Microbiology Commons^™

Psf Sampling In Fluorescence Image Deconvolution, Eric A. Inman

Master's Theses

All microscope imaging is largely affected by inherent resolution limitations because of out-of-focus light and diffraction effects. The traditional approach to restoring the image resolution is to use a deconvolution algorithm to “invert” the effect of convolving the volume with the point spread function. However, these algorithms fall short in several areas such as noise amplification and stopping criterion. In this paper, we try to reconstruct an explicit volumetric representation of the fluorescence density in the sample and fit a neural network to the target z-stack to properly minimize a reconstruction cost function for an optimal result. Additionally, we do …

Identification Of New Substrates From Pseudomonas Aeruginosa Exos, Adam V. Thota

Master's Theses

Pseudomonas aeruginosa is a gram-negative environmental bacterium that uses its intrinsic and acquired antibiotic resistance to infect patients with compromised epithelium, such as cystic fibrosis and burn wounds, and corneal keratitis. In order to cause disease, most strains of P. aeruginosa use the type three secretion system (T3SS) to disrupt cellular signaling and integrity. However, P. aeruginosa may also secrete exotoxins after invading host cells, which prolongs host cell death and maintains an intracellular niche. Of the T3 secreted effector toxins, ExoS, specifically the ADP-ribosyltransferase activity of ExoS, was shown to delay the death of invaded corneal cells, but it …

Role Of Tetraspanins In Sars-Cov-2 Fusion And Entry, Marcos Saul Santiago Figueroa

Master's Theses

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for the pandemic the world has been facing in recent years. Even as new countermeasures are developed, there is still much we don’t know in terms of its entry into host cells. Tetraspanins are transmembrane proteins that are near ubiquitous amongst cell types. They fulfill numerous roles, including that of a viral co-receptor. Here, we considered whether tetraspanins, specifically CD9, CD63, and CD81 influence SARS-CoV-2 fusion and entry. Using ACE2-LgBit and tetraspanin overexpressing EVs and HeLa cells, we find that the presence of excess tetraspanins inhibit fusion and entry. However, we …

Interrogating Antagonistic Differences Among Pseudomonas Aeruginosa Cdia Alleles, Haotian Yang

Master's Theses

Bacteria are complex social organisms that live within mixed communities where microorganisms compete for limited resources. We studied Contact-Dependent Growth Inhibition (CDI), a particular type of competition mediated by a type 5b, or two-partner, secretion system that is widely spread among Gram-negative bacteria. CdiB and CdiA make up the two-partner secretion system that mediates CDI. CdiB translocates CdiA to the cell surface, and CdiA delivers its C-terminal toxin domain to the target cell. The C-terminal toxin (Tox) domain of CdiA is highly diverse. We studied Tox domains of various Pseudomonas aeruginosa clinical isolates. We found different strains that contain unique …

Bisacodyl Limits Chikungunya Virus In Vitro And Is Broadly Antiviral, Natalie June Lomascolo

Master's Theses

Identifying novel antivirals requires significant time and resource investment, and the continuous threat of viruses to human health necessitates commitment to antiviral identification and development. Developing antivirals requires years of research and validation, and recent outbreaks have highlighted the need for preparedness in counteracting pandemics. One way to facilitate development is to repurpose molecules already used clinically. By screening such compounds, we can accelerate antiviral development. Here, we screened compounds from the National Institutes of Health’s Developmental Therapeutic Program for activity against chikungunya virus, an alphavirus that is responsible for a significant outbreak in the Americas in 2013. Using this …

Investigating The Role Of Nagc Reveals It To Be A Global Regulator, Natasha Peterson

Master's Theses

Bacterial colonization of host surfaces can result in symbiotic or pathogenic associations.As bacteria becomes more resistant to antibiotics, pathogenic bacterial colonization poses an increasing risk to human health. Symbiotic associations that have beneficial outcomes for both the host and the microbe can serve as models to investigate how bacterial processes affect colonization of host surfaces. The symbiotic relationship between the Gram-negative, bioluminescent bacterium Vibrio fischeri and the Hawaiian bobtail squid is one such model (E. V. Stabb & Visick, 2013). The study of this exclusive relationship has developed in a way where in vitro phenotypes can predict the ability of …

Calcium-Mediated Induction Of Vibrio Fischeri Es114 Biofilms, Katia Elizabeth Semenchuk

