Life Sciences Commons^™

Investigating The Escape Mechanism Of Sre Bearing Mrna Transcripts During Viral Host Shutoff, Daniel Macveigh-Fierro

Doctoral Dissertations

During viral infection, the virus and host clash for control over gene expression in an evolutionarily arms race that has raged for thousands of years. During lytic replication, Kaposi’s sarcoma associated herpesvirus (KSHV) triggers a massive RNA decay event known as host shut off. This causes over 70% of all RNA to be degraded suppressing the host antiviral response while freeing resources for viral replication. Our lab focuses on a subset of transcripts that escape from this viral degradation event using a cis acting 3’ UTR element known as a “SOX resistant element” or SRE. Although we have identified a …

The Impacts Of Environment And Host Evolutionary Relationships On Lemur Microbiota, Rachel B. Burten

Doctoral Dissertations

Recent studies have shown that the mammal microbiome is modified by environmental conditions, and that reduced microbiome functionality is associated with host health issues. Microbiome data in wild and captive primate populations can therefore be used to assess their health as they encounter a variety of environments. Comparative studies of the microbiome can also inform disease ecology, conservation, and captive management strategies tailored to different primate species. Therefore, this study examines how the hair, oral, and gut microbiota of nine wild and captive lemur species are determined by host phylogenetic relationships and host environment. I found that host species identity …

Exploring Soil Microbial Dynamics In Southern Appalachian Forests: A Systems Biology Approach To Prescribed Fire Impacts, Saad Abd Ar Rafie

Doctoral Dissertations

Prescribed fires in Southern Appalachian forests are vital in ecosystem management and wildfire risk mitigation. However, understanding the intricate dynamics between these fires, soil microbial communities, and overall ecosystem health remains challenging. This dissertation addresses this knowledge gap by exploring selected aspects of this complex relationship across three interconnected chapters.

The first chapter investigates the immediate effects of prescribed fires on soil microbial communities. It reveals subtle shifts in porewater chemistry and significant increases in microbial species richness. These findings offer valuable insights into the interplay between soil properties and microbial responses during the early stages following a prescribed fire. …

The Identification Of Small Molecule Inhibitors To Candida Albicans Phosphatidylserine Synthase, Yue Zhou

Doctoral Dissertations

Candida albicans phosphatidylserine (PS) synthase, encoded by the CHO1 gene, has been identified as a potential drug target for new antifungals against systemic candidiasis due to its importance in virulence, absence in the host and conservation among fungal pathogens. This dissertation is focused on the identification of inhibitors for this membrane enzyme. Cho1 has two substrates: cytidyldiphosphate-diacylglycerol (CDP-DAG) and serine. Previous studies identified a conserved CDP-alcohol phosphotransferase (CAPT) binding motif present within Cho1, and here we revealed that mutations in all but one conserved amino acid within the CAPT motif resulted in decreased Cho1. For serine, we have predicted a …

Impact Of Cadmium Stress On Soil Virus Reproduction And The Persistence Of Viruses Under Abiotic Conditions, Zhibo Cheng

Doctoral Dissertations

Soil viruses are ubiquitous and greatly impact the structure and function of soil microbial communities, with their effects modulated by various environmental factors. This study investigates the inactivation of naturally occurring soil viruses in sterilized soil, as well as the effects of cadmium (Cd) exposure and carbon (C) and nitrogen (N) nutrient availability on the population dynamics of virus-host communities in agricultural soil.

Lab-scale slurry and unsaturated experiments were performed to examine virus inactivation in the absence of host bacteria in sterilized soil. In slurry experiments, virus abundance declined by over 90% after 10 days of incubation. The addition of …

Integration Of Raman Spectroscopy And Python-Based Data Analysis For Advancing Neurobiological Research, Natalie E. Dunn

Doctoral Dissertations

The field of Raman spectroscopy continues to expand into biological applications due to its usefulness as a non-invasive technique that can be utilized qualitatively and quantitatively. However, the inherent weakness of Raman scattering leads to the need for each collected spectra to undergo a preprocessing step to remove noise, background drift, and cosmic rays. Biological research in particular needs large datasets due to the increased variability in samples. As datasets grow, the need to perform preprocessing on each individual spectra becomes daunting. Often, these steps are done by hand with the help of specialized software programs. Preprocessing can be accelerated …

