhelmingly positive feedback. BAQC offered a unique educational opportunity by providing students with applied technical experience preparing them for a fruitful career in the brewing industry. Brewing industry growth between 2012 and 2018 indicated strong growth will be sustained into the future, supplying an expanding number of industry jobs. BAQC also provides students a basic understanding of quality control and analysis objectives needed for other fermented food industry products such as cheese, yogurt, and baked foods. As the brewing industry had grown, a greater focus on quality products has emerged, according to relevant industry sources. Offerings within the Brewers Association, American Society of Brewing Chemists, Masters Brewers Association of the Americas, and Institute of Brewing and Distilling support beer quality control and analysis through publications and forums. These industry sources allow students who have completed BAQC to stay abreast of new developments in the knowledge and skills provided in this course. BAQC is now an integral part of the Fermentation Science and Technology degree offered at CSU. Through course evolution by new instructors BAQC will continue to stay relevant.Item Open Access Effects of cooking methods on antioxidant properties, quality attributes, and sensory characteristics of selected leafy greens(Colorado State University. Libraries, 2014) St. Onge, Matthew Mark, author; Bunning, Marisa, advisor; Stone, Martha, advisor; Newman, Steven, committee memberKale, arugula, spinach, and other types of leafy vegetables are rich sources of vitamins and minerals and well-suited to growing in Colorado's climate. This project focused on the chemical, quality, and sensory assessment of a selection of 6 specialty leafy greens (arugula, cherokee lettuce, mache, pac choi, red kale, and spinach). Total phenolic (TP) content, radical scavenging capacity (1,1-diphenyl-2-picrylhydrazyl, DPPH), color measurement, instrumental texture characteristics, and sensory attributes of raw versus cooked (boiled, microwaved, or steamed) samples of the select taxa were analyzed and evaluated. All 6 varieties were cultivated in a greenhouse using organic planting medium. Samples of the greens were freeze-dried as raw or post-heat treatment, then underwent an extraction procedure, and were analyzed for total phenolics compared to gallic acid standards and radical scavenging using DPPH compared to trolox standards. Fresh and cooked comparisons were tested for color differences using a HunterLab ColorFlex spectrophotometer and changes in texture utilizing a TA-XT2 texture analyzer. Consumer (n=50; n=51) sensory analysis was administered using a 9-point hedonic scale. Fresh, uncooked mache had higher (p<0.05) total phenolics and radical scavenging ability than all other cultivars. Cooked samples revealed that spinach values for each test did not differ (p>0.05) from fresh samples and microwaved and steamed samples of red kale contained more total phenolics than fresh, but less (p<0.05) was observed in boiled samples. Other significant findings included reductions (p<0.05) in lightness of cooked samples and cooked spinach samples after all heating treatments were observed to be more tender than fresh samples (p<0.05). This research helps to fill information gaps which exist in leafy greens research. Many studies focus on one cultivar using one testing method, but little research has been conducted on these types of greens using several analytical testing methods to obtain comparable data.Item Open Access Exploring personal, business, and community barriers and opportunities for food entrepreneurs(Colorado State University. Libraries, 2017) Colpaart, Ashley M., author; Bunning, Marisa, advisor; Thilmany McFadden, Dawn, advisor; Auld, Garry, committee member; Harmon, Alison, committee member; Miller, Jeffery, committee member; Carolan, Michael, committee memberSpecialty food businesses, characterized as local, craft or artisan, produce unique and highly differentiated food items often made in small quantities from high-quality ingredients. Nationally, the increasing market demand for specialty food is simultaneously spurring a growth in food entrepreneurship and food businesses that need access to licensed commercial space. Due to their unique values, a subset of food entrepreneurs may be considered 'social entrepreneurs' who use their business as a catalyst for social, cultural, or environmental change. This dissertation research model and hypotheses were developed as a triangulation of three innovative approaches to various fields of study influencing how the food sector is evolving to address emerging consumer and supply chain dynamics. These include; a) a new management behavioral concept, Perceived Business Effectiveness, b.) previous research on entrepreneur characteristics, and c.) potential experience and opportunities that may influence food entrepreneurs based on the Community Capital Framework. The primary objectives of this research are to determine the unique mission, values or community capital-based attributes of food entrepreneurs and to evaluate how this set of factors may affect a food entrepreneur's interest and key criteria when searching for commercial kitchen space. The primary methods included a national survey of food entrepreneurs (n=140) and a pilot program resulting in 4 case studies from Northern Colorado. Multidisciplinary empirical analysis was applied including gamma correlations to compare and contrast various factors and a 2-step probit regression analysis and the calculation of marginal effects from that model. Survey results found that food entrepreneurs in search of commercial kitchen space had dissatisfaction with finding appropriate space (p=0.04), availability of enough days/time to rent (p=0.00), location (p=0.07), availability of equipment (p=0.02), and parking (p=0.07). Results also found significant gamma correlations for questions related to food