Microbiology Commons^™

Exploring Competitive Dynamics Between Pseudomonas Baetica And Salmonella In A Polymicrobial System, Braxton E.M. Stout

Electronic Theses and Dissertations

The global rise in antimicrobial resistance has catalyzed a greater focus on polymicrobial research aimed at characterizing microbial interactions and uncovering new therapeutic strategies. Though it has long been of interest, environmental soil-derived bacteria remain largely under-represented in research and thus uncharacterized in this field. This project aimed to identify soil microbes that display inhibitory mechanisms to potential nosocomial pathogens. Soil sample S1S3-2 displayed inhibitory phenotypes toward numerous bacteria and was identified as Pseudomonas baetica. In both planktonic and biofilm growth, P. baetica was able to inhibit Salmonella arizonae through a contact-dependent and temperature-dependent manner. Screening of a transposon insertion …

Strategic Enhancement Of Biofuel Production Using Bacillus Subtilis As A Model Organism: A Dual Approach Of Gene Essentiality Mapping And Optimization, Angela A. Zebede

Electronic Theses and Dissertations

With rising global demand for renewable energy, enhancing microbial platforms for biofuel production is vital. This study engineered Bacillus subtilis to improve resilience to solvent stress by targeting membrane lipid biosynthesis pathways essential for integrity. Growth curve analysis (OD₆₀₀) showed that phosphatidylserine (PS) and phosphatidylethanolamine (PE) knockout strains grew significantly slower than wild type (168), indicating the importance of these lipids under stress. Laurdan fluorescence spectroscopy was used to assess membrane properties. Generalized polarization (GP) analysis revealed a concentration-dependent reduction in membrane order upon 1%–2% 1-butanol exposure. GP values declined progressively from control to 2% treatment, reflecting increased membrane fluidity …

Effect Of Histidine Tag On The Enzymatic Activity Of Pseudolysin And Its Interaction With The Streptomyces Metalloprotease Inhibitor (Smpi) Protein, Adewale Adeboye Adewole

Electronic Theses and Dissertations

Pseudolysin (PLN) protein is a major virulent factor in Pseudomonas aeruginosa infection. PLN belongs to the family of zinc-dependent metalloproteases. It is naturally inhibited by a smaller protein called streptomyces metalloprotease inhibitor (SMPI), secreted by Streptomyces nigrescens. Previous and current studies demonstrate that the interaction between PLN and SMPI offers unique mechanistic insights for drug development against infections caused by P. aeruginosa. This study investigated the effect of N-terminal Histidine (His) tag on the enzymatic activity of PLN and its interaction with SMPI. His-tagged PLN and SMPI proteins were expressed in bacteria and purified by affinity chromatography …

A Toi Rna Editing In Three Members Of The Microbotryum Violaceum Fungal Complext And Characterization Of Adar Genes Of Microbotryum Superbum., Shikhi Baruri

Electronic Theses and Dissertations

A-to-I RNA editing is a process that occurs post-transcriptionally. Through this process, adenosine (A) is replaced by inosine (I) in RNAs by adenosine deaminase enzymes that act on the single-stranded RNA. These enzymes, also known as ADARs, act on RNA. The translation and splicing mechanisms subsequently interpret inosine as guanosine (G), which effectively alters genetic information. This kind of RNA alteration can cause both nonsynonymous and synonymous changes in codon, which may have an impact on protein function. A group of over 89 basidiomycete fungal species known as the Microbotryum violaceum complex infects a similarly large group of plant host …

Orchestrated Architecture Dismantling: Borrelia Burgdorferi’S Coordinated Destruction Of Adaptive Immune Responses, Craig A. Land

Electronic Theses and Dissertations

Borrelia burgdorferi represents the most prevalent vector-borne bacteria in North America, affecting ~476,000 Americans annually yet persisting despite robust immune responses. This project reveals spirochetes target adaptive immunity through coordinated lymphatic colonization and precision immune dysregulation.

