Microbiology Commons^™

Harnessing Antagonistic Pleiotropy: Tolc-Dependent Coliphages Can Drive Reduced Antibiotic Resistance In E. Coli, Cooper Mckenna

CMC Senior Theses

The rise of multidrug-resistant (MDR) bacterial pathogens has been a growing concern in healthcare around the world, and bacteriophage (phage) therapy is a promising method to combat MDR infections. One defense mechanism of E. coli against antibiotics is the TolC-AcrAB efflux pump. At the same time, some phages use TolC as a surface receptor to infect the cell. This puts TolC at a crossroads in which it is beneficial to the bacteria to protect against antibiotics but harmful by allowing phage infection, creating the potential for evolutionary trade-offs. Previous research has identified only three TolC-dependent phages and shown phage-resistant E. …

Investigating Antimicrobial Producing Bacteria Within The Soil, Adam Zwawi

A with Honors Projects

Genetic sequencing techniques used to identify soil antibiotic bacteria

Effect Of Dietary Flavonoids On Growth In Saccharomyces Cerevisiae, Vivian Rivera

Mahurin Honors College Capstone Experience/Thesis Projects

Plant foods contain a diversity of biologically active compounds called phytochemicals, produced by plant species as secondary metabolites. A phytochemical group known as flavonoids has received much scientific attention in recent times with respect to their antioxidative and anti-inflammatory roles. This investigation examined how quercetin and naringenin, the most common dietary flavonoids, affect growth potential of yeast under optimal and stress conditions (temperature, glucose, and oxidative stress). Using Saccharomyces cerevisiae as a model, a dose dependency effect of quercetin and naringenin was observed under optimal conditions when liquid cultures were treated with three different concentrations (75 μM, 150 μM, and …

Screening And Analysis Of The Urobiome For The Production Of Antimicrobial Peptides To Target Uropathogenic Escherichia Coli, Jennifer Jones

Theses

Urobiome research is currently at a nascent stage, due to a previously incorrect assumption that the bladder is a sterile environment. Improvements in bacterial culturing and DNA sequencing techniques have revealed a diverse resident bacterial community within the urinary tract (urobiome). As global incidences of multidrug- resistant urinary tract infections (UTIs) continue to increase, so too does the demand for novel antimicrobial therapies. Due to its relatively understudied nature, the urobiome represents a reservoir that could be mined as a potential source of novel antimicrobial compounds (e.g., bacteriocins). The aim of this thesis was to identify urobiome bacteriocins that could …

Contributions Of The Replication Initiator Protein Dnaa To Physiology And Virulence Of The Lyme Disease Spirochete, Andrew Krusenstjerna

Theses and Dissertations--Microbiology, Immunology, and Molecular Genetics

Borrelia burgdorferi, the etiological agent of Lyme disease, is the most prevalent vector-borne disease in the United States. The spirochetal population is naturally maintained by consistently cycling between Ixodid ticks and small vertebrates. To survive within these diverse niches, B. burgdorferi has evolved a unique physiology and regulatory network to sense and respond to environmental fluxes. Replication is a hallmark of the Lyme spirochete’s enzootic life cycle, specifically at the nymphal tick-to-vertebrate interface. The mechanisms and regulatory schema for the basic cellular processes in B. burgdorferi, or indeed any member of the phylum spirochaetota, are largely unknown. The central hypothesis …

Role Of Autophagy-Related Genes In Lipid Transport For Efficient Replication Of Tombusviruses, Yuanrong Kang

Theses and Dissertations--Plant Pathology

Tombusviruses are positive-strand RNA viruses, which utilize numerous co-opted host factors and massive de-novo synthesized membrane resources to modify host membranes and form large viral replication organelles (VROs). Among plenty of host cellular processes, autophagy emerges as a potential membrane resource for VROs formation due to its ability to de-novo generate the double membrane structures, known as autophagosomes. Besides, as an important host defense pathway, autophagy is also a target for tombusviruses to prevent its antiviral activities. In this study, I identify the role of key autophagy related genes, Atg11, Atg8, and Atg2, which are involved in autophagosome formation, and …

Role Of Ph And Plant Age On The Selection Of Rhizobial Endophytes In Soybean Root Nodules, Cole Alexander Roman

