Thesis (M.S.)--Oregon State University, 2007. / Printout. Includes bibliographical references (leaves 59-63). Also available on the World Wide Web.

Thesis (Ph. D.)--University of Oregon, 2006. / Typescript. Includes vita and abstract. Includes bibliographical references (leaves 80-91). Also available for download via the World Wide Web; free to University of Oregon users.

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There are concerns about using synthetic phenolic antioxidants such as butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA) as food additives because negative effects to human health have been reported. thus, a replacement of these synthetics by antioxidant extractions from various foods has been proposed. By the generation of more than eight thousand different phenolics, fruits and vegetables are the prime sources of natural antioxidants. The consumption of fruits and vegetables has been strongly linked with several health benefits, a result of their medicinal properties and high nutritional value. Antioxidants control and reduce the oxidative damage in foods by delaying or inhibiting oxidation caused by reactive oxygen species (ROHs); ultimately increasing the shelf-life and quality of these foods.Beta-carotene, ascorbic acid, and many phenolics play dynamic roles in delaying aging, reducing inflammations, and preventing certain cancers. Increasing the consumption of fruits and vegetables has been recommended by many agencies and health care systems. In order to extract, measure, and identify bioactive compounds from a wide variety of fruits and vegetables, researchers use many different techniques and methods. Ultrasonic assisted technique (UAE) is known to be one of the easiest extraction techniques because it uses common laboratory equipment such as ultrasonic baths and probes. In this technique, a smashed sample is mixed with the suitable solvent and placed into the ultrasonic bath while temperature and extraction time are controlled. Five experiments were conducted to study the effects of different ultrasonic parameters on the polyphenol and antioxidant content and antimicrobial activity of extracts from plants grown in southern Illinois. In the first experiment, ultrasonic treatments at different frequencies,temperatures, power levels, and exposure times were compared for their effects on the yield of total phenolics, total flavonoids and antioxidant activities of spinach extracts. The best conditions for extraction judged by yields were an ultrasonic frequency of 37 kHz, extraction time of 30 min, reaction temperature of 400C, and ultrasonic power of 50%. The mean yield (mg/100g), total phenolics (mg gallic acid equivalence/ g DW), flavonoids (mg/g DW), % DPPH free radical scavenging activity, and % ferric reducing antioxidant power were all high (64.88±21.84, 33.96±11.30, 27.37±11.85 , 64.18±16.69 and 70.25 ±9.68). In the second experiment, the study was designed to optimize ultrasonic-assisted extraction of lutein and β-carotene from spinach. Further, the identities and purity of the natural products in the TLC spots were assayed using matrix-assisted laser desorption/ionization time-off-light mass spectrometry (MALDI-TOF MS) analysis. The optimal UAE settings for output of lutein and β-carotene simultaneously from spinach extracts were an extraction temperature of 40 °C, extraction power of 40% (28 W/cm3) and extraction time of 16 min. UAE assisted extraction of carotenes from spinach can provide a source of lutein and β-carotene for the dietary supplement industry. In the third study, investigations of the effects of extraction parameters for UAE on antioxidant activity of peach and pumpkin extracts; and studying the chemical structures of samples before and after processing by Fourier transformed infrared spectroscopy were made. The optimal conditions for peach extracts were an extraction temperature of 41.53 ̊C, power of 43.99 % and time of 27.86 min for total phenolics. The optimal conditions for extractions of total phenolics from pumpkins were inferred to be a temperature of 41.45 ̊C, a power of 44.60 % and a time of 25.67 min. In the fourth study, TLC followed by Quantity-OneTM (Biorad) image analysis was used as a simple and rapid method for identification and quantification of compounds in complex mixtures. The results were consistent under optimal conditions among the experimental values and their predicted values. The MALDI-TOF MS technique was also applied for result confirmation. In the last experiments, UAE and Box-Wilson design were applied to optimize the extraction of antimicrobial activities from spinach extracts and to test their antimicrobial activities on both Gram-negative and Gram-positive bacteria. The spinach extracts exhibited antimicrobial activities against both bacterial pathogens with MICs in the 60-100 mg.ml-1 range. SEM showed that cells of the treated bacterial strains were damaged and showed structural defects compared to the controls. RAPD analysis of genomic DNA showed that the number and sizes of amplicons were decreased for the treated pathogens compared with non-treated strains.

