Graduate Theses & Dissertations

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Population cycles are regular fluctuations in population densities, however, in recent years many cycles have begun to disappear. With Canada lynx this dampening has also been seen with decreasing latitude corresponding to an increase in prey diversity. My study investigates the role of alternate prey on the stability of the lynx-hare cycle by first comparing the functional responses of two sympatric but ecologically distinct predators on a primary and alternate prey. I then populated a three species predator-prey model to investigate the role of alternate prey on population stability. My results showed that alternate prey can promote stability, though they are unlikely to “stop the cycle”. Furthermore, stability offered by alternate prey is contingent on its ability to increase intraspecific competition. My study highlights that population cycles are not governed by a single factor and that future research needs to be cognizant of interactions between alternate prey and intraspecific competition. Author Keywords: alternate prey, Canis latrans, functional response, Lepus americanus, Lynx canadensis, Tamiasciurus hudsonicus
Stopover Movement Patterns by Blackpoll and Canada Warblers Across Southeastern Canada During Fall Migration
Stopover ecology is a topic that surges in relevancy as choices made by migrants during stationary periods (stopover sites) may not only have important individuals’ fitness consequences but also can affect population dynamics. I used MOTUS automated telemetry array to study fall stopover duration of Blackpoll Warbler (BLPW) and departure decisions of BLPW and Canada Warbler (CAWA) in relation to various predictors. I affixed radio-transmitters on 55 BLPWs and 32 CAWAs at two banding stations in Ontario in September-October 2014-2015. Radio-tagged individuals were tracked through the MOTUS network across southeastern Canada. I developed models relating age class, fat score, Julian date and stopover movement types to Blackpolls’ stopover duration. I also examined whether there were species-related differences of wind selectivity when resuming migration. No explanatory variable significantly influenced BLPW’s stopover duration. Both species tended to depart under increased tailwind assistance, but with no difference in the effect of wind conditions between the two species. This study provides further evidence supporting the relevance of local wind conditions as a key factor affecting the departure likelihood, especially when migrating birds face an ecological barrier. Author Keywords: Cardellina canadensis, departure decisions, minimum stopover length, MOTUS, overland fall migration, Setophaga striata
Assessing the Potential for Contamination of Lakes from Upwelling of Arsenic-Laden Groundwater Through Sediments
A bedrock fracture hosting arsenic (As) contaminated groundwater was suspected to be transported to Ramsey Lake, a drinking water resource for more than 50,000 residents of Sudbury, Ontario. A high resolution, spatial, water quality mapping technique using an underwater towed vehicle (UTV) was used to identify sources of upwelling groundwater into lake water and localize the upwelling As contaminated groundwater vent site. The top 7 cm of lake sediments (in-situ) at this vent site were observed to adsorb 93 % of the dissolved As, thus inhibiting lake water quality degradation from this contaminant source. Sediment samples from this location were used in laboratory experiments to assess the potential for this system to become a source of As contamination to Ramsey Lake water quality and elucidate As(III) fractionation, transformation and redistribution rates and processes during aging. Arsenic speciation is important because As(III) has been shown to be more toxic than As(V). To accomplish this a sequential extraction procedure (SEP) that maintains As(III) and As(V) speciation in (sub)oxic sediments and soils was validated for the operationally defined fractions: easily exchangeable, strongly sorbed, amorphous Fe oxide bound, crystalline Fe oxide bound, and the residual fraction for total As because the characteristics of the reagents required to extract the final fraction do not maintain As species. Batch reaction experiments using sediment spiked with As(III) or As(V) and aged for up to 32 d were sequentially extracted and analysed for As(III) and As(V). Consecutive reaction models illustrate As(III) is first adsorbed to the sediment then oxidized to As(V). Fractionation analyses show As(III) most rapidly adsorbs to the easily exchangeable fraction where it is oxidized and redistributes to the strongly sorbed and amorphous Fe oxide bound fractions. Oxidation of As(III) adsorbed to the amorphous and crystalline Fe oxide bound fractions is less efficient and possibly inhibited. Select samples amended with goethite provide evidence supporting Mn(II) oxidation is catalyzed by the goethite surface, thus increasing As(III) oxidation by Mn(III/IV) complexed with the strongly sorbed fraction. Although As immobilization through groundwater sediment interactions may be inhibited by increased ion activity, particularly phosphate or lake eutrophication, this threat in Ramsey Lake is likely low. Author Keywords: arsenic, fractionation, modelling, redistribution, speciation, water quality mapping
