Microbiology Commons^™

Regulation Of Acetyl Phosphate-Dependent Acetylation And Identification Of Novel Lysine Acetyltransferases In Escherichia Coli, David George Christensen

Dissertations

Over billions of years, organisms have organized chemical reactions into metabolic pathways to sustain life. However, metabolic substrates can undergo many uncatalyzed, extra-metabolic reactions. Acetyl phosphate (AcP), an intermediate of the acetate fermentation pathway in E. coli, is one such metabolite that has been shown to non-enzymatically acetylate hundreds of proteins. This diverse set of targets suggests that acetylation could be a way for the cell to sense its nutritional status and regulate protein activity accordingly. However, how E. coli regulates acetylation, if at all, is unknown.Previous work showed that acetylation becomes pronounced in stationary phase cells. I determined that …

The Role Of Polyamines In Enteroviral Attachment, Thomas Mckenzie Kicmal

Master's Theses

Polyamines are small polycationic molecules with flexible carbon chains that are found in all eukaryotic cells. Polyamines are involved in the regulation of many host processes and have been shown to be implicated in viral replication. Depletion of polyamine pools in cells with FDA approved drugs restricts replication of diverse RNA viruses. Viruses can exploit host polyamines to facilitate packaging, transcription, translation, and protease activity but other mechanisms remain largely unknown. Picornaviruses, including Coxsackievirus B3 (CVB3), are sensitive to depletion of polyamines and remain a significant public health threat. We employed CVB3 as a model system to investigate a potential …

A Possible Role Of Clostridium Difficile Association With The Gi Tract In Relapsing Clostridium Difficile Infection, Asiyya Ashraf

Master's Theses

Clostridium difficile infection (CDI), caused by the anaerobic spore-former C. difficile, is a major cause of health-care associated morbidity. CDI occurs in individuals that have a disrupted GI tract microbiota, typically in patients who are on antibiotic therapy. After recovery from an episode of CDI, approximately, 15-30% of patients experience relapse of disease. The mechanism of relapse is not well understood. It is plausible for relapse to occur if C. difficile cells and spores associate with the GI tract mucosa during infection and persist after infection. However, little is known about the association of cells and spores with the GI …

Assessing The Role Of A Putative Peptidase In The Processing And Transport Of Aip In The Agr Quorum Sensing System Of Staphylococcus Aureus, Chance Joseph Cosgriff

Master's Theses

Staphylococcus aureus infections are a global burden on human health. The wide rangeof infectious diseases caused by S. aureus stem from the production of myriad virulence factors that manipulate host immune responses and promote bacterial survival. These virulence traits, coupled with the rise in antibiotic resistance, demonstrate a need for novel therapeutics to control infection. Many current therapeutic initiatives focus on targeting the virulence factors produced by S. aureus or the regulatory systems that control their expression such as the Accessory gene regulatory system, Agr.Initial characterization of a putative S. aureus peptidase MroQ indicates a major role in regulation of …

Longitudinal Urinary Microbiome Studies: A Need To Transition To Voided Urine, Baylie Hochstedler

Master's Theses

It is now established that the bladder is not sterile; it contains communities of microbes (microbiota). While the healthy bladder microbiota have been defined using expanded quantitative urine culture (EQUC) and urines obtained by transurethral catheterization (TUC), longitudinal and population studies have not previously been possible. These studies cannot be done using TUC urines, as it would be impractical to catheterize participants daily. Instead, we must transition to using voided urines.In order to make this switch, we have addressed three main issues raised by using voided urine. First, we showed that EQUC out performs standard urine culture on voided urines …

Bacterioplankton Biogeography Of The Mississippi River Basin, Jason Taylor Payne

Electronic Theses and Dissertations

Bacterioplankton are important structural and functional components of river networks yet their biogeographical patterns in these systems are largely unknown. Using Illumina sequencing of the 16S rRNA gene we characterized variation in bacterioplankton community alpha diversity (within-sample richness of operational taxonomic units OTUs) and beta diversity (between-sample differences in composition) (1) along a 1 300-km downstream reach of the Mississippi River (2) over a range of temporal scales in the Lower Mississippi River (LMR) and (3) along cross-sectional transects repeated monthly for six months across the LMR floodplain. Particle-associated assemblages were richer in bacterial OTUs and more productive than free-living …

