Theses and Dissertations from UMD
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New submissions to the thesis/dissertation collections are added automatically as they are received from the Graduate School. Currently, the Graduate School deposits all theses and dissertations from a given semester after the official graduation date. This means that there may be up to a 4 month delay in the appearance of a give thesis/dissertation in DRUM
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Item INHIBITION OF TYPE ONE INTERFERON SIGNALING THROUGH CROSSTALK WITH TOLL-LIKE RECEPTOR SIGNALING(2024) Shuster, Michael; Briken, Volker V; Cell Biology & Molecular Genetics; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Interferons (IFNs) are a class of cytokines that play a prominent role in host immunity. Type I IFN is broadly associated with antiviral immunity and susceptibility to bacterial pathogens, but others have shown that type I IFN can be beneficial in some bacterial infections. Additionally, some bacterial infections such as Mycobacterium tuberculosis and Legionella pneumophila can inhibit type I IFN signaling. Questions remain such as how these bacteria inhibit type I IFN signaling as well as if other bacterial pathogens, such as Salmonella enterica, can also inhibit type I IFN signaling. Additionally, type III IFN is a relatively new class of IFN, providing antiviral protection similar to and at times redundant to type I IFN. There are some important non-redundant differences from type I IFN though, such as type III IFN’s broader activity at epithelial surfaces (like those in the lungs) and its reduced proinflammatory effects. The role of type III IFN in bacterial infections as well if bacteria can inhibit this signaling pathway remains poorly understood.Here, we examined if Salmonella enterica can inhibit type I IFN signaling, the specificities of the previously observed inhibition with Mtb infection, and how these bacterial infections are inhibiting this signaling. We demonstrate that Salmonella Typhimurium infection inhibits type I IFN signaling through crosstalk with TLR4 signaling. We establish that TLR4 signaling results in reduced surface level type I IFN receptor, which dampens cellular responsiveness to type I IFN. We show that Mtb does not inhibit type III IFN signaling and that it inhibits type I IFN signaling independently of virulence, specifically EsxA and ESX-5. Additionally, this inhibition of type I IFN signaling seems specific to mouse cells as Mtb-infected human macrophages and dendritic cells did not have inhibited type I IFN signaling. We observed that other TLR signaling pathways result in specifically inhibited type I IFN signaling. Synthesizing a model from our results, there appears to be a mouse-specific crosstalk pathway between TLR signaling and type I IFN signaling, resulting in dampened responsiveness to type I IFN through downregulation of cell surface type I IFN receptor.Item EVALUATING DIFFERENCES IN BODY WEIGHT, GUT MORPHOLOGY, IMMUNE RESPONSE, AND SICKNESS BEHAVIOR IN FAST- AND SLOW- GROWING BROILER CHICKENS WHEN INFECTED WITH SALMONELLA ENTERICA SEROVAR TYPHIMURIUM(2021) Snyder, Ashlyn Marie; Weimer, Shawna; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Fast growth rate in broilers comes with welfare concerns and research is needed to determine if fast- and slow-growing broilers differ in pathogen resistance. The objective of this study was to evaluate differences in fast- (FG) and slow-growing (SG) broilers when challenged with Salmonella Typhimurium or broth (control) 14 days post-hatch. Plasma IgA and IgG, jejunum and ileum histomorphology, and behaviors were measured. FG had greater d12 and d24 body weight and d7 jejunum measures, indicating better absorption, and earlier increases in plasma IgA and IgG, indicating earlier immune development. SG had greater d7 IgG, indicating stronger maternal immunity. Post-challenge, FG gut morphology was more impaired, and SG had greater IgA and reduced sham foraging, indicating a stronger immune response to challenge. The results illustrate fast- and slow-growing broilers differ in Salmonella resistance, which can help breeders make selection decisions to prevent Salmonella transmission into the human food supply.Item SALMONELLA ENTERICA STRATEGIES FOR PERSISTENCE ON TOMATO (SOLANUM LYCOPERSICUM) AND SEROVAR DYNAMICS IN SURFACE AND RECLAIMED WATER(2019) Ferelli, Angela Marie Cecelia; Micallef, Shirley A.; Plant Science and Landscape Architecture (PSLA); Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)While select aspects of Salmonella enterica subspecies enterica persistence in agricultural matrices have been illustrated, serovar specific survival strategies in surface water, transmission, and persistence on plants are multifaceted and remain only partially examined. In the present work, we utilized an interdisciplinary approach to illustrate novel mechanisms by which S. enterica may adapt to plants as an alternative host. Furthermore, we leveraged the wealth of diversity in S. enterica serovars to investigate specific dynamics and drivers of persistence in water and transfer onto produce crops. Through biochemical, gene expression, and plant challenge assays of both tomato (Solanum lycopersicum) vegetative and fruit organs, we found that plant-derived NO was generated in response to S. Newport recognition. Furthermore, bacterial gene expression on both leaves and fruit was indicative of adaptation to a novel environment including upregulation in NO detoxification machinery, indicating plant-derived NO as a novel bacterial stress. NO tolerance of various S. enterica was then evaluated to investigate drivers of “produce associated’ S. enterica adaptation to the plant niche. We identified that plant derived NO can negatively affect titers of all S. enterica serovars tested and that serovar specific tolerance to NO in vitro was apparent in a concentration and exposure time dependent manner. Finally, the survival of various S. enterica in surface and reclaimed water was investigated while evaluating the potential for transition to viable but non-culturable (VBNC) organisms. Furthermore, surface water used for irrigation, a common water environment for S. enterica, was investigated as a priming reservoir for various S. enterica serovars for enhanced transmission onto tomato crops. Persistence in water included VBNC subpopulations and was driven by water type. Transfer success onto tomato was driven by serovar, and prolonged incubation in water increased the transfer ability of serovars that initially transferred poorly onto tomato. Finally, attachment to polystyrene and water oxidation-reduction potential were identified as possible indicators of foodborne pathogen transfer success onto tomato. Moving forward, a greater understanding of the environmental queues used by S. enterica subspecies enterica responding to the agricultural environment will aid researchers in developing S. enterica targeted on-farm management techniques to ensure safe yet sustainable fresh produce cultivation practices.Item The Impact of Private Drinking Water Wells and Animal Feeding Operations on the Incidence of Acute Gastroenteritis in Maryland: A Mixed Methods Approach(2019) Murray, Rianna Teresa Frederika; Wilson, Sacoby M; Maryland Institute for Applied Environmental Health; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)More than 44.5 million people in the United States (approximately 14% of the population) rely on private domestic wells as their primary source of drinking water. The water quality of private wells is not regulated at the state or federal level, leaving homeowners with wells responsible for the safety of their water. Meanwhile, each year, an estimated 48 million people in the U.S. are sickened, 128,000 are hospitalized, and 3,000 die of gastrointestinal (GI) illnesses caused by microorganisms that are typically transmitted through contaminated food. Given that the potential of private wells as a non-foodborne transmission pathway for these microorganisms is not well understood, my dissertation evaluated these relationships, as well as characterized the quality of private well water in Maryland. My objectives were to: 1.) Investigate the water quality of private wells in Maryland and the effect of animal feeding operations on this water quality using fecal indicator bacteria; 2.) Investigate associations between the prevalence of private wells and the incidence of campylobacteriosis in Maryland at the zip code level using data from the FoodNet active surveillance System; and 3.) Investigate associations between the prevalence of private wells and the incidence of salmonellosis in Maryland at the zip code level using FoodNet data. My findings demonstrated that 43.2% of private wells tested in Maryland did not meet at least one federal health-based drinking water standard. Additionally, my data showed that the prevalence of private wells in Maryland is a risk factor for the incidence of campylobacteriosis and salmonellosis in the coastal