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More Than The Sum Of Their Parts: Building A Framework For Understanding Host-Microbe Interactions In Medicago Sativa, Katherine Mackenzie Moccia

Doctoral Dissertations

This dissertation seeks to understand plant-microbe interactions in the agriculturally relevant plant Medicago sativa from three distinct vantage points within microbiology. Within the plant microbiome, we examine how primer usage and the application of peptide nucleic acids impacts 16S and 18S rRNA gene sequencing. In doing so, we design a novel peptide nucleic acid, PNA, and test its impact using multiple primers and sequencing protocols. Once microbial sequencing methodology is established, we generate a synthetic consortium of bacterial isolates from M. sativa leaves and modulate nitrogen levels to better understand microbial structure. Drop out communities, where we remove one member …

Extracting Detailed Metabolic Information And Connections From Mammalian Gut Microbiomes Via Metaproteomics, Jose A. Blakeley-Ruiz

Doctoral Dissertations

A diverse community of bacteria populates the mammalian gastrointestinal tract. These populations exist in a balance with the host assisting with key functions, particularly metabolism of intractable fibers and immune modulation. Disruption of this balance can lead to diseases such as infection, inflammatory bowel syndrome, and obesity. Common symptoms include chronic pain, chronic inflammation, and altered metabolism. Several taxonomic classifications of bacteria have been associated with these diseases, but Recent studies have indicated that these finding are not always statistically valid. An explanation for this is that microbial communities between individuals and even across time can vary substantially even when …

Diverse Community Of Arsenic Resistant Bacteria Display Arsenate Reducing Capabilities, Stephanie Maeda

Undergraduate Honors Theses

Naturally found in soil and water environments arsenic is toxic to many organisms, carcinogenic to humans, and poses a significant public health risk. Yet communities of bacteria found thriving in arsenic ridden environments have evolved mechanisms to tolerate and exploit both oxidation states of this heavy metal (arsenite and arsenate). As the site of an old arsenic mine, Black Mountain Open Space Park in San Diego, California has yielded concentrations of arsenic in the soil between 111-14,800 ppm. Exceeding average arsenic soil concentrations and levels considered safe in the environment by the World Health Organization, we sought to characterize the …

Innovative Approaches In The Discovery Of Aquatic Mycobacteriophages, Janis H. Doss

Biomedical Sciences Theses & Dissertations

Bacteriophages (phages), viruses that infect bacteria, have many applications in medicine, agriculture, molecular biology, and other fields. As antibiotic resistance becomes an increasing problem, interest in phages has grown. The traditional techniques of phage discovery are successful for some phages, but others require modified procedures to achieve detectable host infection.

Mycobacterium is a diverse bacterial genus characterized by a unique cell wall containing mycolic acids, which aids in survival and pathogenesis. The aims of the present research were to isolate mycobacteriophages, use bioinformatics techniques to analyze mycobacterial prophages, and combine genetic analysis with multi-well plate host range studies to identify …

Microbial And Metabolomic Biomarkers Of Feed Efficiency In Angus Cattle, Brooke Ashley Clemmons

Doctoral Dissertations

In Tennessee and the United States, cattle account for a significant proportion of the livestock agricultural sector. In the beef cattle industry, 40-70% of the total cost of production comes from feed costs, resulting in billions of dollars going towards feeding cattle. Thus, identifying methods for improving feed efficiency is imperative for reduced feed input costs and more effective use of environmental and economic resources. The rumen microbiome contributes a vast amount of nutrients for the host ruminant, including approximately 70% of energy precursors as well as vitamins, proteins, and other nutrients. These microbes produce metabolites that are then available …

Whole Genome Sequence Analysis Of A Transmissible Multidrug-Resistance Plasmid Captured Without Cultivation From Poultry Litter, Emma C. Eisemann

Senior Honors Projects, 2020-current

Use of antibiotics in the agricultural industry introduces selective pressure and, consequently, could increase the presence of antibiotic resistant organisms in surrounding environments. One such environment is litter (manure and bedding) produced during large-scale poultry production in the Shenandoah Valley. Litter, with its microorganisms, is commonly applied to fields within the Shenandoah River watershed. Antibiotic resistance (AR) and virulence genes are potentially transmissible between organisms through horizontal gene transfer of genetic mobile elements, for which poultry litter could be a reservoir. The typical, culture-based approach to detecting and analyzing AR plasmids and other mobile genetic elements is limited due to …

Lysogeny And Use Of Mycobacteriophage Pita2, Eleanor Behling, Neocles B. Leontis, Jill Zeilstra-Ryalls

