Life Sciences Commons^™

The Effects Of Temperature And Phasins On Polyhydroxyalkanoic Acid (Pha) Accumulation In Heterologous Hosts Containing A Pha Synthase From Vibrio Species B-18, Edward R. Lyons

Master's Theses, Dissertations, Graduate Research and Major Papers Overview

Polyhydroxyalkanoic acids (PHA) are bacterial carbon reserves with application as biodegradable biocompatible plastics. PHA synthase, encoded by phaC, creates the PHA polymer. Evidence presented by many researchers has emphasized the role of phasin proteins in PHA accumulation. Phasins have been implicated in PHA granule localization, PHA synthase activation, and possibly chaperone and nucleoid binding functions. The phaC gene from Vibrio species B-18 with and without an upstream gene showing homology to phasin genes (orf2) were placed into runaway replication vectors, which use temperature to control gene expression. The aim of this thesis is to explore earlier observations that lower temperatures …

Determining Effects Of Management Practices On Potato Early Dying And Soil Microbiome And Assessing Risk Of Fungicide Resistance In Verticillium Dahliae, Kedi Li

Electronic Theses and Dissertations

Potato early dying (PED) is a yield-constraining soilborne disease of potato, caused by Verticillium spp. with V. dahliae being the predominant causal agent. Since the pathogen inhabits soil for long periods, PED management aims to reduce the population of V. dahliae in soil. Benzovindiflupyr and azoxystrobin are effective chemicals and frequently used in the control of V. dahliae. In this study, field trials were conducted at Aroostook Farm, Presque Isle, ME in 2019 and 2020. Chemical and biological products have been studied for PED control, and fungicide resistance was also examined. To evaluate fungicide resistance, benzovindiflupyr was characterized on …

Analysis Of Spore Shape Determination In Streptomyces, Ning Sun

Electronic Theses and Dissertations

Streptomycetes are Gram-positive, soil-dwelling bacteria that possess a complex life cycle with the alternation of vegetative mycelium, aerial mycelium, and spores. Streptomyces. coelicolor spore maturation is a complex process that involves spore shape metamorphosis from cylindrical pre-spores into ellipsoid spores, but the details of this process have remained enigmatic. Previously, our lab identified a novel gene ssdA that might play a role in spore shape determination using a transposon-based insertion mutagenesis in S. coelicolor. In this study, I isolated a S. coelicolor ssdA-null mutant that showed increased colony hydrophobicity and misshapen spores in sizes and shapes, confirming the …

Retroviral Infection Dynamics In Maine's Wild Turkeys, Stephanie A. Shea

Electronic Theses and Dissertations

Widespread wild turkey reintroductions in the late 1900s have led to increases in population density and geographic distribution across North America. This rapid population expansion has put them into proximity with closely-related wild and domestic avian species, increasing the risks of pathogen transmission. Lymphoproliferative disease virus (LPDV) is an avian oncogenic retrovirus detected in wild turkeys in 2009, and previously known to infect domestic turkeys. Following its initial detection, surveys reported variable LPDV prevalence across eastern North America with most wild turkeys being asymptomatic, however diagnostic cases revealed 10% mortality of LPDV-infected individuals. Given its recent detection, little is known …

Understanding The Impact Of Chlamydia Trachomatis Infection On Host Vesicular Trafficking, Lisa M. Jorgenson

Theses & Dissertations

Chlamydia trachomatis is a highly evolved obligate intracellular bacterial pathogen. During infection, organisms reside within a host epithelial cell in a membrane-bound vacuole called the inclusion. The inclusion membrane contains both pathogen and host components, including unique chlamydial type III effector proteins called Incs and host lipids like cholesterol and sphingomyelin. The inclusion membrane serves as the main barrier between the bacteria and the host cytosol, and thus, functions as the platform supporting host-chlamydial interactions. Via the inclusion membrane, C. trachomatis intercepts components of host vesicular traffic including many host SNARE proteins, which are a conserved family of eukaryotic proteins …

Metabolic Heterogeneity And The Roles Of Cody And Ccpa In Central Metabolism And S. Aureus Biofilm Formation., Logan L. Bulock

Theses & Dissertations

Staphylococcus aureus is a metabolically versatile human pathogen, causing disease in many areas of the body. Its versatility can be attributed to the fact that it utilizes a variety of tools to adapt to many different environments, including toxins to scavenge from the host and multiple transporters to compete for its preferred carbon sources. S. aureus can also survive in harsh conditions through biofilm development, which are notoriously recalcitrant to antibiotics and immune defenses. Biofilms exhibit marked heterogeneity, with division of labor for production of matrix components and differential gene expression among various niches within the biofilm.

