Graduate Theses & Dissertations

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Regulation of Cytokinins During Kernel Development in High and Low Yielding Oat and Barley Lines
Cytokinins (CKs) are a family of plant phytohormones responsible for many areas of plant growth and development. There are four free base types of CKs found in higher plants, trans-zeatin (tZ), N6-(∆2-isopentenyl)adenine (iP), cis-Zeatin (cZ) and dihydrozeatin (DZ). CK biosynthesis is regulated by adenosine phosphate-isopentenyltransferase (IPT), which is encoded by a multi-gene family in many plant species. There are two types of IPT pathways responsible for CK production, the tRNA pathway and the AMP (ATP/ADP) pathway. The tRNA pathway putatively produces cZ and the latter predominantly produces iP type nucleotides. CKs have long been studied for their role in stress tolerance, signal transduction, and involvement in many areas of plant growth and development. This study focuses on the role of CKs and CK biosynthesis by IPT during kernel development and comparisons of its regulation in high and low yielding barley and oat lines. The sequence of a putative IPT encoding gene in barley and oat was identified by a blast search of other known IPT gene fragments in closely related species. Quantitative Real time PCR results based on primers designed for the putative barley and oat IPT gene revealed changes in expression of IPT during different stages of kernel development, but no significance difference was associated with yield. Correlation of IPT gene expression in barley with cZ CK profiles measured by HPLC-MS/MS, confirms a putative IPT gene is a tRNA- IPT. HPLC-MS/MS results reveal some CK types, such as benzyladenine, are more predominant in higher yielding lines. This suggests different types of CKs play a role in yield production. Future studies on more IPT genes in the barley and oat IPT gene family will outline a more clear representation of the role of IPT in barley kernel development. Author Keywords: Benzyladenine, Cereal grain, Cytokinin, Isopentenyl Transferase, Mass Spectrometry, Real Time PCR
Geochemistry and Toxicity of a Large Slag Pile and its Drainage Complex in Sudbury, Ontario
This study was designed to determine the geochemistry and potential toxicity of water draining a large slag pile in Sudbury, Ontario, which runs through a pond complex prior to entering Alice Lake. Slag leaching experiments confirmed slag is a source of sulphate, heavy metals (including Fe, Al, Ni, Co, Cu, Zn, Pb, Cr, Mn) and base cations (Ca, K, Mg, Na). Concentrations of most metals draining through slag in column experiments were similar to metal concentrations measured at the base of the slag pile, although base cations, S and pH were much higher, possibly because of water inputs interacting with the surrounding basic glaciolacustrine landscape. The increase in pH rapidly precipitates metals leading to high accumulation in the surface sediments. Away from the base of the pile, an increase in vegetation cover leads to an increase in DOC and nutrients and transport of metals with strong binding affinities (Cu). Total metal concentration in water and sediment exceed provincial water quality guidelines, particularly near the slag pile, however WHAM7 modeling indicated that the free metal ion concentration in water is very low. Nevertheless, toxicity experiments showed that water with greater concentrations of solutes collected close to the slag pile negatively impacts D. magna suggesting that water draining the slag pile can adversely impact biota in nearby drainage areas. Author Keywords: geochemistry, heavy metals, leaching, non-ferrous slag, precipitation, toxicity
Expression and characterization of cytochrome b5 from Giardia lamblia
Giardia lamblia is an intestinal parasite found globally in freshwater systems that is responsible for endemic outbreaks of infectious diarrhea. As a unicellular parasite that lacks mitochondria, a respiratory chain and lives in the anaerobic environment of its host's intestine, Giardia was assumed for decades to lack heme proteins. However, its genome encodes several putative heme proteins, including three with sequence similarity to the cytochrome b5 family, referred to as Giardia cytochromes b5 (gCYTb5). Recombinant expression of one of these genes (gCYTb5-I), results in a protein (17-kDa) that is isolated with noncovalently bound heme. Resonance Raman and UV-visible spectra of gCYTb5-I in oxidized and reduced states resemble those of microsomal cytochrome b5, while sequence alignment and homology modelling supports a structure in which a pair of invariant histidine residues act as axial ligands to the heme iron. The reduction potential of gCYTb5-I measured by cyclic voltammetry is -165 mV vs the standard hydrogen electrode and is relatively low compared to those of other family members. The amino- and carboxy-terminal sequences that flank the central heme-binding core of the gCYTb5 are highly charged and do not occur in other family members. An 11-kDa core gCYTb5-I variant lacking these flanking sequences was also able to bind heme; however, we observe very poor expression of this truncated protein as compared to the full-length protein. Author Keywords: b-type cytochrome, cytochrome b5, electron transfer protein, Giardia intestinalis, heme/heam protein, spectroelectrochemistry