counties of the state. The presence of broiler chicken operations in a zip code is also a risk factor for campylobacteriosis and salmonellosis in coastal counties. These findings provide evidence for the strengthening of private well water regulations and for improving education and outreach to private well owners on proper maintenance and testing for their wells.Item EVALUATION OF THE BEHAVIOR OF SALMONELLA ENTERICA IN REHYDRATED DRY DOG FOODS(2017) Qu, Yinzhi; Pradhan, Abani K; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Recent human salmonellosis outbreaks associated with dry dog foods have raised concern over these products as potential vehicles for Salmonella. In this study, different behavior (decline or growth) of Salmonella across twenty-six different brand dog foods that were rehydrated to a moisture content of 35% and stored at 30°C for 72 hr were characterized. Decline data were fitted with log-linear model and growth data were fitted by reparameterized Gompertz model. The distributions for the parameters in the fitted reparameterized Gompertz model were obtained. The effects of pH and water activity of rehydrated dog foods on changes in Salmonella levels (Log CFU/g) within 72 hr were modeled by the second order polynomial regression. The results can be implemented in the future quantitative microbial risk assessment studies. This study was useful in providing critical information regarding Salmonella and dog food to develop effective contamination prevention and mitigation strategies.Item Antimicrobial Resistance of Salmonella and E. coli from Pennsylvania Dairy Herds(2015) Cao, Huilin; Pradhan, Abani K.; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)The emergence and dissemination of bacterial antimicrobial resistance has become a major public health concern. A total of 444 manure composite samples were collected from 80 dairy farms in Pennsylvania, representing pre-weaned calves, post-weaned calves, dry cows, and lactating cows. E. coli and Salmonella were isolated, and tested for antimicrobial susceptibility. Salmonella was isolated from at least one sample from 51 (64%) farms and was more prevalent in adult animals than young animals. The predominant serotypes were Cerro, Montevideo and Kentucky. Salmonella isolates were mostly susceptible to all antimicrobials. E. coli were commonly resistant to tetracycline, streptomycin, sulfisoxazole and ampicillin. Resistance of up to 8 classes of antibiotics was observed in E. coli isolated from young animals. The blaCMY- and blaCTX-M-carrying E. coli were detected in 35% and 5% of the farms, respectively. The presence of multi-drug resistant E. coli suggested potential risks to human health associated with dairy farming.Item Investigating Metrics Proposed to Prevent the Harvest of Leafy Green Crops Contaminated by Floodwater(2015) Callahan, Mary Theresa; Buchanan, Robert L; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Pathogens can be transported by water through soil to contaminate distant crops. The California LGMA states that leafy green crops within 30ft of flooded soil should be destroyed due to potential contamination. Previously flooded areas should not be replanted for 60 days. This study investigated the transport of Salmonella enterica and Citrobacter freundii through soil in a model system with a positive slope (uphill). Field trials involving flooding one end of a spinach bed with a negative slope (downhill) with water containing Escherichia coli were also conducted. Soil type, soil moisture content, and slope affected bacterial movement. In field trials, E. coli was quickly transported to the 30ft boundary, and persisted significantly longer in the fall trial than the spring. These data suggest the LGMA metrics need to provide additional parameters to prevent the harvest of leafy green crops potentially contaminated by floodwater.Item Prevalence of Salmonella on Laying Hen Farms and Control of Colonization in Poultry Through Egg Yolk Antibodies(2014) Almario, Jose Alejandro Navarro; Biswas, Debabrata; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)In the United States, rates of foodborne illness caused by Salmonella have not changed significantly. One study in this thesis estimated Salmonella prevalence and antimicrobial resistance of various samples from conventional (n=181) and organic (n=252) farms. Rates of Salmonella contamination were significantly lower on conventional than organic farms. Antimicrobial resistance was significantly higher on isolates from conventional