Exploring The Regulation Of A Bacillus Pumilus Bioplastic Degrading Protein, Elise Keara Phillips

Doctoral Dissertations

Biodegradable plastics are being adopted to reduce the environmental impacts associated with petroleum-based plastics, including long residence times, and reliance on fossil fuels as a feedstock. Microbial degradation of bioplastics is still poorly understood, which leads to inconsistent degradation outcomes and challenges for implementation. Bacillus pumilus B12 degrades the biodegradable bioplastic polylactic acid (PLA) via the protease AprE, but the regulatory mechanisms controlling aprE expression in B. pumilus B12 are largely unknown. To explore this question, we established developed a conjugation- based method to introduce DNA into B. pumilus B12 and diverse Bacillus species that is applicable to environmental and …

Novel Microbial Guilds Implicated In N2o Reduction, Guang He

Doctoral Dissertations

N₂O is a long-recognized greenhouse gas (GHG) with potential in global warming and ozone depletion. Terrestrial ecosystems are a major source of N₂O due to imbalanced N₂O production and consumption. Soil pH is a chief modulating factor controlling net N₂O emissions, and N₂O consumption has been considered negligible under acidic conditions (pH <6). In this dissertation, we obtained solids-free cultures reducing N₂O at pH 4.5. Furthermore, a co-culture (designated culture EV) comprising two interacting bacterial population was acquired via consecutive transfer in mineral salt medium. Integrated phenotypic, metagenomic and metabolomic analysis dictated that the Serratia population excreted certain …

Detection And Control Of Environmentally Transmissible Viruses, Anand R. Soorneedi

Doctoral Dissertations

Viruses, owing to their ubiquitous nature and ability to infect almost every other species, have long been a subject of interest for scientists. Some of the virus species can be very deadly to humans and animals alike and can impose a huge economic and health burden across the world. The recent CoVID-19 pandemic underscores the importance of timely detection for developing effective intervention strategies. Unfortunately, some of the virus species that cause significant health and economic impacts do not have robust and reliable detection methods due to several reasons. In some cases, despite having gold standard methods for detection of …

A Biochemical Approach To Characterize A Divergent Trypanosoma Brucei Mitochondrial Dna Polymerase, Polib, Stephanie B. Delzell

Doctoral Dissertations

Trypanosoma brucei is a single-celled parasitic protist that causes African sleeping sickness in people and nagana in cattle in sub-Saharan Africa. T. brucei and related trypanosomatid parasites contain an unusual catenated mitochondrial genome known as kinetoplast DNA (kDNA) composed of dozens of 23 kb maxicircles and thousands of 1 kb minicircles. The kDNA structure and replication mechanism are divergent from other eukaryotes and essential for parasite survival. POLIB is one of three Family A DNA polymerases that are independently essential to maintain the kDNA network, and has been implicated in minicircle replication. However, the division of labor among the paralogs, …

A Shift In Rna Fate: Investigating The Role Of C19orf66 During Kshv Lytic Replication, William Rodriguez Jr.

Doctoral Dissertations

During viral infection, virus and host clash for control of the cell in a conflict that ultimately drives the evolution of both sides and has lasting consequences in the form of pathogenesis. At the heart of this struggle is a contest for control of cellular gene expression, a struggle epitomized by an evolutionary tug-of-war for supremacy over RNA fate. During lytic replication, Kaposi’s sarcoma-associated herpesvirus (KSHV) triggers a massive RNA decay event known as viral “Host Shutoff” which decimates greater than 70% of the host transcriptome, simultaneously suppressing the cellular anti-viral response and freeing host resources for viral replication. However, …

Roles Of Soil Pores In Determining Water Retention And Microbial Dynamics, Huihui Sun

Doctoral Dissertations

Soil pore provides an essential habitat for microbial communities to participate in various biogeochemical processes. The complex pore spaces, which are defined by the arrangement of particles of varying sizes, govern the distribution of water for microbial dispersal and movement and microbial interaction with one another, such as bacterial cell-to-cell and bacterium-virus interactions. This research focused on exploring how soil pores influence soil water retention and bacterial interactions and quantify the spatial distribution of bacteria and viruses in fine-scale of soil pores. Using a mathematical model, this study simulated a soil water retention curve based on the relationship between soil …

Induction And Evasion Of Neutrophil Extracellular Traps By Campylobacter Jejuni And Its Implication In Disease, Sean M. Callahan