origin and ecological function of an ion inducing anti-predator behaviour in Lithobates tadpoles
Chemical cues are used commonly by prey to identify predation risk in aquatic environments. Previous work has indicated that negatively-charged ions of m/z 501 are possibly a kairomone that induces anti-predator responses in tadpoles. This thesis found that this ion species: (i) is produced by injured tadpoles; (ii) exhibits increased spectral intensity with higher tadpole biomass; and (iii) is not produced by starved predators. These results refute the hypothesis that the ion is a kairomone, and rather support its role as an alarm cue released from tadpoles. High resolution mass spectrometry (HR-MS) revealed a unique elemental composition for [M-H]-, m/z 501.2886, of C26H45O7S-. Collision induced dissociation (CID) of ion m/z 501 formed product ions of m/z 97 and m/z 80, HSO4- and SO3-, respectively, indicating the presence of sulfate. Green frog (Lithobates clamitans) tadpoles exposed to m/z 501, and an industrial analogue, sodium dodecyl sulphate (NaC12H25O4S), exhibited similar anti-predator responses, thereby suggesting the potential role of organic sulfate as a tadpole behavioural alterant. Author Keywords: Alarm cue, Amphibian, Chemical Ecology, Mass spectrometry, Predator-prey interactions
Groundwater Recharge in a Managed Forest on the Oak Ridges Moraine, Southern Ontario
Groundwater recharge was estimated and compared in two open grasslands, three mixed deciduous forest stands (100+ years in age), three young red pine plantations (27 ¨C 29 years in age) and two old red pine plantations (62 ¨C 63 years in age) on the Oak Ridges Moraine, southern Ontario, Canada. Recharge was estimated using a 1-d water balance with measured precipitation, throughfall, stemflow, snowpack water equivalent and soil water storage, and modelled evapotranspiration. Throughfall distribution beneath red pine canopies showed no consistent variation with distance from the tree boles. Old red pines were not major stemflow producers and although the young red pines showed a slight tendency to focus stemflow (focussing ratio > 1), the inclusion of focussed stemflow when calculating recharge at the stand scale made little difference. Conversely, sugar maple (the predominant species in the mixed deciduous stands) showed a strong tendency to focus throughfall proximal to tree boles and produce large quantities of stemflow, resulting in relatively high soil moisture contents and enhanced opportunities for recharge within ~ 0.5 m of tree boles. Inclusion of these focussed inputs resulted in a ~ 11 ¨C 18 % increase in stand scale recharge estimates. The interpretation of land cover control on recharge was complicated by variations in soil texture between sites. Soil texture and its influence on soil water storage capacity resulted in temporal variations in recharge, with sites exhibiting large storage capacities producing less recharge in the fall and greater recharge in the spring than sites with limited storage capacities. Recharge estimates for the entire study period or seasonal values for sites grouped on the basis of soil water storage capacities showed a general trend of increasing recharge in the order: old red pine ¡Ö young red pine ¡ú mixed deciduous forest ¡Ö open grasslands. The disparity between the red pine plantations and the other sites was driven in large part by greater modelled evapotranspiration in the red pine plantations. The similarity in recharge between mixed deciduous forests and open grasslands was the result of focused inputs and less soil evaporation and transpiration in the mixed deciduous forests compared to the open grasslands. The results of this study suggest planting red pine on grasslands on the Oak Ridges Moraine will initially decrease recharge and this decrease will continue as the red pines mature. However, as the red pine plantations are succeeded by mixed hardwood stands recharge will recover to that of the initial grasslands. Author Keywords: Groundwater Recharge, Land Cover Type, Oak Ridges Moraine, Stemflow, Throughfall, Water Balance
Utilizing Class-Specific Thresholds Discovered by Outlier Detection
We investigated if the performance of selected supervised machine-learning techniques could be improved by combining univariate outlier-detection techniques and machine-learning methods. We developed a framework to discover class-specific thresholds in class probability estimates using univariate outlier detection and proposed two novel techniques to utilize these class-specific thresholds. These proposed techniques were applied to various data sets and the results were evaluated. Our experimental results suggest that some of our techniques may improve recall in the base learner. Additional results suggest that one technique may produce higher accuracy and precision than AdaBoost.M1, while another may produce higher recall. Finally, our results suggest that we can achieve higher accuracy, precision, or recall when AdaBoost.M1 fails to produce higher metric values than the base learner. Author Keywords: AdaBoost, Boosting, Classification, Class-Specific Thresholds, Machine Learning, Outliers
