Molecular Biology Commons^™

Next-Generation Sequencing Shows Increasing Temperatures Affect Stream Fungal Communities, Regan Hodgson

Electronic Theses and Dissertations

Under climate change predictions, surface air temperature is expected to continue to rise and will likely affect functioning of stream ecosystems and microbial communities. Aquatic hyphomycetes are key fungal decomposers of plant litter in headwater streams controlling carbon, energy, and nutrient flows to higher trophic levels. This project addressed the effects of temperature on stream fungal communities in two recently conducted experiments (microcosm and streamside channel studies) using Illumina sequencing. The objectives were to determine (1) if temperature affects community structure of stream fungi based on relative abundances of ITS2 rDNA sequences, (2) if the effects of temperature on aquatic …

Pacific Black Duck Ecology And Habitat Assessment In Aunu'u, American Samoa, And Their Relationship To Other Mallard-Like Ducks Of Oceania, Greater Indonesia, And The Philippines, Marissa Kaminski

Open Access Theses & Dissertations

Determining a species’ ecological needs, assessing the quality of their habitat, and determining genetic differentiation and connectivity among populations is essential to their conservation. My dissertation focuses on obtaining such a holistic view for a population of Pacific Black Ducks recently established on the Island of Aunu’u, American Samoa. Specifically, I present the first evaluation of the ecology and habitat of a recently established population of Pacific Black Ducks on the Island of Aunu’u, American Samoa, in Chapter 1, while I assess the genetic connectivity and relationship of this population to other Mallard-like ducks found in Greater Indonesia, Oceania, and …

Polycyclic Aromatic Hydrocarbons And The Microbiomes Of Two Benthic Species, Samantha Ells

Master's Theses

The presence of oil has been shown to affect the microbiomes of the water column, sediments, and organisms, both by altering the diversity and the composition of those microbial communities. If the microbiome is altered it may no longer provide benefits to its host organism, impacting its ability to survive. Thus, it is important to understand the effects of large-scale contamination events including the Deepwater Horizon oil spill. This study set out to examine the effects of oil exposure on the microbiome of two benthic species, southern flounder (Paralichthys lethostigma) and eastern oysters (Crassostrea virginica) to further understand the effects …

Selection And Demography Drive Range-Wide Patterns Of Mhc Variation In Mule Deer (Odocoileus Hemionus), Rachel M. Cook

Theses and Dissertations

Variation at functional genes involved in immune response is of increasing concern as wildlife diseases continue to emerge and threaten populations. The amount of standing genetic variation in a population is directly associated with its potential for rapid adaptation to novel environments. For genes in the major histocompatibility complex (MHC), which are crucial in activating the immune response and which have extremely high levels of polymorphism, the genetic variation has been shown to be influenced by both parasite-mediated selection and historical population demography. To better understand the relative roles of parasite-mediated selection and demography on MHC evolution in large populations, …

Not Too Hot, Not Too Cold, But Moderately Variable: The Influence Of Environmental Variability On Coral Thermal Tolerance, Courtney Nicole Klepac

Biological Sciences Theses & Dissertations

Anthropogenic climate change is causing an increase in the frequency and severity of marine heat waves, resulting in declining health of coral reef ecosystems worldwide. Coral bleaching events – the breakdown in symbiosis between the coral host and their intracellular photosynthetic algae – are increasingly common in recent years and contribute to widespread losses in coral cover. However, bleaching and heat stress responses vary across spatial scales both within and among coral species. Coral populations native to highly variable environments can have greater bleaching resistance than corals from more stable habitats and corals transplanted into these variable reef sites can …

1,4-Dioxane Biodegradation In Propanotrophs: Molecular Foundations And Implications For Environmental Remediation, Li Fei

Dissertations

1,4-Dioxane (dioxane) has emerged with an escalating concern given its human carcinogenicity and widespread occurrence in groundwater. Bioremediation is promising as an effective and cost-efficient treatment alternative for in situ or ex situ cleanup of dioxane and co-existing pollutants in the field. Soluble di-iron monooxygenases (SDIMOs) are reputed for their essential roles in initiating the cleavage of dioxane and other pollutants. In this doctoral dissertation, molecular foundations for SDIMOs-mediated dioxane biodegradation are untangled to promote the development and implication of site-specific bioremediation and natural attenuation strategies. This dissertation focused on propanotrophic bacteria given their pivotal roles in dioxane metabolism and …