safety, social fairness, and resource mobilization indicators like sourcing locally and participating in the sharing economy. Further, respondents looking for commercial space were 9% more likely than those not looking for space to use a theoretical sharing economy technology to help them find and access commercial kitchen space. The three-month pilot program successfully placed four food entrepreneurs searching for production space in four different commercial kitchens in Northern Colorado. The kitchens included a school district, church, commissary kitchen, and functioning pizza parlor. A major contribution of this work is in the identification of key drivers for food entrepreneurs in the emerging access economy, suggesting that "access" to goods and services may becoming more desirable than "ownership" of them.Item Open Access Farm food safety plans: customizing educational materials for small-scale and campus-based farms(Colorado State University. Libraries, 2017) Morris, Rachael E., author; Bunning, Marisa, advisor; Uchanski, Mark, committee member; Stone, Martha, committee member; Sullins, Martha, committee memberA comprehensive farm food safety plan is an integral part of all growing operations, regardless of size, output, or production practice. As small-scale and campus-based, student-run farms grow in popularity, there is an increasing need to establish the concept of comprehensive produce safety practices as a vital part of the farming process. Moreover, the passing of the Food Safety Modernization Act (FSMA) in 2011 addresses mandated accountability for large-scale agriculture within the Produce Safety Rule, but leaves many small farms exempt from regulation, including the types of farms targeted within this study. A need for a curriculum addressing produce safety on campus farms and small-scale operations, and lack of literature surrounding food safety training curriculum development drove the research design for this project. The components of this study were three fold: 1) university and college campus farm managers (n=12) from 14 states were surveyed on current practices; 2) classroom and on-farm presentations were developed and delivered to Colorado State University horticulture students (n=54), and a pre-and post- questionnaire was delivered to assess learning objectives; and 3) three on-line modules and a 27-page farm plan template were developed for Colorado growers, detailed during a webinar for Colorado Fruit and Vegetable Growers Association (CFVGA) members (participants n=33). Results offered positive a relationship between resource delivery and food safety knowledge for both students and growers. The pre-and post-questionnaire responses showed change (p≤0.05), including Likert scale questions stating that farm workers (p=0.001) and volunteers (p<0.0005) should receive food safety training. Post-webinar polling questions revealed that 100% (n=21) of voluntary respondents learned something new during the webinar, which focused on utilization of resources to build and modify farm food safety plans for any growing operation. With the national movement toward a prevention-focused food safety strategy, the need for implementing better produce safety practices has been identified as a top priority. Review and feedback from this study will aid in the continued development of materials for both campus and small-scale growers to expand their food safety practices.Item Open Access Farm-to-table food safety for Colorado produce crops: a web-based approach for promoting good agricultural and handling practices(Colorado State University. Libraries, 2011) Wall, Gretchen Lynn, author; Bunning, Marisa, advisor; Kendall, Patricia, committee member; Lamm, Dennis, committee memberFresh fruits and vegetables have been increasingly associated with cases of foodborne illness (CDC, 2010; FDA, 1998). Direct links of these outbreaks to farms highlights the need to employ strategies to reduce pathogenic microbial contamination of fresh produce at this stage of the food system (Bihn & Gravani, 2006). One approach to fresh produce safety on the farm involves participation in voluntary audits based on Good Agricultural Practices (GAPs) and Good Handling Practices (GHPs). Currently, only a small percentage of Colorado producers employ this strategy. In response to new regulations and guidelines, changing food consumption patterns, and improved market access, Colorado producers need to have the flexibility to adopt programs that can help them meet the demands of providing a safer food supply (Rejesus, 2009). In response to these issues, Farm to Table Food Safety for Colorado Producers, a series of three web-based GAPs/GHPs trainings, was developed following the guidelines of FDA, USDA, and the National GAPs program. These trainings were implemented and evaluated targeting small farm producers of fresh fruits and vegetables based on an initial needs assessment to identify potential barriers and drivers for adopting these types of programs. In addition, a series of consumer-friendly, down-loadable fact sheets with nutrition, safe food handling, and recommended storage guidelines was developed to help promote ten selected Colorado specialty crops: apples, berries, broccoli, leafy greens, melons, peaches, peppers, potatoes, squash, and tomatoes. The approach of developing technology-based training modules and educational materials allowed for improved accessibility to growers and produce consumers across the state while accommodating their variable time schedules and need for convenient, reliable information. Pre and post questionnaires were used to measure self-assessed knowledge outcomes along with overall course evaluation items. As a result of participating in the webinar series, producers, buyers, and Extension professionals indicated they planned to utilize the information and resources. Specific topics related to GAPs such as irrigation water quality, management of manure and compost, and food