Microbiology Commons^™

Optimizing Crispri Gene Silencing Of Putative Drug Targets In Mycobacterium Abscessus, Max Ivanov

Honors Undergraduate Theses

Among nontuberculous mycobacteria (NTM), Mycobacterium abscessus (Mab) poses a particular threat to patients with cystic fibrosis and chronic obstructive pulmonary disease due to the lung damage and higher mortality rate it causes. Current treatments involve prolonged multi-antibiotic regimens that are often ineffective, highlighting the need for new drug targets.

The CRISPR Interference (CRISPRi) system uses a deactivated Cas9 protein (dCas9) to enable inducible gene silencing, offering a potential approach to identifying new targets. Genes that lead to a loss of Mab viability when silenced are considered essential, an important attribute of potential drug targets. While CRISPRi has shown …

Elucidation Of The Post-Transcriptional Regulation And Role In Iron Homeostasis Of The Outer Membrane Vesicle Protein Ynce In Salmonella, Mira J. Schmidt

Honors Undergraduate Theses

Iron is an essential element for most bacteria as it is a crucial cofactor for enzymes involved in fundamental reactions such as DNA synthesis. Despite being the fourth most abundant element on Earth, bacteria face challenges acquiring iron under aerobic conditions due to the poor solubility of its oxidized form (Fe³⁺). To overcome this, bacteria have developed various strategies of iron acquisition, such as the production of siderophores. Siderophores are small molecules that scavenge free Fe³⁺ in iron-deprived environments. Iron acquisition is tightly regulated due to its capability of promoting hydrogen peroxide formation via the Fenton reaction. In Salmonella Typhimurium, …

Development Of A Proposed Selective Medium For The Isolation And Differentiation Of P. Sordellii, Dimyana Bottros

Honors Undergraduate Theses

Clostridium, a genus of anaerobic, spore-forming bacteria, includes clinically and ecologically significant species like Clostridium difficile and Paeniclostridium sordellii. C. difficile causes antibiotic-associated colitis, while P. sordellii is linked to fatal infections and spontaneous abortions. Recent studies found both species in marine sediments, suggesting their role in environmental and wildlife health disturbances, including the 2013 mass mortality of over 300 manatees in the Indian River Lagoon. However, isolating and differentiating P. sordellii is challenging due to the lack of selective media optimized for its growth. This thesis proposes a novel selective medium tailored to P. sordellii’s physiological …

Cultures For Acid-Fast Bacilli: Are We Being Good Stewards?, Jennifer Hanrahan, Julie D. Sill, Patricia Ver Schneider, Jessica F. Copeland, Elsie Frimpongmaa Amoako-Kissi, Ogochukwu O. Ilobi, Angela J. Toepp

Department of Medicine Faculty Publications

Objective: To describe the use of cultures for acid-fast bacilli (AFB) and situations in which AFB cultures are unlikely to be of clinical benefit.

Design: Retrospective descriptive study of AFB cultures submitted to Sentara Health microbiology laboratory from December 1, 2021, to December 1, 2023. Data were collected from the electronic medical record and included patient demographics, the service line that ordered the culture, specimen source, and culture results.

Setting: Sentara Healthcare System.

Patients: All patients who had specimens submitted to the microbiology laboratory during the study period were included.

Results: A total of 13,944 AFB cultures from 8,243 …

Oral Microbiome And Inflammatory Bowel Disease: New Understanding And Call To Action, Michael Saadeh, Sydney Donohue, Shaina Ailawadi, Gordon Hong, David A. Johnson

Department of Medicine Faculty Publications

The oral microbiome is the second largest microbial community in the human body after the gut microbiome. It includes an array of fungi, bacteria, amoebae, flagellates, archaea, and viruses, all of which are potential pathogens. This microbiome can act as a facilitator not only for protection but also for aggravation when dysbiosis occurs. Although conventional thought is this is primarily in terms of oral health issues, such as dental caries and gingival disease. The systemic effects of the oral microbiome however, are relevant to both gastrointestinal (GI) disease and non-GI disease. These systemic risks occur for several reasons, including upregulation …