In this study, …

Investigating Antimicrobial Properties Of Snake Venoms Against B. Cereus, B. Subtilis, E. Coli, And P. Vulgaris, Savannah Berger

Honors Program Theses and Research Projects

The increasing incidence of antibiotic resistant bacterial infections has provoked the attention of health officials and scientists as a major threat to global public health. Antibiotic resistance is the product of overprescription by doctors, ineffective or shortened dosing by the patient, and more. When a bacterial infection is ineffectively treated as such, persistent pathogenic cells are given the opportunity to proliferate and spread their resistance to other cells. With bacteria utilizing such tools to fight and ultimately resist our current treatment methods, investigation towards the next novel mechanism of inhibition is essential. Venom is of particular interest to many scientists …

Virology: Mutagenizing The Proteolytic Cleavage Site In The Major Capsid Protein In Ssv1, Thejanee Liyanaarachchi

University Honors Theses

SSV1, spindle-shaped virus 1, is an archaeal virus with a unique shape that belongs to the family of fuselloviruses and infects hyperthermophilic archaea that thrive at 80°C and pH 3. SSV1 has two capsid proteins, VP1 and VP3, that presumably produce the lemon-shaped virion structure characteristic of fuselloviruses. The VP1 amino acid sequence is highly conserved among SSVs. Here, we investigate the importance of the proteolytic cleavage site in position E66 in VP1 by changing its glutamic acid to the other 19 amino acids. We have developed a new protocol for creating SSV1 mutants through HiFi Assembly (NEB) to increase …

Human Plcg2 Haploinsufficiency Results In A Novel Immunodeficiency, Joshua Brandon Alinger

Arts & Sciences Electronic Theses and Dissertations

NK cells are critical for the recognition and lysis of herpesvirus-infected cells. Patients with NK cell immunodeficiency may suffer from unusually severe and/or recurrent herpesvirus infections; however, the genetic cause is frequently unknown. PLCG2 encodes a signaling protein in NK cell and B cell receptor signaling, in which dominant-negative or gain-of-function mutations may cause cold urticaria, antibody deficiency, or autoinflammation. However, loss-of-function mutations and PLCG2 haploinsufficiency have never been reported in human disease. We examined 2 families with autosomal dominant NK cell immunodeficiency with dual high-dimensional techniques, mass cytometry and whole-exome sequencing, to identify the cause of disease. We identified …

Quantitative Characterization Of Microbial Ecologies In Dysbiosis And Infection, Eric Keen

Arts & Sciences Electronic Theses and Dissertations

In 1973, Theodosius Dobzhansky famously wrote that nothing in biology makes sense except in the light of evolution. Today, nearly 50 years later, little in microbiology – or in biology, for that matter – makes sense except in the light of genomics. Microbial genomics populates the field with innumerable testable hypotheses for evaluation in vitro and in vivo, allows us to monitor microbial populations in real time and at a massive scale, and underpins our approach to entire domains of microbiology, including microbial evolution. In this Thesis, I present three studies from my graduate research united by their common theme …

The Effects Of Hydrologic Heterogeneity On Harmful Algal Blooms In Freshwater Reservoir, Lake Sinclair, Georgia, Margaret Blackledge

Biology Theses

Aquatic habitats are frequently studied after a major water quality problem like the occurrence of an algal bloom. In this study, A proactive rather than a reactive response was considered, where the complexity of conditions conducive to uncontrolled cell growth were studied before a bloom took place by sampling regularly. This study aimed to monitor water quality by monthly sampling of algal communities for approximately one year. As the base of the aquatic food web, algae are a highly diverse group of organisms with varying sensitivity to physical and chemical changes in the environment. Four shallow sites were monitored at …

Genetic Characterization Of Antimicrobial Activities Of Endophytic Bacteria Burkholderia Strains Ms455 And Ms389, Jiayuan Jia

Theses and Dissertations

Strains MS455 and MS389, endophytic bacteria, were isolated from healthy soybean plant growing adjacent to a patch of plants affected by charcoal rot disease, caused by the fungal pathogen Macrophomina phaseolina. The complete genomes of both strains were sequenced and identified as Burkholderia species Strain MS455 exhibits broad-spectrum antifungal activities against economically important pathogens, including Aspergillus flavus. Random and site-specific mutations were employed in discovery of the genes that share high homology to the ocf gene cluster of Burkholderia contaminans strain MS14, which is responsible for production of the antifungal compound occidiofungin. RNA-seq analysis demonstrated ORF1, a …