safety legislation were of most interest to the participants and had the most significant increases (p<0.001) in self-reported knowledge, pre to post webinar. The course evaluation showed that produce buyers, Extension professionals, and other webinar attendees intend to utilize the material presented in the webinar series directly or indirectly in their professions, with average mean scores on a 5- point Likert scale (1= Very Unlikely and 5= Very Likely), of 4.2, 4.2, and 3.6, respectively. Overall, reviewers rated the produce fact sheets with a mean score of 4.34 for usefulness (1=Not Useful and 5= Extremely Useful). Feedback gained from the webinar and produce fact sheet evaluations will be a helpful tool in making improvements for future web-based on-farm food safety educational materials.Item Open Access High elevation food preparation: consumer assessment and toolkit development(Colorado State University. Libraries, 2021) Engelhardt, Heidi, author; Bunning, Marisa, advisor; Van Buiten, Charlene, committee member; Hyatt, Doreene, committee memberAt higher elevations, reduced air pressure and dry conditions impact food preparation in a multitude of ways. The boiling point of water decreases, the rate of evaporation is higher, and the functionality of leavening agents can be altered. Cooks, bakers, and food scientists alike face challenges in adjusting processing methods and ingredients to ensure desirable results of recipes at various elevations. Current information on food preparation at high elevations lacks consistency and accessibility and often requires using multiple sources that may not be reliable. This leaves the home cook vulnerable to failed recipes and in some cases, foodborne illness. An assessment of consumer cooking, baking, and food preserving practices was needed to identify and prioritize information that could contribute to successful and safe food preparation at higher elevations. To assess these needs, a survey was developed, conducted, and results were analyzed to guide resource development for a high elevation food preparation toolkit. The purpose of the project was to construct useful materials as part of a set of tools to empower home cooks to apply research-based knowledge in Colorado and other high elevation locations in the United States. Developed resources included eight ingredient information sheets, a troubleshooting guide with suggestions for nine food products or methods. A set of presentation slides and two activities with pre- and post-evaluations to measure behavior change are included for county extension agents to use while engaging with their communities. Expanding awareness related to the impacts that higher elevations have on food preparation connects home cooks with food science as well as food safety. In addition to nutritional needs, food related pastimes often serve a greater purpose providing comfort and a rewarding way to cope with stress, promoting general well-being. Success in a high elevation kitchen would include recipes that do not fail as often, have desired taste and texture, and appropriately address food safety. This toolkit can be utilized in many different ways with the goal of helping consumers become more knowledgeable and successful when safely preparing foods at high elevation. We expect these materials to have national usefulness and aid in the development of skills that can be routinely incorporated in food preparation at higher elevations.Item Open Access Influence of barley genetics on beer chemistry, flavor, and flavor stability(Colorado State University. Libraries, 2017) Bettenhausen, Harmonie M., author; Bunning, Marisa, advisor; Heuberger, Adam, advisor; Avens, Jack, committee member; Broeckling, Corey, committee member; Stone, Martha, committee memberIn the brewing industry, identifying superior ingredients that provide distinct flavors is an important area of research. While the contribution of raw ingredients such as yeast and hops to flavor is well understood, it is currently unclear if different genotypes of barley provide unique flavor to beer. In brewing, barley is malted to provide saccharides and enzymes for fermentation, however the malt also contains thousands of metabolites that may influence flavor. The goals of this study were to determine (i) if there would be metabolite differences among six commercial barley genotypes, (ii) if differences in barley chemistry are reflected in the chemistry of the beer, (iii) if the differences in the beer chemistry impact sensory attributes of beer, through flavor and flavor stability, and (iv) if there are barley and/or malt metabolites that can be markers for beer flavor and/or flavor stability. Six distinct malts were brewed into six beers using a recipe designed to evaluate differences in flavor. The malts were derived from the barley genotypes: Copeland, Expedition, Full Pint, Meredith, Metcalfe and PolarStar were grown and malted in either Canada or the U.S. Metabolomics was used to characterize chemical variation among the six malts and beers using RP-UHPLC-MS, HILIC-MS (non-volatile metabolites), HS/SPME-GC-MS (volatiles), and ICP-MS (metals). The metabolomics analysis detected 5,042 compounds in malt, and 217 were annotated as known metabolites and included amines (20 metabolites), amino acids (36), fatty acids/lipids (40), sugars (11), phenols (30), and others (80). A total of 4,568 compounds were detected in beer and included 246 annotated metabolites and included amines (9), amino acids (37), fatty acids/lipids/fatty acyls (28), sugars (10), phenols (20), esters (89), aldehydes (21), others (31). The chemical profiles of the six malts and beers were evaluated for metabolite variation using principal component analysis (PCA) and analysis of variance (ANOVA). Principal component analysis was conducted on the annotated metabolites and demonstrated that each of the six malts and beers contained unique chemical profiles. ANOVA characterized 150/217 malt metabolites (69.1%) and 150/246 beer metabolites (60.9%) varied among genotype (ANOVA, FDR adjusted p < 0.05). The six beers were evaluated for flavor using a modified Quantitative Descriptive Analysis® (QDA) for 45 sensory traits at 0, 4, and 8 weeks of storage at 13 °C. PCA characterized flavor differences among the six beers at 8 weeks and Full Pint was described as fruity and Meredith as corn chip. The metabolite and sensory data were integrated using two approaches: Spearman's correlation and two-way orthogonal projection to latent structures (O2PLS). The analyses revealed associations between fruity or corn chip flavor in beer with beer purines/pyrimidines, volatile ketones, amines, and phenolics; and malt lipids, saccharides, phenols, amines, and alkaloids. Taken together, these data support a role of barley metabolites in beer flavor and flavor stability. As a raw ingredient, malted barley genotypes should be evaluated for a contribution to flavor, and this may be a future target for plant breeding efforts to selectively improve flavor and flavor stability quality in beer.Item Open Access Longitudinal study of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes in a small and very small fresh meat processing plant environment(Colorado State University. Libraries, 2012) Orellana, Eva Danira Borjas, author; Bunning, Marisa, advisor; Nightingale, Kendra, committee member; Kendall, Patricia, committee memberSmall and very small fresh meat processing facilities have scarce resources to monitor foodborne pathogen contamination patterns and transmission dynamics in their premises. Environmental control of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes, is important to prevent cross-contamination of meat products by pathogens that may persist in a facility. Although Listeria spp. are non-pathogenic, a high prevalence in a meat processing environment indicates failures in the cleaning and sanitation procedures. The purpose of this study was to conduct a six month longitudinal study to monitor Escherichia coli O157:H7, S. enterica, L. monocytogenes and other Listeria spp. contamination patterns, and to identify potential harborage sites in a small and very small fresh meat plant. Additionally, in order to gain insights about the facilities, manufacturing practices, and other relevant practices, managers from the two participating plants were asked to complete a questionnaire. Feedback of the study results was given to plant staff in a bilingual session, along with a basic training in food safety topics. Both plants were sampled during mid-shift operation on a monthly basis. Environmental site (n ≤ 54) and beef carcass composite samples were collected mid-shift. Samples collected included food contact surfaces (e.g., tables, scales, bins), and non-food contact surfaces (e.g., walls, drains, sinks). Overall, 1,979 environmental sponge samples were collected and microbiologically analyzed to detect and isolate S. enterica, E. coli O157:H7 and L. monocytogenes. Further characterization of the recovered pathogen isolates by molecular subtyping (e.g. PFGE, ribotyping) was performed to gain insight in contamination transmission within the facilities. S. enterica was isolated from 15 (4.5%) and 8 (2.4%) samples from Plant 1 and Plant 2, respectively. Characterization by PFGE using XbaI generated 6 different patterns in Plant 1, whereas all isolates from Plant 2 had the same pattern. S. enterica was recovered more than once from two sites in Plant 1, but only 2 isolates recovered from a drain in the slaughter area yielded the same PFGE pattern. E. coli O157:H7 was detected in 1.2% of samples in Plant 1; PFGE using XbaI generated 2 different patterns, and none was recurrently isolated from a single site. E. coli O157:H7 was not isolated from Plant 2. Over the course of the study, roughly 28% and 6% of the samples tested positive for Listeria spp. other than L. monocytogenes, in Plant 1 and 2, respectively. Listeria innocua was the predominant Listeria spp. in both plants. L. innocua allelic type AT-1 was recovered from 15% of samples collected across Plant 1, whereas in Plant 2, type AT-6 was found mostly contained in the slaughter area. L. monocytogenes was isolated from 17% of the samples from Plant 1 and 1.2% of samples from Plant 2. Roughly 97% (54/56) of L. monocytogenes isolates recovered from Plant 1 belonged to ribotype DUP-1042B, which was recovered up to five times from 15 different sampling sites across the facility; the remaining two isolates belonged to ribotype DUP-1057B. Noteworthy, ribotype DUP-1042B belongs to a major human outbreak-associated clonal group known as Epidemic Clone I, posing a high risk for meat product contamination in this facility. Conversely, in Plant 2, L. monocytogenes DUP-1030B was recovered from three samples and only one isolate belonged to DUP-1030A; no single site in Plant 2 repeatedly tested positive for L. monocytogenes. In light of the bacteriological results from this study, good manufacturing practices for the control of environmental contamination practiced by Plant 2 seem to be effective in the prevention of contamination spread and pathogen persistence; e.g. sanitizer dip stations at entry points, use of foam to clean equipment, use of quaternary ammonium compounds (QAC) or chlorine for sanitation of floors and other food contact surfaces, and QAC for sanitation of drains. Some high risk procedures practiced by the cleaning crew in Plant 1 may contribute to contamination spread; e.g., use of high pressure water for daily cleaning of drains, no sanitation step after cleaning drains, and lack of designated cleaning tools for drains. Although economic resources may be limited, microbiological monitoring of the plant environment is useful from a risk assessment standpoint. In this study, the prevalence of L. monocytogenes was high and widespread in Plant 1, and a predominant strain belonging to an Epidemic Clone group was elucidated. This information increases