Doctoral Dissertations

Campylobacteriosis, the foodborne disease caused by Campylobacter spp., infects one out of 10 individuals every year. C. jejuni accounts for 90% of these infections resulting in numerous postinfectious disorders including the development of colorectal cancer, Guillain-Barré syndrome, irritable bowel syndrome, and reactive arthritis. Despite its large impact on human health, the host immune response to the bacterium is largely uncharacterized. Chapter two of this dissertation addresses the development of neutrophil extracellular traps (NETs) within human and ferret campylobacteriosis. We observed NET-associated proteins increase in the feces of C. jejuni-infected patients and that in vitro C. jejuni induces NETs, which …

Investigating Drivers Of Algal Bloom Succession In Lake Erie, Brittany Zepernick

Doctoral Dissertations

Harmful Algal Blooms (HABs) are algae undergoing prolific, unregulated growth. A well-documented HAB taxa is the cyanobacterium Microcystis spp., which induces anthropogenic, ecological, and economic consequences due to the production of toxins and biomass which results in lake hypoxia. Microcystis spp. blooms are globally distributed in freshwater systems, with climate change and the aquatic continuum serving to further exacerbate bloom distribution, duration, and frequency. Thus, there is a need to elucidate the factors driving the ecological success of Microcystis spp., and the ecological “failures” of their competitors, such as diatoms. In Lake Erie, a seasonal pattern of algal bloom succession …

Unraveling Plant-Microbe-Microbe Interactions: Host And Microbial Colonization Determinants Across Experimental Scales, Bridget O'Banion

Doctoral Dissertations

Plant microbiomes are assembled and modified through a complex milieu of biotic and abiotic factors. Despite dynamic and fluctuating contributing variables, specific host and microbial mechanisms are likely important mediators of interactions. We combine information from large-scale datasets across diverse plant hosts with experimental genetic manipulation assays in model Arabidopsis seedlings to converge on a conserved role for metabolite production and transport in mediating host-microbe interactions. Applying a diverse set of analytical tools to microbial communities at various complexity levels will advance our knowledge of the scalability of observed phenotypes and, ultimately, help to decipher the mechanisms dictating plant–microbe interactions …

Bifidobacterial Metabolism Of Fucosylated Human Milk Oligosaccharides Influences Structure And Function Of The Infant Gut Microbiome, Liv R. Dedon

Doctoral Dissertations

Human milk contains human milk oligosaccharides (HMOs) that are indigestible and pass intact through the infant gastrointestinal tract where they are available for microbial metabolism. HMOs incorporate the same monosaccharide building blocks but vary structurally in primary sequence of monomeric components. Primary sequences are further diversified by degree of polymerization, branching, and secondary modifications such as fucosylation. Fucosylated HMOs (fHMOs) are highly abundant and can account for over 30% of total HMOs. Infant-colonizing Bifidobacterium longum subsp. infantis (B. infantis) possesses a specialized gene cluster conveying the ability to metabolize fHMOs. This study presents an in-depth analysis of B. …

Identification Of The Type Eleven Secretion System (T11ss) And Characterization Of T11ss-Dependent Effector Proteins, Alex S. Grossman

Doctoral Dissertations

Host-associated microbes live in dangerous environments as a result of host immune killing, nutrient provisioning, and physiological conditions. Bacteria have evolved a host of surface and secreted proteins to help interact with this host environment and overcome nutrient limitation. The studies included within this dissertation describe the identification of a novel bacterial secretion system which has evolved to transport these symbiosis mediating proteins. This system, termed the type eleven secretion system (T11SS), is present throughout the Gram negative phylum Proteobacteria, including many human pathogens such as Neisseria meningitidis, Acinetobacter baumanii, Haemophilus haemolyticus, and Proteus vulgaris. Furthermore, …

The Sos Response In Escherichia Coli K12: An Exploration Of Mutations In Lexa And Reca Using Fluorescence Microscopy, Steven Van Alstine

Doctoral Dissertations

Faithful replication of the genome is paramount for maintaining the fitness of an organism. Therefore, life has evolved inducible mechanisms to be able to repair damaged DNA and maintain evolutionary fitness. The SOS response is a highly conserved DNA damage inducible response that is tightly regulated. Multiple factors contribute to the ability of the cell to perform proper DNA repair and induction of the SOS response including the amount of RecA, mutations in RecA that affect competition for DNA, and other proteins that interact with the RecA filament. The complex relationship between RecA and LexA is the subject of this …