Identification and Quantification of Organic Selenium Species Produced by Microbiological Activity in Freshwater Environments
Despite being an essential nutrient at trace levels, selenium can be devastating to aquatic environments when present in excess. There is no apparent correlation between total aqueous selenium concentrations and observed toxic effects because bioaccumulation varies over several orders of magnitude depending on the chemical species of selenium and the biological species present in the lowest trophic level of the aquatic food chain. Despite being used in toxicity models due to its high bioavailability, free selenomethionine had not been found previously in the environment outside of a biological entity. Here, it is confirmed that selenomethionine is produced during the biological treatment of selenium-contaminated wastewater, and released in the effluent along with other discrete organic selenium species, including selenomethionine oxide. This identification followed the development of a rigorous preconcentration and cleanup procedure, allowing for the analysis of these organic selenium species in high-ionic strength matrices. A newly optimized anion-exchange chromatographic separation was coupled to inductively-coupled plasma mass spectrometry for the simultaneous quantification of these organic selenium species along with the more ubiquitous selenium oxyanions, selenite and selenate. This separation method was also coupled to electrospray tandem mass spectrometry for structural confirmation of selenomethionine and selenomethionine oxide. High resolution orbitrap mass spectrometry was used to identify another oxidation product of selenomethionine – a cyclic species which was tentatively identified, by coelution, in a selenium-contaminated river water sample. The production and release of selenomethionine, selenomethionine oxide, Se-(methyl) selenocysteine, and methyl selenic acid were observed for various laboratory algal cultures. Once the presence of free selenomethionine in a water system was confirmed, factors affecting its uptake into algal cultures were examined. The uptake of selenomethionine into Scenedesmus obliquus was noted to be significantly higher under low nitrate conditions, where it was incorporated into selenium-containing proteins more readily than at higher nitrate conditions where other metabolites were produced. With the increasing popularity of biological treatment systems for the remediation of selenium-contaminated waters, these observations, combined with existing knowledge, could be used to make predictions regarding the potential toxicity of selenium in various environmental scenarios. Author Keywords: bioremediation, electrospray mass spectrometry, inductively-coupled plasma mass spectrometry, selenium, selenoamino acids, selenomethionine
Carbon Exchange along a Natural Gradient of Deciduous Shrub Coverage in the Low-Arctic
Arctic terrestrial ecosystems have experienced substantial structural and compositional changes in response to warming climate in recent decades, especially the expansion of shrub species in Arctic tundra. Climatic and vegetation changes could feedback to the global climate by changing the carbon balance of Arctic tundra. The objective of this thesis was to investigate the influence of increased shrub coverage on carbon exchange processes between atmosphere and the Arctic tundra ecosystem. In this study a space-for-time substitution was used, referred to as a shrub expansion “chronosequence”, with three sites along a natural gradient of deciduous shrub coverage in the Canadian low Arctic. Leaf-level photosynthetic capacity (Amax) of dominating birch shrub Betula glandulosa (Michx.) was significantly higher (P<0.05) at the site where shrubs were more abundant and taller than at the other sites. For all sites, mean Amax in 2014 was significantly lower than in 2013, in part potentially due to differences in precipitation distribution. Bulk soil respiration (RS) rate was significantly higher (P<0.05) at the site with more shrubs compared with the other sites. The differences in RS across sites appeared to be driven by differences in soil physiochemical properties, such as soil nitrogen and soil bulk density rather than soil microclimate factors (e.g. soil temperature, moisture). The three sites were either annual CO2 sources (NEP<0) to the atmosphere or CO2 neutral, with strongest annual CO2 sources (-44.1±7.0 g C m-2) at the site with most shrubs. Overall this study suggests that shrubs tundra carbon balance will change with shrub expansion and that shrub ecosystems in the Arctic currently act as annual carbon sources or neutral to the atmospheric CO2 and further shrub expansion might strengthen the CO2 emissions, causing a positive feedback to the warming climate. Author Keywords: arctic tundra, carbon exchange, climate change, photosynthetic capacity, shrub expansion, soil respiration