Pyocyanin, A Virulence Factor Produced By Sepsis-Causing Pseudomonas Aeruginosa, Promotes Adipose Wasting And Cachexia, Nika Larian

Theses and Dissertations--Pharmacology and Nutritional Sciences

Sepsis is a leading cause of death among critically ill patients that results in metabolic alterations including hypercatabolism, lipoatrophy, and muscle wasting, contributing to the development of cachexia. Septic cachexia is associated with loss of body weight, fat mass, and lean mass and dysregulated immune function. There are currently no efficacious treatment strategies for septic cachexia, and nutritional interventions have limited success in preventing hypercatabolic wasting. Pyocyanin is a virulence factor produced by sepsis-causing Pseudomonas aeruginosa that has been shown to activate the aryl hydrocarbon receptor (AhR), increase inflammation, and produce reactive oxygen species. Thus, pyocyanin represents a novel mechanistic …

Development And Evaluation Of Nonradioactive Methods For Monitoring T Lymphocyte Response To Equine Arteritis Virus (Eav) In Horses, Annet Kyomuhangi

Theses and Dissertations--Veterinary Science

Target cell lysis is the hallmark of immune effector responses of cytotoxic T lymphocytes (CTL), natural killer (NK) cells, and monocytes. The most commonly used assay to measure target cell lysis is the 51Cr release assay and is considered the ‘gold standard’. However, this assay has many disadvantages that limit its use by most laboratories. Thus, several alternative assays have been developed. Some of these alternative assays are more sensitive, easy to perform and do not use radioactive elements.

In this study, four of these assays were evaluated for their ability to detect antigen- specific CTL responses in equine blood. …

Modulation Of Inflammatory Cytokine, Chemokine, And Toll-Like Receptor Genes And Transcriptome Analysis Of Equine Endothelial Cells Following Infection With Equid Herpesvirus-1, And Equine Arteritis Virus., Saranajith Wangisa Dunuwille

Theses and Dissertations--Veterinary Science

EHV-1 is a double-stranded DNA virus whereas EAV is a positive sense, single-stranded RNA virus. Therefore, genetically, they are very different from one another. However, both these viruses are endotheliotropic and thus, infect and replicates in equine endothelial cells resulting in vasculitis. Vasculitis is central to the pathogenesis of these two viruses. Thus, the main objective of this thesis was to investigate the inflammatory and innate immune responses of EECs that contribute towards the development of vasculitis following infection with EHV-1 and EAV in-vitro. Since proinflammatory cytokines and chemokines produced by endothelial cells play a significant role in the …

Mechanisms Of Type-I Ifn Inhibition: Equine Herpesvirus-1 Escape From The Antiviral Effect Of Type-1 Interferon Response In Host Cell, Fatai S. Oladunni

Theses and Dissertations--Veterinary Science

Equine herpesvirus-1 (EHV-1) is one of the most important and prevalent viral pathogens of horses causing a major threat to the equine industry throughout most of the world. EHV-1 primarily causes respiratory disease but viral spread to distant organs enables the development of more severe sequelae; abortion and neurologic disease. In order to produce disease, EHV-1 has to overcome the innate barrier of the type-I interferon (IFN) system in host cells. However, the underlying mechanisms employed by EHV-1 to circumvent the type-I IFN response in host cells are not well understood. In this project study, using molecular techniques, we explored …

A Novel Sparse Compositional Technique Reveals Microbial Perturbations, Cameron Martino, James T. Morton, Clarisse A. Martoz, Luke R. Thompson, Anupriya Tripathi, Rob Knight, Karsten Zengler

Faculty Publications

The central aims of many host or environmental microbiome studies are to elucidate factors associated with microbial community compositions and to relate microbial features to outcomes. However, these aims are often complicated by difficulties stemming from high-dimensionality, non-normality, sparsity, and the compositional nature of microbiome data sets. A key tool in microbiome analysis is beta diversity, defined by the distances between microbial samples. Many different distance metrics have been proposed, all with varying discriminatory power on data with differing characteristics. Here, we propose a compositional beta diversity metric rooted in a centered log-ratio transformation and matrix completion called robust Aitchison …