Honors Projects

The CDC has classified antibiotic resistance as the biggest health challenge of our era; every year 2 million lives are impacted and even lost due to resistant bacteria. Bacteriophages provide an alternative route to fighting infections that does not further the development of antibiotic resistance among bacterial species. A bacteriophage replicates inside a bacterial cell and then causes that cell to lyse, an event that kills the bacterial host. However, some phage can integrate their genomes into the host chromosome without causing lysis. The HHMI SEA-PHAGES program has generated a collection of bacteriophage that infect Actinobacteria species. Over 13,000 phages …

Microbiota Characterization Of Poultry Processing Systems And Associated Microbiological Sampling Materials Collected At Commercial Processing Facilities, Jennifer A. Wages

Graduate Theses and Dissertations

The poultry industry and associated regulatory bodies use whole bird carcass (WBC) rinsates to evaluate different stages of broiler processing systems for the prevalence of food-borne pathogens, including Salmonella spp. and Campylobacter spp. Within industry and research groups, the same sample collections are enumerated to determine E. coli, Enterobacteriaceae (EB), and Aerobic Plate Count (APC) microorganisms. Analysis of these indicator microorganisms provides numerical data that can be used to demonstrate the effects of specific process control steps where low occurrences of target pathogens hinder the exclusive use of prevalence data. With the utilization of next generation sequencing (NGS), including analysis …

Evaluation Of Microbiome On Chicken Necrotic Enteritis And Growth Performance, Mussie Abraha

Graduate Theses and Dissertations

Necrotic enteritis (NE) has re-emerged following restriction of antimicrobial usage and costs $6 billion every year worldwide. The primary objective of the studies was to evaluate prevention and treatment of NE using a microbiota metabolic product, secondary bile acid deoxycholic acid (DCA), in drinking water. Day-old birds were tagged and placed in floor pens. In experiments 1 and 2, the birds were infected with Eimeria maxima (Em) at d 18 and C. perfringens at d 23 and 24 and the birds were euthanized at d 26. In experiment 3, birds were infected with Eimeria at d 20 and C. perfringens …

Characterization Of The Broad-Spectrum Inhibitory Capability Of Alcaligenes Faecalis And A. Viscolactis Against Potential Pathogenic Microorganisms, Andrew Fuqua

Undergraduate Honors Theses

The recent rise of multidrug resistant microorganisms has grown from an isolated concern to a massive public health crisis. It has become imperative that scientists look for new ways to combat this issue. Due to the selective pressures of competition, bacteria and other microbes possess a host of defenses and weapons designed to exploit vulnerabilities in other microorganisms. Consequently, the study of these systems and microbial interactions has much to reveal in the search for novel antimicrobial treatments. Previous research from our laboratory has discovered that both Alcaligenes faecalis and Alcaligenes viscolactis, two rarely studied and generally non-virulent bacteria, …

Development Of In Vitro Models To Study The Rapid Extraintestinal Dissemination Of Salmonella., Adarsh Gopinath

Electronic Theses and Dissertations

Salmonella appears in the bloodstream of mice in as little as 15 minutes after oral inoculation and establishes persistent colonies in the spleen and liver. While its pathway to blood is undetermined, this phenomenon is dependent on the activity of Salmonella pathogenicity island 2 (SPI-2) coded type III secretion system (T3SS) and CD18+ phagocytes. We hypothesize that dendritic cells associated with the basal face of the gut epithelium, that are naturally migratory and known to sample for luminal antigens directly transport Salmonella to the bloodstream. This process comprises of at least two phases, dissociation and reverse transmigration. We define dissociation …

Antimicrobial Properties Of An Unknown Microorganism Isolated From The Local Environment, Danielle Duryea

Undergraduate Honors Thesis Projects

Bacterial species that cause disease can usually be combatted with antibiotics; but as the years pass, more and more bacterial pathogens are becoming resistant to these treatments. In fact, the Center for Disease Control has identified eighteen classes of bacteria ranging from urgent to concerning threats due antibiotic resistance (2019), warning the advance of an antibiotic resistance crisis in which untreatable bacterial infections will become a leading cause of death (Bennadi, 2014). The Small World Initiative is a program created in 2012 at Yale University to address the antibiotic crisis through a crowdsourcing effort where undergraduate students are encouraged to …

The Use Of Polymer-Coated Magnetic Nanoparticles For Amelioration Oil Toxicity And Enhancement Microbial Remediation, Amjed Alabresm

