Graduate Theses & Dissertations


Flavohemoglobin expression in Giardia intestinalis exposed to nitrosative stress
The parasitic protist Giardia intestinalis lacks most heme proteins yet encodes a flavohemoglobin (gFlHb) that converts nitric oxide to nitrate and likely protects the cell from nitrosative stress. In this work an antibody raised against gFlHb was used to examine both changes in gFlHb expression levels and intracellular localization in Giardia in response to nitrosative stress. Giardia trophozoites exposed to stressors which either directly release nitric oxide (diethyltriamine NONOate, 1 mM) or are sources of other reactive nitrogen intermediates (sodium nitrite 20 mM or S-nitrosoglutathione, 1 or 5 mM) exhibited a 2 to 9-fold increase of gFlHb after 24 hours. Increased expression levels of gFlHb were detectable by 8 hours in S-nitrosoglutathione and diethyltriamine-NONOate-treated trophozoites, and by 12 hours after sodium nitrite exposure; these differences were likely due to differences in the rates of release of RNS from these compounds. In addition to a band of the expected size for gFlHb (52 kDa), western blots detected a second, higher molecular weight band (72 kDa) with comparable or higher intensity upon treatment with these RNS donors, which is consistent with sumoylation of gFlHb. Immunofluorescence microscopy of Giardia trophozoites detected gFlHb diffused throughout the cytoplasm and more punctuated staining along the cell membrane and between the nuclei. The punctuated staining may be due to the association of gFlHb with either peripheral vacuoles or basal bodies. Author Keywords: Flavohemoglobin, Giardia intestinalis, Nitrosative stress
Effects of flooding on nutrient budgets and ecosystem services
Increases in flooding due to anthropogenic influences such as climate change and reservoir creation will undoubtedly impact aquatic ecosystems, affecting physical, chemical, and biological processes. We used two approaches to study these impacts: a whole-ecosystem reservoir flooding experiment and a systematic literature review. In the whole-ecosystem experiment, we analyzed the impact of flooding on nutrient release from stored organic matter in an upland forest. We found that flooded organic matter produced N (nitrogen) and P (phosphorus), but that more N was released relative to P, increasing the N:P ratio over time. In the systematic literature review, we linked small (<10 year recurrence interval) and extreme (>100 year recurrence interval) floods to changes in 10 aquatic ecosystem services. Generally, extreme floods negatively impacted aquatic ecosystem service provisioning, while small floods contributed positively. Overall, we found that flood impacts vary depending on ecosystem properties (organic matter content) and flood characteristics (magnitude). Author Keywords: ecosystem services, flooding, nutrients, reservoirs, rivers
Isotopes of the Caribbean
This research represents the first stable carbon and nitrogen isotope analysis of human bone collagen (n = 29) from the Escape Site (AD 300 - 1000), Saint Vincent. As a two-pronged investigation, this research had the following goals: (1) determining the ideal pretreatment for poorly preserved bones and (2) reconstructing the Escape Site sample population diet. By incorporating powdered specimens, shorter demineralizations and increased acid:sample exposure, higher collagen yields were produced, thereby expanding the sample size for isotopic analysis. Notably, the elemental data suggests that not all isolated collagen was biogenic and was perhaps contaminated by non-collagenous proteins. This highlighted the importance of using multiple criteria to rigorously evaluate collagen based on the full quality indicator profile. In the end, 5 individuals yielded useable isotope data which was consistent with a broad spectrum diet relying primarily on C3 plants as well as terrestrial, reef, nearshore and freshwater fauna. Within the broad region, the Escape Site data was comparable to other islands from the Lesser Antilles and Cuba emphasizing the influence of regional biodiversity as well as the likelihood that the studied population contributed and benefitted from the extensive Saladoid trade networks which existed at the time. Author Keywords: Caribbean, Collagen, Escape Site, Human diet, Saladoid, Stable isotope analysis
"Multimodal Contrast" from the Multivariate Analysis of Hyperspectral CARS Images