Master's Theses

Vibrio fischeri bacteria form biofilms that facilitate symbiotic colonization of the Hawaiian bobtail squid, Euprymna scolopes. These host-associated biofilms require production of the SYP polysaccharide. However, it has been difficult to observe robust biofilm formation in vitro in a laboratory setting using wild-type strain ES114; instead, most work investigating syp-dependent biofilm formation has relied on genetically altered strains. Guided by recent findings that increasing calcium induces c-di-GMP (Tischler et al., 2021), a molecule known in other systems to promote biofilm formation, the question was posed as to whether high levels of calcium alone could promote syp-dependent biofilm formation by ES114. …

Polycyclic Aromatic Hydrocarbons And The Microbiomes Of Two Benthic Species, Samantha Ells

Master's Theses

The presence of oil has been shown to affect the microbiomes of the water column, sediments, and organisms, both by altering the diversity and the composition of those microbial communities. If the microbiome is altered it may no longer provide benefits to its host organism, impacting its ability to survive. Thus, it is important to understand the effects of large-scale contamination events including the Deepwater Horizon oil spill. This study set out to examine the effects of oil exposure on the microbiome of two benthic species, southern flounder (Paralichthys lethostigma) and eastern oysters (Crassostrea virginica) to further understand the effects …

Development Of A Portable Low-Moisture Food Pasteurization Device Using Rf Heating, Eric Jason Ohata

Master's Theses

Bacterial presence in low-moisture foods such as flour, cereals, baby formula, and spices, have become a concern due to sanitizing challenges. The food industry currently focuses on wet food sanitation as opposed to low-moisture foods because of bacteria’s inability to reproduce in low water activity media. Traditionally, food processing RF heating pasteurizes in mass quantities while an equivalent consumer device does not exist the market today. A consumer product would help eliminate food waste by providing an easy way to sanitize food and extend shelf life. The Portable Food Pasteurization (PFP) project is an interdisciplinary project involving the Electrical Engineering, …

The Role Of Med31 And Med12 In Directing Adipogenesis Of Human Adult-Derived Mesenchymal Stem Cells, Joseph Straub

Master's Theses

Selective gene expression is crucial in maintaining the self-renewing and multipotent properties of stem cells. Mediator is a large, evolutionarily conserved, multisubunit protein complex that modulates gene expression by relaying signals from cell type-specific transcription factors to RNA polymerase II. In humans, this complex consists of 30 subunits arranged in four modules: head, middle, tail, and kinase. In our introduction, we show the state of the field of Mediator study with a focus on the critical kinase module. In the following chapters, we used siRNA knockdowns to investigate the roles of the highly-conserved core subunit MED31 and the kinase module …

How Historic Shipwrecks Influence Dispersal Of Deep-Sea Microbiomes, Rachel Moseley

Master's Theses

This thesis investigates how historic shipwrecks potentially shape dispersal of deep-sea microbiomes. Processes impacting dispersal of microbiomes around artificial structures in the ocean and how they connect to other deep-sea habitats is unknown. Dispersal processes are explained for some macroorganisms by theories in ecology, specifically the Theory of Island Biogeography and the Wooden Stepping Stone hypothesis. These have not been investigated for microorganisms, and thus this work will probe if they are applicable to microbial biogeography in this habitat. Experiments were conducted to establish new “island-like” features in near proximity (25-125 m) to wooden-hulled historic shipwrecks in the northern Gulf …

The Response Of Marine Synechococcus To A Landscape Of Environmental Stressors: A Proteomic Exploration, Dana E. Michels

Master's Theses

In the field of marine microbial ecology, many questions remain unanswered with regards to the physiological trade-offs made by phytoplankton to maximize growth (e.g., nutrient acquisition) and minimize loss (e.g., predation defenses). These tradeoffs, which occur at the cellular level, have wide reaching impacts on food web dynamics and global biogeochemical cycles. In the first chapter, we explored the use of a non-canonical amino acid (NCAA) technique, bioorthogonal non-canonical amino-acid tagging (BONCAT), in phytoplankton model systems. This technique has potential to work well in natural systems by enabling isolation of only newly synthesized proteins during an incubation period with the …

Characterizing A Novel Cocksackievirus B3 Protease Mutant And Its Response To Polyamine Depletion, Bridget Hulsebosch