Evaluating The Effects Of Pine And Miscanthus Biochar On Water Activity And Escherichia Coli Populations In Commercial Broiler Litter, Christopher Austin Marty

Theses and Dissertations

The decrease in subtherapeutic antibiotic administration in poultry has increased the need to address production challenges caused by pathogens, such as E. coli. One potential way to improve bird health and reduce bacterial infection is through the addition of litter amendments that absorb moisture. Biochar (BC) has previously been shown to increase water holding capacity in poultry litter, but its effects on E. coli mitigation are unknown. The objectives of this research were to 1) evaluate water activity of poultry litter amended with pine and miscanthus BC, and 2) determine the effects of different BC inclusion rates on litter …

A Temporal Analysis Of The Microbiota And Biofouling Development On Artificial Substrates In The Port Everglades Inlet, Florida, Denise Swack

All HCAS Student Capstones, Theses, and Dissertations

A pilot project was deployed in Port Everglades Inlet, Florida that aimed to evaluate the biofilm composing the microbiome on ecologically engineered artificial substrates used to build Coastal Marine Infrastructure. In April of 2017, an Articulated Concrete Block Mattress comprised of an ecological engineered concrete substrate and a standard smooth surface control substrates were compared. This study will provide a profile on the microbiome community on artificial substrates within Port Everglades Inlet on bio-enhancing concrete-based solutions in our Coastal Marine Infrastructure. To study the microbial community, the 16s rRNA technology was used in Illumina’s high-throughput DNA sequencing. Samples were collected …

Response Of Soil Viral And Microbial Functional Diversity To Long-Term Agricultural Management In Jackson, West Tennessee, Ning Duan

Doctoral Dissertations

Soil microbial communities are a critical component for ecosystem stability and function. Viruses, as an important biotic controller, have the potential to regulate the abundance and diversity of bacterial communities through infection. Soil is known to harbor abundant and diverse viral assemblages but their ecological role and influence on microbial processes has not been fully elucidated. Microbes can be influenced by viruses not only from infection but though biogeochemical feedbacks of the “microbial (bacterium–phage–DOC) loop” or “viral shunt”. However, we know relatively little about the microbial community and function under the regulation of viruses in soil and how they respond …

Microbial Community Dynamics Of A Microcystis Bloom, Helena Pound

Doctoral Dissertations

Harmful algal bloom events are notoriously associated with massive economic and environmental consequences, causing wildlife and human health risks. As these blooms increase in occurrence, duration, and severity around the world, it is essential to understand conditions leading to bloom formation and why they persist. Abiotic factors such as nutrients are commonly considered in bloom dynamics, but biotic interactions with co-occurring microbial species and viruses must also be taken into account. Harmful algal blooms dominated by the cyanobacterial genus Microcystis occur in bodies of water around the world and provide an ideal system in which to study top-down controls on …

Determining The Ecological And Physiological Factors Contributing To The Competitive Success Of Prochlorococcus In The Oligotrophic Ocean, Benjamin C. Calfee

Doctoral Dissertations

Prochlorococcus is a genus of extremely successful marine cyanobacteria. This success is realized through its pervasive biogeographical range and presence in almost all open ocean environments where it usually it the dominant phytoplankton. Limited capabilities of culturing and genetic manipulation of this organism have resulted in assumptions about this success overwhelmingly based on field observations. These studies have assumed adaptations for resource uptake and utilization in nutrient limited environments to cause dominance of Prochlorococcus over other photosynthetic microbes. In an attempt to definitively explain this through laboratory culture, we developed a culturing system to assay questions of nutrient limitation effects …

Perturbing Fatty Acid Metabolism In Enterococcus Faecalis Disrupts Responses To Exogenous Fatty Acids And The Antibiotic Daptomycin, Rachel D. Johnston

Doctoral Dissertations

Enterococcus faecalis is an opportunistic pathogen. In the host, it is exposed to fatty acids which impact cellular physiology and induce tolerance to the antibiotic daptomycin. To determine the requirements for induction of daptomycin tolerance, I examined the impacts of blocking de novo fatty acid synthesis or protein synthesis and removing cell wall. I observed that removal of the cell wall induced daptomycin tolerance, indicating that peptidoglycan is necessary for daptomycin to function. As specific exogenous fatty acids induce protection against daptomycin in E. faecalis, I also opted to examine whether incorporation of these free fatty acids was necessary …

High Arctic Permafrost Microbial Characterizations: Siberian And Svalbard Microbiology Of Ancient And Active Layer Permafrost, Katie Sipes

