Microbiology Commons^™

Marine Heatwaves And Fish Reproduction: Endocrine Responses In The Eurythermal Sheepshead Minnow (Cyprinodon Variegatus), Isabel Villafuerte

Master's Theses

Marine heatwaves associated with climate change are increasing in frequency and severity, posing significant threats to fish populations in shallow habitats such as estuaries and salt marshes. Because temperature is a primary regulator of fish reproduction, understanding how heatwaves affect reproductive physiology is critical for anticipating the biological consequences of ongoing climate change for coastal fishes. Here we examined how simulated heatwave conditions affect the reproductive hypothalamic-pituitary-gonadal (HPG) endocrine axis in the estuarine sheepshead minnow (Cyprinodon variegatus), a eurythermal fish well suited to studying thermal stress responses in estuarine environments. Adult sheepshead minnows acclimated to a daily oscillating …

Examining The Ligands, Location, And Function Of Toll-Like Receptor 10, Kiefer Keanu Tam

Master's Theses

Toll-like receptors are a class of innate immune pattern recognition receptors that detect pathogen-associated molecular patterns and regulate downstream inflammatory responses. Toll-like receptor 10 remains the only human TLR whose ligand specificity, cellular localization, and immunological function are not fully defined. Emerging evidence suggests that TLR10 may play an anti-inflammatory role and can heterodimerize with TLR2 to modulate immune signaling. Previous studies from our laboratory demonstrated that stimulation with heat-killed Mycobacterium tuberculosis promotes TLR2-TLR10 heterodimerization and trafficking to the cell surface. In this study, we expanded our ligand panel to include additional gram-positive bacteria, particularly anti-inflammatory probiotic species, to further …

Enhancing Preparedness: A Genetic Study Of Sar-Cov-2 And Other Microbial Public Health Threats In Wastewater From A College Town, Christian N. Hutchinson

Master's Theses

Problem

The SARS-CoV-2 (COVID-19) pandemic revealed both the systemic vulnerabilities of our healthcare system as well as the importance of timely, localized data for informing public health responses. Traditional testing approaches were essential but often failed to capture the full extent of viral spread, especially in those with mild or asymptomatic conditions that relied on at home testing. This underreporting hindered effective allocation of resources and made developing effective intervention strategies difficult. We also realized the potential for wastewater surveillance to be applied to tracking antimicrobial resistance gene markers. Taken together, this approach should have important implications for improving public …

Impact Of Temperature And Microbial Infection On The Honey Bee Hemocytes, Michael E. Oeth

Master's Theses

Introduction

Honey bees (Apis mellifera) rely on hemocytes for immunity, wound healing, and hemolymph homeostasis. While 32°C reflects optimal hive conditions, suboptimal temperatures (22°C) and stressors like pathogens (Varroa destructor, bacteria) or pesticides impair hemocyte function. Using clodronate liposomes (CLD) to deplete hemocytes, we assessed how immune suppression interacts with these stressors.

Methods

Newly emerged (5-day-old) and older (15-day-old) bees were assigned to: CLD, control liposomes (LP), PBS, injection wound, or no treatment. Cohorts were exposed to 32°C (optimal) or 22°C (stressful for non-immunosuppressed bees). Fifteen-day-old bees were challenged with Escherichia coli or Staphylococcus aureus. Survival, hemolymph volume, and hemocyte …

Induced Systemic Resistance Against Nematodes By Plant Growth-Promoting Microbes In Potatoes, Cole Grannan

Master's Theses

Biofertilizers consist of plant growth-promoting microorganisms that establish symbiotic relationships with plant roots and stems. These beneficial microbes play a crucial role in enhancing plant defenses against various threats including insects, nematodes, and microbial pathogens. The purpose of this study was to investigate the effectiveness of two fungal biofertilizers (an endophytic strain of Beauveria bassiana (BA) and a mixture of arbuscular mycorrhizal (AM) fungi), specifically their ability to induce systemic resistance against plant- parasitic nematodes (PPNs) in potatoes (Solanum tuberosum cv. Masquerade) under controlled greenhouse conditions. The study utilized soil collected from Colorado State University’s San Luis Valley (SLV) Research …