Using comparative host models (C57BL/6 vs. reservoir Peromyscus leucopus), we demonstrate lymph node colonization within 48-hours, with spirochetes exploiting LYVE-1-positive lymphatic structures as dissemination highways while avoiding the interior of follicular regions. Critically, we identify targeted CD4+ T-cell elimination in germinal center border zones at 21-DPI, precisely when these cells should be coordinating protective immunity. Concurrent upregulation of inhibitory receptor CD32b creates molecular …

Sars-Cov-2 Modulates Pla2g2d/Pgd2 Signaling To Manipulate Host Immune Response To Increase Disease Severity., Dante L. Hamiel

Electronic Theses and Dissertations

Background: Previous studies suggested that SARS-CoV-2 infection could cause diverse immune and metabolic dysregulations, although the underlying mechanisms remain poorly understood. In addition, viruses may escape the immune response by modulating cellular metabolism, mitochondria, and protein functions indicating that immune or metabolic components may represent novel targets for anti-viral strategies. However, whether immune and metabolic disorders interact or affect each other waits to be determined. On the other hand, previous investigations identified the critical roles of a cell membrane-bound phospholipase PLA₂G2D and its downstream products PGD₂ in regulating the metabolism and inflammation in respiratory system. More importantly, …

Exploring The Antagonistic Polymicrobial Interaction Of Alcaligenes Faecalis With Prokaryotic And Eukaryotic Microorganisms, Aly Hargrave

Electronic Theses and Dissertations

Alcaligenes faecalis, a generally nonpathogenic, Gram-negative bacterium, has been observed by our laboratory to possess antagonistic activity against antimicrobial-resistant genera Staphylococcus, Candida, and Bacillus. This study focused on exploring the inhibition mechanism of A. faecalis against the three genera, with an emphasis on S. aureus. A library of A. faecalis transposon mutants was constructed and identified fourteen possible genetic factors linked to this inhibitory activity. Characterization of this inhibition indicated that A. faecalis suppressed S. aureus planktonic and biofilm growth via a contact-dependent mechanism, with pronounced effects during biofilm attachment phase. In a Caenorhabditis elegans …

The Impacts Of Management Practices On Industrial Hemp Chemistry, Physiology, And Microbiome Ecology., Allison Marie Rietman

Electronic Theses and Dissertations

Agroecology incorporates ecology and agronomy to inform crop management. Industrial hemp (Cannabis sativa L.) is a prime system to investigate agroecological questions because it has only recently been authorized for commercial cultivation and its sensitivity to biotic and abiotic stressors is unresolved. Here, I investigate how some conventional management choices, including the application of fungicide, plastic mulch, and irrigation, impact the physiology, chemistry, and microbial ecology of industrial hemp. Overall, I found that industrial hemp can be both resilient and resistant to these stressors, an important development for the industry. In Chapter II, I tested the impacts of fungicide application …

Integration Of Multi-Omics Datasets Evaluating Structural And Functional Features Of The Gut Microbiome In People With Hiv (Pwh)., Aakarsha Vijayakumar Rao

Electronic Theses and Dissertations

Gut dysbiosis characterized by reduced abundance of beneficial butyrate-producing bacteria has been independently linked to HIV-1 infection and heavy alcohol drinking. Further, gut dysbiosis results in loss of gut barrier integrity, microbial translocation and host-specific systemic inflammation. Therefore, to evaluate the functional consequences of structural changes in the gut microbiome, integrated data analysis is imperative. In this dissertation, we perform integrated analyses using data from multi-omics platforms to examine the structural and functional features of the gut microbiome of PWH. Gut microbiome composition was evaluated by sequencing V4 region of 16S rDNA, concentrations of metabolites and host-specific immune markers were …

Isolation, Characterization & Identification Of Cultivable Field Pea Seed Bacteria In South Dakota, Diana Wairimu Njuguna

Electronic Theses and Dissertations

Pea (Pisum sativum L.) is a legume, cultivated on over 25 million acres, particularly in Canada, France, China, Russia, and the United States. Its seeds harbor microbiota including bacteria and fungi, either on the surface (epiphytes) or within seed tissues (endophytes). These microbes can be pathogenic or beneficial, enhancing seedling metabolism and protection from environmental stresses. Characterizing beneficial seed microbiota enables development of microbial inoculants for seed treatment. Seeds from 12 varieties (green or yellow seed coats) were harvested from Sturgis and Dakota Lakes in 2021. Epiphytes were extracted using sonication followed by grinding the seeds to obtain endophytes. Isolates …

The Formation And Degradation Of Carboxysomes During Heterocyst Development In Filamentous Cyanobacteria, Maxwell Anton Jakubiak