Detecting anti-estrogens and anti-androgens in surface waters impacted by municipal wastewater discharges and agricultural runoff
This study focused on detecting 22 target anti-estrogenic and anti-androgenic compounds in surface waters influenced by both discharges of municipal wastewater and agricultural runoff in Canada and Argentina. Polar organic chemical integrative samplers (POCIS) were used to monitor the target compounds in surface waters. The removals of the target compounds in a municipal wastewater treatment plant (WWTP) in Canada were also evaluated. In both Canada and Argentina pesticides with potential anti-estrogenic and anti-androgenic activities were detected in the surface waters. The highest concentrations were found in Argentina (up to 1010 ng L-1) in areas impacted by heavy agricultural practices. Cyproterone acetate and bicalutamide were the only two anti-cancer drugs detected only at the Canadian study site, the Speed River, ON. In the Guelph WWTP, that discharges into the Speed River, these target compounds were not all efficiently removed (>70%) during treatment. Overall, this study provides insight to possible anti-estrogenic and anti-androgenic compounds that may be contributing to endocrine disrupting activities in surface waters. Author Keywords: Anti-androgens, Anti-estrogens, Cancer Therapy Drugs, Current use pesticides, Pharmaceuticals and Personal Care Products, Polar Organic Chemical Integrative Samplers
Incidental Take and Population Dynamics of Nesting Birds in a Red Pine (Pinus resinosa) Plantation Under Single-Tree Selection Harvesting
I determined the direct influence of single-tree selection harvesting on the daily nest survival rates and nest success of 5 focal bird species within a monotypic red pine (Pinus resinosa) plantation on the western edge of the Oak Ridges Moraine in southern Ontario, Canada. I located and monitored 290 nests during the 2012 and 2013 breeding season. I used the logistic-exposure method to evaluate the daily nest survival rates of American Robin (Turdus migratorius), Eastern Wood-pewee (Contopus virens), Ovenbird (Seiurus aurocapilla), Rose-breasted Grosbeak (Pheucticus ludovicianus), and Red-eyed Vireo (Vireo olivaceus). Only five nests were destroyed as a result of forestry activity over the study period. Neither daily nest survival rates nor nest success of these focal species were substantially affected by single-tree selection harvesting. I also monitored the impact of single-tree selection harvesting on the density and territory size of 4 of 5 focal species. Ovenbird had a significantly smaller territory size but decreased density in the harvested areas. Although not significant, Eastern Wood-pewee and Red-eyed Vireo tended to have higher densities and larger territory sizes in harvested areas, whereas Rose-breasted Grosbeak showed a mixed effect as density was higher while territory size was smaller. Single-tree selection produces minor to moderate disturbance that takes place locally over a short period of time. As a result, nests that are indirectly disturbed by nearby harvesting, felling trees and mechanical operations and are not destroyed remain and adults do not appear to abandon eggs or young from the disturbance. Habitat alteration from harvesting of the general forest structure and especially the forest floor must be minimized in order to conserve forest bird species diversity. Further research examining incidental take using various intensities of single-tree selection harvesting would provide important insight into maintaining avian and forest diversity by means of forest management. Author Keywords: daily nest survival rates, forest management, Incidental Take, nest success, red pine monotypic forest, single-tree selection harvesting
Machine Learning Using Topology Signatures For Associative Memory
This thesis presents a technique to produce signatures from topologies generated by the Growing Neural Gas algorithm. The generated signatures have the following characteristics: The signature's memory footprint is smaller than the "real object" and it represents a point in the n x m multidimensional space. Signatures can be compared based on Euclidean distance and distances between signatures provide measurements of differences between models. Signatures can be associated with a concept and then be used as a learning step for a classification algorithm. The signatures are normalized and vectorized to be used in a multidimensional space clustering. Although the technique is generic in essence, it was tested by classifying alphabet and numerical handwritten characters and 2D figures obtaining a good accuracy and precision. It can be used for many other purposes related to shapes and abstract typologies classification and associative memory. Future work could incorporate other classifiers. Author Keywords: Associative memory, Character recognition, Machine learning, Neural gas, Topological signatures, Unsupervised learning
