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 …

Donor And Recipient Microbiome Interactions Determine Functional Outcomes Of Fecal Microbiota Transplantation For Clostridioides Difficile Infection, Sophie A. Millard

All Dissertations

Clostridioides difficile is a major cause of healthcare-associated diarrhea. Infection typically occurs after antibiotic treatment disrupts the gut microbiome, allowing C. difficile to colonize and produce toxins leading to disease (CDI). Standard antibiotic treatments can resolve acute infection but frequently lead to recurrence, highlighting the need for alternative strategies that restore the gut ecosystem. Fecal microbiota transplantation (FMT), the transfer of stool from a healthy donor, has emerged as an effective therapy for recurrent C. difficile infection (rCDI). Understanding the mechanisms underlying the success of FMT for treatment of rCDI could lead to safer, more defined microbial mixtures.

This dissertation …

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 …

Distribution, Roles And Environmental Drivers Of Bacterial Communities In The Chesapeake And Delaware Bays, Mir Alvee Ahmed

All Dissertations

In this dissertation, water samples from the Chesapeake and Delaware Bay estuaries were analyzed to see how estuarine environmental gradients drive distributions, functional potential, growth estimates and gene expressions of bacteria. To answer this, the abundant and versatile taxa– Rhodobacterales and Actinobacteriota, along with broader particle-attached (PA) vs. free-living (FL) bacterial assemblages of these Bays were analyzed using a multifaceted ‘omics approach.

The PA fraction, differing from the FL in taxonomy, distribution, functional potential and estimated growth rates across salinities, and bays harbored genes degrading complex aggregates, and surface attachment. The FL counterpart presented genes involved in nutrient uptake …

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 …

Elucidating The Roles Of Septin Proteins In Thermotolerance And Cell Wall Integrity In Cryptococcus Neoformans, Stephani S. Martinez Barrera

All Dissertations

Cryptococcus neoformans is a globally distributed fungal pathogen responsible for causing cryptococcal meningitis in immunocompromised individuals. This pathogenic yeast must adapt to changes in temperature upon entering the human host. Septin proteins are conserved filament-forming GTPases that assemble as higher-order complexes at the cell cortex to support cytokinesis and morphogenesis in fungal and animal cells. In C. neoformans, four septin homologs (Cdc3, Cdc10, Cdc11, and Cdc12) assemble at the mother-bud neck, contributing to cytokinesis through poorly understood mechanisms. C. neoformans strains lacking the septins Cdc3 or Cdc12 are viable at 25°C, but fail to proliferate at 37°C, and are …

Assessment Of Tce And Chiral Pcb Dechlorination Rate, Congener Diversity, And Enantioselectivity In Town Creek, Sc, Usa Sediment Microcosms, Catherine P. Sumner

All Dissertations

Polychlorinated biphenyls (PCBs) and trichloroethene (TCE) are ubiquitous contaminants and are recognized as persistent organic pollutants due to their extreme chemical stability. PCBs were manufactured by chlorinating biphenyls that created 209 congeners with various structures, of which 19 are chiral and can exist as a pair of stable atropisomers. PCBs have been known to cause developmental and neurological toxicity in humans and wildlife; they can act as endocrine disrupters, carcinogens, and teratogens. Sangamo Weston Inc. was an industrial plant located near Town Creek in Pickens Country, South Carolina, that manufactured capacitors and used Aroclors 1016 and 1254 as dialectic fluids …

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. …

Exploring Novel Nutrient Acquisition Mechanisms For Maintaining Energy Metabolism In Toxoplasma Gondii, Melanie Key

All Dissertations

Toxoplasma gondii is an infectious intracellular parasite that establishes infection within various mammalian hosts, including humans. Toxoplasma infects approximately one-third of the global population, making it an impactful pathogen of study. T. gondii is the causative agent of toxoplasmosis, an opportunistic infection that causes severe illness or fatality in immunocompromised individuals or pregnant women. The parasite possesses a very effective invasion strategy, allowing it to cross different tissue types to disseminate infection across the placental and blood-brain barrier. Although there are a few treatments for acute toxoplasmosis, these drugs can produce adverse side effects. These options are not effective against …

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 …

Developing Disinfection Strategies For Controlling Human Norovirus, Sars-Cov-2, And Clostridioides Difficile Endospores In Long-Term Care Facilities, Jinge Huang

All Dissertations

Long-term care facilities (LTCFs) provide an environment favorable for the transmission of three critical human pathogens: human norovirus (HuNoV), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and Clostridioides difficile. Given residents in LTCFs are susceptible to infections due to their advanced ages and compromised immune systems, effective environmental surface disinfection plays a crucial role in controlling the spread of human pathogens within these settings and, therefore, mitigates the risk of infections caused by these pathogens. This dissertation aimed to assess the efficacy of various types of disinfectants against two HuNoV surrogates [feline calicivirus (FCV) and Tulane virus (TuV)], two …

Elucidating Acetate Metabolism: Identification Of Transporters And Enzymes Required For Acetate Utilization In The Fungal Pathogen Cryptococcus Neoformans, Perry L. Kezh

All Dissertations

Cryptococcus neoformans is the leading cause of fungal meningitis world-wide. While exposure to this environmental sporophyte is common during childhood, those who are immune compromised are at risk of infection. Following inhalation, this basidiomycetous fungus subsequently colonizes other organs though hematogenous dissemination, eventually crossing the blood brain barrier and colonizing the brain where it causes as cryptococcal meningitis. Changes in the availability of carbon sources stemming from the movement from soil to the lungs induce changes in fungal metabolism. Specifically, alveolar macrophages, which present a first line of defense against infection, provide a glucose-/amino acid-poor environment. As such, the use …

Effect Of Authentic Kefir And Nigella Sativa On Broilers Challenged By Coccidia And Clostridium Perfringens, Julian E. Nixon

All Dissertations

Black seed oil concentrations of 0%, 0.1%, 1% and 5% were added to milk inoculated with kefir grains and incubated at 25°C for 22 h. The pH and microbial count indicated 1% black seed oil caused low inhibition (P > 0.05) of fermentation, but 5% black seed oil caused significant inhibition of the kefir microorganisms (P < 0.05).

