An Investigation Of Entamoeba Histolytica Chitin Synthases And Other Encystation Factors In Early Encystation, John N. Talledo

All Theses

Entamoeba histolytica is a food-and-water-borne human protozoan pathogen that relies on a two-stage life cycle for successful proliferation outside of the host. E. histolytica exists in its dormant cyst form while outside of its host and converts to the motile trophozoite (amoeba) form in the small intestine. The trophozoites colonize the large intestine to cause symptomatic disease in ~10% of infections, resulting in ~100 million cases of amoebic dysentery each year. Trophozoites can convert back to the cyst form, which is shed in feces to continue the infection cycle.

Currently, there is a lack of understanding of the exact molecular …

Investigating The Impacts Of Azole Antifungals On Growth Of Cryptococcus Neoformans, Gnanambika Sindhe

All Theses

Cryptococcus neoformans, an opportunistic fungal pathogen is responsible for cryptococcosis, leading to approximately 181,000 deaths annually, predominantly in regions such as Sub-Saharan Africa. Fluconazole, a widely used fungistatic azole, is often administered due to its relative affordability and lower toxicity. However, resistance to fluconazole in C. neoformans can arise through aneuploidy, complicating treatment efforts. This study aims to deepen our understanding of resistance mechanisms to azole drugs by comparing fluconazole with other azoles, including both clinical and agricultural azole antifungals. Disk diffusion assays were performed to evaluate the ability of C. neoformans to develop resistance to these compounds, focusing …

Determining The Impact Of Surface Roughness On Disinfection Efficacy Of Commercial Products Against Pseudomonas Aeruginosa And Staphylococcus Aureus Wet Surface Biofilms, Kayley Stallings

All Theses

Biofilms are resistant to many stressors including disinfectant treatments. Disinfection may be further complicated by biofilms formed on damaged surfaces, as the damaged parts provide bacteria a better place to attach and grow. To better understand the risks, a new method to standardize surface scratching, to emulate worn surfaces via surface roughness changes was developed. These surfaces were used to grow biofilms with an adapted standardized EPA method to determine disinfection efficacy differences on scratched and non-scratched high-density polyethylene (HDPE) and stainless steel (SS) coupons treated with commercial disinfectants [hypochlorite (CL)-based, hydrogen peroxide (HP)-based, and quaternary ammonium (QA)-based]. Post-80 grit …

Impact Of Shear Flow On Marine Biofilms Using Hyperspectral Imaging, Shantanu Mahesh Kore

All Theses

Various microbial communities in marine biofilms cause biofouling on submerged surfaces, posing challenges to marine industries. Despite their ecological and economic importance, biofilms' biomechanical and biochemical responses to hydrodynamic shear stress are insufficiently understood, particularly in dynamic flow conditions. To fill this gap, our study uses an innovative fiber-optic hyperspectral imaging (HSI) system and a supercontinuum laser source to examine marine biofilms' structure, composition, and detachment behavior at different shear stress levels.

To detect spectral and spatial heterogeneity in live biofilms without labeling, we created a custom imaging pipeline that captures reflectance spectra in the 600-850 nm range, targeting microbial …

Thymosin Beta-4: A Novel Treatment For Acanthamoeba Keratitis, Victoria A. Murphey

All Theses

Acanthamoeba keratitis (AK) is a severe corneal infection caused by the free-living amoeba, Acanthamoeba. Both stages of its life cycle (trophozoite and cyst) cause infection, primarily in contact lens wearers. It is often misdiagnosed, leading to delayed treatment and poor clinical outcomes. Current treatment options for AK rely on anti-amoebic agents that are toxic to corneal epithelial cells. Further, no single drug can eliminate both forms of the amoeba. Approximately 30% of all AK infections require a corneal transplant.

Thymosin beta-4 (Tβ4) is a G-actin binding protein synthesized in almost all human cells, including corneal epithelial cells. Tβ4 exhibits …

Functional Characterization Of A Lncrna Afu-254 In Stress Response And Pathogenesis Of Aspergillus Fumigatus, Ritu Devkota

All Theses

Aspergillus fumigatus is a saprophytic fungus that can cause a collection of diseases termed aspergillosis; the most severe is invasive pulmonary aspergillosis (IPA). Azole is a major class of antifungal drugs used to treat invasive pulmonary aspergillosis. However, in recent years, there has been an increase in fungal resistance to azole drugs, making the treatment of IPA even more challenging. It is becoming increasingly clear that long non-coding RNAs (lncRNAs), i.e., the RNAs that do not translate into proteins, mediated regulation is vital in stress response and pathogenesis; however, the mechanisms of how lncRNA contribute to fungal stress resistance and …