Graduate Theses/Dissertations

Soybeans form symbiotic relationships with nitrogen-fixing bacteria in their root nodules to acquire most of their nitrogen requirement; nitrogen is often one of the limiting nutrients in the soil. Members of the Bradyrhizobium, Sinorhizobium, Mesorhizobium, and Rhizobium genera can nodulate soybeans. Bradyrhizobium is preferentially selected over other types of rhizobia in varying environmental conditions and is often found in acidic soils. Sinorhizobium is often identified in the root nodules of soybeans in neutral and alkaline soil. So far, the potential role of soil pH on the selection of rhizobial endophytes with soybean root nodules is unknown. Likewise, …

The Effect Of Nanoconfined Liquid Properties On The Water-Responsive Behavior Of Bacterial Cell Walls, Seungri Kim

Dissertations and Theses

Water-responsive (WR) materials have the ability to mechanically swell and shrink in response to changes in relative humidity (RH). These WR materials are used by many biological systems to perform essential tasks; for example, pinecones use WR materials to release their seeds in dry environments, and wheat awns open and close to propel seeds into the soil, driven by daily RH changes. The WR actuation of some biomaterials is extremely powerful, for example Bacillus subtilis cell walls display record-high actuation energy and power densities of 72 MJ m^-3 and 9.1 MW m^-3, surpassing those of all existing …

The Identification And Characterization Of Novel And Improved Antigens For Next-Generation Pertussis Vaccines, Gage Michael Pyles

Graduate Theses, Dissertations, and Problem Reports (ETD)

Whooping cough, or pertussis, is a highly infectious respiratory disease caused by infection with the Gram-negative bacterial pathogen Bordetella pertussis. Pertussis affects all age groups and populations; however, it is most severe in infants who are too young to receive vaccinations and remains a global health concern with more than­­­­ 200,000 cases reported globally in 2023. Vaccines against pertussis have been available in the United States since the 1940’s with the development and widespread use of whole cell pertussis vaccines. Following their introduction, the incidence of pertussis was controlled and limited to fewer than 5,000 cases per year. Despite …

Endogenous Glucocorticoid Signaling Is Required For Normal Macrophage Response To H. Pylori Infection, Stuti Khadka

Graduate Theses, Dissertations, and Problem Reports (ETD)

Gastric cancer is the fifth leading cause of cancer-related deaths worldwide, affecting millions of people. H. pylori is the strongest risk factor for gastric cancer, with about 90% of the cases strongly associated with H. pylori infection. Half of the world's population is infected with this bacterium. However, only a small subset of the infected population (1-3 %) go on to develop gastric cancer. Diagnosis of the disease at an early stage is the biggest challenge because of the widespread prevalence of H. pylori infection and a long asymptomatic phase before the advanced stages. Patients are left with limited treatment …

Examining The Role Of Microbial Communities In Soil Carbon Cycling Across Levels Of Complexity, Chansotheary Dang

Graduate Theses, Dissertations, and Problem Reports (ETD)

Microorganisms provide the foundation for many ecosystem processes and are important drivers of carbon and nutrient cycling. The advancement of next generation sequencing has allowed researchers to lift open the lid on the ‘black box’ in microbial ecology. While previously unculturable microbes can now be characterized with the 16S rRNA gene, answering ‘who’s there?’ is only part of the challenge. Hidden inside the ‘black box’ remains the question, ‘who’s doing what?’. Recent development in trait-based methods like quantitative stable isotope probing (qSIP) equipped microbial ecologists with the tool to connect taxon-specific traits to ecosystem processes. Despite the growing body of …

Social Interactions And Spatial Structure Drive Community Assembly Of Bacterial Biofilm, Matthew C. Bond

Dartmouth College Ph.D Dissertations

Despite the critical nature which microbial communities play in the natural world and human civilization, the breadth of understanding remains shallow. Challenged by scale, high variability between environments, and extensive diversity, microbial ecologists strive to understand connections between a community’s structure and function, as well as the ecological and evolutionary mechanisms underlying observed natural patterning of communities. Biofilms are the predominate mode of growth for microbial communities—characterized by cellular attachment to a surface via a self-produced matrix and heterogeneous structure, often resulting in a primary growth front along the biofilm surface due to differential access to bulk nutrients (cite). This …

Batch Culture Models Of The Murine Gut Microbiome & The Impact Of Simple Dormancy On Dormancy-Capable Microorganisms Models, Ana C. Mendez

Mathematics Dissertations - Archive

The proposed mathematical biology research utilizes mathematical models to gain insight into biological systems. These systems of ordinary differential equations model diverse topics, ranging from gut microbiomes to harmful algal blooms. A complete stability analysis, supporting phase plane portraits, bifurcation diagrams, and numerical simulations will accompany the models presented. In Chapter 2, the murine gut microbiome is modeled to match laboratory experiments in the literature. In these experiments, mice eat plasmid-carrying “donor” bacteria and naturally carry plasmid-free “resident” bacteria in their gut. The models aim to capture the behavior of plasmids, donor bacteria, and resident bacteria. Chapter 3 explores dormancy …