Metaproteomics Reveal That Rapid Perturbations In Organic Matter Prioritize Functional Restructuring Over Taxonomy In Western Arctic Ocean Microbiomes, Molly P. Mikan, H. Rodger Harvey, Emma Timmins-Schiffman, Michael Riffle, Damon H. May, Ian Salter, William S. Noble, Brook L. Nunn

OES Faculty Publications

We examined metaproteome profiles from two Arctic microbiomes during 10-day shipboard incubations to directly track early functional and taxonomic responses to a simulated algal bloom and an oligotrophic control. Using a novel peptide-based enrichment analysis, significant changes (p-value < 0.01) in biological and molecular functions associated with carbon and nitrogen recycling were observed. Within the first day under both organic matter conditions, Bering Strait surface microbiomes increased protein synthesis, carbohydrate degradation, and cellular redox processes while decreasing C1 metabolism. Taxonomic assignments revealed that the core microbiome collectively responded to algal substrates by assimilating carbon before select taxa utilize and metabolize nitrogen intracellularly. Incubations of Chukchi Sea bottom water microbiomes showed similar, but delayed functional responses to identical treatments. Although 24 functional terms were shared between experimental treatments, the timing, and degree of the remaining responses were highly variable, showing that organic matter perturbation directs community functionality prior to alterations to the taxonomic distribution at the microbiome class level. The dynamic responses of these two oceanic microbial communities have important implications for timing and magnitude of responses to organic perturbations within the Arctic Ocean and how community-level functions may forecast biogeochemical gradients in oceans.

Relationship Between The Cervical Microbiome, Hiv Status, And Precancerous Lesions, Cameron Klein, Daniela Gonzalez, Kandali Samwel, Crispin Kahesa, Julius Mwaiselage, Nirosh Aluthge, Samodha Fernando, John T. West, Charles Wood, Peter C. Angeletti

Nebraska Center for Virology: Faculty Publications

Nearly all cervical cancers are causally associated with human papillomavirus (HPV). The burden of HPV-associated dysplasias in sub-Saharan Africa is influenced by HIV. To investigate the role of the bacterial microbiome in cervical dysplasia, cytobrush samples were collected directly from cervical lesions of 144 Tanzanian women. The V4 hypervariable region of the 16S rRNA gene was amplified and deep sequenced. Alpha diversity metrics (Chao1, PD whole tree, and operational taxonomic unit [OTU] estimates) displayed significantly higher bacterial richness in HIV-positive patients (P = 0.01) than in HIV-negative patients. In HIV-positive patients, there was higher bacterial richness in patients with …

Nitrogen Assimilation In Picocyanobacteria Inhabiting The Oxygen-Deficient Waters Of The Eastern Tropical North And South Pacific, Montserrat Aldunate, Carlos Henriquez-Castillo, Qixing Ji, Jessica Lueders-Dumont, Margaret R. Mulholland, Bess B. Ward, Peter Von Dassow, Osvaldo Ulloa

OES Faculty Publications

Prochlorococcus and Synechococcus are the most abundant free-living photosynthetic microorganisms in the ocean. Uncultivated lineages of these picocyanobacteria also thrive in the dimly illuminated upper part of oxygen-deficient zones (ODZs), where an important portion of ocean nitrogen (N) loss takes place via denitrification and anaerobic ammonium oxidation. Recent metagenomic studies revealed that ODZ Prochlorococcus have the genetic potential for using different N forms, including nitrate and nitrite, uncommon N sources for Prochlorococcus, but common for Synechococcus. To determine which N sources ODZ picocyanobacteria are actually using in nature, the cellular 15N natural abundance (δ15N) and assimilation rates of …

A Full-Ocean-Depth Rated Modular Lander And Pressure-Retaining Sampler Capable Of Collecting Hadal-Endemic Microbes Under In Situ Conditions, Logan M. Peoples, Matthew Norenberg, David Price, Madeline Mcgoldrick, Mark Novotny, Alexander Bochdansky, Douglas H. Bartlett

