Microbiology Commons^™

Adenoviral-Vectored Epigraph Vaccine Elicits Robust, Durable, And Protective Immunity Against H3 Influenza A Virus In Swine, Erika M. Petro-Turnquist, Matthew J. Pekarek, Nicholas Jeanjaquet, Cedric Wooledge, David J. Steffen, Hiep Vu, Eric A. Weaver

Nebraska Center for Virology: Faculty Publications

Current methods of vaccination against swine Influenza A Virus (IAV-S) in pigs are infrequently updated, induce strain-specific responses, and have a limited duration of protection. Here, we characterize the onset and duration of adaptive immune responses after vaccination with an adenoviral-vectored Epigraph vaccine. In this longitudinal study we observed robust and durable antibody responses that remained above protective titers six months after vaccination. We further identified stable levels of antigen-specific T cell responses that remained detectable in the absence of antigen stimulation. Antibody isotyping revealed robust class switching from IgM to IgG induced by Epigraph vaccination, while the commercial comparator …

Single-Molecule Super-Resolution Imaging Of Geobacter Sulfurreducens Under Anaerobic Conditions, Ziyi Hu

McKelvey School of Engineering Graduate Student Theses & Dissertations

Geobacter sulfurreducens are anaerobic bacteria capable of making electrical contacts with other organisms and extracellular electron acceptors. The challenge of imaging live Geobacter bacteria is maintaining anaerobic conditions during the imaging process. In this thesis, we augment a single-molecule localization microscope (SMLM) with a home-built anaerobic imaging chamber and use constant argon bubbling to maintain oxygen-free imaging conditions. To validate the imaging protocol, we use the transient binding of Nile red to resolve the spherical morphology of lipid-coated glass spheres with nanoscale resolution. However, when imaging Geobacter, the distribution of Nile red localizations is non-uniform, both between different cells …

Characterization Of Lps As A Virulence Factor In Burkholderia Cepacia During Plant And Human Infection, Jack Klahr, Elizabeth Danka

Biology Senior Theses

Burkholderia cepacia is a gram-negative bacterium first characterized as the causative agent of sour skin in onion crops. More recently, B. cepacia has become a clinical concern as an opportunistic pathogen that can colonize the upper respiratory tract of cystic fibrosis patients and increase mortality in these patients. Infection is exacerbated by the intrinsic resistance to antibiotics found in this genus of organisms. Additional virulence factors help the bacteria persist in the host during infection. However, few of these factors have been described. In this work, we characterized a putative virulence factor that was first identified through an onion infection …

The Effects Of Aloe Vera On The Human Gut Microbiome, Samantha Leon

Honors Theses

The human gut microbiome is a diverse community of bacteria influenced by multiple factors including host age, geographical location, and diet. This study observed the effects of aloe vera juice on the human gut microbiome. Fecal samples were taken every two weeks over a period of one year; the subject consumed aloe vera juice in alternating cycles of consumption “on” and abstaining “off” with each cycle lasting for eight weeks. The bacterial community in each sample was characterized using 16S rRNA gene sequencing. Sequences were identified and their relative abundances were used to look for patterns distinguishing “on” and “off” …

An Analysis Of The Sensory Characteristics And Microbial Constituents Of Kefir, Katherine Quay Robinson

Honors Theses

Kefir is a fermented beverage produced using sugar water or milk, kefir grains, either water kefir grains or milk kefir grains, respectively, and microbial aggregates consisting of diverse bacteria and yeasts. As kefir and fermented beverages in general are increasing in popularity, research related to optimal sensory characteristics and consumer-approval is vital. Desirable sensory characteristics for kefir beverages include low sour taste/aroma, high sweetness, carbonation, and high viscosity (Surja et al., 2019; Margareth et al., 2020). Production methods of kefir can alter sensory characteristics, but sensory characteristics are likely strongly influenced by the microbial composition of the grains/product. Thus, an …

The Development Of A Primer Payload With Microparticles For Uti Pathogen Identification Using Polythymidine- Modified Lamp Primers In Droplet Lamp, Jonas Otoo