Inflammation And Tumor Progression: The Differential Impact Of Saa In Breast Cancer Models, Daniel Wilhelm Olivier, Carla Eksteen, Manisha Du Plessis, Louis De Jager, Lize Engelbrecht, Nathaniel Wade Mcgregor, Preetha Shridas, Frederick C. De Beer, Willem J. S. De Villiers, Etheresia Pretorius, Anna-Mart Engelbrecht

Saha Cardiovascular Research Center Faculty Publications

Background: Previous research has shown that the Serum Amyloid A (SAA) protein family is intricately involved in inflammatory signaling and various disease pathologies. We have previously demonstrated that SAA is associated with increased colitis disease severity and the promotion of tumorigenesis. However, the specific role of SAA proteins in breast cancer pathology remains unclear. Therefore, we investigated the role of systemic SAA1 and SAA2 (SAA1/2) in a triple-negative breast cancer mouse model.

Methods: Syngeneic breast tumors were established in wild-type mice, and mice lacking the SAA1/2 (SAADKO). Subsequently, tumor volume was monitored, species survival determined, the inflammatory profiles of mice …

Poor Water Quality As A Trigger Of Harmful Algal Blooms In Lake Manzala, Mohammed A. Hussein, Alaa Eldin Eissa, Reham M. Eltarabily, Mohamed F. Abdelghany, Mahmoud A. Elnakeeb, Eman M. Ismail, Reham H. Ragab, Amina A. Dessouki

Mansoura Veterinary Medical Journal

Objective: The current study aimed to assess the direct effects of poor water quality on the eruption of heavy harmful algal blooms and to assess the physicochemical properties of water in Lake Manzala, as well as the molecular identification of the retrieved green algae.

Procedures: Subsurface water samples were collected from three different sites in September 2023 for water quality assessment. Moreover, morphological and molecular identification of suspected harmful algae is the most reliable diagnostic tool for this eruption.

Results: This study was conducted in the northern and northeastern regions of Lake Manzala. These regions receive large amounts of municipal, …

Bloom: A Microbial Self-Portrait, Emily Lauren Mulvaney

Graduate Student Theses, Dissertations, & Professional Papers

No abstract provided.

Synthetic Microbial Communities For Plastic Upcycling, Isabel B. A. Valencia

Dissertations, Master's Theses and Master's Reports

Plastic waste and human hunger pose major threats to the health and wellbeing of populations world-wide. Using microbial methods to upcycle polyethylene terephthalate (PET) plastic into value-added compounds such as single cell protein (SCP) for human consumption is a unique solution to both these issues. Two monomers of chemically deconstructed PET (DCPET), terephthalate (TPA) and ethylene glycol (EG), have been previously shown to be biodegradable by enriched microbial communities. Using this top-down knowledge to inform the reconstruction of a minimal microbial community constructed from isolated members of these communities is a novel way to efficiently process these monomers via a …

Listeria Adhesion Protein Orchestrates Caveolae-Mediated Apical Junctional Remodeling Of Epithelial Barrier For Listeria Monocytogenes Translocation, Rishi Drolia, Donald B. Bryant, Shivendra Tenguria, Zuri A. Jules-Culver, Jessie Thind, Breanna Amelunke, Donqi Liu, Nicholas L. F. Gallina, Krishna K. Mishra, Manalee Samaddar, Manoj R. Sawale, Dharmendra K. Mishra, Abigail D. Cox, Arun K. Bhunia

Biological Sciences Faculty Publications

The cellular junctional architecture remodeling by Listeria adhesion protein-heat shock protein 60 (LAP-Hsp60) interaction for Listeria monocytogenes (Lm) passage through the epithelial barrier is incompletely understood. Here, using the gerbil model, permissive to internalin (Inl) A/B-mediated pathways like in humans, we demonstrate that Lm crosses the intestinal villi at 48 h post-infection. In contrast, the single isogenic (lap− or ΔinlA) or double (lap−ΔinlA) mutant strains show significant defects. LAP promotes Lm translocation via endocytosis of cell-cell junctional complex in enterocytes that do not display luminal E-cadherin. In comparison, InlA facilitates …

Optimization Of Buffers Used In Phycobiliprotein Extraction From The Cyanobacterium Acaryochloris Marina, Rinah Fleisher