awareness and provides the plant management with valuable information for decision making, and motivates the implementation of new policies and targeted interventions in problematic areas. Our findings suggest that L. monocytogenes and L. innocua have higher prevalence than S. enterica and E. coli O157:H7 in the fresh meat processing plant environments. While L. monocytogenes may persistently contaminate the environment of fresh meat processing plants, E. coli O157:H7 and S. enterica contamination appears to be mostly sporadic.Item Open Access Microbial quality of mixed salad greens and selected fresh and dried herbs(Colorado State University. Libraries, 2010) Woo, Daniel Lee, author; Bunning, Marisa, advisor; Goodridge, Larry, committee member; Stone, Martha, committee memberDirect marketing has been growing in the Western U.S., with 2007 sales of direct-marketed agricultural products totaling nearly $142.6 million in Colorado, New Mexico, Oregon, Idaho, Nevada and Washington-more than twice the 1997 sales level for the region. In addition, the number of farms engaging in direct sales to consumers grew by more than 20% to 18,274 in 2007. With increasing foodborne outbreaks linked to produce consumption, more research is needed to fill in the gaps of knowledge on the microbiological quality of leafy salad greens. Limited research has been conducted on the microbial safety and quality of farmers' market leafy salad greens. This study surveyed the microbial quality of salad greens from Larimer County farmers' markets in conjunction with salad mixes from a local supermarket. Leafy salad greens were obtained weekly for a period of four weeks from September to October, 2009, from both farmers' markets and supermarkets. Total aerobic and coliform counts were assessed within 48 hours of obtaining the samples via plating onto Petrifilm plates, then following one week of storage at refrigeration temperatures to observe changes in the microbial load. In addition, handling methods and temperatures were also recorded. Gloves and tongs were not used by vendors when handling salad greens. The vendors surveyed also lacked adequate refrigeration or ice for holding salad greens. Farmers' market salad greens were lower than supermarket salad greens in terms of initial aerobic plate counts. Both farmers' market and supermarket salad greens had no detectable levels of Escherichia coli and low levels of coliforms. After 1 week of storage, aerobic counts were higher in all samples (P<0.05). Coliform counts tended to decrease in all samples but no significant differences were observed (P>0.05). The results reiterate the need for consumers to thoroughly wash their salad greens. Further research should be conducted to assess the microbial quality o~ other produce at local farmers' markets. Herbs are often used in cooking to add aroma and flavor to foods. Consumers may choose to dry herbs from their garden or purchased from the market. Herbs, like other agricultural produce, may be exposed to a wide range of potential microbial contamination. There is currently little research on safe drying practices of herbs at home. Microwaving herbs is a potentially popular and time-saving approach for drying herbs at home. This research project investigated the impact of three drying methods (microwave, dehydrator, and conventional air drying) for improving the microbial quality of dried parsley and cilantro. Herb samples were obtained weekly from a local supermarket for a 2 month period from January to March, 2010. Standard guidelines from Oregon State University Extension service were followed for microwave drying of herbs. The manufacturer's drying temperatures/times were used for drying herbs in a dehydrator. Herbs were also air-dried for 1 week. Microbial testing was performed using plating onto 3M Petrifilm and when counts were below the detection limit, via the Most Probable Number (MPN) method. Microwave drying provided the greatest reduction in aerobic counts of bacteria in herbs. All drying methods reduced coliform counts to undetectable amounts; however, the determination of the effectiveness of each drying method in comparison with others for reducing the coliform count was complicated by the low initial load of coliforms in herbs used in the study.Item Open Access Promoting the consumption of beans and other pulses for public health: a translational approach to address consumer barriers to intake(Colorado State University. Libraries, 2023) Didinger, Chelsea, author; Foster, Michelle, advisor; Bunning, Marisa, advisor; Thompson, Henry, committee member; Jablonski, Becca, committee memberBeans and other pulses (i.e., the dry, edible seeds of non-oilseed legumes like chickpeas, cowpeas, dry beans, dry peas, and lentils) are linked to a myriad of positive impacts on human and environmental health, including promotion of gut health and healthy weight management, reduction of chronic disease risk, mitigation of greenhouse gas emissions, improvements in soil health, conservation of water resources, and more. Moreover, pulses are highly nutrient-dense, have a long shelf-life, demonstrate wide culinary versatility, and are relatively affordable compared to other foods. Accordingly, pulses offer the chance to consume a healthful diet based on sustainable food choices, all at an economical price point. Although this combination of positive assets may make pulses seem like a natural choice for consumers to include as a dietary staple, global consumption of pulses has stagnated at around 21 g/day/capita. This research focuses on United States consumers, who eat less than 1 cup of cooked pulses per week, which is below the recommendation. According to the Dietary Guidelines for Americans (DGA), there are only four dietary components of public health concern (i.e., nutrients or other dietary components – like fiber – for which low intakes are associated with health concerns) in the United States. Among these four dietary components