Doctoral Dissertations

Permafrost is soil that has remained frozen for at least two years. The active layer is a surface portion above the permafrost that experiences seasonal thaw and refreezing. The environmental characteristics of permafrost and active layer are different but are directly related to each other. As the climate continues to warm, the active layer will expand into the permafrost and the continuously frozen soil will be subjected to seasonal thawing. The organisms that inhabit both the active layer and the permafrost soil will respond differently to the climate based on where in the soil they are present and the soil …

Kshv Susceptibility And Transmission Within Tonsillar Specimens, Farizeh Aalam

Pharmaceutical Sciences (PhD) Dissertations

Despite nearly three decades of research, not much is known regarding the early stages of development for KSHV lymphoproliferative disorders, hindering our ability to develop prophylactic measures or effective treatments. This dissertation will focus on the host and viral factors influencing the magnitude and dynamics of KSHV infection in the human tonsil to pave the way for future interventions directed at limiting person-to-person transmission of KSHV.

To understand the contribution of host factors to KSHV susceptibility in B lymphocytes, we generated a library of 40 tonsillar specimens. Our results indicate that the immunological composition of tonsillar lymphocytes varies across our …

Deciphering The Role Of Hsp110 Chaperones In Diseases Of Protein Misfolding, Unekwu M. Yakubu

Dissertations & Theses (Open Access)

Molecular chaperones maintain protein homeostasis (proteostasis) by ensuring the proper folding of polypeptides. Loss of proteostasis has been linked to the onset of numerous neurodegenerative disorders including Alzheimer’s, Parkinson’s, and Huntington’s disease. Hsp110 is a member of the Hsp70 class of molecular chaperones and acts as a nucleotide exchange factor (NEF) for Hsp70, the preeminent Hsp70-family protein folding chaperone. Hsp110 promotes rapid cycling of ADP for ATP, allowing Hsp70 to properly fold nascent or unfolded polypeptides in iterative cycles. In addition to its NEF activity, Hsp110 possesses an Hsp70-like substrate binding domain (SBD) whose biological roles are undefined. Previous work …

Examination Of The Association Between C - Reactive Protein(Crp) And Covid-19 Infection Severity And Length Of Hospitalization, Abdulahi Aremu Ayanwale

Open Access Theses & Dissertations

Chronic stress can lead to many systemic complications and low-grade systemic inflammation including increased levels of inflammatory cytokines, such as C-reactive protein (CRP). CRP is a marker of systemic inflammation and is associated with depression and perceived stress. Elevations can result in ineffective immune responses, thereby increasing the risk of complications and mortality from infections. Recent evidence suggests that uncontrolled inflammatory responses associated with COVID 19 are a major determinant of disease severity.  The COVID 19 is transmitted from one person to another through droplets from coughing, sneezing, talking, touching droplets on surfaces and contamination by hand-to-mouth routes. The mechanism …

Microbial Communities Hosted By Carnivorous Pitcher Plants: Diversity, Recruitment, Functions And Succession In Sarracenia Purpurea Microbiomes, Jacob Jeffrey Grothjan

Theses and Dissertations

The pitcher plant Sarracenia purpurea supplements nutrient acquisition through carnivory, capturing insect prey which are digested by a food web community of eukaryotes and bacteria. Analysis of both bacterial and eukaryotic diversity, and an understanding of bacterial recruitment into pitchers and succession of bacterial and eukaryotic communities over time have not been well explored. This thesis presents three studies designed to address these gaps using field sampling and manipulative greenhouse experiments. Study I compared bacterial and eukaryotic composition and diversity of pitcher communities within and between populations of plants in two distinct wetland habitats. Genetic sequence analysis revealed an underappreciated …

Prospecting For And Isolation Of Microalgae In South Texas, Mauricio Alejandro Peña

Theses and Dissertations

Microalgae is a very important microorganism that can be used for beneficial purposes for humans. One way to show the importance of algae in this planet is the contribution of Oxygen it provides to the atmosphere, which many organisms rely on (Medeiros et al., 2015). Carbon footprint has been one of the greatest causes for global warming and it is due to the use of fossils to create diesel. One of the ways to avoid contamination is to replace those fuels that are used on daily basis, such as for transportation. The microalgae are able to produce lipids that can …

From Microbes To Management: Spatial And Temporal Variations In Available Vegetation Affect The Gut Microbiota In A Population Of Free-Ranging Generalist Herbivores, Olivia K. Rodríguez