Passive sampling of indoor and outdoor atmospheric nitrogen dioxide in the greater Toronto area
The reliability and performance of four passive sampler membrane coatings specific to nitrogen dioxide (NO2) were evaluated through co-exposure at multiple Ontario Ministry of Environment and Climate Change (OMOECC) active monitoring stations. All four coatings performed relatively similar under a wide range of meteorological conditions, notably showing exposure-specific atmospheric uptake rates. Further, indoor and outdoor atmospheric concentrations of NO2 (a marker of traffic-related air pollution) were evaluated at multiple elementary schools in a high-density traffic region of Toronto, Ontario, using a Triethanolamine based passive sampler membrane coating. Samplers were also co-exposed at OMOECC active monitoring stations to facilitate calibration of exposure-specific atmospheric uptake rates. Indoor NO2 atmospheric concentrations were 40 to 50% lower than outdoor concentrations during the spring−summer and autumn−winter periods, respectively. In large cities such as Toronto (Population 2,700,000), the influence of a single major road on outdoor and indoor NO2 concentrations is predominantly masked by spatially-extensive high-density traffic. Author Keywords: active sampler, membrane coating type, nitrogen dioxide, passive sampler, Toronto, traffic density
Mitogenome characterization of the shortnose sturgeon (Acipenser brevirostrum) for international trade validation of aquaculture-reared caviar
Identifying the population origin of aquaculture-reared caviar is crucial for both conservation and management strategies of farmed fish but could also facilitate international trade of a CITES regulated product. Shortnose sturgeon (Acipenser brevirostrum) is the main source of caviar production in Atlantic Canada, from Breviro Caviar Inc. aquaculture facility. Shortnose sturgeon are also listed as a species-at-risk under the Species At Risk Act. Currently there is no genetic method for delineating wild from aquaculture-reared caviar. By targeting the mitochondrial genome (mitogenome) using novel long-range PCR primers and next-generation sequencing (NGS) methods we have successfully sequenced the full mitogenome of 37 shortnose sturgeon. The purpose of this study was to increase the resolution of diagnostic variation among populations and to validate Canadian aquaculture-reared stock from wild US populations. Results provided a previously unobserved novel control region haplotype in high frequency within both the aquaculture-reared and Saint John River wild sample sets. Similar frequencies were observed with whole mitogenome haplotypes. Diagnostic mitochondrial lineage found in high frequency within the captive Breviro Caviar Inc. population has the potential to allow caviar product from Breviro Caviar Inc. to be distinguished from protected US shortnose sturgeon populations. The application of full mitogenomic characterization provides the potential to further resolve differences between aquaculture and natural Canadian shortnose sturgeon stocks, US/Canadian populations and to contribute to future conservation strategies. Future research identifying signatures of selection on the mitogenome between captive and wild populations and across latitudinal gradients found within the species range. These novel methods have produced a proof-of-concept to provide a "farm-to-fork" validation and ecobrand of Breviro Caviar Inc. product and its aquaculture origin to support importation into US caviar markets. Author Keywords: aquaculture, mitogenome, next-generation sequencing, species-at-risk, sturgeon
It’s a Mad, Mad, Mad, Mad World
The purpose of this study was to understand the lived experiences of mental health and illness in Kuwait. Twenty-six participants were interviewed, including mental health professionals, family physicians, and service users. Findings suggest that inequality, oppression, and human rights violations may drive mental health issues in Kuwait. However, rather than addressing these factors, many healthcare providers are endorsing psychological testing and psychiatric medication, which may be resulting in the same iatrogenic (physician-induced) drug dependence that is seen in North America. An analysis of mainstream psychological theory, research and practice is provided, along with a bioethical critique of the World Health Organization’s efforts to reduce the global ‘burden’ of mental disorders. This study cuts across disciplinary boundaries and 1) supports medical anthropologists’ criticisms of the ‘advancement’ of global mental healthcare; 2) provides participant-driven, community-based alternatives that are specific to Kuwait; and 3) informs culturally defined notions of ‘care’ and ‘ethics’. Author Keywords: Clinicians' narratives, Critical Psychology, Human rights, Kuwait, Qualitative research, Transcultural psychiatry

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