of public health concern are dietary fiber and potassium. Pulses are rich in both, thus increasing intake across the population could contribute to raising levels of these critical dietary components. Although it depends on age group and gender, the levels of pulse intake recommended by the DGA are around 1 – 3 cups per week. Increasing intake to meet this recommendation would help provide adequate levels of dietary components of public health concern. Raising it even further, to around 1 cup per day, has been associated with numerous human health benefits, as discussed in the Introduction. Reversing the current trend of dramatically low consumption would allow the public to better capitalize on all the benefits that pulses have to offer. To achieve this, it is vital to address the potential barriers to pulse intake that consumers face, paramount among them being unfamiliarity with how to cook and prepare pulses, long cooking times, and concerns over flatulence. Simultaneously, it is key to highlight the many motivating factors to eat pulses, including taste and culinary versatility, nutrition and health, and environmental benefits. Through engaging in a translational approach that addresses barriers and emphasizes motivators, not only is knowledge disseminated, but consumers can be motivated to engage in behavior change and increase their pulse consumption. This research is comprised of three primary efforts designed to directly target motivators and mitigate barriers to pulse consumption in the United States: 1.) creation of an Extension Bean Toolkit, which includes various consumer resources and an online 1-hour class; 2.) development of the Bean Cuisine and engagement with citizen scientists to improve the cuisine and monitor impacts of participation; and 3.) testing of the effects of elevation and soaking conditions on bean cooking time to address the concern over long cooking times and provide consumers with accessible cooking tips. Before designing the Extension Bean Toolkit, a Food Habits Survey was conducted to better understand preparation and consumption habits, barriers, motivators, and potential points of consumer interest related to pulses. Based on results and in conjunction with review of the literature, resources were developed for the toolkit, as well as the 1-hour class, titled Beans: Good for You, Good for the Planet. Participation in the class resulted in significant gains in knowledge about pulses, an increase in the importance of motivators, a decrease in how much barriers discourage consumers, and consumption frequency also appeared to increase. See Chapter 2 for details. The Bean Cuisine is a 2-week cuisine (i.e., meal plan) with 56 pulse-centric recipes that correspond to 14 unique breakfast, lunch, snack, and dinner ideas. The Bean Cuisine was designed to have 35% of dietary protein from pulses, due to recent preclinical findings that suggest this level of consumption is when benefits for gut health and weight maintenance are attained. The main barrier addressed was lack of awareness of how to cook and prepare pulses, and culinary versatility was a main motivator. Fifty-six citizen scientists were recruited and provided feedback on one day (i.e., four recipes) of the Bean Cuisine, and impacts of participation were monitored. The Bean Cuisine was modified based on their feedback to improve the recipes to ensure they were clearly written and the taste would appeal to a wider audience. Participation in the project resulted in significant gains in knowledge about pulse health benefits, versatility, and how to cook dry pulses. Moreover, common themes in free response data demonstrated that citizen scientists had increased awareness of pulse variety and versatility, they changed the frequency of and ways in which they ate pulses, they had a positive experience and thus a good perception of citizen science, and that some became pulse advocates, sharing the benefits of pulses with their communities. More details can be found in Chapter 3. Not knowing how to cook dry pulses and the long cooking times pose barriers to regular pulse intake. Therefore, to be able to provide consumers with better information regarding the effects of cooking conditions and elevation on cooking time, a Mattson cooker was used to assess the cooking time of pinto beans at four locations, ranging from around sea level to over 3,000 meters. Seven different cooking conditions were evaluated in replicate at each location: an overnight soak or a quick soak in only water or in a 1% solution of sodium chloride or sodium bicarbonate, with a no soak and no salt added comparison. Cooking time increased with elevation, and both the soaking of beans and the addition of salt shortened cooking time. A handout was created to equip consumers with information and practical, accessible tips to facilitate faster, better experiences when cooking dry pulses. Refer to Chapter 4 for details. Through the Extension Bean Toolkit and Bean Cuisine citizen science work, a translational approach was adopted to reach the public with current research findings that aligned with areas in which they expressed interest. The mitigation of potential barriers to pulse intake and the highlighting of the numerous benefits of pulses was a primary focus in this work. After participating in the Extension class or citizen science project, participants demonstrated greater knowledge about pulses. More importantly, they expressed a greater intention to eat pulses, indicating that participation in these translational projects helped motivate them to change their behavior and regularly integrate more pulses into their diets.Item Open Access Rapid detection of viable escherichia coli O157:H7 by antimicrobial incorporated multi-angle light scattering spectroscopy(Colorado State University. Libraries, 2010) León, Juan Carlos, author; Goodridge, Lawrence D., advisor; Bunning, Marisa, advisor; Nightingale, Kendra K., committee memberEscherichia coli 