Multisociety Guidance For Infection Prevention And Control In Nursing Homes, Lona Mody, Sonali D. Advani, Muhammad Salman Ashraf, Allison H. Bartlett, Suzanne F. Bradley, Deborah P. Burdsall, Jennifer A. Hanrahan, Susan S. Huang, Robert L.P. Jump, Lindsay Nicolle, Mary-Claire Roghmann, Patricia Stone, Rekha K. Murthy

Department of Medicine Faculty Publications

This multisociety guidance was endorsed by SHEA, APIC, IDSA, PALTmed, and AGS. It provides recommendations for infection prevention and control (IPC) in the context of the complexity of nursing home care in the United States: increased medical acuity of residents, the spread of multidrug-resistant organisms, and the threat of emerging pathogens. Recommendations and implementation suggestions address IPC leadership, staffing, and resources, healthcare personnel and residents‘ adherence to precautions and effective hand hygiene, outbreak preparedness, training, occupational health, cleaning and disinfection in the care environment, and the involvement of IPC in the facility. The guidance also addresses the challenges of maintaining …

Microbiological Examination Of Yellow Mealworm (Tenebrio Molitor) Larvae Under Various Processing And Storage Conditions As Potential For Human Consumption, Rachel Midkiff

Graduate Theses, Dissertations, and Problem Reports (ETD)

Yellow mealworm (Tenebrio molitor) larvae have gained attention as a sustainable and nutritionally balanced food source, yet their high natural bacterial load raises food safety concerns. This study investigates the impact of pre-processing, processing, and storage conditions on microbial load and diversity in mealworms, comparing thermal (hot) and non-thermal (cold) treatments. These treatments include freeze-drying, wet heat treatments, dry heat treatments, and the combination of wet and dry heat treatments against untreated controls. Microbial loads were quantified by plate counts after day 0, day 10, and day 28 of vacuum-sealed refrigerated (4°C) storage. Water activity (Aw), pH, and moisture content …

Molecular Characterization Of Fungus-Arthropod Associations, Angie M. Martin

Graduate Theses, Dissertations, and Problem Reports (ETD)

Arthropods, or phylum Arthropoda, are the largest group of animals, accounting for over three quarters of all animal records on NCBI Taxonomy. And yet, only 40% of all deposited sequences on NCBI Nucleotide come from arthropods. Clearly, arthropods are an understudied group of animals: they are small, often drab, and lead lives very alien to our own. Arthropods have chitinous exoskeletons, segmented bodies, and jointed appendages, and include crabs, barnacles, insects, spiders, and millipedes. Since the era of DNA based studies took off in the 1980s, DNA and other small molecules have been integral to our understanding of the Earth …

The Chemical Ecology Of A Caribbean Boulder Coral During Restoration And The Interplay Between Fish Predation And Microbiome Variation, Catherine E. Raker

Open Access Dissertations

Coral reefs are one of the most biodiverse and productive ecosystems on the planet. In recent decades, however, climate change and other global threats have caused severe declines in live coral cover worldwide. Coral propagation, or “gardening,” is a popular strategy to attempt to restore healthy corals to degraded reefs. Slow-growing boulder corals are often restored using microfragmentation, where cutting coral colonies into roughly 1 cm² fragments can increase their growth rates up to 5x. However, extreme early mortality of fragments has been observed in the field, which has been at least partially attributed to predation. It has been …

Characterizing Agroecosystem Effects Of Industrial Hemp On A Rotational Row Cropping System And Microbial Communities, Kent D. Pham