KGI Theses and Dissertations

Nucleic acid amplification tests (NAATs) are among the diagnostic tests with the highest sensitivity and specificity. However, they are more complex to develop than other diagnostic tests such as biochemical tests and lateral flow immunoassay tests. Polymerase chain reaction (PCR) is the gold standard for NAATs. PCR requires thermal cycling to achieve clonal amplification of the target pathogen DNA for diagnosis. Thermal cycling poses a challenge in the development of PCR diagnostics for point-of-care (POC) settings. Loop-mediated isothermal amplification (LAMP) offers an isothermal method for NAATs diagnostics. The advancement of the microfluidics field significantly enhances the development of LAMP diagnostics …

Eco-Interoception: What Plants, Fungi And Protista Have Taught My Body, Sara Riley Dotterer

Art Theses and Dissertations

To me, ecology is the relational, full-body awareness that I am made up of and deeply connected to everything around me; and for better or worse, this is reciprocal. I form ecotones, an ecological transitional zone between two ecosystems, with the world around me. I use this ecotonal lens to blur binaries and dissolve boundaries between me and the world “outside my body.” During my Masters of Fine Arts at Southern Methodist University, I have continuously explored and represented the lives of various more-than-human species outside of my body, including plants, fungi and protista through an ecotonal lens. Although these …

Development Of Recombinant Varicella-Zoster Virus Vaccines Expressing Sars-Cov-2 Antigens, Cody Wilson

Honors Theses

The COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus (SARS-CoV-2) continues to cause infections and deaths around the globe. Since its emergence, scientists have worked diligently to discover and produce vaccines to battle the virus. While they successfully created and distributed vaccines, the vaccines are not without flaw. These vaccines do not grant long term immunity, require multiple booster shots, and have not prevented the emergence of viral variants. In this study, the SARS-CoV-2 receptor binding domain (RBD), which is a key part of the spike protein (S), and nucleocapsid (N) genetic sequences were inserted into the live varicella …

Mechanistic Examination Of Protist-Mediated Plant Growth Through The Comparative Development Of Medicago Truncatula, Shane Connolly

University Scholar Projects

Protists are known to increase plant growth through two main mechanisms: the microbial loop and the alteration of the root microbiome. The microbial loop is a nutrient recycling method in which protists provide inorganic nitrogen ions to the plant. Alteration of root microbiome leads to the removal of plant pathogens and shifting communities towards plant growth-promoting bacteria (PGPB). This study aimed to elicit which mechanism could produce the largest boost in shoot weight for Medicago truncatula. A series of microcosm experiments were explored in which M. truncatula was grown with variable microbiome structures to allow for mechanism differentiation. The …

Comparative Analysis Of Rhizosphere And Endosphere Microbiome Of Different Blueberry Species (Vaccinium Sp.), Niladri Bhowmik

Master's Theses

Blueberries are an important agricultural commodity in all over the United States. Due to its health benefits, there is a huge demand globally, thus expanding the industry. Breeding programs are essential to maintain such industries. Challenges that play a role in contemporary breeding programs are various biotic and abiotic stress factors. Studies have shown that microorganisms are recruited by plants to alleviate them during stressful conditions. Though blueberries have been cultivated for about 100 years, how the microbiome has been affected due to this is poorly understood. We hypothesized that interspecific crosses and artificial selection have significantly changed the microbiome …

The Response Of Microcystis And The Microbiome To Exogenous Hydrogen Peroxide, Bryan A. Puma

Masters Theses, 2020-current

Microcystis spp. produce prolific annual blooms in freshwater systems worldwide. The success of these blooms depends heavily on the Microcystis spp. overcoming environmental factors such as oxidative stress caused by reactive oxygen species (ROS) like hydrogen peroxide. Most Microcystis genomes do not contain many genes for responding to oxidative stress, including those that encode the enzyme catalase. Other cyanobacteria such as Prochlorococcus that also lack hydrogen peroxide degrading enzymes have been shown to benefit from partnerships with heterotrophic bacteria. Microcystis spp. may also receive similar benefits from heterotrophic bacteria in the phycosphere. To test this hypothesis, we examined the …