0157:H7 continues to cause outbreaks of produce-associated foodborne illness. Contaminated water has been shown to be a vehicle for transfer of E. coli 0157:H7 to leafy greens during irrigation. Timely detection of this pathogen in irrigation water can prevent contamination of the final product. Current detection methods which are culture-based can be labor intensive and require several days to produce results. There is a need for development of rapid detection methods, which can detect E. coli 0157:H7 in irrigation water. These methods have to be sensitive, robust and ideally should be able to differentiate between viable and non-viable microorganisms. Multi-angle light scattering spectroscopy (MAES) is a powerful technique that has been applied to qualitatively and quantitatively distinguish internal structural changes in cells upon perturbation by chemical/biological agents. We hypothesized that combining bacteriophage (phage) infection, which occurs only in viable bacterial cells, with MAES would allow for detection of the target bacteria and distinguish between viable and nonviable bacterial cells. The objective o f this study was to use E. co//-specific bacteriophages in conjunction with immunomagnetic separation and MAES to develop an assay for rapid detection of viable E. coli 0157:H7 in irrigation water. We have termed this new method Antimicrobial Incorporated Multi-angle Light Scattering (ANIMALS) to highlight the combination of a specific antimicrobial agent (phages) with the multi-angle light scattering technique to form a sensitive, rapid and specific assay for detection of E. coli 0157;H7. E. coli 0157:H7 and Salmonella Typhimurium strains were diluted in lambda buffer to form concentrations of 10*^ to lO"^ CFU/ml and 10*, respectively. The samples were subjected to immunomagnetic separation (IMS) using E. coli 0157-specific IMS beads. Following IMS, the beads (and attached bacteria) were resuspended in 1 ml of tryptic soy broth (TSB) and one half (500 pi) of each sample was added to 10 mis of TSB that contained 1 ml of phage ARl (lO'** PFU/ml). The other half of the samples were added to TSB that did not contain phage AR l, but contained 1 ml of lambda buffer (to maintain constant volume), and these samples served as a reference. The samples were incubated at 37° C, with shaking, for 15 hours. Following incubation, 100 pi aliquots were removed from each sample, and separately assayed using a SpectraPoint light scattering spectrometer (Spectra Digital Corp., Toronto, Ontario, Canada). E. coli 0157:H7 was detected in 100% of pure culture samples in TSB containing various concentration (10*^ to 10^) of the bacterium within 15 hours. An algorithm was developed to evaluate the area under the curve of each spectra. When compared to the light scattering spectra of the non-phage treated reference, the spectra o f phage infected E. coli 0157:H7 cells differed markedly. In contrast, the spectra of samples that contained Salmonella Typhimurium and the negative controls (containing no cells) were almost identical, because phage ARl does not infect Salmonella Typhimurim and there was no growth in the negative control samples, respectively. Using this method, E. coli 0157:H7 could be detected following 15 hours of incubation in samples spiked at an initial concentration of 10** CFU/ml. The sensitivity and specificity of the ANIMALS assay was determined using environmental water samples. River water samples were artificially contaminated with a 3-strain cocktail of E. coli 0157:H7, followed by IMS and incubation in TSB supplemented with 20 mg/ml novobiocin at 42° C. MALS measurements were taken at 4, 6, 8 and 10 hours. The means and standard deviations of the ratios o f the differences between the test and reference spectra for the four concentrations (10‘^ to 10^ CFU/ml) o f E. coli 0157;H7 were analyzed as well as for negative controls (samples o f river water that were not inoculated with E. coli 0157:H7). One concentration (10^ CFU/ml) of E. coli 0157:H7 was detected consistently after only 6 hours of enrichment. E. coli 0157:1-17 was consistently detected at concentrations from 10‘ to 10^ CFU/ml in spiked river water within 8 hours. All concentrations (10° to 10^ CFU/ml) were detected after 10 hours of enrichment. The presence of the pathogen in positive river water samples was confirmed by using lateral flow devices specific for E. coli 0157:H7. The negative controls resulted in negative readings. Additionally, reservoir water samples were prepared similarly to river water samples and MALS measurements were taken at 4, 6, 8 and 10 hours. The test and reference spectra for the four concentrations (10° to 10^ CFU/ml) of E. coli 0157:H7 were analyzed as well as for negative controls (samples o f reservoir water that were not inoculated with E. coli 0157:H7). Similar results to experiments on river water were obtained with 10^ concentrations of E. coli 0157:H7 detected consistently at 6 hours. This demonstrated that the ANIMALS assay worked consistently across water samples with different physical and chemical characteristics. Regardless, E. coli 0157:H7 was consistently detected in 100% of concentrations (10 to 10 CFU/ml) within 8 hours even against background levels of bacteria as high as 10^ CFU/ml. Lateral flow devices specific for E. coli 0157:H7 confirmed the presence of the pathogen in positive reservoir water samples. The negative controls resulted in negative readings. These results demonstrate the ability of ANIMALS to rapidly and sensitively detect the presence and viability of E. coli 0157:H7 following phage infection. This method has the potential to allow for rapid detection o f viable E. coli 0157:H7 in irrigation water against a high background o f non-target microorganisms if coupled with selective enrichment.Item Open Access Science of food fermentation: development of a university curriculum and outreach educational materials(Colorado State University. Libraries, 2015) Bauer, Laura