Boise State University Theses and Dissertations

The effects of extrinsic environmental factors that shape ecological systems are not only seen at the macroscopic level, but additionally influence and govern the host-associated microbial communities of their mammalian hosts. These microbial communities are susceptible to the fluctuation of abiotic and biotic factors which affect their host organisms. The surge in the research of microbiota–communities of archaea, bacteria, fungi, and viruses residing in various environmental systems–has shown that these communities can profoundly influence animal health. As such, monitoring microbiota has allowed for a new approach to study animal health and physiology. This is of particular benefit in the conservation …

The Study Of The Phytopathogen Dickeya Dadantii 3937 Cpx Signaling On The Regulation Of Virulence And Antimicrobial Resistance, Daqing Jiang

Theses and Dissertations

Bacteria respond to environmental cues through a variety of mechanisms. Two-component systems (TCSs) are a conserved mechanism used by bacteria to accurately respond and sense environmental changes. The monitoring of envelope perturbance is linked to TCS CpxA/CpxR in animal-infecting pathogens. The study of the TCS response regulator (RR) CpxR is largely unexplored in phytopathogens. This work focuses on the genetic linkage between cpxR and T3SS expression regulation. We identified the multiple roles of CpxR on several T3SS regulators and its participation in the bacterial second messenger (c-di-GMP) signaling cascade. Moreover, a compound library screening revealed a novel Cpx inducer CHIR-090, …

Glycan Functionalization Of Iron Oxide Nanoparticles (Ionps) As A Method Of Inactivating Antibiotic Resistant Neisseria Gonorrhoeae Via A Magnetically Mediated Energy Delivery (Magmed) System, Cedric Taylor

All Theses

With the increase of antibiotic resistant bacteria, current established therapeutic treatment methods are rapidly demonstrating decreasing efficacy. For this project, we are studying a novel alternate mechanism of treating antibiotic resistant bacteria by utilizing magnetic iron oxide nanoparticles (IONPs) coated with a heterobifunctional polyethylene glycol polymer (PEG) and functionalized with glycan structures that can interact with Neisseria gonorrhoeae FA1090. Interaction between the IONPs and bacteria is manipulated by an external alternating magnetic field (AMF) in a process known as magnetically mediated energy delivery (MagMED). Magnetic IONPs were synthesized and reacted with PEG (PEG-IONPs) and functionalized with glycans Gal(α1-3)Gal(β1-4)Glc(β1-1) (common name: …

Enzymatic Degradation Of Microcystin-Lr By Microcystinase (Mlra), Faisal Alqhtani

Graduate Theses and Dissertations

Microcystin-LR (MC-LR) is affecting the water supply worldwide. Hence, a way to eliminate this toxin is an essential target. In this study, successful cloning of the mlrA gene and producing MlrA enzyme that can degrade the cyclic MC-LR to linearized MC-LR was done. MlrA protein was expressed in Escherichia coli BL-21 (E. coli). Also, enhancing the MlrA yield by adding nickel to LB media was a success in producing more MlrA enzyme from the same volume. Even though the enzyme showed no activity after adding Ni, the enzyme was expressed at a higher yield. Furthermore, it was to investigate adding …

Toward Lignin Valorization: Development Of Rhodococcus Opacus Pd630 As A Chassis For Triacylglycerol (Tag) Production From Recalcitrant Aromatic Feedstocks, Rhiannon R. Carr

McKelvey School of Engineering Theses & Dissertations

The advent of the industrial era was precipitated by the discovery of fossil fuels, and ushered in unprecedented changes for humanity included but not limited to the development of rapid transit and communications, improvements to food distribution and preservation, the mass production of goods, and a radical rearrangement of communities from relatively small enclaves to metropolises. With all the benefits, however, come considerable costs, especially to the global environment. Greenhouse gas emissions, built up over centuries of unregulated combustion, have precipitated a rate of global temperature change unparalleled in the 4.5 billion-year history of this planet. In order to preserve …

Genetically Engineered Thermosynechococcus Elongatus Bp1: Assessment Of Potential Biorisks And Biofuel Production, Thu Ho Anh Nguyen-Jones

Chemistry & Biochemistry Theses & Dissertations

According to the International Energy Outlook 2019, released by the U.S. Energy Information Administration, it is projected that the energy consumption will increase up to 50% between 2018 and 2050 worldwide. As fossil fuel being a finite source of energy with the risk of depletion, many countries are now facing an energy security crisis. Therefore, it is important to develop other renewable and sustainable energy sources that will allow countries to shift away from depending on fossil fuels. Among several types of renewable energy, biofuel production using genetically engineered cyanobacteria is capturing much interest due to its many advantages. Different …