Williams Honors College, Honors Research Projects

Optimization of buffers used in phycocyanin extraction from A. marina was explored including acetate, phosphate, and citrate buffers as well as deionized water as a negative control. Samples were extracted in triplicates for each trial with two extraction trials completed overall for citrate buffer (n=6), phosphate buffer (n=6), acetate buffer (n=6), and deionized water (n=6). Results reflect no statistical difference in phycocyanin content (μg/mL) between each buffer or deionized water control. Though no statistically significant differences between treatment groups were found, phosphate buffer yielded the greatest phycocyanin content and the deionized water negative control yielded the lowest phycocyanin content overall. …

Determination Of Spore Viability In Concrete Across Several Factors Using Most Probable Number, Samuel Boyer

Williams Honors College, Honors Research Projects

To determine the lowest concentration of spore added to polyurethane-cement composite (PUCCO) particles that can still germinate after curing in concrete. This research project is a small addition to the larger research project being undertaken by Mirza Mohammed Rashiduzzaman for his Masters. The larger project involves the use of fungal spores added in concrete to act as a self-healing component when cracks form in the concrete structure over time. These spores are suspended in a protective oil and loaded into small, hardened sponge-like PUCCO cubes to act as growth points when water and air can reach the PUCCO in the …

Epitope Mapping Novel Monoclonal Antibodies Against Borrelia Burgdorferi Surface Proteins Ospc And Bmpa, Eliza M. Doyle

Honors Undergraduate Theses

In response to the progressive nature of Lyme disease, direct diagnostic testing of Borrelia burgdorferi surface proteins was proposed as a quicker and more sensitive alternative to the current indirect detection methods. To determine the ability of novel monoclonal antibodies to detect the surface lipoproteins BmpA and OspC, recombinant fragments of these proteins were created. These fragments were tested against the putative diagnostic antibodies via western blotting as a means to map the epitope of each of the proteins that the respective antibodies recognize. All BmpA fragments with deletions on the N-terminal end exhibited reactivity, indicating that the epitope is …

A Molecular-Informed Species Inventory Of The Order Ceramiales (Rhodophyta) In The Narragansett Bay Area (Rhode Island And Massachusetts), Usa, Thomas Irvine, Brian Wysor, Alicia Beauvais

Arts & Sciences Faculty Publications

Narragansett Bay is an estuarine system in the western North Atlantic Ocean that harbors a diverse marine flora, providing structure, habitat, and food for native biodiversity. This area has been the center of numerous environmental, biological, ecological, and oceanographic studies; however, marine macroalgae have not been extensively examined using modern molecular methods. Here, we document the biodiversity of the red algal order Ceramiales based on DNA sequence comparisons of the 3′ end of the RuBisCo large subunit (rbcL-3P) and the universal plastid amplicon (UPA). Thirty-seven distinct species of this order were identified and validated with molecular data, including five new …

Application Of Environmental Dna Metabarcoding To Differentiate Algal Communities By Littoral Zonation And Detect Unreported Algal Species, Sergei Bombin, Andrei Bombin, Brian Wysor, Juan M. Lopez-Bautista

Arts & Sciences Faculty Publications

Coastal areas are the most biologically productive and undoubtedly among the most complex ecosystems. Algae are responsible for most of the gross primary production in these coastal regions. However, despite the critical importance of algae for the global ecosystem, the biodiversity of many algal groups is understudied, partially due to the high complexity of morphologically identifying algal species. The current study aimed to take advantage of the recently developed technology for biotic community assessment through the high-throughput sequencing (HTS) of environmental DNA (eDNA), known as the “eDNA metabarcoding”, to characterize littoral algal communities in the Northern Gulf of Mexico (NGoM). …

The Development And Evaluation Of A Method For Complete Collagenase I Digestion Of Bos Taurus Bone Samples, Reilly Price

Master of Science in Forensic Science Directed Research Projects

Current methods for processing aged forensic bone samples for DNA analysis involve pulverization and organic extraction, which further destroys already degraded DNA and uses large amounts of limited samples. Additionally, the success rates for this type of analysis vary widely as homogenization incorporates PCR inhibitors into the sample. Little research has explored how pulverization results in DNA damage in profiles obtained from skeletal remains. This study proposes an enzymatic digestion method to separate osteocytes from PCR inhibitors in the extracellular bone matrix to improve the quality and quantity of DNA isolated from aged bone samples. The technique described here used …

Using The Microbiome To Assess The Health Status Of The Individuals Recovered From The East Marshall Street Well, Arian Karim