The typical contrast mechanism employed in multimodal CARS microscopy involves the use of other nonlinear imaging modalities such as two-photon excitation fluorescence (TPEF) microscopy and second harmonic generation (SHG) microscopy to produce a molecule-specific pseudocolor image. In this work, I explore the use of unsupervised multivariate statistical analysis tools such as Principal Component Analysis (PCA) and Vertex Component Analysis (VCA) to provide better contrast using the hyperspectral CARS data alone. Using simulated CARS images, I investigate the effects of the quadratic dependence of CARS signal on concentration on the pixel clustering and classification and I find that a normalization step is necessary to improve pixel color assignment. Using an atherosclerotic rabbit aorta test image, I show that the VCA algorithm provides pseudocolor contrast that is comparable to multimodal imaging, thus showing that much of the information gleaned from a multimodal approach can be sufficiently extracted from the CARS hyperspectral stack itself. Author Keywords: Coherent Anti-Stokes Raman Scattering Microscopy, Hyperspectral Imaging, Multimodal Imaging, Multivariate Analysis, Principal Component Analysis, Vertex Component Analysis
Understanding the establishment of Typha spp. in North America using population genetics and common garden studies
There are three cattail (Typha) taxa in Canada: T. latifolia (native), T. angustifolia (introduced), and their hybrid T. x glauca. The latter is invasive in regions around the Laurentian Great Lakes, and I investigated the potential role that commercial suppliers may be playing in the introduction of non-native Typha by comparing genotypes of North American, European, and commercially available plants. I found that Ontario garden centres are importing both hybrids and non-native lineages of T. angustifolia into Canada, but was unable to identify the provenance of T. latifolia. I also investigated the possibility that the hybrid cattail leaf litter shade and leachate influences germination and early growth of the parental species of the hybrids. Using three common garden experiments, I found that T. x glauca leaf litter suppresses germination rates of the three taxa. In the early seedling growth experiment, plant performance varied by taxa, and for the competition experiment there were no intra- or interspecific competition or treatment effects on the performance of plants. Overall, my research identified a potential mechanism allowing T. x glauca to dominate wetlands, and also shows that non-native lineages are being introduced into Canada through commercial trade Author Keywords: Competition, Germination, Non-native lineages, Plant nurseries, Seedling Growth, Typha spp.
Range-Based Component Models for Conditional Volatility and Dynamic Correlations
Volatility modelling is an important task in the financial markets. This paper first evaluates the range-based DCC-CARR model of Chou et al. (2009) in modelling larger systems of assets, vis-à-vis the traditional return-based DCC-GARCH. Extending Colacito, Engle and Ghysels (2011), range-based volatility specifications are then employed in the first-stage of DCC-MIDAS conditional covariance estimation, including the CARR model of Chou et al. (2005). A range-based analog to the GARCH-MIDAS model of Engle, Ghysels and Sohn (2013) is also proposed and tested - which decomposes volatility into short- and long-run components and corrects for microstructure biases inherent to high-frequency price-range data. Estimator forecasts are evaluated and compared in a minimum-variance portfolio allocation experiment following the methodology of Engle and Colacito (2006). Some consistent inferences are drawn from the results, supporting the models proposed here as empirically relevant alternatives. Range-based DCC-MIDAS estimates produce efficiency gains over DCC-CARR which increase with portfolio size. Author Keywords: asset allocation, DCC MIDAS, dynamic correlations, forecasting, portfolio risk management, volatility
Prescription Drugs
Medication used to treat human illness is one of the greatest developments in human history. In Canada, prescription drugs have been developed and made available to treat a wide variety of illnesses, from infections to heart disease and so on. Records of prescription drug fulfillment at coarse Canadian geographic scales were obtained from Health Canada in order to track the use of these drugs by the Canadian population. The obtained prescription drug fulfillment records were in a variety of inconsistent formats, including a large selection of years for which only paper tabular records were available (hard copies). In this work, we organize, digitize, proof and synthesize the full available data set of prescription drug records, from paper to final database. Extensive quality control was performed