Cobb 500 male chicks (n = 256) were distributed in a randomized block design and received one of four treatments: CTRL1 (Non-medicated, no kefir, no Clostridium perfringens), CTRL2 (Non-medicated, no kefir, C. perfringens inoculated), CTRL3 (BMD medicated, no kefir, C. perfringens inoculated), KTRT (Non-medicated, …

Investigating The Role Of Cytochrome P450 Enzymes In Triazole Drug Efficacy And Toxicity In Whole Organism Zebrafish Model, Heather Cosh

Honors College Theses

Invasive fungal infections account for nearly 1.5 million deaths a year and therefore are considered one of the leading causes of death worldwide. The first line of therapy towards these infections are antifungal drugs in the triazole class such as voriconazole, posaconazole, itraconazole, and isavuconazole. However, these antifungal drug therapies fail in a large number of patients and can produce toxic side effects, therefore increasing the risk of mortality worldwide. While prior research has demonstrated the efficacy of these drugs in mitigating fungal growth in vitro, it is unclear why these drugs fail to protect patients infected with fungal …

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 …

Diagnosis Of Urinary Tract Infections And Rapid Molecular Characterization Of Antibiotic Resistance, Mohammed Harris

All Dissertations

Urinary tract infections (UTIs) are one of the most common infectious clinical entities in both community and hospital settings. They have a broad range of clinical severity yet inflict large epidemiological burden of morbidity and mortality on patients and the healthcare system with billions of dollars in cost of treatment. Understanding what methods are optimal for diagnosing UTIs are critical to mitigate the marked impact and cost of these infections.

Chapter 1 and 2 in this work surveys the broad array of diagnostic modalities for UTIs and highlights their advantages and limitations in the context of the current standard of …

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 …

The Cryptococcus Neoformans Flc1 Homologue Controls Calcium Homeostasis And Confers Fungal Pathogenicity In The Infected Hosts, Piotr R. Stempinski, Kristie D. Goughenour, Lukas M. Du Plooy, J. Andrew Alspaugh, Michal A. Olszewski, Lukasz Kozubowski

Open Access Publishing Fund

Cryptococcus neoformans, an opportunistic yeast pathogen, relies on a complex network of stress response pathways that allow for proliferation in the host. In Saccharomyces cerevisiae, stress responses are regulated by integral membrane proteins containing a transient receptor potential (TRP) domain, including the flavin carrier protein 1 (Flc1), which regulates calcium homeostasis and flavin transport. Here, we report that deletion of C. neoformans FLC1 results in cytosolic calcium elevation and increased nuclear content of calcineurin-dependent transcription factor Crz1, which is associated with an aberrant cell wall chitin overaccumulation observed in the flc1Δ mutant. Absence of Flc1 or inhibition …

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 …

Microbial, Chemical, And Functional Components In Kefir, Natto, And Feed Ingredients, Dana Mccurdy

All Dissertations

Kefir grains were purchased from online, home kitchen vendors in the United States (n = 22), inoculated into UHT milk, and incubated at 25°C for 22 hours. Results indicated lactobacilli, lactococci and yeast in all samples. Six out of 11 samples contained coliform, and all samples contained pseudomonal and staphylococcal species. Using targeted genome sequencing (16S) of kefir grain samples 1, 2, and 3 revealed that lactobacilli were the predominant genera of bacteria. ITS sequencing revealed different fungal species in each grain. Results indicated potential presence of menaquinone-4 and menaquinone-7 by UPLC-MS/MS analysis.

The fermentation of whole and ground soybeans, …

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 …

Dynamics Of The Supragingival Dental Plaque Microbiome And Its Implications In Early Childhood Caries (Ecc), Lauren O'Connell

All Dissertations

Early childhood caries (ECC) is one of the most common chronic diseases worldwide and remains a serious public health crisis in developing countries and socioeconomically disadvantaged communities. Caries is a complex polymicrobial, sugar-driven disease caused by interactions between the biofilm and tooth surface. While there is a general understanding of the microbial processes driving dental caries development and progression, the microbiota associated with the initiation and progression of dental caries have yet to be fully characterized. The most-studied species associated with tooth health and disease are the oral streptococci. Nevertheless, there are still large gaps in our knowledge of the …

Effects Of Continuous In Situ Low-Dose Ionizing Radiation On Microorganisms, Molly E. Wintenberg

All Dissertations

Precise detection and monitoring of nuclear fuel cycle, enrichment, and weapon development activities are critical for supporting warfighter preparation in chemical, biological, radiological, nuclear, and explosives (CBRNE) operations, clandestine activities, and nuclear compliance. A biological sensing system could serve as an alternative to traditional detection methods by using organic material naturally present in the environment to discreetly detect residual trace nuclear material. Microorganisms provide an optimal platform for an alternative sensing system; however, their response to low levels of ionizing radiation is poorly characterized. Combining the power of next-generation sequencing and transcriptomic analysis, this dissertation takes an approach to obtain …