Causes And Consequences Of Lasr Mutant Selection In Pseudomonas Aeruginosa Populations, Dallas L. Mould

Dartmouth College Ph.D Dissertations

Change is the only constant in life, and these changes, though random in nature, can have consequences. Quorum sensing is heterogeneous in phenotype and prone to negative selection. In P. aeruginosa, the regulator LasR is frequently non-functional in phylogenetically diverse isolates. Through repeated experimental evolution and mathematical modeling, we show that differences in growth enable lasR mutant evolutionary success and this requires a system enabling metabolic choices, known as carbon catabolite repression (or catabolite repression). The differences in catabolite repression between wild type and lasR mutants enable altered metabolite preferences, and the resulting differences in metabolic states enable intraspecies …

Elucidating The Impact Of Sos-Response Timing In On Escherichia Coli Survival Following Treatment With Fluoroquinolone Topoisomerase Inhibitors, Stephanie Schofield

Honors Scholar Theses

Antibiotic treatment failure is a public health crisis, with a 2019 report stating that roughly 35,000 deaths occur in the United States yearly due to bacterial infections that are unresponsive to antibiotics (1). One complication in the treatment of bacterial infection is antibiotic persistence which further compromises our battle to effectively treat infection. Bacterial persisters can exist in clonal bacterial cultures and can tolerate antibiotic treatment by undergoing reversible phenotypic changes. They can survive drug concentrations that their genetically identical kin cannot. Some persisters remain in a slow growing state and are difficult to target with current antibiotics. A specific …

A Critical Assessment Of The Role Of Clpxp-Mediated Proteolysis In The Development And Differentiation Of Chlamydia Trachomatis, Nicholas A. Wood

Theses & Dissertations

Chlamydia trachomatis, an obligate intracellular human pathogen, is the etiological agent of the most commonly reported bacterial sexually transmitted infection and the leading cause of preventable infectious blindness worldwide. In adapting to an intracellular niche, Chlamydia has evolved to eliminate superfluous genes from its chromosome. Despite its highly reduced genome, Chlamydia undergoes a complex developmental cycle in which the bacteria differentiate between two functionally and morphologically distinct forms: the infectious, non-replicative elementary body (EB) and the non-infectious, replicative reticulate body (RB). An EB initiates infection by binding to and inducing uptake into a host cell. Within the cell, the EB …

The Role Of Sinorhizobium Meliloti Type Ivb Pili Protein On Legume Nodule Morphology And Infection, Alexis Chun

Honors Thesis

The symbiotic relationship between legumes and rhizobial soil bacteria leads to the conversion of nitrogen gas to biologically available forms of nitrogen. A critical step in this interaction is bacterial attachment to the host root. A variety of factors are involved in attachment, such as the Type IV pili (Tfp). In pathogens, Tfpb mediates tight adherence to host cells, promoting colonization, however in symbiotic interactions, the role of Tfpb–encoded by pilA genes–is not well understood. To assess the role of Tfpb pili during symbiosis, Melilotus alba plants were inoculated with wild type Sinorhizobium meliloti or ΔpilA deletion mutants; roots were …

The Future Is Now In Twisted Coil Polymer Actuators (Tcpa), Ryan Ronquillo

Electronic Theses and Dissertations

This thesis aimed to fabricate and test twisted coiled polymer actuators (TCPA) to understand the mechanical and thermal aspects of this artificial muscle fiber. The purpose of this thesis was to find a linear relationship using the LVDT sensor, fabricating TCPA fibers, and interpreting the data. The project tested whether nylon/polymer could be used as a better artificial muscle fiber.

This research accomplished three goals: (1) designing and fabricating a system capable of creating supercoiled muscle fibers consistently, (2) calibrating the Linear Variable Differential Transformer (LVDT) and Core, and (3) analyzing/interpreting the data of the Twisted Coiled Polymer Actuators (TCPA) …

On The Resurrection Of Microbes: An Eco-Christological Approach To The Resurrection, Denys Janiga Osb

Obsculta

This article attempts to bring ecology and Christology into conversation through a transdisciplinary approach. It looks at the pastoral implications of eco-anxiety, resurrection ecology in the field of biotechnology, Church teaching on the Resurrection, and concludes by initiating an interpretation of the resurrection appearances in the Gospel of Luke through the lens of the microbiome.