on the data before use. This data was then analyzed for temporal and spatial changes in prescription drug use across Canada from 1990-2013. In addition, one of major research areas in environmental epidemiological studies is the study of population health risk associated with exposure to ambient air pollution. Prescription drugs can moderate public health risk, by reducing the drug user's physiological symptoms and preventing acute health effects (e.g., strokes, heart attacks, etc.). The cleaned prescription drug data was considered in the context of a common model to examine its influence on the association between air pollution exposure and various health outcomes. Since, prescription drug data were available only at the provincial level, a Bayesian hierarchical model was employed to include the prescription drugs as a covariate at regional level, which were then combined to estimate the association at national level. Although further investigations are required, the study results suggest that the prescription drugs influenced the air pollution related public health risk. Author Keywords: Data, Error checking, Population health, Prescriptions
Modelling Request Access Patterns for Information on the World Wide Web
In this thesis, we present a framework to model user object-level request patterns in the World Wide Web.This framework consists of three sub-models: one for file access, one for Web pages, and one for storage sites. Web Pages are modelled to be made up of different types and sizes of objects, which are characterized by way of categories. We developed a discrete event simulation to investigate the performance of systems that utilize our model.Using this simulation, we established parameters that produce a wide range of conditions that serve as a basis for generating a variety of user request patterns. We demonstrated that with our framework, we can affect the mean response time (our performance metric of choice) by varying the composition of Web pages using our categories. To further test our framework, it was applied to a Web caching system, for which our results showed improved mean response time and server load. Author Keywords: discrete event simulation (DES), Internet, performance modelling, Web caching, World Wide Web
Enhanced weathering and carbonation of kimberlite residues from South African diamond mines
Mafic and ultramafic mine wastes have the potential to sequester atmospheric carbon dioxide (CO2) through enhanced weathering and CO2 mineralization. In this study, kimberlite residues from South African diamond mines were investigated to understand how weathering of these wastes leads to the formation of secondary carbonate minerals, a stable sink for CO2. Residues from Venetia Diamond Mine were fine-grained with high surface areas, and contained major abundances of lizardite, diopside, and clinochlore providing a maximum CO2 sequestration capacity of 3–6% of the mines emissions. Experiments utilized flux chambers to measure CO2 drawdown within residues and unweathered kimberlite exhibited greater negative fluxes (-790 g CO2/m2/year) compared to residues previously exposed to process waters (-190 g CO2/m2/year). Long-term weathering of kimberlite residues was explored using automated wet-dry cycles (4/day) over one year. Increases in the δ13C and δ18O values of carbonate minerals and unchanged amount of inorganic carbon indicate CO2 cycling as opposed to a net increase in carbon. Kimberlite collected at Voorspoed Diamond Mine contained twice as much carbonate in yellow ground (weathered) compared to blue ground, demonstrating the ability of kimberlite to store CO2 through prolonged weathering. This research is contributing towards the utilization of kimberlite residues and waste rock for CO2 sequestration. Author Keywords: CO2 fluxes, CO2 mineralization, CO2 sequestration, Enhanced weathering, Kimberlite, Passive carbonation
Phosphoric Acid Chemically Activated Waste Wood
Activated Carbon (AC) is commonly produced by gasification, but there has been increasing interest in chemical activation due to its lower activation temperatures and higher yields. Phosphoric acid, in particular, succeeds in both these areas. Phosphoric acid activated carbon (PAC) can be environmentally sustainable, and economically favourable, when the phosphoric acid used in the activation is recycled. This thesis describes the digestion and activation of waste wood using phosphoric acid, as well as methods used to recover phosphoric acid, functionalize the produced activated carbon with iron salts and then test their efficacy on the adsorption of target analytes, selenite and selenate. In