Made available in DSpace on 2014-06-11T19:27:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-07-03Bitstream added on 2014-06-13T18:31:13Z : No. of bitstreams: 1 nespolo_nm_me_jabo.pdf: 317079 bytes, checksum: b5a59555adeaded005ddc00b0895ba68 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Tem sido evidente o aumento no consumo de pescado, especialmente do salmão (Salmo salar) sob a forma “in natura”, em pratos da cozinha oriental. Como conseqüência, tem havido maior preocupação quanto às suas características higiênico-sanitárias, tendo em vista a facilidade que microrganismos encontram para se desenvolverem em sua carne, o que pode expor os consumidores a agentes que causam desde uma simples gastrenterite até o óbito. Diante desta preocupação, desenvolveu-se este estudo com objetivos de avaliar características microbiológicas do salmão por meio da quantificação de microrganismos heterotróficos mesófilos, coliformes totais e termotolerantes, o perigo de veiculação de Vibrio parahaemolyticus, Staphylococcus aureus, Salmonella sp., Escherichia coli e Aeromonas sp. através da carne e contribuir com subsídios técnicos para criar uma legislação brasileira com padrões microbiológicos específicos para o pescado consumido cru. Foram colhidas 31 amostras de salmão, 16 refrigeradas e 15 congeladas, no comércio varejista de cidades da região nordeste do estado de São Paulo. Os resultados obtidos mostram populações de microrganismos heterotróficos mesófilos variando entre 1,0 x 10 e 3,9 x 106 UFC/g, coliformes totais e termotolerantes em, respectivamente, 32,24% e 19,33% das amostras e Aeromonas sp. em 35,48% das amostras com variação populacional de 2,0 x 102 a 8,0 x 103 UFC/g. Ainda houve a presença de Staphylococcus aureus em uma amostra e ausência de Vibrio parahaemolyticus, Salmonella sp. e Escherichia coli. Os resultados obtidos podem servir de parâmetro para a criação de um padrão microbiológico específico para o pescado consumido cru e servem também de alerta para os consumidores do produto tendo em vista a veiculação de microrganismos potencialmente patogênicos. / The increasing of seafood consumption has become evident especially in the use of salmon (Salmo salar) consumed raw in oriental dishes. Consequently, it has risen up the concern related to their hygienic-sanitary characteristics due to the facility that microorganisms multiply in the meat which can expose consumers to the causative agents of a mild gastroenteritis until the death. Regarding such informations, this study was aimed to evaluate microbiological characteristics of salmon by quantifying microrganisms heterotrophic mesophiles, total coliforms and thermotolerant. It was also evaluated the danger of transmission of Vibrio parahaemolyticus, Staphylococcus aureus, Salmonella sp., Escherichia coli and Aeromonas sp. on the fish muscle and contributed to technical informations to create a Brazilian regulations about specific microbiological standards for consumption of raw seafood. Thirty-one samples of salmon were collected, 16 chilled and 15 frozen, from the retail market in cities of the northeast region of São Paulo State. The results show populations of mesophilic heterotrophic microorganisms ranging from 1.0 x 10 and 3.9 x 106 CFU/g, in total and fecal coliforms, respectively, 32.24% and 19.33% of samples and Aeromonas sp. in 35.48% of samples ranging population of 2.0 x 102 to 8.0 x 103 CFU/g. Staphylococcus aureus was present in one sample and were not found Vibrio parahaemolyticus, Salmonella sp. and Escherichia coli. The results may serve as a parameter for the establishment of a microbiological standard for the consumption of raw seafood and also as a warning to consumers of the product for the presence of potentially pathogenic microorganisms.

5 p. / Growers of organic produce in the Southwestern United States face many challenges, including variation in water and temperature, and exposure to insects and disease. As a result, smallholder organic farmers are increasingly relying on soil additives such as compost tea that improve product quality, use less water, deter pests, and reduce reliance on chemical additives (Diver, 2002). But what exactly is compost tea? Do the benefits of using compost tea outweigh any concerns? For example, can it contain pathogens, and if so, do applicators have to worry about coming into contact with pathogens? This publication provides facts about making compost tea, and reviews both the benefits and potential disadvantages to help smallholder farmers to make educated decisions regarding the use of compost tea.