Effect Of Temperature On The Microbiome Of A Laboratory Reared Colony Of Haemaphysalis Longicornis Ticks, Brianna Mitchell

Poster Presentations

Honors research poster.

Haemaphysalis longicornis is a species of tick native to eastern Asia, including eastern China, Japan, eastern Russia, and Korea. To date, it has invaded and has now established its existence in Australia, New Zealand, and several of the Pacific Islands. This species of tick has recently been introduced to the United States, having a confirmed sighting in November 2017 on a sheep farm in New Jersey and since establishing populations in 18 states along the east coast and Appalachia. Based on the existing locations of H. longicornis in its native regions, as well as in the United …

Immunomodulatory Effects Of Resolvin D2 In A Model Of Infection, Prem Yugandhar Kadiyam Sundarasivarao

Graduate School of Biomedical Sciences Theses and Dissertations

Dysregulated hyperinflammatory host immune response to underlying bacterial infections is a characteristic of sepsis. In sepsis, bacteria often trigger abnormal hyperinflammatory responses which can cause multiple organ failure and if sustained can lead to an immunosuppressive phase where the host is susceptible to secondary infections caused by opportunistic bacteria like Pseudomonas aeruginosa (P. aeruginosa). In our studies, we used a 2-hit model of cecal ligation and puncture (CLP) followed by P. aeruginosa secondary lung infection to investigate cellular and molecular mechanisms in the beneficial action of resolvin D2 (RvD2). Resolvins of the D-series are a group of fatty acids known …

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 …

The Alphaviral Capsid Protein Inhibits Irak1-Dependent Tlr Signaling To Promote Pathogenesis., V. "Trey" Landers

Electronic Theses and Dissertations

Alphaviruses are positive-sense RNA viruses spread by mosquitos. They can cause a severe multi-joint febrile arthritis or encephalitis resulting in death or life-long cognitive impairments. To date there are no approved antiviral therapeutics or vaccine strategies for the treatment of alphaviruses creating a critical need to better understand the host pathogen interactions of alphaviruses that enable pathogenesis. It is known that alphaviruses evade innate immune responses by shutting down host transcription and translation, but these methods are dependent on viral gene expression and leave a critical time frame during early infection before viral gene expression has begun that the virus …

Roles For Mycobacterial Dinb2 In Frameshift And Substitution Mutagenesis, Pierre Dupuy, Shreya Ghosh, Allison Fay, Oyindamola Adefisayo, Richa Gupta, Stewart Shuman, Michael S. Glickman

Publications and Research

Translesion synthesis by translesion polymerases is a conserved mechanism of DNA damage tolerance. In bacteria, DinB enzymes are the widely distributed promutagenic translesion polymerases. The role of DinBs in mycobacterial mutagenesis was unclear until recent studies revealed a role for mycobacterial DinB1 in substitution and frameshift mutagenesis, overlapping with that of translesion polymerase DnaE2. Mycobacterium smegmatis encodes two additional DinBs (DinB2 and DinB3) and Mycobacterium tuberculosis encodes DinB2, but the roles of these polymerases in mycobacterial damage tolerance and mutagenesis is unknown. The biochemical properties of DinB2, including facile utilization of ribonucleotides and 8-oxo- guanine, suggest that DinB2 could be …

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 …

Exploring Extreme Viruses With Ken Stedman, Kenneth Stedman

PDXPLORES Podcast

In this episode of PDXPLORES, Ken Stedman, Professor of Biology at Portland State University, discusses his distinguished virology career. From discovering his love of extreme viruses at Max Planck Institute for Biochemistry in Munich, Germany, to heading the Advanced Molecular and Cell Biology Research Laboratory and the eXtreme Virus Lab, Steadman has investigated extreme virus structures, function, genetics, and evolution for more than 25 years. He is the co-founder of the research lab Center for Life in Extreme Environments as well as the Chief Scientific Officer of StoneStable, Inc. Adding to his scientific accomplishments and accolades, Professor Stedman …