order to achieve an efficient phosphoric acid based chemical activation, further understanding of the activation process is needed. A two-step phosphoric acid activation process with waste wood feed stock was examined. The filtrate washes of the crude product and the surface composition of the produced PAC were characterized using X-ray Photoelectron Spectroscopy (XPS), Fourier Transform-Infrared spectroscopy (FT-IR), Ion Chromatography (IC), and 31P Nuclear Magnetic Resonance (NMR). XPS of the unwashed PAC contained 13.3 atomic percent phosphorous, as phosphoric acid, while the washed sample contained 1.4 atomic percent phosphorous as PO43-, and P2O74-. Using 31P NMR, phosphoric acid was identified as the primary phosphorous species in the acidic 0.1 M HCl washings, with pyrophosphates also appearing in the second 0.1 M NaOH neutralizing wash, and finally a weak signal from phosphates with an alkyl component also appearing in the DI wash. IC showed high concentrations of phosphoric acid in the 0.1 M HCl wash with progressively lower concentrations in both the NaOH and DI washes. Total phosphoric acid recovery was 96.7 % for waste wood activated with 25 % phosphoric acid, which is higher than previous literature findings for phosphoric acid activation. The surface areas of the PAC were in the 1500-1900 m2g-1 range. Both pre and post activation impregnation of iron salts resulted in iron uptake. Pre-activation resulted in only iron(III) speciation while post-activation impregnation of iron(II)chloride did result in iron(II) forming on the PAC surface. The pre-activated impregnated PAC showed little to no adsorption of selenite and selenate. The post-activation impregnated iron(II)chloride removed up to 12.45 ± 0.025 mg selenium per g Iron-PAC. Competitive ions such as sulfate and nitrate had little effect on selenium adsorption. Phosphate concentration did affect the uptake. At 250 ppm approximately 75 % of adsorption capacity of both the selenate and the selenite solutions was lost, although selenium was still preferentially adsorbed. Peak adsorption occurred between a pH of 4 and 11, with a complete loss of adsorption at a pH of 13. Author Keywords: Activated Carbon, doping, Iron, phosphoric acid, selenium
Regional Assessment of Soil Calcium Weathering Rates and the Factors that Influence Lake Calcium in the Muskoka River Catchment, Central Ontario
(MRC) in central Ontario was carried out to determine the range and spatial distribution of soil Ca weathering rates, and investigate the relationships between lake Ca and soil and catchment attributes. The MRC is acid-sensitive, and has a long history of impacts from industrial emission sources in Ontario and the United States. Small headwater catchments were sampled for soil and landscape attributes (e.g. elevation, slope, catchment area) at 84 sites. Soil Ca weathering rates, estimated with the PROFILE model, were low throughout the region (average: 188 eq/(ha·yr)) compared to global averages, and lower than Ca deposition (average: 292 eq/(ha·yr)). Multiple linear regression models of lake Ca (n= 306) were dominated by landscape variables such as elevation, which suggests that on a regional scale, landscape variables are better predictors of lake Ca than catchment soil variables. Author Keywords: Calcium, Lakes, Regional assessment, Regression, Soils, Weathering
Fungal pathogen emergence
The emergence of fungal hybrid pathogens threatens sustainable crop production worldwide. To investigate hybridization, the related smut fungi, Ustilago maydis and Sporisorium reilianum, were selected because they infect a common host (Zea mays), can hybridize, and tools are available for their analysis. Hybrid dikaryons exhibited filamentous growth on plates but reduced virulence and limited colonization in Z. mays. Select virulence genes in the hybrid had similar transcript levels on plates and altered levels during infection of Z. mays relative to each parental dikaryon. Virulence genes were constitutively expressed in the hybrid to determine if its pathogenic development could be influenced. Little impact was observed in hybrids with increased expression of effectors known to modify host response and metabolism. However, increased expression of transcriptional regulators of stage specific pathogenic development increased the hybrid’s capacity to induce symptoms. These results establish a base for investigating molecular aspects of fungal hybrid pathogen emergence. Author Keywords: effectors, hybrid pathogenesis assays, Sporisorium reilianum, transcription factors, Ustilago maydis, virulence factors


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