Principles Of Aaa+ Proteases, Samar Mahmoud

Doctoral Dissertations

ATPases associated with diverse cellular activities (AAA+) proteases in bacteria help maintain protein homeostasis by degrading misfolded and regulatory proteins. While a handful of protein targets for these proteases have been identified in Caulobacter crescentus and other organisms, more research is needed to elucidate mechanisms that govern substrate specificity. In the second chapter of this thesis, I will elaborate on how AAA+ substrate specificity is less rigid than previous work has suggested and how limiting ATP or mutations can alter substrate preferences of the ClpXP protease. In the third chapter, I will highlight our efforts to use a quantitative proteomics …

Biomedical Applications Of Protein Films And Polymeric Nanomaterials, Sanjana Gopalakrishnan

Doctoral Dissertations

Biomaterials are widely applied for the diagnosis and treatment of numerous diseases. In addition to fulfilling specific biological functions, biomaterials must also be non-toxic, biocompatible, and sterilizable to be regarded as safe-for-use. Polymers are excellent candidates for fabricating functional biomaterials due to their wide availability and varied properties and may be natural or synthetic. Polymer precursors are fabricated into coatings, foams, scaffolds, gels, composites, and nanomaterials for several biomedical applications. This dissertation focuses on two types of polymeric biomaterials – protein-based materials and synthetic polymeric nanoparticles. Proteins are biopolymers that naturally occur with a variety of structural and functional properties. …

An Investigation Of Intrinsic And Extrinsic Factors That Influence Soil Microbial Succession During Human Decomposition, Allison R. Mason

Doctoral Dissertations

Decomposer organisms play a vital role in terrestrial nutrient cycling, breaking down complex organic compounds and providing nutrients for primary producers. Microbial communities and other decomposers associated with a vertebrate carcass, or the “necrobiome”, are critical for degradation and recycling of soft tissues following vertebrate mortality. The impact of microbial decomposers in vertebrate decomposition has been shown with regard to soil microbial communities, where the presence of soil microbes impact decomposition rate and undergo succession. However, current assessments of microbial ecology within decomposition-impacted soils have primarily focused on one aspect of microbial succession: succession of bacterial taxa for forensic applications. …

Enterobactin-Based Immune Interventions Against Gram-Negative Bacterial Infections, Huiwen Wang

Doctoral Dissertations

Enterobactin (Ent)-mediated iron acquisition is critical for many Gram-negative bacteria to survive in the host. Given the bacteriostatic effect of lipocalin-2 resulting from its potent Ent-binding ability, immune interventions directly targeting Ent is a promising antimicrobial strategy against Gram-negative bacterial infections. Moreover, hyperimmune egg yolk antibody is an emerging passive immune agent for the control of bacterial infections. Oral administration of anti-Ent egg yolk antibody may confer passive immune protection against Ent-dependent enteric pathogens. In this dissertation study, multidisciplinary approaches in conjunction with different model systems (in vitro, in vivo, and ex vivo) were used to …

Metabolism In The Colonocyte: Where Bacteria And Diet Meet, Bohye Park

Doctoral Dissertations

The host-microbiome interaction and the functions of microbial-derived metabolites, including butyrate, are important in colon health. Butyrate is the preferred energy source in colonocytes and has multiple physiological functions in the colon. A metabolic shift in colonocytes toward increased glucose utilization from butyrate oxidation is followed by several critical genetic modifications in cancerous colonocytes. Moreover, it has been suggested that the gut-microbiota composition is influenced by environmental factors, such as diets and the host's physiological status. Therefore, an understanding of the role of colonocyte metabolism toward impacting the host-microbiota commensal relationship would be an important step in understanding the functional …

Cat Covid, Cmv And Chemokines, Oh My!, Trevor Hancock

Doctoral Dissertations

Cytomegalovirus (CMV) is an important pathogen infecting most humans worldwide. CMV infection within immunocompromised individuals can cause severe morbidity and potential mortality. Disease during CMV infection is due to virus dissemination and subsequent inflammation. Host immune cells lie at the intersection potentially mediating both. The CMV-encoded viral chemokine vCXCL-1 is a proposed virulence factor in mouse models increasing immune cell recruitment and disease. However, the primary immune cell mediator is undetermined. To identify targets, Chapter 2 examines CXCR2 expression (receptor for vCXCL-1) among various mouse tissues and human peripheral blood under steady-state conditions. In vitro, isoforms of HCMV’s vCXCL-1 …