The Stringent Response In Pseudomonas Aeruginosa Influences The Phenotypes Controlled By The Gac/Rsm System, Michael Shawn Hooker

Electronic Theses and Dissertations

Pseudomonas aeruginosa is a ubiquitous, opportunistic pathogen that causes acute and chronic infections. Infection is typically initiated via motile and virulent strains. After exposure to stressors, acute infections make both genotypic and phenotypic switches to a chronic, sessile strain. This is due to intricate regulatory networks directing gene expression in response to stressors. One network, GacA/GacS, has been established to control virulence factors. The stringent response of bacteria is mediated by alarmones produced primarily by RelA which responds to starvation.

To study the effect of the stringent response on the virulence switch. A series of experiments were run in both …

Cyclic-Di-Gmp Regulates Salmonella Typhimurium Infection Of Epithelial Cells And Macrophages, Abdulafiz Musa

Electronic Theses and Dissertations

Regulation of the bacterial second messenger cyclic-di-GMP in Salmonella Typhimurium allows it to delicately alter phenotypes to optimize invasion and survive intracellularly in epithelial cells and macrophages to become virulent and cause infection. Cyclic-di-GMP concentration is regulated by the presence of external stimuli, sensory diguanylate cyclases (DGCs) and phosphodiesterases (PDEs), and cyclic-di-GMP binding effectors. Previous studies established that maintenance of low cyclic-di-GMP concentrations is required for survival in macrophages, and that deletion of 3 active PDEs reduces this survival. Here I showed that these 3 PDEs also influenced the infection of epithelial cells. Further studies re-established the decreased survival in …

Inhibiting Survival Of Salmonella During Desiccation Through The Use Of Naturally Occurring Signals, Joseph Headrick

Electronic Theses and Dissertations

A rising problem in agriculture is the increase of antibiotic-resistant Salmonella cases associated with chicken eggs, which transmit infection to humans. To counter this, new approaches to combat Salmonella in chickens and desiccated on eggshells are vital in the prevention of human foodborne illness. Disrupting signaling pathways with naturally occurring compounds provides a potential novel avenue for prevention of Salmonella infections, as this would disrupt sensing of these environments and inhibit subsequent optimal gene expression. Starting with signals identified in previous studies, salicylic acid was found to inhibit Salmonella desiccation survival on both eggshells and plastic. To expand upon this, …

Role Of Pseudomonas Aeruginosa Exoenzymes S, T, And Y In The Modulation Of Intrinsic Apoptosis In Pulmonary Microvascular Endothelial Cells, Andrea M. Vavrinek

Honors Theses

P. aeruginosa is the most common cause of ventilator-associated pneumonia in ICU patients. P. aeruginosa uses a type III secretion system to inject exoenzymes S, T, U, and Y into host cells, exhibiting unique effects. Pulmonary microvascular endothelial cells (PMVECs) are used to evaluate cellular changes caused by exoenzymes. ExoY infection of PMVECs leads to cell rounding but does not lead to cell death, unlike ExoS/T. The current study evaluated the exoenzyme-induced modulation of intrinsic apoptosis in endothelial cells. Strain PAK, producing ExoS, T, and Y, was compared to isogenic strains expressing either S, SY, T, TY, Y, or none …

Impacts Of Bacterial Evolution On Host Lethality In Drosophila, Andrew Preston

College of Arts & Sciences Senior Theses

Evolution is the process by which species change their genetic traits, such as the pathogenicity of bacteria, over time in response to changes in their environment. Although the genetic mechanisms underlying many evolutionary processes have been revealed, it is still not well understood how opportunistic pathogens, such as Pseudomonas aeruginosa, become virulent. The overall goal of this thesis is to test the Coincidental Evolution Hypothesis, which proposes that the virulence of opportunistic pathogens evolves coincidentally as a by-product of their interaction with their natural predators. I hypothesized that the virulence of ancestral Pseudomonas aeruginosa changes over time if it co-evolves …

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 …