Theses and Dissertations--Plant and Soil Sciences

The implementation of crop rotation on a farm can have many benefits for agroecosystem health and production. This practice can potentially reduce nutrient amendments, disrupt pest and disease cycles, and increase crop yields. In Kentucky, one common rotation consists of either corn or soybean grown the first year, followed by the other crop, repeated every two years. Industrial hemp, grown for fiber or grain, is an emerging crop that has shown promise as a rotation alternative, although not much is known about its effects on soil health, agroecosystem functioning, and microbial communities. To better characterize hemp’s role within American agriculture, …

Schistosoma Mansoni Infection Causes Consistent Changes To The Fecal Bacterial Microbiota Of Mice Across And Within Sites, Mariam A. Mhanna, David T. Gauthier, Lisa M. Shollenberger

Biological Sciences Faculty Publications

Eggs of Schistosoma mansoni are produced by adult female worms in mesenteries of infected hosts. Eggs can cross the intestinal barrier and form granulomas in the tissue or breach and exit the host through fecal excretion. These interactions may affect the host microbiome assemblages. Given the potential for schistosomal alteration of host gut microbiome and subsequent effects on the fecal bacterial composition, it is important to conduct controlled microbiome studies on model animals. While pursuing these studies, it is important to take into account the different conditions in which microbiome studies are conducted and their consequent impacts on variability and …

Analyzing Microbial Communities In Porites Astreoides, Sanaa Caldwell, Katie Parker, Daniel Barshis

Biological Sciences Faculty Publications

Coral reefs are vital marine ecosystems that support many different species and ecosystem services through protecting coastlines from storms and erosion, providing raw materials for medication, and facilitating diversity amongst marine species. Anthropogenic climate change is causing ocean temperatures to exceed the range of coral thermal tolerance resulting in more coral bleaching events globally. However, some corals have the capacity to withstand high temperatures through different thermal tolerance strategies. A balanced microbial community plays a critical role in coral health and recent studies have indicated that a balanced microbiome is associated with temperature-stress tolerant corals. Yet, there is a severe …

Exploring The Role Of Host Immune Responses And Bacterial Sensory Pathways In The Development Of Treponema Denticola-Mediated Periodontal Disease, Annie N. Hinson

Theses and Dissertations

Periodontitis is a chronic inflammatory disease driven by dysbiotic microbial communities and maladaptive host responses. Treponema denticola, a spirochete pathobiont, is strongly associated with disease severity. However, the molecular mechanisms underlying its interaction with host tissues and regulation of virulence remain incompletely understood. This work examines both the epithelial immune responses and bacterial transduction systems that shape T. denticola pathogenesis. Transcriptomic analysis of gingival keratinocytes revealed IL-36γ as the most highly induced cytokine following infection with T. denticola. Functional studies demonstrated that IL-36γ induction occurs in a dose- and time-dependent manner through TLR2/6 signaling, requiring the NF-κB, PI3K/Akt, and …

Functional Analysis Of Ligand Binding Domain 2 Of Outer Surface Protein C Of Borreliella Burgdorferi, Gavin Z. Chambers

Theses and Dissertations

Lyme disease, caused by the spirochete Borreliella burgdorferi, is the most common vector-borne infection in North America. A critical determinant of early mammalian infection is outer surface protein C (OspC), a highly variable lipoprotein that is indispensable for spirochete survival but whose precise function remains unresolved. OspC has been proposed to mediate diverse roles, including complement evasion, adhesion, tissue dissemination, and antiphagocytic activity, supporting the view that it is a multifunctional virulence factor. Structural studies have identified two putative ligand-binding domains (LBD1 and LBD2), with LBD2 forming a solvent-exposed crown at the membrane-distal end of the OspC dimer.