Characterization Of Ato Family Transporters In The Fungal Pathogen Cryptococcus Neoformans, Will Betsill

All Theses

Fungal pathogens are a significant threat to public health as they are becoming increasingly common and more resistant to treatment. Cryptococcus neoformans contributes greatly to this threat annually by causing an estimated 278,000 cases of cryptococcal meningitis resulting in approximately 181,000 deaths globally according to the CDC. C. neoformans is ubiquitous across most of the globe and can be found in such places as in trees or soil. Exposure to this fungus is especially dangerous to individuals who are immunocompromised or immunosuppressed. In these cases, inhalation of spores can lead to infection in the lungs. Once in the lungs, C. …

Identification And Characterization Of Geotrichum Candidum Isolates From Peach With Reduced Sensitivity To Propiconazole, Sara Price

All Theses

Geotrichum candidum Link (1809) is a yeast-like fungus that causes sour rot of stone fruits, including peach (Prunus persica). The disease caused damage between 2021 and 2023 to the peach crop kept in cold storage at a commercial farm in South Carolina despite preharvest and postharvest applications of fungicides. Isolates from 41 symptomatic fruit, from a South Carolina (SC) commercial packing house, as well as other isolates obtained from South Carolina were identified to be G. candidum based on species-specific PCR amplifications of the β tubulin gene. Commonly used postharvest fungicide fludioxonil had no effect on G. candidum mycelial growth …

Disinfectant Efficacy Against Sars-Cov-2 Surrogates, Bovine Coronavirus And Human Coronavirus Oc43, On Hard And Soft Non-Porous Surfaces, Breanna Kimbrell

All Theses

The novel human coronavirus (HCoV), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), emerged from Wuhan, China in the latter part of 2019. The rapid spread of SARS-CoV-2 amid the most recent COVID-19 pandemic forced countless foodservice establishments (FSEs) across the United States to close, taking a tremendous toll on the foodservice industry. Although primarily transmitted person-to-person, previous studies indicate that the duration of SARS-CoV-2 survival on different environmental surfaces provides adequate time for secondary transmission to occur. To prevent the further spread of SARS-CoV-2, effective disinfection of surfaces in FSEs is necessary. While disinfectants approved for use against SARS-CoV-2 are …

Development Of A Novel Fluorescent Reporter Baculovirus Vector For Testing Promoters In Lepidopterans, Daniel Howard

All Theses

Testing gene expression patterns is an important process in learning more about an organism. The standard methods of rtPCR and RNASeq provide highly detailed data on specific expression patterns, but can be resource consuming if many different samples including genes or organisms must be tested. A potential alternative for studying expression is the use of a reporter system carried by a vector system that encodes fluorescent proteins under the control of promoters of interest. Many lepidopterans (moths) are pestiferous and better understanding of gene expression levels in lepidopterans, both endogenous and exogenous including from their viruses, would be beneficial. Baculoviruses …

Generation And Characterization Of A Multi-Functional Panel Of Monoclonal Antibodies For Sars-Cov-2 Research And Treatment, Lila D. Patterson

All Theses

The Coronavirus disease 2019 (COVID19) pandemic caused by SARS-CoV-2 is an ongoing threat to global public health. To this end, intense efforts are underway to develop reagents to aid in diagnostics, enhance preventative measures, and provide therapeutics for managing COVID-19. The recent emergence of SARS-CoV-2 Omicron strains with enhanced transmissibility, altered antigenicity, and significant escape of existing monoclonal antibodies and vaccines underlines the importance of the continued development of such agents. The SARS-CoV-2 spike protein and its receptor binding domain (RBD) are critical to viral attachment and host cell entry and are primary targets for antibodies elicited from both vaccination …

The Effects Of Tubulin Post-Translational Modifications On The Flagellar Motility Of Trypanosoma Brucei, Katherine Wentworth

All Theses

Trypanosoma brucei is a parasitic kinetoplastid that causes African trypanosomiasis and is transmitted to a mammalian host by the tsetse fly (Glossina spp.). T. brucei relies on its flagellar motility to carry out its morphological changes from the procyclic form (predominant in the fly vector) to the bloodstream form (infectious form in mammals) and navigate the bloodstream of its host. The driving structure within the flagellum is the axoneme, which is composed of microtubules and dynein motor proteins. The tubulin code hypothesis suggests that cells regulate microtubule motor protein activity through post-translational modifications (PTMs) of alpha and beta …