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

The bacterial flagellum is a rotary motor that propels motile bacteria through their surroundings via swimming motility, or on surfaces via swarming motility. The flagellum is a key virulence factor for motile pathogenic bacteria. Viruses that infect bacteria via this appendage are known as flagellotropic or flagellum-dependent bacteriophages. Much like other phages, flagellotropic phages are of interest for clinical applications as antibacterial agents, particularly against multidrug resistant (MDR) bacteria. Bacteriophage χ is a flagellotropic phage that infects multiple species of motile pathogens. In the projects described below, we characterized several aspects of the complex interactions between χ and two of its hosts: Salmonella enterica and Serratia marcescens. In Chapter I, we describe in detail the existing knowledge on flagellum-dependent bacteriophages, pathogenic bacteria, and the flagellar motility system. We also expand significantly on flagellotropic phage χ. In Chapter II, we describe our discovery of S. enterica cellular components other than motility that are crucial for bacteriophage χ infection, making the key discovery that the AcrABZ-TolC multi-drug efflux system is required for infection to proceed. We additionally found that the host molecular chaperone trigger factor is important for the χ phage lifecycle. In Chapter III, we outline our characterization of the initial binding interaction between χ and the flagellum, determining that of flagellin's seven domains, C-terminal domain D2 is the most important for χ adsorption. In Chapter IV, we expand on this by discussing our work that determined that the χ tail fiber protein is encoded by the gene CHI_31, purification of this recombinantly-expressed protein, and demonstration of its direct interaction with the flagellar filament. Lastly, in Chapter V, our findings indicate that S. marcescens is able to detect χ infection and lysis in the surroundings and alter gene expression, resulting in an increase in the production of the red pigment prodigiosin. Overall, our hypothetical model for χ infection is as follows: χ binds to the flagellum of its host using its single tail fiber, composed of monomers of the CHI_31 gene product gp31. This tail fiber interacts with CTD2 of flagellin, and the rotation of the flagellum brings the phage to the cell surface, where it interacts with AcrABZ-TolC to inject its genetic material into the host cytoplasm. At some point during the process of production of phage particles and subsequent cell lysis, the host molecular chaperone trigger factor likely assists with proper folding of χ proteins. After cell lysis, cells in the surroundings are capable of detecting lysis and responding accordingly, at least in the case of S. marcescens. This research is clinically relevant for a number of reasons. Phage therapy, the use of bacteriophages as antibacterial agents, requires knowledge of phage infection pathways for optimal implementation. The fact that the flagellum and a complex mediating MDR are both essential for χ infection leads to particular interest in χ for this application. Knowledge of the host-determining factors between χ and Salmonella may lead to the ability to alter the χ phage genome to target specific pathogenic Salmonella or Escherichia coli strains while avoiding disruption of beneficial bacterial communities. / Doctor of Philosophy / Bacteriophages (phages) are viruses that only infect bacteria. They do not harm animal cells or the human body, despite being highly effective predators of bacteria. As such, they have applications in the medical field as antibacterial agents, similar to antibiotics. Phages that infect pathogenic bacteria like Salmonella are of particular interest for scientific research. Bacteriophage χ (Chi) infects bacteria by binding to their flagella, propeller-like appendages that a bacterial cell uses to swim through its surroundings. In many bacterial species, flagella and the ability to swim are closely involved in human infection. Due to this, flagellotropic (flagellum-dependent) phages like χ may be particularly useful as antibiotics. Throughout this project, we characterized the χ phage infection process, including exploring how it attaches to flagella, interactions it has on the surface of and inside Salmonella cells, and the largely unexplored relationship with Serratia marcescens, another bacterial species that causes illness in humans and is highly antibiotic resistant. Overall, our research contributes to the medical field, and indicates that χ may serve as a highly effective antibacterial treatment.

Submitted in partial compliance with the requirements for a Master Degree in Technology: Chiropractic at the Durban University of Technology, 2009. / Background: Research has indicated the majority of bacteria on chiropractic adjustment beds (beds), can persist on dry inanimate surfaces for months. Thus, insufficient disinfection procedures create continuous sources of pathogens endangering patients and healthcare workers alike. This research study aimed to assess the beds as reservoirs for micro-organisms, at a chiropractic teaching clinic (clinic) in South Africa. Method: A selection of samples obtained from the headrests and armrests of the beds were serially diluted, plated in duplicate (using the spread plate technique) and incubated for 24-48 hours at 37°C. After inspection for the presence of micro-organisms, those present were enumerated to determine their quantities, the microbial build-up throughout the day, as well as the degree of the transmission from the patients to the beds during treatment. The incidence of the micro-organisms was established, along with their identities, using microscopic and macroscopic characteristics. These micro-organisms were also used to assess the efficacy of the disinfectant currently in use by the clinic. Results: Microbial growth was present on 89.4% of the beds sampled. The quantities of the micro-organisms increased significantly (p=0,027) from 7:30 am to 16:30 pm, with the median increasing from 25 colony forming units (cfu) / cm2 to 714 792 cfu/ cm2. The microbial build-up was highly significant (p<0.001), with a median of 346 cfu/ cm2 at 7:30 am and 10:30 am; increasing to 162 291 cfu/ cm2 by 13:30 pm and 250 million cfu/ cm2 by 16:30 pm. There was also a significant increase (p<0.001) in the quantity of micro-organisms during treatment with a median of 0 cfu/ cm2 before treatment that rose to 23 479 cfu/cm2 after treatment, indicating that the micro-organisms present on the beds were being deposited by the patient`s skin during the treatment. The most prevalent micro-organisms identified were Staphylococci and Serratia, with an average of 59% and 40% of colonies; while Micrococci and Bacilli were relatively uncommon. No growth was evident after 5 minutes of exposure to the disinfectant during the growth inhibition test. For the Kirby Bauer test, the average size of the zone of inhibition increased as the dilution decreased. The disinfectant is effective but more so against the Gram-positive than the Gram-negative bacteria. The disinfectant was 5,0, 5,5 and 5,6 times more effective than phenol in eradicating Staphylococci, Serratia and Bacilli, respectively. Conclusions and Recommendations: This study showed that micro-organisms were present on the beds. Staphylococci and Serratia have been implicated in many healthcare associated infections. The present disinfectant is effective, but should be used in between every patient. A different or additional disinfectant that is more effective against the Gram-negative bacteria should be considered for future use.