Lack Of Inflammatory Response In Mouse Embryonic Cells To Listeria Infection, Damilola Oyebode

Master's Theses

Approximately 1 in 5 pregnancies in the United States results in miscarriage, with a significant percentage of these losses attributed to failed implantation due to immunological challenges encountered by the early embryo. Recent research has shown that key cell types within mouse blastocysts, including embryonic stem cells (ESCs) and trophoblast stem cells (TSCs), exhibit remarkable resilience against the cytotoxic effects associated with viral infections and embryotoxic cytokines, such as TNFα and IFNγ, both of which are known to hinder blastocyst development. However, the immune properties of the preimplantation stage embryo, specifically the blastocyst, remain poorly understood. Using mouse cell model, …

Development Of A Urothelial Cell Line Expressing A Caspase-1 Biosensor To Measure Cellular Inflammatory Responses To Potential Uropathogens, Nikita Riya Kesav

Master's Theses

Urinary tract infections are one of the most common bacterial infections, historically implicating Escherichia coli as the causative pathogen. However, it is now known that there are a variety of bacteria that exist within the urinary tract, acting as either beneficial uroprotectives or opportunistic uropathogens. Furthermore, there are significant gaps in our understanding as to how these bacteria interact with the urothelium and what role each plays in maintaining or disrupting homeostasis. Research has shown that Toll-like receptor 4 (TLR4) signaling and Nod-like Receptor Protein 3 play a role in inflammasome activation during infection, triggering caspase-1 activation and inducing release …

Investigating The Roles Of Sirtuins During Cvb3 Infection And Innate Immunity, Bridget M. Hulsebosch

Master's Theses

Coxsackievirus B3 (CVB3) is an enterovirus that chronically infects heart tissue, leading to dilated cardiomyopathy. However, no vaccines or antivirals exist for CVB3 infection or to treat CVB3-induced dilated cardiomyopathy, except for a heart transplant. Sirtuins (SIRTs), class III lysine deacetylases, have diverse and complex roles in cellular functions, contributing to energy production, NF-κB signaling, cell cycle regulation, and inflammation. Previous studies showed that SIRTs are essential for hepatitis B and A viral replication and regulate the IFN response during viral infection. However, the role of SIRT1 and 2 during CVB3 infection is entirely unknown. CVB3-infected SIRT1 and SIRT2 knockout …

Investigation Of The Microbiome Of A Recirculating Aquaculture System (Ras) For Crassostrea Virginica Larvae, Isabelle Townsend

Master's Theses

Recirculating Aquaculture Systems (RAS) offer control over physicochemical and biological factors impacting aquaculture success. One understudied facet of RASs is the water microbiome. Oyster larval microbiomes are shaped by their aquatic microbiomes. Understanding the dynamics of and the factors shaping RAS microbiome may provide insights to how the microbiome can be a tool for optimizing larval production. This study’s goals were to investigate the microbiome stability of a RAS rearing oyster larvae across time and space, determine impacts of larval presence on the microbiome, and examine connections between larvae production outcomes and microbiome stability. Water samples were collected weekly from …

Antimicrobials Produced By Ants: Chemical Properties And Microbe Specificity, Katy Chon

Master's Theses

Antibiotic resistance is a growing public health concern worldwide, and for over a decade, we have been witnessing the growth of difficult to treat infectious diseases caused by bacteria that are becoming resistant to antibiotics. This resistance can be mostly related to the misusing and improper use of antibiotics in both humans and animals. To restore this problem, humans have turned to new sources for the production of antibiotics. In this study, we focused on using ants. Social insects, including ants and bees, have faced strong disease pressures during their evolution and have developed a range of methods to fight …

Novel Antivirals Unveiled Against Acute And Persistent Coxsackievirus B3 Infection Via Rapid Screening, Maria Del Mar Villanueva Guzman