Master's Theses

Enteroviruses, including Coxsackievirus B3 (CVB3), are pervasive pathogens that cause significant disease, including cardiomyopathies. Unfortunately, no treatments or vaccines are available for infected individuals. We identified the host polyamine pathway as a potential drug target, as inhibiting polyamine biosynthesis significantly reduces enterovirus replication in vitro and in vivo. Here, we show that CVB3 is sensitive to polyamine depletion through the polyamine analog diethylnorspermidine (DENSpm) which enhances polyamine catabolism through induction of polyamine acetylation. We demonstrate that CVB3 acquires resistance to DENSpm via mutation of the 2A protease, which enhances proteolytic activity in the presence of DENSpm. Resistance to DENSpm occurred …

Assessing The Role Of The Interaction Between Polyamines And Heparan Sulfate In Coxsackievirus B3 Viral Infection, Oreoluwa Omoba

Master's Theses

Non-polioenteroviruses are highly infectious viruses that typically cause mild asymptomatic cases but can cause severe disease such as aseptic meningitis and dilated cardiomyopathy. Coxsackievirus B3 (CVB3) is a member of this group and is prevalent in the United States and Eastern Asia. Viruses like CVB3 rely on the host cell for many molecules in order to replicate. One such set of molecules is the polyamines. Polyamines are small, positively charged molecules that play a role in a multitude of cellular processes including RNA/DNA stabilization, gene expression, translation, and regulating membrane fluidity. Previous studies have shown CVB3 relies on polyamines for …

Elucidating Regulatory Connections Between The Second Messenger Cyclic-Di-Gmp With Biofilm And Motility Phenotypes Of Vibrio Fischeri, Ali Razvi

Master's Theses

The symbiosis between the gram-negative marine bacterium Vibrio fischeri and its host Euprymna Scolopes occurs when the bacteria reaches and colonizes the light organ. This pathway to colonization begins in the outer marine environment and finishes within the deep crypts of the light organ, with states of biofilm formation and dispersal in between. This switch between biofilm formation to dispersal is important as the bacteria transitions from the surface on the light organ into a pore leading to the interior of the light organ. This dispersal is regulated by LAP system which is dependent on the second messenger c-di-GMP. Through …

Chga-Depleted Urothelial Amp And Immune Response During Urinary Tract Infection, Theadora Jane Ceccarelli

Master's Theses

Women are 8 times more likely than men to experience a urinary tract infection (UTI) with up to 60% of women reporting a UTI in their lifetime. This significant healthcare burden is caused by the infection of uropathogenic Escherichia coli in the urinary tract. At this time, treatment is limited to antibiotic therapy, which is challenged by antibiotic-resistance. These factors make UTIs an NIH priority and are motivation for our research.CgA is a protein of the granin family co-released with catecholamines from neuroendocrine cells throughout the body. Pro-hormone convertases post-translationally cleave CgA into peptides, like catestatin, which can have antimicrobial …

Study Of Plants Used Against Infections By California Native American Tribes, Maria J. Rojas

Master's Theses

The objectives of this research were to evaluate the antibacterial activity and to determine the chemical composition of a list of medicinal plants used by Native Americans in California. Artemisia californica, Mimulus aurantiacus, Equisetum telmateia, Equisetum hyemale, and Marah fabacea were selected from a list of plants reported as having been used for ailments related to infections by tribes located in California. The extracts obtained through steam distillation from E. telmateia, E. hyemale and M. fabacea were assayed for in vitro antibacterial activity against 16 Gram-negative and 6 Gram-positive bacteria using disk diffusion assays and measuring the diameters of inhibition …

Microplastic Abundance, Distribution And Impacts On Sargassum-Associated Juvenile Fishes In The Gulf Of Mexico, Olivia Lestrade

Master's Theses

Microplastics are a concern in marine environments because they are highly durable, ubiquitous, and can be mistaken for food and ingested by small organisms. Pelagic Sargassum, an important habitat for larval and juvenile stages of many fish species, is found in large surface aggregations, and may provide complex structure in which microplastics become trapped. This could lead to greater risk of microplastic ingestion by fish early life stages associated with Sargassum habitats. To better understand the impacts of microplastics within Sargassum communities, this study examined 1) microplastic concentrations and ingestion by juvenile fishes associated with Sargassum; 2) the microbial …

Analysis Of Biofilm Remediation Capacity For Octenyl Succinic Anhydride (Osa), A Bioactive Food Starch Modifier Compound, Matthew R. Borglin