OES Faculty Publications

The hadal zone remains one of the least studied environments because of its inaccessibility, in part because of hydrostatic pressures extending to 110 MPa. Few instruments are capable of sampling from such great depths. We have developed a full-ocean-depth-capable lander that can be fit with sampling packages for the collection of still images, video, motile megafauna, and hadal seawater. One payload includes a pressure-retaining sampler (PRS) able to maintain seawater samples under in situ pressure during recovery. We describe the technical specifications of the lander and the PRS and preliminary results from three deployments at depths in excess of 10,700 …

The Role Of Cody In Hydrogen Peroxide Tolerance And Biofilm Formation Of Streptococcus Mutans, Kaitlin M. Rioux, Brandaliz Santiago-Narvaez

Honors Program Theses

Streptococcus mutans is the major cariogenic bacterium in the oral cavity. S. mutans causes disease by three main virulence factors: acid production, acid tolerance, and biofilm formation. S. mutans outcompetes various other species in the oral cavity, however it is affected by hydrogen peroxide production by these species. Based on previous findings and this research, hydrogen peroxide is shown to extend the lag phase of S. mutans but does not kill it unless present in extremely high amounts that are generally not produced by commensal species like S. sanguinis and S. gordonii in the oral cavity. CodY is a global …

Assessing The Temporal Dynamics Of The Lower Urinary Tract Microbiota And The Effects Of Lifestyle, Travis Kyle Price

Dissertations

Urinary tract infections (UTIs) are among the most common bacterial infections in humans, accounting for $3.5 billion in health care expenditures yearly in the United States alone. Yet, treatments for UTI have seen little innovation over the past decade. As demonstrated in other body sites, such as the vagina and gastrointestinal (GI) tract, acute and infectious diseases often have indirect microbial contributions which serve as intriguing new targets for therapies. The recent discovery of the existence of a resident community of bacteria (i.e., microbiota) in the bladders of both women and men, represents a novel avenue for targeting UTIs. However, …

Long-Term Land Management Practices And Their Effect On Soil Health And Crop Productivity, Thomas Joseph Muratore Jr.

Theses and Dissertations--Plant and Soil Sciences

Agricultural intensification reliant on monocrops could change soil health in a way that does not support maximum crop productivity. Twenty-nine-year-old no-till field plots at the University of Kentucky Spindletop research farm showed a significant reduction in corn yields from continuous corn plots compared to those from plots in various types of rotation. The objective of this study was to determine what role soil microbes might play in yield reduction and how management and time effects microbial community structure. Samples were collected from the following treatments: continuous corn (CC), continuous soybean (SS), a 2-year corn/soybean rotation (CCSS), Corn in rotation with …

Improvement Of Dried Distillers’ Grains With Solubles Utilizing Pretreatments And Fungal Fermentation, Burgandy Zschetzsche

Electronic Theses and Dissertations

Limited inclusion of distillers’ grains in animal feed is primarily due to low protein and high fiber content. Other elements, such as phytate phosphorus levels in DDGS are still an issue for monogastric animals such as swine, poultry, and fish. Furthermore, unabsorbed phytic acid in manure poses a high risk for environmental pollution, because bacteria can hydrolyze phytic acid into free phosphorus that can result in algal blooms and eutrophication of surface or ground water. The objective of this thesis was to improve the nutritional value of corn dried distillers’ grains with solubles (DDGS). This was done utilizing submerged fungal …

Dissecting A Role For Polyamines In Rift Valley Fever Virus Infection, Vincent Mastrodomenico

Master's Theses

Bunyaviruses are emerging viral pathogens that cause encephalitis, hemorrhagic fevers, and meningitis. Rift Valley fever virus is a particularly devastating bunyavirus, infecting both humans and livestock with significant morbidity and mortality. By coordinating several host and viral processes Rift Valley fever virus is able to produce infectious virions. Polyamines are small, positively-charged host-derived molecules that play diverse roles in human cells and in infection. We previously demonstrated that polyamines are crucial for RNA viruses; however, the mechanisms by which polyamines function remain unknown. Here, we investigated polyamines' role in the replication of the Rift Valley fever virus (vaccine strain MP-12). …