Mechanisms By Which Xenorhabdus Nematophila Interacts With Hosts Using Integrated -Omics Approaches, Nicholas C. Mucci

Doctoral Dissertations

Nearly all organisms exist in proximity to microbes. These microbes perform most of the essential metabolic processes necessary for homeostasis, forming the nearly hidden support system of Earth. Microbial symbiosis, which is defined as the long-term physical association between host and microbes, relies on communication between the microbial community and their host organism. These interactions among higher order organisms (such as animals, plants, and fungi) and their bacteria links metabolic processes between interkingdom consortia. Many questions on microbial behavior within a host remain poorly understood, such as the colonization efficiency among different microbial species, or how environmental context changes their …

The Spatial, Temporal, And Ecological Constraints Of Plant-Host Associated Microbial Communities, Jonathan Dickey

Doctoral Dissertations

With the development of next generation sequencing technology, ecologists have recently been able to describe microbial communities across a wide array of niches at an accelerated pace. De-novo-based patterns in richness and relative abundance have been described for bacterial and fungal communities in terrestrial, aquatic, and host-associated microhabitats. A recent synthesis has shown that these communities exhibit similar geographical patterns that have been traditionally described for plant and animals. Yet, there is a lack of hypothesis-based research for host-associated microbial communities. Throughout this dissertation, I will address how spatial scale, sequencing resolution, and manipulative rainfall exclusion govern host-associated microbial communities …

Making Sense Of Soil Microbiome Complexity For Plant And Ecosystem Function In A Changing World, Kendall K. Beals

Doctoral Dissertations

Soils contain the highest biodiversity on Earth. While the importance of the soil microbiome for larger-scale ecological phenomena such as nutrient and carbon cycling, plant growth and plant community dynamics is well-established, the fundamental question of the ecological and evolutionary function of this immense belowground microbial diversity for plant and ecosystem function still remains a great challenge in microbial ecology research. The objective of this dissertation is to understand how the importance of soil microbial community composition for plant and ecosystem function and how changes to soil microbial community composition from climate change-induced disturbance events, specifically fire, influence plant and …

Multi-Omic Systems Biological Analysis Of Host-Microbe Interactions, Piet Jones

Doctoral Dissertations

Systems biology offers the opportunity to understand the complex mechanisms of various biological phenomena. The wealth of data that is produced, at an increasing rate, provides the potential to meet this opportunity. Here we take an applied approach to integrate multiple omic level data sources in order to generate biologically relevant hypotheses. We apply a novel analysis pipeline to model both, in concert, the microbial and transcriptomic signature from COVID-19 positive patients. We show patients may suffer from an increased microbial burden, with an increased pathogen potential. Gene expression evidence further shows patients may exhibit a compromised barrier immunity, owing …

Size Progression Of Oxygenic Photogranules (Opgs) And Its Effect On Opg Wastewater Treatment, Ahmed S.A. Abouhend

Doctoral Dissertations

In recent years, the oxygenic photogranule (OPG) process has gained increasing interest because of its potential to treat wastewater without supplemental aeration. Oxygenic photogranules (OPGs) are dense spherical aggregates comprised of phototrophic and nonphototrophic microorganisms. In OPG wastewater treatment reactors, photogranules grow in number as well as in size. The primary goal of this dissertation was to investigate how OPGs grow in size and how the growth affects their structure and functions. We found that OPGs undergo structural changes as they grow bigger in size. As OPGs grow larger, filamentous cyanobacteria become enriched while other phototrophic microbes diminish significantly. OPGs …

Community Assembly And Stress Response Of Grassland Phyllosphere Bacteria, Emily Bechtold

Doctoral Dissertations

Grasslands are an important ecosystem with potential to help stabilize food security and reduce greenhouse gas levels. As global temperatures rise, weather patterns are predicted to drastically change. The resulting increase in intensity, duration, and frequency of drought in important grassland areas will not only affect agricultural production, but also increase grassland susceptibility to fire, disease, and soil erosion. Thus, developing ways to sustainably promote grassland health and production is essential to increase food security and reduce environmental strain. Microbes in the phyllosphere, or aerial surface of plants, promote host fitness through phytohormone and nutrient production, increased stress tolerance, and …