Reproductive Ecology And Microbial Communities From Wood Duck Nest Boxes In Georgia And Florida, Jake Shurba

All Theses

Wood ducks (Aix sponsa) are an important game species throughout the Atlantic Flyway that was nearly extirpated by the early 20^th century due to overhunting and the loss of habitat. Wood ducks are secondary cavity-nesters that utilize artificial nest boxes and natural cavities. It is reported that the use of nest boxes is likely what led to re-establishment of the species in North America. Where boxes are numerous, overuse of boxes by multiple hens throughout a nesting season can occur and result in a buildup of bacteria, parasites and other potentially detrimental pathogens that can impact egg …

Application Of A 14c-Assay To Assess Methanotrophic Biodegradation Of Tce In Low Ph Groundwater, Evan Groome

All Theses

Current biological strategies for remediating trichloroethylene (TCE) in low pH aquifers (i.e., pH14C-TCE assay was developed to determine pseudo first-order rate constants for the degradation of TCE in microcosms containing soil and groundwater from the Boeing Michigan Aeronautical Research Center (BOMARC) superfund site, where the pH ranges from 4.1 to 4.9. The ¹⁴C-TCE assay was also adapted to calculate soil-normalized rate constants for data from this site, as well as data that Szwast²¹ collected from the Beltsville Agricultural Research Center (BARC). In addition to natural attenuation, biostimulation through amendments of methane and nutrients were also assessed. This treatment …

The Effects Of Short Chain Fatty Acids On Clostridioides Difficile Growth, Sporulation, And Toxin Production, Michelle Baldassare

All Theses

Short chain fatty acids (SCFAs) are products of bacterial fermentation that help maintain important gut functions such as the intestinal barrier, signaling, and immune homeostasis. The main SCFAs acetate, propionate, and butyrate have demonstrated beneficial effects for the host, including importance in combatting infections caused by pathogens such as Clostridioides difficile. C. difficile infection (CDI) is a public health concern and causes 30,000 deaths each year mainly due to the overuse of antibiotics. Despite the potential role of SCFAs in mitigating C. difficile infection, their direct effect on C. difficile remains unclear. Through a set of in-vitro experiments, we …

Identification Of Critical Points For Bacterial Contamination In The Microbrewery Environment, Alex Ryan Thompson

All Theses

There are 8,884 craft breweries producing over 23 million barrels of beer in the United States as of 2020. These 23 million barrels of craft beer account for 12.3% of the United States beer consumption in 2020. The American craft beer industry is substantial and needs to protect its product from bacterial contamination.

Overall, beer is a microbially stable product. Beers pH, ethanol levels, CO₂ concentrations, the presence of hop-derived antimicrobial compounds, and low levels of O₂ make beer a highly unfavorable environment for most bacterial species. Furthermore, the brewing process, which involves heat treatments and chemical sanitizers …

Glycan Functionalization Of Iron Oxide Nanoparticles (Ionps) As A Method Of Inactivating Antibiotic Resistant Neisseria Gonorrhoeae Via A Magnetically Mediated Energy Delivery (Magmed) System, Cedric Taylor

All Theses

With the increase of antibiotic resistant bacteria, current established therapeutic treatment methods are rapidly demonstrating decreasing efficacy. For this project, we are studying a novel alternate mechanism of treating antibiotic resistant bacteria by utilizing magnetic iron oxide nanoparticles (IONPs) coated with a heterobifunctional polyethylene glycol polymer (PEG) and functionalized with glycan structures that can interact with Neisseria gonorrhoeae FA1090. Interaction between the IONPs and bacteria is manipulated by an external alternating magnetic field (AMF) in a process known as magnetically mediated energy delivery (MagMED). Magnetic IONPs were synthesized and reacted with PEG (PEG-IONPs) and functionalized with glycans Gal(α1-3)Gal(β1-4)Glc(β1-1) (common name: …

Investigation Of Dissimilatory Iron Reduction By Exophiala Hkrs030, Zachary Burton

All Theses

Iron (Fe) is the fourth most abundant element in the Earth’s crust and is an essential component in many biological processes. While highly abundant, free iron is not readily available to biological systems since most iron is present in the earth’s crust in the form of ferrous (Fe2+) and ferric (Fe3+) iron minerals. Because of this limited availability, bacteria, plants and fungi have developed diverse ways to safely assimilate and store iron. Bacteria and some fungi have the ability to dissimilate iron. Dissimilatory iron reduction (DIR) is where Fe3+ is reduced to Fe2+ without the cell assimilating the Fe2+ and …