Master's Theses

Matthew R. Borglin

This thesis demonstrates efficacy of Octenyl Succinic Anhydride (OSA), as a biofilm sanitizer. Biofilms allow bacteria to adhere to solid surfaces with the use of excreted polymeric compounds. For example, surfaces found in food production or processing facilities such as the interior of a raw milk holding tank, are some of the most susceptible to biofilm contamination. When present, biofilms can cause a variety of negative effects, which include; reduction of product shelf life, corrosion, and outbreaks of foodborne illnesses. The close association of biofilms with the majority of foodborne illness cases led the US Environmental Protection …

Analysis And Optimization Of Colorimetric Nanosensors For Rapid Detection Of Microbes In Water, Ruby A. Lang

Master's Theses

Access to safe water is a basic human right recognized by the United Nations General Assembly in 2010 (WHO, 2020). However, a least 2.2 billion people globally still are without safely managed water services meaning they use a drinking water source that can be contaminated with faeces (WHO, 2020). With such a pressing global health issue, it is clear that improvement to water systems is important and required in the Agenda 2030 Sustainable Development Goals (SDGs). However, to improve water systems and prove they are safe water sources, water quality testing must occur. A solution to this issue is the …

Elucidating The Molecular Pathways Involved In The Fitness And Water Stress Response Of Pseudomonas Synxantha 2-79 In The Plant Rhizosphere, Clint Henry D. Pablo

Master's Theses

Pseudomonas synxantha 2-79 is a biocontrol agent that represents beneficial indigenous rhizobacteria that are broadly distributed in the Pacific Northwest, USA and flourish in the rhizosphere of commercially grown wheat under surprisingly arid conditions. The molecular adaptation of 2-79-like bacteria to plants growing in dry soils is poorly understood. We hypothesized that the ability of 2-79 to colonize and persist in the rhizosphere of water-stressed plants is underpinned by the formation of hydrating biofilms and the utilization of root exudates that contain plant-derived osmoprotectants called quaternary ammonium compounds (QACs). We tested this hypothesis by identifying waters stress response pathways in …

Development Of An Attenuated Zika Virus By Editing The 5’ Untranslated Region, E. Ashley Thompson

Master's Theses

Zika Virus (ZIKV) is a mosquito-transmitted flavivirus that usually causes no symptoms to mild febrile in humans, and it has been regarded as an insignificant pathogen to public health. However, recent outbreaks of ZIKV infection have revealed that ZIKV can cause severe neurological effects in adults, such as Guillain-Barre Syndrome (GBS), and in infants whose mothers acquired the virus during pregnancy, causing Congenital Zika Syndrome (CZS). Currently, no approved vaccine is available and there is a critical need to develop an effective and safe vaccine. While most vaccine developmental strategies target the viral prM-E protein of ZIKV, we aimed to …

Investigating The Role Of Hypusine In Viral Translation And Infection, Jeremy Joseph Esin

Master's Theses

Polyamines are small, positively charged, molecules found in eukaryotic cells with roles in cellular function and viral infection. in previous work we have shown RNa viruses, including bunyaviruses, to be sensitive to polyamine depletion, but exact mechanisms are unknown. Bunyaviruses are emerging pathogens that cause encephalitis, hemorrhagic fevers, and meningitis. Rift Valley Fever Virus is a bunyavirus that infects both humans and livestock with severe mortality and morbidity. in previous work done with Rift Valley Fever Virus polyamine depleted results in noninfectious interfering particles without affecting the shape or size of the particles. Here, I investigate a specific metabolic pathway …

Zika Virus Inactivates Polyamine Catabolism Via Alternative Splicing To Enhance Infection, Marion Lea Graham

Master's Theses

During viral infection, the virus and host must compete for resources inside the cell. One of these resources is polyamines. Polyamines are small, positively charged molecules that are found in all eukaryotic cells. They play a key role in several cellular functions including growth and proliferation, transcription and translation, and membrane stability. Viruses also rely polyamines for productive replication, utilizing them during DNA/RNa polymerization, nucleic acid packaging, and protein synthesis. in response to a virus infecting a host cell, the host cell will begin to regulate polyamine levels as a way to combat the infection. Polyamine levels are regulated by …

A Susceptibility Study Of Urinary Isolates To Methenamine Hippurate And Its Metabolites, Nancy Sloan