Assessment Of Soil Protein And Refractory Soil Organic Matter Across Two Chronosequences Of Newly Developing Marshes In Coastal Louisiana, Usa, Stuart Alexander Mcclellan

LSU Doctoral Dissertations

The impacts of sea-level rise and hydrologic manipulation are threatening the stability of coastal marshes throughout the world, thereby increasing the potential for re-mineralization of soil organic matter (SOM) in these systems. Such threats have prompted marsh restoration efforts, particularly in coastal Louisiana, yet it is unclear how the slowly decomposing (refractory) and quickly decomposing (labile) fractions of SOM may be differentially affected by different approaches to marsh restoration. Additionally, otherwise labile compounds may accumulate in the soil via a range of protective mechanisms, including rapid burial and association with organic compounds that are thought to enhance soil aggregation, such …

Engineering Of Escherichia Coli 2-Oxoglutarate Dehydrogenase Complex With Mechanistic And Synthetic Goals, Joydeep Chakraborty

Dissertations

The Escherichia coli 2-oxoglutarate dehydrogenase complex (OGDHc) compromises multiple copies of three enzymes - 2-oxoglutarate dehydrogenase (E1o), dihydrolipoyl succinyltransferase (E2o), and dihydrolipoyl dehydrogenase (E3). OGDHc is found in the Krebs cycle and catalyzes the formation of the all-important succinyl-Coenzyme A (succinyl-CoA). OGDHc was engineered to understand the catalytic mechanism and optimized for chemical synthetic goals.

Succinyl-CoA formation takes place within the catalytic domain of E2o via a transesterification reaction. The succinyl group from the thiol ester of S8-succinyldihydrolipoyl-E2o is transferred to the thiol group of CoA. Mechanistic studies were designed to investigate enzymatic transthioesterification. His375 and Asp374 was shown to …

Metabolic Engineering Of Cyanobacteria For Production Of Chemicals, Po-Cheng Lin

McKelvey School of Engineering Theses & Dissertations

Concerns over the impact of climate change caused by CO2 emission have driven the research and development of renewable energies. Microbial production of chemicals is being viewed as a feasible approach to reduce the use of fossil fuels and minimize the impact of climate change. With recent advances in synthetic biology, microorganisms can be engineered to synthesize petroleum-based chemicals and plant-derived compounds. Cyanobacteria are photosynthetic prokaryotes that use only sunlight, CO2, and trace minerals for growth. Compared to other microbial hosts, cyanobacteria are attractive platforms for sustainable bioproduction, because they can directly convert CO2 into products. However, the major challenge …

Metabolic Engineering Of Cyanobacteria For Production Of Chemicals, Po-Cheng Lin

McKelvey School of Engineering Theses & Dissertations

Concerns over the impact of climate change caused by CO2 emission have driven the research and development of renewable energies. Microbial production of chemicals is being viewed as a feasible approach to reduce the use of fossil fuels and minimize the impact of climate change. With recent advances in synthetic biology, microorganisms can be engineered to synthesize petroleum-based chemicals and plant-derived compounds. Cyanobacteria are photosynthetic prokaryotes that use only sunlight, CO2, and trace minerals for growth. Compared to other microbial hosts, cyanobacteria are attractive platforms for sustainable bioproduction, because they can directly convert CO2 into products. However, the major challenge …

Development Of Indicators For Human Fecal Pollution Using Deep-Sequencing Of Microbial Communities, Shuchen Feng

Theses and Dissertations

The gut microbiome is shaped by both host physiology and environmental factors, which results in unique communities that contain certain members specific to a host. Microbial source tracking (MST) methods that rely on host-specific fecal microorganisms have been applied to detect human fecal pollution over the past two decades. The most common approach uses quantitative polymerase chain reaction (qPCR) to amplify certain sequences of these microorganisms, or human fecal markers. To date, most bacterial human fecal markers have focused on the HF183 cluster within the genus Bacteroides. However, cross-reactions with animals or variable Bacteroides abundance in humans have been found. …

Environmental And Genetic Factors Affecting Antibiotic Resistance Of Extended Spectrum Î?-Lactamase Bacteria From The Rio Grande River In El Paso, Tx And Cd. Juarez, Mexico, Maria D. Fuentes