Relationships Between Growth Rate And Gene Expression In Ruegeria Pomeroyi Dss-3, A Model Marine Alphaproteobacterium, Nattasha Vinas

All Theses

Microbes are important contributors to ecosystem processes such as biogeochemical cycling. Their activities vary, depending on environmental parameters such as carbon type and concentration, temperature, and salinity. Current estimates of in situ microbial growth is limited to gross estimates or rely on incubation based methods that disturb the natural state of the community. The goal of this master’s thesis is to develop molecular methods to directly assess microbial growth rates in the environment at the level of a taxonomic group. Here we grew Ruegeria pomeroyi DSS-3 under different temperatures (15 °C or 30 °C) and different carbon sources (yeast extract/tryptone, …

Characterization Of Biofilms On Medical Device Materials With Application To Reusable Surgical Instruments, Amanda Macaluso

All Theses

Reusable medical devices or reprocessed single-use devices are original medical devices that have been used once and then are cleaned, sterilized, and remanufactured for the purpose of an additional single use on a single patient [4]. Improperly reprocessed devices are a significant contributor to hospital-associated infections [4]. Challenges that hinder reprocessing are related to the complexity of reusable medical device design, the necessary validation of cleaning protocols required by the U.S Food and Drug Administration (FDA), the impact of human factors throughout the reprocessing cycle, as well as economic factors within new business models that are centered on reprocessing. Current …

Determining The Fate Of Non-O157 Shiga Toxin-Producing Escherichia Coli In Dairy Compost During Storage Using The Optimized Detection Method, Hongye Wang

All Theses

Shiga toxin-producing Escherichia coli (STEC), a member of Enterobacteriaceae family, has been recognized as emerging pathogens. Dairy compost is commonly applied to farmland as a soil amendment. Despite the agricultural benefit of manure-based soil amendment, the inadequately treated compost can contribute to fresh produce contamination on the farm. Moreover, the epidemiological results showed that the non-O157 STEC cases have surpassed those of E. coli O157. Therefore, it is critical to evaluate the behavior of non-O157 STEC strains in the dairy manure-based compost. The objectives of this study were to: 1) optimize a culturing method for detecting non-O157 STEC from dairy …

The Effects Of Probiotics Supplementation On Health Using Caenorhabditis Elegans As A Model System, Miranda Klees

All Theses

The 'Western Diet,' prominent among developed nations, often refers to a diet rich in meat proteins and refined sugars. Western society may in part be plagued with obesity and obesity related diseases due to a diet enriched with glucose. With an increasing glycemic index observed in Western society, it comes to no surprise that obesity, diabetes, heart disease, and other obesity related diseases are on the rise. The Centers for Disease Control and Prevention describes obesity as an epidemic, affecting more than 35% of U.S. citizens. Shortened lifespan and increased susceptibility to pathogens are associated with these diseases and linked …

Inhibition Of Vibrio Cholerae Biofilm Formation By Water-Soluble Cranberry Extracts, Daniel Pederson

All Theses

Vibrio cholerae, a Gram-negative bacterium found in natural aquatic environments, is the causative agent of cholera, a severe diarrheal disease most commonly spread through drinking water. An essential component to this pathogen's success and persistence in the environment is its ability to attach to both biotic and abiotic surfaces via biofilm formation. Biofilms not only aid in surface attachment, but also provide a barrier that protects and enhances survival. Water-soluble extracts from the North American cranberry (Vaccinium macrocarpon) were found to dramatically inhibit V. cholerae biofilm formation at a low concentration of 2mg/ml. This inhibition was not due to acidification …

A Comparison Of Saponin And Tween 80 On The Solubility And The Degradation Enhancement Of Fluoranthene And The Influence Of Laccase On Fluoranthene Degradation, Stacia Ballew

All Theses

The ubiquitous contamination of polycyclic aromatic hydrocarbons (PAHs) in the environment require an economical approach as well as effective remediation techniques in the removal of these contaminants. Through the use of more eco-friendly and economically available biosurfactants, such as saponin, there is a possibility they could prove to be more useful surfactants than conventional synthetic surfactants for the solubilization of fluoranthene (FLA). Enhanced solubility would potentially facilitate the increased degradation by S. paucimobilis EPA505. The addition of laccase produced by Pleurotus ostreatus, a white rot fungus that produces enzymes known to facilitate or assist in the degradation of fluoranthene (FLA), …