To …

Bio-Inspired Airfoil Design Based On The Aerodynamic Analysis Of A Bird Feather Microstructure, Jack C. Ihm

College of Graduate Studies: Theses & Dissertations

This study investigates the aerodynamic effects of a bio-inspired airfoil design based on the microstructure of a secondary feather from a female wood duck. The research hypothesized that incorporating barbs and barbules into an airfoil design would modify boundary layer behavior by introducing localized flow disturbances. To test this, a bio-inspired airfoil was developed by integrating the microstructure into a NACA 0012 airfoil and analyzed using computational fluid dynamics (CFD) simulations at angles of attack of 0°, 5°, 10°, and 15°. The results showed that the bio-inspired airfoil influenced boundary layer behavior, producing localized flow disturbances near x/c = 0.7 …

Molecular Mechanisms Of Kynurenine-Mediated Lifespan Regulation In Yeast, Ryan E. Counterman Jobb

Theses and Dissertations

The kynurenine pathway is a highly conserved pathway that acts as the primary pathway for tryptophan catabolism and has been found to be involved in lifespan regulation across many diverse organisms. Caloric restriction (CR) is the only known dietary intervention to date that both increases the mean lifespan and the health span of multiple organisms, although the molecular mechanisms of these effects are not fully understood. There are known and sometimes contradictory relationships between the kynurenine pathway activity and the life- extending effect of CR but the crosstalk between the two is not well characterized. Understanding these relationships could lead …

Soils From Different Solidago Altissima Clones Mediate Intraspecific Interactions In Schizachyrium Scoparium, Sarah Alharbi

Masters Theses

While previous studies have examined plant intraspecific competition and soil microbial effects separately, fewer studies have investigated how these factors interact, particularly in the context of genotype-specific soil conditioning effects. Plant-soil feedbacks can significantly alter interspecific competitive outcomes, yet few studies have examined how local variation in soil conditioning influences plant responses to density-dependent competition. I investigated how soil inocula derived from four Solidago altissima clones that were previously shown to differ in their soil conditioning effects (Foster et al., 2022) affected the growth and root traits of Schizachyrium scoparium across a gradient of intraspecific competition (1, 2, or 4 …

Influence Of Thermophilic Biofilms On Proliferation Of Opportunistic Pathogens In Domestic Water Systems, Olajumoke Akinbami

Masters Theses

Domestic hot-water systems quietly grow biofilms under elevated temperatures that can serve as shelter to Opportunistic Premise Plumbing Pathogens. Although focus has been placed on microbes in domestic systems, there has been no clear conclusion on the mechanism of survival and persistence of opportunistic pathogens despite various disinfection strategies. In this study, we determined the distinction between showerhead and drip-flow biofilms at the whole-community level and analyzed the coexistence of thermotolerant organisms with opportunistic genera, such as Legionella. We sampled eight showerheads and simulated four biofilm samples in the laboratory using the drip-flow biofilm reactor.

We observed a strong …

Identification Of Gut Bacterial Symbionts From Atlantic Salmon That Are Involved In Metabolizing Feed Ingredients Derived From Soybean Meal, Emily Fowler

Electronic Theses and Dissertations

In the aquaculture industry, plant-based protein sources have emerged as an economically and environmentally sustainable replacement for traditional animal protein sources. Yet, due to reduced digestibility, the presence of anti-nutritional factors, as well as an increased risk of inflammation of the digestive system, the inclusion of plant-based protein ingredients must be limited, especially in the diets of carnivorous fishes. To gain further insight on how to adapt the fish digestive tract to more effectively utilize or tolerate plant-based protein ingredients, slurry from cultured Atlantic Salmon (Salmo salar) was used as a proxy in an in vitro model system to identify …

Quantification Of Auxin-Cytokinin Response During Lateral Root And Nodule Development In Medicago, Aniket Singh

Electronic Theses and Dissertations

Lateral roots (LRs) and nodules are post-embryonic root organs in legumes formed through the activity of de novo meristem. LRs, which are present in all vascular plants, are developed primarily via intrinsic developmental cues, whereas root nodules, found only in a small group of plants known as legumes, are formed in response to legume–rhizobium symbiosis. LRs arise from the pericycle layer of the root, whereas nodule initiation can begin either in the pericycle (indeterminate nodules with persistent meristem, as in Medicago) or in the cortex (determinate nodules with transient meristem, as in soybean), pointing toward a conserved developmental origin, especially …