Master's Theses

Enteroviruses cause 10 to 15 million infections annually in the United States, and Coxsackievirus B3 is one of the most commonly reported. Coxsackieviruses may become persistent, characterized as a viral infection that is not cleared from host cells and that generates a continuous infection. Patients who develop persistent CVB3 infection may not respond to the same antivirals as an acute infection, which may be detrimental. Therefore, there is a need for broad-range antiviral drugs to combat acute and persistent CVB3 infection, as there is no well-accepted treatment available. We developed a model system to study persistent CVB3 using a pancreatic …

Novel Antivirals Unveiled Against Acute And Persistent Coxsackievirus B3 Infection Via Rapid Screening, Maria Del Mar Villanueva Guzman

Master's Theses

Enteroviruses cause 10 to 15 million infections annually in the United States, and Coxsackievirus B3 is one of the most commonly reported. Coxsackieviruses may become persistent, characterized as a viral infection that is not cleared from host cells and that generates a continuous infection. Patients who develop persistent CVB3 infection may not respond to the same antivirals as an acute infection, which may be detrimental. Therefore, there is a need for broad-range antiviral drugs to combat acute and persistent CVB3 infection, as there is no well-accepted treatment available. We developed a model system to study persistent CVB3 using a pancreatic …

Characterization And Evaluation Of Bacteriophages Φpa-C And Φpa-Tc Infecting Pseudomonas Aeruginosa, Joanne Harrison

Master's Theses

Pseudomonas aeruginosa are prevalent opportunistic bacterial pathogens that cause nosocomial infections in at-risk individuals. The rise of multidrug-resistant P. aeruginosa poses an immense threat to hospitals. Additionally, P. aeruginosa are common food spoilage organisms that contribute to food waste. Alternative effective treatment methods are needed to improve public health and reduce food waste. Recent studies have shown that bacteriophages, viruses that only kill host-specific bacteria, are promising alternatives. This study characterizes two lytic P. aeruginosa phages, ΦPa-C and ΦPa-Tc, and evaluates their efficacy against P. aeruginosa in two model food systems. The morphology, genome, structural proteins, host range, growth kinetics, …

Characterization And Evaluation Of Bacteriophages Φpa-C And Φpa-Tc Infecting Pseudomonas Aeruginosa, Joanne Harrison

Master's Theses

Pseudomonas aeruginosa are prevalent opportunistic bacterial pathogens that cause nosocomial infections in at-risk individuals. The rise of multidrug-resistant P. aeruginosa poses an immense threat to hospitals. Additionally, P. aeruginosa are common food spoilage organisms that contribute to food waste. Alternative effective treatment methods are needed to improve public health and reduce food waste. Recent studies have shown that bacteriophages, viruses that only kill host-specific bacteria, are promising alternatives. This study characterizes two lytic P. aeruginosa phages, ΦPa-C and ΦPa-Tc, and evaluates their efficacy against P. aeruginosa in two model food systems. The morphology, genome, structural proteins, host range, growth kinetics, …

Impact Of Toll-Like Receptor 10 Mutations On Tuberculosis Infections, Adriana Nicole Rafael, Sangman Kim

Master's Theses

Tuberculosis (TB), caused by the intracellular pathogen Mycobacterium tuberculosis (M. tb), is a leading global health crisis causing millions of cases and deaths each year. Though it affects individuals of any race, ethnicity, or cultural group, research shows the disease disproportionately affects individuals with Asian ancestry. Immunity to M. tb starts with its recognition by pattern recognition receptors such as Toll-like receptors (TLRs), which are key players in immune responses. TLR-10, an understudied toll-like receptor due to its inability to be studied through mouse models, is thought to play a key role in the clearance of inflammatory diseases. SNP I775V, …

The Immune Contribution To Postpartum Remodeling, Jessica Caroline Ming

Master's Theses

Following parturition, the uterus must undergo significant changes in size and structure through a process called postpartum remodeling. This process is known to have both pro- and anti- inflammatory components. However, how various immune cells contribute to remodeling and the pathways that promote inflammation are not well understood. We hypothesize that immune cells that infiltrate the uterus postpartum modulate inflammation to promote postpartum remodeling. Our preliminary data suggests that inflammatory cell death may contribute to the inflammation present in the early postpartum period, and thus further studies will analyze caspase-1, gasdermin D, and IL-1β and IL-18. To analyze the immune …