Open Access Theses & Dissertations

Background: The Rio Grande River provides a major source of potable and agricultural water for the population of the Texas/Mexico border region. Cattle farming and ranching are the most prevalent activities, which may contribute to the microbial burden of pharmaceuticals into our state's water resources. Antibiotics, presumably released into the environment by discharges originating from waste-water treatment plants, septic disposal systems, animal feeding operations and urban runoff have a definite impact on the ecosystem and may contribute to an increase in antibiotic resistance. We hypothesized that waters of the Rio Grande River contained Multi Drug Resistant Organisms (MDRO) and mobile …

Marine Reserves Promote Coral Reef Resilience By Mitigating Human Impacts Through The Restoration Of Parrotfish Populations, Increasing Their Reproductive Output And Seeding Neighboring Overfished Reefs, Brian L. Stockwell

Biological Sciences Theses & Dissertations

Coral reefs are some of the most productive ecosystems on the planet, providing fisheries resources for over a billion people with billions of dollars in revenue from tourism for developing nations. Coral reefs are under threat from overfishing and water pollution, resulting in less productive algae dominated reefs. Marine reserves are widely expected to promote the resilience of reefs by protecting and increasing the abundance of herbivorous fishes that can graze on algae, therefore directly or indirectly preventing coral to algal phase shifts. However, the ability of marine reserves to mitigate human impacts, restore herbivorous fish populations and seed nearby …

Transport Mechanisms For Human Fecal Indicator Bacteria In An Urban Stormwater Basin In Southeastern Wisconsin, Chelsea M. Corson

Theses and Dissertations

Discharge of stormwater runoff to receiving waters is a known source of human pathogens; however the primary mechanisms by which these pathogens enter the stormwater system have yet to be quantified. This study builds upon and utilizes prior research findings in an attempt to explain the influence of the age of the pipes within stormwater and sanitary conveyance systems, rainfall and hydrogeological characteristics, and select infrastructure variables that contribute to the observed contamination of an urban stormwater basin in Southeastern Wisconsin.

Over the course of approximately two years from 2012 to 2014, a total of 260 samples from 22 stormwater …

The Presence Of Clostridium Difficile On Environmental Surfaces In Healthcare Facilities Pre- And Post-Decontamination Of Patient Rooms, Theresa Trice

UNLV Theses, Dissertations, Professional Papers, and Capstones

Healthcare-associated infections (HAIs) are infections related to receiving medical care. HAIs are responsible for an excess of morbidity and mortality among hospitalized patients. Though most HAIs rates are on the decline, Clostridium difficile infection rates are at an all-time high, primarily due to the persistence of C. difficile spores in the environment. In the United States, Clostridium difficile-related mortality rates per million have increased from 5.7 in 1999 to 23.7 in 2004, with an estimated 26,642 deaths due to Clostridium difficile infections (CDIs). Clostridium difficile is transmitted via the fecal-oral route or aerosolized endospores, but it can also be transmitted …

Pharmaceuticals And Personal Care Products: Emerging Contaminants In Aquatic Ecosystems, Jenny E. Zenobio

Open Access Theses

In recent years, the presence of pharmaceuticals and personal care products (PPCPs) in aquatic systems has led to research on their fate and effects. PPCPs have been found in mixture in wastewater effluents, surface, ground, and drinking water at low concentrations from areas of intense urbanization. Although adverse effects to human health from the current environmental concentrations are unlikely, the impacts to ecological receptors are not clear. We performed field and laboratory studies to quantify and evaluate effects of PPCPs on fish. First, a field study was conducted at the Baca National Wildlife Refuge, Colorado (2010-2012) because a portion of …

Dephosphorylation Of Iqg1 By Cdc14 Temporally Regulates Actin Ring Formation, Daniel Patrick Miller

Masters Theses

"Cytokinesis is the final step in cell division when the cell separates the cytoplasm by contracting a ring composed of filamentous actin (F-actin) and type II myosin. Iqg1, an IQGAP family member, is an essential scaffolding protein in budding yeast (S. cerevisiae) required for actin recruitment to, and contraction of, the actomyosin ring. Actin is recruited by the calponin homology domain (CHD) in anaphase after Iqg1 is localized to the bud neck. Consensus sites for the cyclin-dependent kinase (CDK) Cdc28 were identified flanking the CHD. This led us to the hypothesis that phosphorylation of Iqg1 by Cdc28 negatively regulates actin …

Transcending Microbial Source Tracking Techniques Across Geographic Borders: An Examination Of Human And Animal Microbiomes And The Integration Of Molecular Approaches In Pathogen Surveillance In Brazil And The United States, Amber Mae Koskey