Fungal Biotransformation Of Thin Stillage And Soybean Hulls And Bacterial Production Of Lysine In Ethanol Industry Co-Products, Bhawana Khadka

Electronic Theses and Dissertations

Thin stillage (TS) and whole stillage (WS) are major by-products of the corn ethanol industry. These by-products are generated in large quantities and contain nutrients that remain after yeast fermentation. Current industrial utilization of these by-products involves a series of downstream processing steps such as centrifugation and evaporation, which are energy-intensive and often result in nutrient losses. Traditionally, these by-products are dried to produce distillers dried grains with solubles. In this study, microbial fermentation was investigated as an alternative bioprocessing tool to expand the utilization of these by-products. The first objective was to explore the upcycling of TS, both alone …

Sars-Cov-2 Variants Of Concern: Biological Implications Of Immunogenicity And Comorbidity, Jacob Nichols

Theses and Dissertations (ETD)

When coronavirus disease 2019 (COVID-19) emerged in human populations in late 2019, there was nothing known about the virus, SARS-CoV-2, besides that it had a high degree of similarity to severe acute respiratory syndrome coronavirus (SARS-CoV) and middle east respiratory syndrome coronavirus (MERS-CoV). The prior published research on these viruses set a strong foundation for the research approaches needed to quickly stand-up medical countermeasures for treatment of COVID-19. However, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) soon revealed a high propensity to evolve new variants that could engage host cell receptors with greater affinity and escape host immunity. With each wave …

Characterization Of Novel Hantaviruses In Paraguay, Ifebuche Nnamani

Theses and Dissertations (ETD)

The genus Orthohantavirus, is a group of tripartite, negative-sense RNA viruses of which some members are the etiologic agents for hemorrhagic fever with renal syndrome (HFRS) or orthohantavirus cardiopulmonary syndrome (HCPS) in the Old and New Worlds, respectively. Sampling of wild rodents for these viruses has increased our knowledge of their host range and continues to lead to the discovery of new orthohantaviruses species and strains. HFRS and HPS have case fatality rates of up to 15% and 40%, respectively. And in some years, morbidity rates of up to 100,000 cases can be reached, globally. The epidemiological evidence of human-to-human …

Reducing Stress: The Role Of The Rcl Operon In Bacterial Stress Response To Hypothiocyanite, Julia Meredith

All ETDs from UAB

The human immune system utilizes a variety of weapons against bacterial communities within the body, which allow it to control the overall microbial population. Understanding how some bacterial species are able to better deal with and survive damage by the immune system while others are not, can help us elucidate the complex relationship between us and our microbiome. One tool that immune cells use for microbial control is the potent oxidant hypothiocyanite/hypothiocyanous acid (-OSCN/HOSCN). The response mounted by Escherichia coli and other bacterial species in order to survive damage caused by HOSCN is the focus of this dissertation. In the …

Role Of Coronavirus Envelope Protein On Infectivity, Garrett E. Zaagman

Dissertations

Coronaviruses infect a wide range of hosts and have caused recent human pandemics. While much has been learned, gaps remain in understanding viral assembly and egress, particularly the role of the envelope (E) protein in specific infectivity. To study this, recombinant MHV viruses were created with E gene deletions and HiBiT-tagged spike (S) proteins to monitor virion release. In mouse cells, MHV∆E S-HiBiT secreted less spike and fewer infectious particles, indicating reduced specific infectivity. Comparisons between human HeLa-CEACAM and mouse DBT cells showed species-specific support for virus production. TMED10, a host membrane protein, was tested as a potential E-interacting partner …

Investigation Of Multiple Factors Influencing The Vector-Borne Transmission Of Salmonella Typhimurium Via Blattella Germanica, Landen Sean Van Hulzen