Electronic Theses and Dissertations

Photosynthetic carbon (CO2) fixation and solar-powered nitrogen (N2) fixation are foundational biochemical processes that introduce essential elements into the biosphere. While atmospheric dinitrogen gas (N2) is unusable to most organisms, diazotrophs utilize the nitrogenase enzyme to reduce N2 to ammonia. Because all known nitrogenases are highly susceptible to irreversible oxygen inactivation, N2 fixation is generally incompatible with oxygenic photosynthesis. However, several cyanobacteria are both photosynthetic and diazotrophic, balancing these opposing metabolisms by temporally or spatially separating the respective enzymes. Anabaena sp. PCC 7120 is a model multicellular cyanobacterium that grows in filaments of photosynthetic vegetative cells, which respond to nitrogen …

Comparative Evaluation Of Fungal Strains For The Bioprocessing Of Soybean Hulls Using Cheese Whey As Fermentation Medium, Ruchita Bhattarai

Electronic Theses and Dissertations

Globally, soybean is the most widely cultivated oilseed crops, and serve as the primary source of cooking oil, contributing to around 50% of global oilseed production. During the industrial processing of soybean oil, a significant amount of soybean hull (SH) is produced, which makes up about 8% of the grain yield, resulting in approximately 18–20 million tons of hulls, one of the largest by-products in the soybean industry. The hulls have limited commercial value and are primarily used as fiber additives in dairy cattle feed, but a large portion is still discarded as agricultural waste. Similarly, cheese whey (CW), a …

Isolation And Characterization Of Rhizobia Bacteria From Midwest Soils, Jr Loya

Electronic Theses and Dissertations

Symbiotic bacteria, known as rhizobia, can colonize the roots of legume plants and then convert atmospheric nitrogen into a usable form by the host through the process called biological nitrogen fixation (BNF). Many of these bacteria produce other plant-growth-promoting benefits, such as nutrient acquisition, phytohormone production, and the modification of plant stress responses. Agricultural inoculants made from these bacteria have helped reduce the need for man-made nitrogenous fertilizers, known causes of pollution and eutrophication. There are additional benefits in developing bacteria inoculants local to regional soils. The ability to survive and compete in Midwest soil and climates present an opportunity …

Deciphering Genetic Contributions To Antibiotic Persistence: Insights From Multi-Gene Knockout Studies, Elise K. Schweer

Electronic Theses and Dissertations

Antibiotic resistance is a critical global health challenge, complicating efforts to treat bacterial infections effectively. Antibiotic persistence plays a significant role in driving resistance. Persisters are a subpopulation of non-dividing bacterial cells that often evade lethal antibiotic treatment. Once antibiotics are removed, these cells "resurrect," resuming growth and may lead to relapsing infections. Notably, the progeny of persister cells exhibit a heightened tendency to acquire mutations or incorporate antibiotic resistance genes, further compounding the challenge of antibiotic resistance. In our recent study, we knocked out seven hypothetical genes and observed a significant reduction in persister levels. Compared to the wild …

Biotransformation Of Camelina Meal Via Pretreatment And Fermentation For Functional And Nutritional Upgrading, Ram Kumar Bhattarai

Electronic Theses and Dissertations

Camelina sativa, an oilseed crop of the Brassicaceae family, has seen increased cultivation in North America in response to the rising demand for biofuels over the past two decades. With this growth, camelina meal, a byproduct of oil extraction, has emerged as a promising yet underutilized resource, particularly for applications in animal nutrition. This study aimed to evaluate the potential of camelina meal as a substrate for submerged fungal fermentation and to examine how this bioconversion affects its physicochemical and nutritional properties. The influence of mild pretreatments (unwashed, washed only, and washed/dried and ground) on fermentation outcomes was also assessed. …

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 …

Interactions Of The Sars-Cov-2 Viral Genome 3’-Untranslated Region With Viral And Host Rnas, Caleb Frye, Mihaela Rita Mihailescu

Electronic Theses and Dissertations

This dissertation focuses on the characterization of RNA-RNA interactions within the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome and with host microRNAs. As the causative agent of coronavirus disease 2019 (COVID-19), SARS-CoV-2 has evolved rapidly since its appearance. This has warranted prompt characterization of the virus particularly of its single stranded RNA (ssRNA) genome. By using a combination of bioinformatics, biophysics, and/or biological assays, we analyzed the SARS-CoV-2 viral genomic RNA and uncovered interactions of genomic RNA with host RNAs, highlighting an underutilized method of targeting RNA viruses. We showed here that the conserved elements in the viral genomic …