Master's Theses

Urinary Tract Infection (UTI) is one of the most common bacterial infections in adult women. Treatment is one of the most common reasons for antibiotic prescription. the rise of antibiotic-resistant infections coupled with the shortage of new antibiotics creates an urgent need for alternative non-antibiotic treatments. One alternative is methenamine hippurate (MH), which is a non-antibiotic prophylaxis. It is reported to be active against some common uropathogens including Escherichia coli and Enterococcus species, but its effects on the newly discovered bladder microbiota (urobiome) are undocumented. Given this knowledge gap, this study aimed to evaluate the drug's spectrum of susceptibility on …

Determining Interactions Between Bacteriophage And Urinary Microbiome Lysogens, Michael Bochert

Master's Theses

Bacteriophages (phages) are viruses that infect bacteria. They are the most abundant life forms on earth. It is estimated that there are approximately 10³⁰ phages on the planet (Chibani-Chennoufi et al. 2004). They outnumber their bacterial hosts approximately 10 to 1. Bacteria inhabit most niches on the planet including the human body, where they play critical roles in health and disease. Since phages shape bacterial populations the same way that viruses, such as smallpox and measles, shaped human populations, it is important to understand how they interact with their bacterial hosts. The majority of the phages that we know of …

The Thiol Specific Antioxidant (Tsa1) Gene Is Required For Survival In Macrophages And Oxidative Stress Resistance In Histoplasma Capsulatum, Lauren Kennedy

Master's Theses

Histoplasma capsulatum(Hc) is a pathogenic fungus that causes one of the most common invasive fungal respiratory diseases, Histoplasmosis. Histoplasmaundergoes a dimorphic shift from mold to yeast which is crucial to pathogenesis of the organism.

The thiol specific antioxidant gene, Tsa1,is strongly upregulated in the yeast (pathogenic) morphotype. This data led to the hypothesis that this gene plays a role in protecting Hcfrom host mediated oxidative attack. To characterize Tsa1 function, a knockdown strain (tsa1-RNAi) was created by RNAi gene silencing. Expression of Tsa1in the tsa1-RNAistrain was reduced to 10% that …

The Effect Of Historic Shipwrecks On Sediment Microbiomes In The Northern Gulf Of Mexico, Melissa Brock

Master's Theses

Microorganisms are ubiquitously distributed across all aquatic habitats, but it is the environmental conditions of a habitat that determine which microorganisms can thrive in terms of abundance or metabolic activity. Habitats that experience consistent physical and chemical environmental conditions repeatedly favor specific taxonomic groups which may result in a microbial assemblage that is commonly associated with that habitat (i.e., a core microbiome). Core microbiomes have been identified for a variety of natural marine habitats including methane seeps, wood falls, octocoral, and deep-sea sediments. However, it was unknown if the presence of man-made structures on the seafloor, including historic shipwrecks, also …

The Evolutionary Diversity And Biological Function Of Phenazine Metabolite Biosynthesis In Burkholderia Spp, Samuel Hendry

Master's Theses

Burkholderia encompass a group of ubiquitous Gram-negative bacteria that include numerous saprophytes, as well as several species that cause infections in animals, immunocompromised patients, and plants. Some species of Burkholderia produce colored redox-active secondary metabolites called phenazines (Phz). In the model opportunistic pathogen Pseudomonas aeruginosa, phenazines strongly contribute to the competitiveness, formation of biofilms, and virulence in multiple models of infection. Similar depth of knowledge on the diversity, biosynthesis, and biological functions of phenazines in Burkholderiais missing. This project aimed to bridge this gap in knowledge by focusing on phenazine pathways of B. lata and closely related …

Development Of A Prolyl Endopeptidase Expression System In Lactobacillus Reuteri To Reduce The Clinical Manifestation Of Celiac Disease, Kara Lynn Jew

Master's Theses

Celiac Disease (CD) is an autoimmune disorder that emerges due to the ingestion of gluten, a protein found in a variety of common grains such as wheat, rye, and barley. Approximately 1 in 100 individuals in the US suffer from CD, making it the most commonly diagnosed gastrointestinal disorder (Ciclitira et. al., 2005). These proline-rich gluten peptides are resistant to proteolysis and accumulate in the duodenum of the small intestine. Once in the duodenum, these peptides illicit an autoimmune response resulting in villous atrophy. Current treatment for CD requires a rigorous adherence to a gluten-free diet. Nevertheless, gluten-containing grains are …