The Role Of Human Norovirus Surrogates, Feline Calicivirus And Murine Norovirus, On Non-Porous And Soft Porous Surfaces, Thomas Yeargin

All Theses

Human Noroviruses (HuNoV) are the leading cause of acute gastroenteritis worldwide as well as the leading cause of foodborne disease in the U.S. HuNoV can persist in the environment even after proper disinfection, making preventing HuNoV infections and controlling subsequent outbreaks extremely challenging. Epidemiological evidence suggests that soft surfaces may be a relevant source of HuNoV due to the inability to effectively decontaminate. The objectives of this study were to: 1) review the current published literature on prevalence, transmission, and disinfection pertaining to HuNoV and surrogates with an emphasis on soft surfaces as fomites, 2) optimize a recovery method capable …

Investigating The Impact Of Novel Iron Oxide Nanoparticles On Legionella Pneumophila Biofilms And Trophic Interactions, Brennen Jenkins

All Theses

Microbial biofilms serve as the base of food webs and are important for nutrient cycling in aquatic ecosystems. Nanoparticles (NPs) that enter into these aquatic systems have the potential to settle and become trapped within biofilms. As NPs become further integrated into consumer products, understanding their fate and effects on aquatic ecosystems is of paramount importance. Previous studies from our lab show that gold NPs induce dispersal of Legionella pneumophila biofilms. NPs with platinum and iron oxide core chemistries also lead to similar dispersal events, however, silver core NPs do not seem to induce these events due to NP aggregation. …

Production Of Biodiesel-Like Components By The Type I Methanotroph Methylomonas Methanica, Megan Diane Burdette

All Theses

Bacteria that utilize methane as a sole source of carbon and energy are referred to as methanotrophs. Industrial uses of these types of organisms include the production of poly-hydroxybutyrate as well as the degradation of some chlorinated hydrocarbons that are considered pollutants in the environment. Methanotrophs also play an important role in methane cycling in the environment. Methane gas has the ability to trap 20 times more heat than carbon dioxide, which makes it a potent greenhouse gas. A defining characteristic of methanotrophs is the production of extensive intracytoplasmic membranes composed of lipids that are 16 or 18 carbons in …

Bioelectrochemical Reduction Of Carbon Dioxide To Acetate Using A Microbial Consortium Derived From The Cow Rumen, Ryan Hammonds

All Theses

Pure cultures of chemoautotrophic microorganisms have previously been reported to produce acetate from carbon dioxide using reductive electric current. We hypothesized that electrofuel precursors could be produced in a similar manner from a consortium consisting of microbial populations acting as a stable community. This approach might offer improved efficiency and selectivity, greater sustainability, a more diverse group of end products, and more opportunities for community optimization in electrofuel production than is possible with pure cultures. To evaluate this concept, bovine rumen contents were enriched for autotrophic, anaerobic microbes for several generations using hydrogen and carbon dioxide for growth and energy. …

Effects Of Larval Exposure To Bacterial Species In Three Genera On Life History Traits And Oviposition Behavior In The Southern House Mosquito, Culex Quinquefasciatus, Jonathan Gravgaard

All Theses

The biofilm is the most common niche for microorganisms living in aquatic environments. Forming a biofilm provides the means to resist shear forces, adverse osmolarity, chemical agents, and other environmental stressors. Biofilms also provide a major food source for aquatic arthropods like mosquito larvae. Culex quinquefasciatus larvae are found in freshwater pools with high organic content, and have the ability to graze on bacterial biofilms. To study how C. quinquefasciatus mosquitoes interact with bacteria in their environment I developed an experimental system that allows mosquito larvae to freely graze on bacteria in either the planktonic or biofilm state. The efficacy …

Influence Of Environmental Conditions On Biofilm Formation Of Listeria Monocytogenes, Jing Li

All Theses

L. monocytogenes is a Gram-positive bacterium, ubiquitously distributed in the environment. It is the etiologic agent of listeriosis, a food-borne disease affecting humans and a variety of vertebrates. L. monocytogenes can form biofilm which is a multi-cellular layer of adherent bacteria surrounded by an extracellular polysaccharide matrix. Biofilm bacteria are extremely resistant to sanitizers and are difficult to be eradicated, therefore posing a big challenge to food processing industry and food service department.
In this work, we tested the biofilm formation in a number of L. monocytogenes serotypes under different environmental conditions, such as temperatures, pH and media (BHI, LB, …