Theses and Dissertations

Oil spills from different sources such as natural seeps, waste waters, runoffs and accidental oil spills can have serious effects on the environment while also resulting in potentially major economic damage. Due to the harmful effects of crude oil components on ecosystems, a rapid response to sequestrate oil is required to minimize any resulting environmental impact. Currently used oil removal methods, in addition to their advantages, also have severe limitations. New techniques have been developed including nanotechnology to address these limitations. A new type of nanoparticle (NP), composed of polyvinylpyrrolidone (PVP) coated iron oxide (magnetite), has been successfully used to …

Characterization Of Ph-Based Inactivation Of Respiratory Syncytial Virus (Rsv), Jenna Nosek

Undergraduate Honors Thesis Collection

Respiratory syncytial virus (RSV) is a negative-sense, single-strand RNA virus that affects the upper and lower respiratory system in humans. Currently, RSV is the leading cause of infant mortality worldwide, while also infecting target populations of the elderly and immunocompromised. Significant efforts towards vaccine development have been made due to limited therapeutic options, however, physical instabilities of RSV may hinder movement in vaccine development. We hypothesize that strain-specific differences in stability likely attribute to differences in RSV F protein, as proposed by previous studies about the intrinsically instability of RSV. A panel of recombinant RSV viruses were analyzed for strain-specific …

Bioinformatic Anlayses Of Identified But Uncultivated Microbial Dark Matter Organisms Using Archival Single Amplified Genomes (Sags), Daniel Adam Nuccio

Graduate Research Theses & Dissertations

This project characterizes two independent, previously undescribed microbial dark matter organisms from the candidate phyla JL-ETNP-Z39 and NBK19. These characterizations are based on seven single amplified genomes (SAGs) for JL-ETNP-Z39 and six SAGs for NBK19. All SAGs used were provided by the Bigelow Laboratory for Ocean Sciences Single-Cell Genomics Center (scgc.bigelow.org). Bioinformatic analyses using a combination of publicly available software and custom scripting written in the computer programming languages Python and R were carried out to determine the basic genomic properties, evolutionary relatedness, representative environmental pangenomes, and metabolic properties of these organisms. Analyses indicated that the examined JL-ETNP-Z39 SAGs are …

Group B Streptococcus: Molecular Epidemiology, Pathogenic Profiling And Control Strategies, Katherine Mary Hayes

Theses

Group B Streptococcus (GBS) are frequent colonisers of the gastrointestinal tract of healthy adults with worldwide rates of 18% colonisation; however, they are also opportunistic pathogens capable of causing invasive disease, particularly in neonates and adults with underlying conditions. Indeed, GBS is the leading cause of invasive neonatal disease worldwide with a global incidence of 0.49 per 1000 live births and GBS disease in adults is increasing globally, with estimated rates of 10.9 cases per 100,000 people. While GBS remain largely susceptible to penicillin, there have been reports of reduced beta-lactam susceptibility in certain countries and resistance to other antibiotic …

Analysis Of Bacterial Dna And Water Quality: Surface Water Sampling At Gardner-Webb University, Celsea Reeder

Undergraduate Honors Theses

Part 1 of this experiment was to explore the bacterial types within the surface water on the campus of Gardner-Webb University, located in Boiling Springs, NC. Two water samples were collected from four locations on campus, including the boiling spring, Lake Hollifield, and an adjacent creek. Using the Zymo Research “Quick-DNA^™ Fungal/Bacterial Miniprep Kit,” the bacterial DNA within these samples was isolated and sent to Psomagen, Inc. for analysis. The resulting DNA sequences were analyzed through BLAST, and subsequently interpreted. For the second part of this experiment, one surface water sample was taken from the boiling spring, and another …

Applications Of Hydrodynamic Cavitation In Dairy Manufacturing: Process Development And Standardization, Jae Young Sim

Electronic Theses and Dissertations

The phenomenon of hydrodynamic cavitation involves the formation, growth, and subsequent collapse of bubbles when a given liquid experienced a reduction of pressure below its vapor pressure. The presence of cavitation limits the performance and the safe operation of many machinery and pumps. However, innovation in the design of the hydrodynamic cavitation devices has offered promising applications in the food and dairy industry. Upon collapse of the cavities, the fluid experiences significant mechanical effects (shear and turbulence) as well as instantaneously elevation of the fluid temperature. All these effects can be put to work for mixing, dispersion, particle size reduction, …

Antifungal Defense Molecules From Bacterial Symbionts Of North American Trachymyrmex Ants, Georgia Scherer

CMC Senior Theses

Defensive symbioses, in which microbes provide molecular defenses for an animal host, hold great potential as untapped sources of therapeutically useful antibiotics. Fungus-growing ants use antifungal defenses from bacterial symbionts to suppress pathogenic fungi in their nests. Preliminary chemical investigations of symbiotic bacteria from this large family of ants have uncovered novel antifungal molecules with therapeutic potential, such as dentigerumycin and selvamicin.