Investigating The Role Of The Metabolite Je As An Inflammatory Modulator, Rim Ahmed Noufal

Master's Theses

Innate immunity represents the first line of defense against pathogens that invade our bodies. It greatly relies on a family of abundant receptors called pathogen recognition receptors (PRRs). These are germ-line encoded proteins abundant in immune cells like macrophages, dendritic cells, and natural killer cells. PRRs encompass five categories, one of which includes toll-like receptors (TLRs). TLRs could either be located on the surface of the cells or intracellularly. The mechanism by which TLRs act is through recognizing a myriad of foreign structures derived from microbes, such as proteins, lipids, and carbohydrates as well as their genetic material. Generally, these …

Investigating Putative Toxin-Immunity Pairs From A New Class Of Rhs Proteins In Pseudomonas Aeruginosa, Syed Faieq Imam

Master's Theses

Pseudomonas aeruginosa is a gram-negative, opportunistic pathogen that is ubiquitous in nature, especially in human/animal impacted environments. One of the reasons for this is the arsenal of protein systems and toxins that P. aeruginosa utilizes to mold its environment to suit its needs. Recently, we discovered a novel locus in the P. aeruginosa genome, which resembles other loci that are known to mediate interbacterial competition. Our bioinformatic analysis found this locus to be enriched in a rearrangement hotspot (rhs) motif which led us to ad-interim term the three ORFs of the loci as rhsB, rhsC, and rhsI. Furthermore, we have …

​ Soilborne Pathogens Of Strawberry In The Central Coast Region Of California: ​ Survey And Cover Cropping With Wheat For Management Of Macrophomina Phaseolina, Mary Steele

Master's Theses

Surveys of the four major soilborne pathogens of strawberry (Fusarium oxysporum f. sp. fragariae, Macrophomina phaseolina, Phytophthora spp., and Verticillium dahliae) to determine their relative prevalence were conducted in Watsonville-Salinas, CA in 2021 and in Santa Maria, CA in 2022. All four major pathogens were detected at relatively similar prevalence in Watsonville-Salinas, between 22% and 31% of sampled fields. In Santa Maria, M. phaseolina was far more prevalent at 52% of sampled fields, the other three falling between 14% and 17%. Additionally replicated greenhouse and field trials were conducted to evaluate the effects of wheat as …

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 …

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 …

Investigating Putative Toxin-Immunity Pairs From A New Class Of Rhs Proteins In Pseudomonas Aeruginosa, Syed Faieq Imam

Master's Theses

Pseudomonas aeruginosa is a gram-negative, opportunistic pathogen that is ubiquitous in nature, especially in human/animal impacted environments. One of the reasons for this is the arsenal of protein systems and toxins that P. aeruginosa utilizes to mold its environment to suit its needs. Recently, we discovered a novel locus in the P. aeruginosa genome, which resembles other loci that are known to mediate interbacterial competition. Our bioinformatic analysis found this locus to be enriched in a rearrangement hotspot (rhs) motif which led us to ad-interim term the three ORFs of the loci as rhsB, rhsC, and rhsI. Furthermore, we have …

Investigating Structural And Biochemical Characteristics Of A Peptidase Required For Staphylococcus Aureus Virulence, Sagar Bhusal

Master's Theses

Staphylococcus aureus is an opportunistic Gram-positive pathogen that transiently colonizes 30% of the population at any given time. Upon breaching host skin or mucosal barriers, S. aureus can infect and cause disease in nearly all host tissues. Once inside the host, S. aureus expresses a plethora of virulence factors, mostly regulated by the Accessory Gene Regulatory (Agr) quorum sensing system, which facilitates the evasion of immune responses or directly promotes pathogenesis. The S. aureus Agr system relies on the production of a small peptide known as an auto-inducing peptide (AIP) to control virulence gene expression in response to cell density. …

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 …

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 …

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 …

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 …