Master of Science in Forensic Science Directed Research Projects

In 1994, during the construction of the Kontos Building on the Medical College of Virginia Campus, a well on East Marshall Street was discovered to contain artifacts and human remains of African descent. Upon archaeological analysis by the Smithsonian, the ancestral remains can be dated back to the mid-19th century, when grave robbing and the use of cadavers for anatomical purposes [BS1] in medical schools were common. The human oral microbiome is the second largest human microbiota, containing over 600 species of organisms. Dysbiosis of the oral microbiome can lead to the overgrowth of pathogenic species, resulting in illness and …

Role Of Interleukin 2 Receptor Alpha In The Cellular Response To Dna Damage, Shristi Khanal

Browse all Theses and Dissertations

The interleukin 2 receptor alpha (IL-2Rα), which is integral to the functioning of the cytokine interleukin 2, has recently been found to be present in different cell types besides just lymphocytes. Previous studies done by our laboratory showed that several other non-lymphoid cells including smooth muscle cells express IL-2Rα. Vascular smooth muscle cell (VSMC) deficient in IL-2Rα exhibited increased proliferation, decreased size, and hypodiploid DNA content when compared to wild type (WT) cells. Previous studies also suggested that VSMC proliferate in response to IL-2 and that IL-2 increases following interventions to stimulate intimal hyperplasia in vivo. These findings, in total, …

Studies Of Selective And Reversible Inhibitors Of Calcium Entry In Immune Cells, Naga Lakshmi Gandi

Browse all Theses and Dissertations

Calcium signaling is a crucial regulator in many cellular processes, from immune response modulation to gene expression and apoptosis. Here, I have investigated the dynamics of calcium entry pathways, namely, store-operated calcium entry and store-independent calcium entry using a variety of cell types such as Jurkat T cells, HEK-293 cells and murine macrophages. Using fluorescence calcium imaging and selective inhibitor compounds, I have investigated the contributions of Orai and related calcium channels to calcium entry in both immune and non-immune cell types. The results demonstrate the pharmacological modulation of store-operated and store-independent calcium entry, showing the ability of compounds such …

Co-Divergence Of Lemur Hosts And Gut Microbiomes Across Multiple Evolutionary Scales, Mariah E. Donohue

Theses and Dissertations--Biology

This dissertation bridges micro- and macroevolutionary approaches to understand the co-diversification of animal hosts and their gut microbiomes at different evolutionary timescales. I examined these dynamics using wild lemurs, a diverse Primate clade endemic to the island of Madagascar.

First, I tested the relative effects of host phylogenetic relationships, diet, and habitat in shaping the gut microbiome through deep evolutionary time and across distinct microbial kingdoms. I designed a natural experiment sampling feces from rainforest and dry forest lemur communities with similar phylogenetic assemblages and considerable variation in dietary strategy. Then, I amplified and sequenced two metabarcoding genes, the V4 …

Interactions Between Multiple Stressors, Nanomaterials And A Pathogen, In Caenorhabditis Elegans, Jarad Cochran

Theses and Dissertations--Plant and Soil Sciences

Metal and metal oxide nanoparticles (NP), which have received interest for use in agriculture, show in vitro and in vivo toxicity at relatively low concentrations. Silver nanoparticles (Ag NP) and zinc oxide nanoparticles (ZnO NP) cause adverse effects in model soil organisms under environmentally relevant conditions. In most toxicity assessments of nanomaterials, model organisms are exposed to a single stressor. Though single stressor experiments are essential to understanding the fate and effects of released NP, in nature, organisms are affected by multiple stressors, including pathogens, which might exacerbate or mitigate negative effects of NP exposure. There is limited research on …

Ribosome Hibernation Factor Induces Antibiotic Persisters Of Mycobacterium Tuberculosis In Neutrophil-Rich Lung Lesions, Jamie Corro

Electronic Theses & Dissertations (2024 - present)

Mycobacterium tuberculosis (Mtb), the etiological agent of tuberculosis, is a prescient global health threat and the leading cause of death by an infectious agent. Treatment of Mtb infection involves administering 2-4 antibiotics for a minimum of 6 months. This extended drug regimen is required to target a subpopulation of persistent bacilli. As their name implies, these bacteria can “persist” by acquiring nonhereditary and transient antibiotic tolerance. Although the precise mechanism is unknown, it’s often attributed to slowing cellular growth and metabolic processes. During zinc deprivation, bacteria can remodel their ribosomes by replacing their zinc-binding paralogues containing the CXXC motif (C+) …