The Relationship Between Salvinia Molesta Infestation And The Microbial Ecology Of Wetlands At Caddo Lake, Tx, Usa, Martin Hadsell

Electronic Theses and Dissertations

Wetlands are known to host many beneficial geochemical processes such as nutrient cycling and carbon sequestration. These processes are primarily driven by the microbial communities within a wetland. The establishment of non-native invasive species within wetlands has the potential to alter the once beneficial functionality of wetland microbial communities due to the sensitivity microbes have towards environmental changes. Because of this, several metagenomic studies have been conducted to assess microbial community response to aquatic plant invasion; however, there haven’t been any studies researching the influence of Salvinia molesta (D.S. Mitch.), commonly known as Giant Salvinia, on microbial communities. The goal …

Analysis Of Microbiome In Various Human Medical Conditions Using Next-Generation Sequencing Of The 16s Rrna Gene, Pallavi Sharma

Electronic Theses and Dissertations

Studies of human microbiomics, spanning diverse anatomical sites, offer a unique perspective into the complex microbial communities within the body. Influenced by age, gender, and disease, it mirrors broader microbial ecosystems. This study specifically delves into the ocular and gastrointestinal tract microbiome, comparing its composition in 43 healthy and diseased patients using 16S rRNA gene sequencing. We identified a rich microbial landscape, identifying over 338 bacterial taxa. Notably, Acinetobacter, Cutibacterium, Acidovorax and Herbaspirillum were prevalent in the ocular samples and Akkermansia, Faecalibacterium, and Anaerostipes were mostly prevalent in gut samples. Moreover, dry eye disease conditions were characterized by …

Differential Responses Of Human Neutrophils To Legionella Species., Hannah E. Hanford

Electronic Theses and Dissertations

Legionella species are Gram-negative facultative intracellular bacterial pathogens commonly found in freshwater or soil environments where they survive and replicate within their natural protozoan hosts. Humans serve as an accidental host for Legionella species after inhalation of contaminated aerosolized water droplets or soil particles enabling the bacteria to enter the lungs, infect alveolar macrophages, and result in severe pneumonia. Evasion of degradation and proliferation within alveolar macrophages by Legionella species and manifestation of “Legionnaires’ disease” is dependent on the Dot/Icm type IV secretion system (T4SS). The T4SS apparatus is a needle-like channel for translocation of over ~350 effector proteins, depending …

Sequence Specificity Of Non-Structural Protein 15 From Sars-Cov-2, John Vincent Mcguire

Electronic Theses and Dissertations

The SARS-CoV-2 virus was recently able to cause a global pandemic, largely due to the asymptomatic period that occurs during initial infection. During this period, patients are infectious but do not exhibit symptoms, resulting in the unintentional spread of the virus. This occurs because the virus effectively avoids detection by the immune system during the early stages of infection, and as symptoms are the result of immune system activation, they never manifest. Non-structural Protein 15 (Nsp15) is one of the tools the virus uses to remain undetected, via its degradation of Pathogen-Associated Molecular Patterns (PAMPs) that would otherwise be recognized …

Rna Secondary Structures: Structure-Function Studies In Viral And Neurodegenerative Diseases, Caylee Cunningham

Electronic Theses and Dissertations

The studies within this dissertation highlight the importance of RNA structure and function, highlighting potential mechanisms in the pathogenesis of the SARS-CoV-2 virus and hallmarks of ALS. Specifically, the s2m, a 41-nt structure found within the hyper variable region of the 3’ UTR of the SARS-CoV-2 virus, was investigated as this element has no definitive role in viruses but its high conservation in several viral families warrants investigation. We used various biophysical techniques to determine if a palindromic sequence in the terminal loop could participate in dimerization activities, which may have implications in recombination and the longevity of the virus. …

Manipulation Of Cellular Proliferating Cell Nuclear Antigen (Pcna) During Herpes Simplex Virus Type 1 Infection, Jessica Packard, Jill A. Dembowski

Electronic Theses and Dissertations

Herpes simplex virus type 1 (HSV-1) is a double stranded DNA virus that replicates within the nucleus of host cells. Using accelerated native isolation of proteins on nascent DNA (aniPOND), cellular proteins were found to associate with HSV-1 replication forks. One host factor of interest is proliferating cell nuclear antigen (PCNA), the DNA sliding clamp that tethers DNA polymerases and DNA damage response proteins to replicating cellular DNA. Although HSV-1 encodes its own DNA polymerase processivity factor (UL42), PCNA plays a key role in HSV-1 infection. We used commercially available PCNA inhibitors and short hairpin RNAs (shRNA) to investigate how …