M., author; Bunning, Marisa, advisor; Miller, Jeff, advisor; Cunningham-Sabo, Leslie, committee member; Balgopal, Meena, committee memberWhile food and beverage fermentation is rooted in thousands of years of global traditions, today it is experiencing a revitalization by consumers interested in the health benefits and organoleptic qualities. A research-based, learner-centered, introductory-level, undergraduate academic curriculum was developed to address the need for understanding biochemical processes related to fermented food and developing critical thinking skills. Course findings, supporting research, and demand from consumers and outreach educators, guided complementary outreach materials development on related fermentation topics. The curriculum for FTEC 210, Science of Food Fermentation, was designed, implemented, and evaluated for outcomes in Year 1 (n=15) and Year 2 (n=22). The course focused on science, history, culture, gastronomy, safety, health, and nutrition aspects of fermented foods and beverages, while addressing core food science competencies in food chemistry, microbiology, food processing, and applied food science. Curriculum development was designed around students’ initial knowledge level, and then gaps were addressed for deeper understanding. Learning events alternated between direct instruction and experiential learning to engage diverse undergraduate learners in problem solving and application and inspire cognitive growth through evaluation and creation. Student change in knowledge, attitudes, and behaviors was assessed via pre-course and post-course questionnaires compared to a control group. Qualitative interviews, conducted mid-semester in Year 1, and course survey comments from Years 1 and 2, clarified quantitative data and provided feedback on curriculum usefulness and course satisfaction. Areas of analysis included demographics and grades, for reflection on student comprehension, content delivery, and assessments. Complementary teaching materials requested by and created for Extension educators and consumer use included an online training, a hands-on workshop, and related outreach publications. Students enrolled in FTEC 210 significantly increased knowledge from pre to post course (P<.0001), and their mean increase in knowledge was significant in comparison to the control group (P<.005). All students reported increased consumption of fermented food and drinks, and indicated active learning, in laboratory exercises, furthered opportunities for positive impacts on their education including self-studies and home practices. Curriculum content and informal hands-on student yogurt variation trials led to outreach publication development. Outreach trainings integrated experiential learning with research-based information to nurture retention and confidence. Requests from the public health community for the dissemination of credible fermentation information continues to guide fermentation outreach material development beyond the scope of this project. In this mixed methods study, results showed the curriculum had impact and relevance for cognitive development of university students and food knowledge application across disciplines, including integration of cultural diversity education and incorporation of biochemical processes. The use of experiential learning in course design enabled students to construct new knowledge through integrating base knowledge from lecture, collective interaction with lab partners, and hands-on learning practices in this course. This food fermentation curriculum met academic demands, increased student understanding and confidence, and supported material development for outreach dissemination.Item Open Access Species specific detection of adulteration of water buffalo milk with cow milk using high resolution melting analysis(Colorado State University. Libraries, 2015) Musetti, James, author; Stone, Martha, advisor; Bunning, Marisa, advisor; Hyatt, Doreene, committee memberEconomically motivated adulteration, a subset category of food fraud, was defined in 2009 by the Food & Drug Administration (FDA) as "the fraudulent, intentional substitution or addition of a substance in a product for the purpose of increasing the apparent value of the product or reducing the cost of its production," and can often encompass effects of public safety through the unknown addition of allergens, toxins, and hygienic risks (Wheatley & Spink, 2013). According to the FDA database which tracks scholarly records of adulterated foods, dairy products are the second most reported adulterated food products (Moore, Spink, & Lipp, 2012). Water buffalo milk represents the second most produced milk worldwide and higher value dairy raw products make it subject to economically motivated adulteration by means of dilution with cow milk (Bonfatti, Giantin, Rostellato, Dacasto, & Carnier, 2013). It is of interest to consumers, manufacturers and governing bodies to have a simple, fast, accurate, and sensitive method to detect adulteration in water buffalo milk. Several methods have been developed to detect species adulteration in dairy products including immunological, chromatography, and molecular methods (Mayer, 2005). In this investigation a real-time PCR assay was developed to specifically detect and relatively quantify cow milk adulteration in water buffalo milk utilizing high resolution melting analysis. A duplex real-time PCR reaction was performed targeting mitochondrial DNA in both species of interest and the resulting amplified products were analyzed after a melt cycle was performed. Results indicated that the assay was specific, efficient, and validated the use of real-time PCR for analysis. The use of high resolution melting analysis allowed for the discrimination of dilution of water buffalo milk with cow milk down to 0.1%. This study indicates the feasibility of real-time PCR and high resolution melting analysis in the detection of adulteration in dairy products.