versus organic farms. These findings suggest that poultry production practices may have significant effects on prevalence and antibiotic resistance patterns of Salmonella. The other study assessed the efficacy of a Salmonella control strategy using anti-Salmonella antibodies, two chicken cell lines, an HD-11 macrophage and a DF-1 fibroblast line, and Salmonella serovars Typhimurium and Enteritidis. In DF-1 cells, treatment showed decrease adherence of the pathogen. However, in HD-11 cells, treatment showed an increase in pathogen adherence, indicating a more detailed understanding of chicken response to treatment with the antibodies is needed before full-scale implementation.Item ALTERNATIVE APPROACHES IN MOLECULAR CHARACTERIZATION OF FOODBORNE PATHOGENS: SHIGA TOXIN-PRODUCING Escherichia coli AND Salmonella SEROTYPES(2014) Toro Ibaceta, Magaly Alejandra; Meng, Jianghong; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Shiga toxin-producing E. coli (STEC) and Salmonella enterica subspecies enterica (S. enterica) are two major foodborne pathogens. They cause almost 1.5 million of cases of disease each year in the US. Due to their public health impact, development of new methods for their detection and identification are top priority. This research focused on identifying alternative molecular methods and markers for the identification of STEC and Salmonella. First, a suspension array was developed to simultaneously identify the seven most prevalent STEC (O26, O45, O103, O111, O121, O145, and O157) in the US. The panel targeted genes wzx or wzy and Shigatoxin genes. Testing and optimization employed four to eleven isolates of each serotype in the panel. STEC fluorescence values were 30 to >270 times greater than those of negative controls, demonstrating the method's effectiveness for the molecular serotyping of STEC. STEC strains (n=194) of 43 serotypes were examined for clustered regularly interspaced short palindromic repeats (CRISPR) arrays to study relatedness among serotypes. A subset of strains (n=81) was analyzed for cas and virulence genes to determine a possible relationship. CRISPR spacer content correlated well with serotypes, although some strains with different serogroup but the same H type shared identical arrays (O26:H11, O103:H11, and O111:H11). cas and virulence genes were not associated, but strains with greater probability of causing outbreaks and disease showed fewer spacers than those less likely to cause them (p<0.05). Therefore, CRISPR array content correlated well with STEC serotype, and CRISPR-cas systems were inversely related to strain virulence potential. Finally, the CRISPR arrays of 221 S. enterica of 53 serotypes were analyzed to define their relationship. CRISPR-cas systems of 50 S. enterica serotype Bareilly (S. Bareilly) were analyzed to resolve intra-serotype variations. CRISPR arrays correlated well with serotypes, although some serotypes displayed more than one type of array (e.g. S. Bareilly). Additionally, CRISPR-cas system elements reflected S. Bareilly phylogeny, but the array content was not linked to food vehicle or isolate's geographical origin. In conclusion, CRISPR array are useful for designing molecular serotyping assays, but a range of strains should be included to account for variation in S. enterica.Item The Role of Organic Matrices (dried turkey manure) in the Contamination and Survival of Salmonella spp. on Baby Spinach Leaves(2012) Oni, Ruth Adeola; Buchanan, Robert L; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Microbiological safety of fresh produce has attracted attention due to constant produce outbreaks. Manure contaminated with enteric pathogens has been identified as a major pre-harvest contamination source. This study investigated the survival of Salmonella in dust particles generated from dehydrated turkey manure, and how this could serve as an airborne contamination of leafy greens. Laboratory trials of the UV resistance of Salmonella in manure dust were also conducted to assess its protective effect; along with greenhouse studies to determine the survival of Salmonella in manure dust on spinach leaves. Results indicate survival times were inversely related to moisture content and particle size of manure dust. The presence of manure particles substantially enhanced the pathogen's survival under UV and reduced its inactivation on spinach leaves. This study presents data that could be used to assess the potential role of aerosolized manure as a pre-harvest risk factor for contamination of leafy greens.