Pilz Domain-Containing Proteins Regulate Motility In Acinetobacter Baumannii, Gabriel Smith

Electronic Theses and Dissertations

Acinetobacter baumannii is an increasingly multidrug-resistant pathogen contributing to hospital-acquired infections, necessitating a greater understanding of how it interacts with its surroundings. Many bacteria utilize different methods of bacterial motility to move about and interact with these surroundings. A bacterial second messenger, cyclic diguanosine monophosphate (c-di-GMP), can regulate various motility factors that are potentially advantageous for survival in and adaptation to their environment. Concentrations of c-di-GMP are regulated by specific synthesizing and degrading enzymes. Controlled levels of c-di-GMP allow interaction between the c-di-GMP and its binding effectors that induce changes in bacterial phenotypes such as biofilm formation and motility. A …

Characterizing Enterobacter Cloacae Genetic Elements Responsible For Interactions With Candida Albicans, Abigail Suarez

Electronic Theses and Dissertations

Polymicrobial interactions are an important, yet understudied area of research. Candida albicans is the most common human fungal pathogen. The bacterial genus, Enterobacter, is a source of nosocomial acquired infections and increased drug resistance. Our lab has previously discovered that Enterobacter preferentially adheres to C. albicans hyphae. From an E. cloacae transposon library screen, six candidates displayed reduction in C. albicans attachment. These candidates were identified genetically and characterized for involvement in attachment to C. albicans. A fluorescent plasmid was introduced into E. cloacae to measure and observe adherence to C. albicans in planktonic and biofilm growth. In vivo …

Characterization Of A Novel Metalloid Reductase Outer Membrane Porin Rara From Sulfurospirillum Barnesii Ses-3, Narthana Jeganathar Kanmanii

Electronic Theses and Dissertations

Sulfurospirillum barnesii SES-3 is a gram-negative epsilon proteobacterium that grows anaerobically by coupling the oxidation of lactate to the reduction of arsenate. Previous work on metal and metalloid metabolism in S. barnesii had identified a redox active protein, designated RarA that when provided an artificial electron donor (i.e., reduced methyl viologen, FMN, or NADH), could reduce a variety of substrates, including arsenate, selenate, selenite, nitrite, and even phosphate. The enzyme’s inferred amino acid sequence showed weak similarity (30%) to outer membrane porins with homologs found in only certain species of Sulfurospirillum. Further, the enzyme did not contain cysteine residues …

Advancing Biological Applications Through Microfluidic-Based Tool Development, K.M. Taufiqur Rahman

Electronic Theses and Dissertations

This research undertook an interdisciplinary approach, integrating bioengineering, microbiology, molecular biology, and systems biology to investigate bacterial dynamics behavior. Specifically, it delved into the development of microfluidic devices for biological applications such as bacterial cell counts, real-time observation of plant roots (here, specialized lectin-coated microbeads are used that mimic root characteristics), and soil microbe interactions. Furthermore, Next-Generation Sequencing and systems biology methodologies were employed to explore the intricate, multifaceted survival mechanisms of Escherichia coli persister population. Studying and quantifying persisters or testing for the existence of VBNC (viable but nonculturable) is challenging. These experiments require precise counts. It has been …

Role Of Interferon Induced Transmembrane Protein 3 (Ifitm3) And Neural Precursor Cell Expressed Developmentally Down Regulated Protein 4 (Nedd4) In Seneca Virus A (Sva) Replication, Shamiq Aftab

Electronic Theses and Dissertations

In this study, we studied the role of IFITM3 in SVA replication in vitro by silencing the endogenous IFITM3 in NCI-H1299 cells. An average of 1.94-fold increase in SVA VP2 expression and an average of 4.4-fold increase in supernatant virus titer was observed after 24h SVA isolate SD15-26 infection. We have indeed observed an increase in viral mRNA copies by an average of 5-fold in IFITM3 silenced NCI-H1299 cells at 8 hpi indicating that IFITM3 restricts SVA replication post virus attachment and entry. A positive correlation between interferon-alpha induced IFITM3 upregulation and reduced SVA replication was observed. To further confirm …