Theses and Dissertations

Waterborne illnesses, attributed to the ingestion or contact with contaminated water, present a significant global health concern. Surface water sources can be impacted by wide array of pollution inputs, but fecal pollution generates the most significant and acute threat to human health. Therefore, the detection of fecal bacteria in surface water sources remains an important public health objective. Current surface water monitoring employs the use of fecal indicator bacteria (FIB) including E. coli and enterococci as proxies for pathogenic organisms carried in fecal pollution. These traditional indicators, detected by culture-based microbiological methods, do not discriminate fecal sources from another. New …

Refining Molecular Approaches For Bacterial Source Tracking (Bst) In Texas, Joy Truesdale

Open Access Theses & Dissertations

Bacterial source tracking (BST) is a new science that aids in identifying sources of human and animal fecal contamination of environmental water. There are currently two approaches used in BST, library-dependent and library-independent methods. The fecal bacterium E. coli is often used in library dependent methods because it indicates fecal contamination, has been used in human health risk assessments, and has water quality regulatory significance. A disadvantage of E. coli library dependent BST methods is that identification libraries of E. coli isolated from known fecal sources are needed to identify the sources of E. coli isolated from water samples. Further, …

Environmental Stress In Hard Coral: Evaluating Lipid As An Indicator Of Sublethal Stress On Short Time Scales, David Harold Niebuhr

Dissertations, Theses, and Masters Projects

Lipid quality was evaluated in Montastrea spp. under sediment- and heat-stressed conditions to evaluate lipid ratio as an indicator of sub-lethal stress on short time scales. The ratio of storage lipid (wax ester + triacylglyceride) to structural lipid (sterol esters + phospholipid) decreased significantly (0.25 to 0.14, p < 0.01) after experimental sedimentation. FAME analysis of colonies exposed to experimental sedimentation showed a reduction of the algal, 18:3(n-6) and 18:4(n-3), polyunsaturated fatty acid (PUFA) in the polar lipid fraction. This loss of PUFA suggests a loss of algal membrane in sediment-stressed colonies. Lipid quality was similarly measured in M. faveolata over a 10-day period. Mean (n = 20) ratio of storage to structural lipid in M. faveolata dropped from 2.43 to a level of 0.98 immediately following a natural sedimentation event before recovering to levels of 1.4 and 2.9 on post-storm days 2 and 4, respectively. Colonies of M. annularis subject to heat-stress (35??C) exhibited no significant change in storage lipid ratio, while levels of Free Fatty Acids increased significantly from 0.012 (n = 22) to 0.156 mg lipid/g dry tissue (n = 22)(p < 0.05). FAME analysis of tissue lipids extracted from the heat-stressed colonies showed changes in the polar fraction, with significant decreases in the 18:3(n-6), 18:3(n-3), 18:4(n-3), 20:4(n-6) and 20:5(n-3) (p < 0.05) PUFA and subsequent significant increases in the saturated fatty acids, 16:0 and 18:0 (p < 0.05). These changes in lipid quantity and quality indicate possible oxidation and preferential digestion of zooxanthellar membranes. Stress experiments were repeated in M. annularis using VacutainerRTM blood collection tubes to collect micro-tissue samples without destroying skeleton of the sample colonies. A significant decrease in storage: structural lipid ratio after sedimentation was also detected using the micro-tissue technique. This study indicates that the relative abundance of lipid subclass components can indicate sub-lethal environmental stress, on short time scales, in M. annularis and M. faveolata. Furthermore, micro-tissue collection techniques permit repeated monitoring coral colonies to assess the manifestation of stress from first detection of impact at the cellular level to changes in community to changes in community structure detectable over longer time scales.

Expression Of The Xenobiotic Transporter P-Glycoprotein In The Mummichog (Fundulus Heteroclitus), Peter S. Cooper

Dissertations, Theses, and Masters Projects

The mammalian mdr1 P-glycoprotein (Pgp) has been implicated in xenobiotic resistance of drug-resistant cell lines and tumors, and may function in excretion or exclusion of toxic xenobiotics at several sites within the body. Pgp gene family members are expressed in excretory epithelia of several aquatic taxa and may contribute to the survival of pollution-tolerant aquatic animals in contaminated environments. For this reason the expression of Pgp family members was investigated in the livers and liver tumors of creosote-resistant mummichog (Fundulus heteroclitus) inhabiting a creosote-contaminated site (Atlantic Wood) in the Elizabeth River, Virginia. Expression of members of the Pgp gene family …