Dissertations and Theses

The German cockroach (Blattella germanica) is a common urban pest that lives ubiquitously in human structures around the world. German cockroaches are a severe public health threat as they are known carriers and vectors of human pathogens including Salmonella enterica, Escherichia coli, Shigella spp, and more. German cockroach infestations have been associated with Salmonellosis outbreaks. While mechanical transmission by the German cockroach vector is well known, new research suggests that more nuanced biological transmission has a role in maintaining and transmitting Salmonella enterica serovar Typhimurium (S. Typhimurium). Despite this recent finding, many biological variables influencing vector-borne transmission of S. Typhimurium …

Antibody-Induced Multicellular Behaviors Of Salmonella Typhimurium, Samantha K. Lindberg

Electronic Theses & Dissertations (2024 - present)

Salmonella enterica is a globally distributed bacterial pathogen that is commonly acquired through consumption of contaminated food and water. The enterica subspecies includes over 1500 unique subvariants, or serovars, that are classified by antigen presentation. Due to its global environmental prevalence, broad host range, and increasing pathogenicity, serovar Typhimurium is of particular concern for human and animal health, as well as food safety. Once in the gastrointestinal tract, Salmonella Typhimurium (STm) exploits host immune mechanisms, implements an arsenal of virulence factors, and outcompetes the microbiota to invade the intestinal epithelium. While STm infections are generally self-limiting and relatively mild, emerging …

Wasting Disease Of A Marine Foundation Species Links Community Interactions To Disease Dynamics, Lillian R. Aoki, Olivia J. Graham, Reyn M. Yoshioka, Rebecca Maher, Lindsay Alma, Eileen E. Hofmann, Colleen A. Burge, Drew Harvell, Maya L. Groner

CCPO Publications

Infectious disease plays a key role in shaping marine communities, including in seagrass meadows, which form biodiverse coastal habitats. Eelgrass (Zostera marina) is the most widespread seagrass species and is susceptible to seagrass wasting disease, caused by the protist Labyrinthula zosterae. As a foundation species, eelgrass strongly influences ecosystem structure, function and services; recent work has begun to explore the links between critical community interactions and seagrass wasting disease. Here, we highlight recent advances about how the eelgrass community regulates and responds to seagrass wasting disease, from the microbiome to herbivores and filter feeders. We further show …

Spatial And Temporal Shifts In Bacterial Communities And Nutrient Removal In Two Springfield Wastewater Facilities, Rubina Sherchan

Graduate Theses/Dissertations

Bacterial communities play a central role in the biological nutrient removal and overall performance of wastewater treatment plants (WWTPs). However, very few is known about the spatial and seasonal dynamics of these communities. This study explores the spatial and seasonal variations in bacterial community composition across multiple treatment stages in two municipal WWTPs—Southwest (SWTP) and Northwest (NWTP)—in Springfield, Missouri. Using 16S rRNA gene amplicon sequencing, I analyzed 87 samples collected during summer and winter of 2024 from influent, sludge, aeration basins, clarifiers, and final effluent stages. I retrieved 16.4 million (summer) and 3.2 million (winter) sequences. Proteobacteria dominated both seasons …

Characterization Of Adaptive Quorum Sensing In Pseudomonas Aeruginosa, Kayla Simanek

Electronic Theses & Dissertations (2024 - present)

Pseudomonas aeruginosa is an opportunistic pathogen that is highly antibiotic resistant and causes tens of thousands of infections each year. It persists in hospitals by forming biofilms on medical equipment like ventilators and catheters, making it difficult to eradicate. Biofilm formation, among other virulence factors of P. aeruginosa, is regulated through quorum sensing, a mechanism of bacterial communication. Quorum sensing is primarily coordinated through LuxI-R type systems comprised of an autoinducer synthase and receptor, respectively. Canonically, the first system to be expressed is LasI-R, and LasR upregulates the second arm of the quorum sensing network, the RhlI-R system. Research …