In this study, the bacterial symbionts of North American Trachymyrmex fungus-growing ants are investigated for antifungal molecules. Plate-based bioassays using ecologically-relevant fungal pathogens confirmed that these bacteria have antifungal activity. In order to purify and identify the antifungal …

The Persisting Threats Of Cholera: A Cyclical Public Health Problem In Ghana, Rita Laryea Amediavor

Browse all Theses and Dissertations

The prevalence of communicable diseases continues to be one of the continent's leading causes of deaths. Cholera is a waterborne disease triggered by toxigenic strains of the Gram-negative bacteria Vibrio Cholerae O1 strain and less common O139 strain. with symptoms such as severe acute watery diarrhea and vomiting leading to dehydration, progressing to hypovolemic shock and death if not treated timely due to its short incubation period (Pasetto et al., 2018). West African countries are largely portrayed as endemic to cholera, though the dynamics of outbreaks in these developing countries remain largely uncertain. The purpose of the study is to …

Invasive Species Shift Fungal Driven Decomposition In Midwestern Forests, Adam M. Reed

Browse all Theses and Dissertations

Midwestern forests are currently impacted by two prominent invaders, Agrilus planipennis and Lonicera maackii. The Ag. planipennis induced loss of Fraxinus spp. trees can facilitate Lo. maackii invasion, which is likely altering microbial driven forest nutrient cycling. To assess these changes in microbial processes, I conducted litter bag and culture-based decomposition experiments using leaf litter from Acer spp., Quercus spp., F. nigra, F. pennsylvanica, Lindera benzoin, and Lo. maackii. For the culture-based decomposition experiment, I inoculated six species of fungi separately onto both single species and multispecies (half Lo. maackii and half native spp.) leaf litter and measured decomposition rate, …

Comparison Of Fe(Iii) Reduction Rates By Iron-Reducing Bacteria Within Sub Muros Samples From Quadrilátero Ferrífero Iron-Ore Caves, Brazil, Summer Ellis

Williams Honors College, Honors Research Projects

Previous research investigating the speleogenesis of iron-ore caves (IOC) in the Quadrilátero Ferrífero, or “Iron Quadrangle,” of Brazil suggests that microbial iron reduction and subsequent dissolution of the surrounding Fe(III) rich rocks is responsible for cave formation. A soft intra-wall substance (sub muros) containing iron-reducing bacteria (FeRB) was discovered underneath the durable crusts of cave walls. The goal of the study was to determine if reduction rates were comparable between sub muros samples, while additionally observing how different electron donors affect microbial iron reduction. To do this, I compared Fe(III) reduction rates between sub muros samples collected from …

The Role Of Streptococci In Oral Periodontitis: Growth And Disruption Of Yellow Complex Bacteria, Megan Gordon

Theses and Dissertations (Comprehensive)

Periodontitis is described as the inflammation of the periodontium and affects millions of people worldwide. This disease is caused by biofilm growth of certain bacteria on the tooth surface, adjacent to the gum tissue in the oral cavity. There are currently treatments against severe periodontitis, such as oral surgery and use of broad-spectrum antibiotics, but they fail to target the specific bacteria associated with periodontitis. Socransky et al., (1998), identified the preponderance for specific organisms in distinct areas of the subgingival pockets to associate with different bacteria forming consortia and complexes. The yellow complex, containing facultatively anaerobic, Gram positive …

Genomic Insights And Ecological Adaptations Of Deep-Subsurface And Near Subsurface Thermococcus Isolates, Lilja Caitlin Strang

WWU Graduate School Collection

Members of the Archaeal genus Thermococcus are sulfur-dependent hyperthermophiles found in hydrothermal vents throughout the world. Previous analysis of a Thermococcus culture collection containing isolates from the Juan de Fuca Ridge, Gorda Ridge, and South East Pacific Rise using amplified fragment length polymorphism analysis and multilocus sequence typing revealed a distinct clade of Thermococcus isolated from the 1996 megaplume event at Gorda Ridge, indicating that they originated from a deep-subsurface habitat. The aim of this study was to elucidate the functional adaptations that allow for the survival of the Gorda Ridge clade in a deepsubsurface habitat as compared to representative …