Investigation of a Model for Stain Selection and a Robust Estimation for Area of Origin in Bloodstain Pattern Analysis
Bloodstain pattern analysis is fundamental to crime scene investigations by providing insight into the estimation of the area of origin of a blood source. The selection of bloodstains from an impact pattern is the foundation from which the estimation of the area of origin is derived. The purpose of this research was to develop a theoretical model for stain selection and establish a more robust estimation for the area of origin in the analysis of impact patterns. This model was developed by using quantitative data from marked zones on the impact pattern target surface. The minimum distance between the known blood source location and a single bloodstain line of trajectory was calculated for every bloodstain within each pattern investigated. It has been determined that the zone location of a bloodstain is relevant for improving accuracy, and that stains with all alpha angles can be used to calculate an area of origin estimation. Finally, research within this thesis also validated directional analysis and investigated the number of stains that may be required for a precise area of origin estimation. Keywords: Forensic science, Bloodstain, Impact pattern, Bloodstain pattern analysis, Statistics, Forensic Physics
Elemental Variation in Daphnia
Environmental variation can affect consumer trait expression and alter ecological and evolutionary dynamics in natural populations. However, although dietary nutrient content can vary by an order of magnitude in natural ecosystems, intra-specific differences in consumer responses to food quality have not been thoroughly investigated. Therefore, the purpose of my dissertation was to examine the influence of dietary nutrition and other environmental factors on consumer phenotypic variation using the freshwater cladoceran Daphnia. I conducted a series of complementary laboratory and field studies where I examined the effects of dietary phosphorus (P) content and additional biological/environmental variables (multi-elemental limitation, genetic variation, and temperature) on daphnid life-history, biochemistry, body elemental composition, and population growth. In general, phenotypic expression within a species varied significantly in response to all experimental variables, but the relative influence of each was highly context dependent. In my first chapter, I found that dietary P content and environmental calcium (Ca) concentrations both altered Daphnia body Ca:P ratios and growth rates of individuals and affected intrinsic rates of increase at the population level. However, food quality appeared to have a much larger effect on trait expression, and body Ca:P ratios were highly sensitive to other forms of dietary nutrient limitation. Next, I documented significant quantitative genetic variation and phenotypic plasticity in daphnid P content, growth, and P use efficiency of field collected animals grown across dietary P gradients. Trait expression was also influenced by genotype X diet interactions suggesting that consumer responses to dietary nutrient limitation can be heritable and may be adaptive in different nutrient environments. Finally, I found that temperature appeared to override food quality effects and decouple P metabolism in natural Daphnia populations, but total biomass production was affected by both dietary P content and temperature, depending on the nutrient content of the lake. Overall, my dissertation shows that consumer responses to nutrient limitation can vary significantly within a species and that changes in trait expression may be modified by other environmental variables. These results should be incorporated into existing stoichiometric models and used to investigate the eco-evolutionary consequences of consumer phenotypic variation in response to nutritional stress. Author Keywords: ecological stoichiometry, evolution, life-history, nutrient limitation, nutrient metabolism, zooplankton
Mutation of the B10 Tyrosine and E11 Leucine in Giardia intestinalis Flavohemoglobin
The flavohemoglobin in Giardia intestinalis (gFlHb) is the only known protozoan member of a protein class typically associated with detoxifying nitric oxide (by oxidation to nitrate) in bacteria and yeast. Mutants of the B10 tyrosine (Y30F) and E11 leucine (L58A), conserved residues thought to influence ligand binding, were expressed and studied using Resonance Raman (RR) spectroscopy. In the wild type protein, RR conducted using a carbon monoxide probe detects two distinct Fe-CO stretches associated with two different active site configurations. In the open configuration, CO does not interact with any polar side chains, while in the closed configuration, CO strongly interacts with one or more distal residues. Analysis of the Y30F mutant provided direct evidence of this tyrosine’s role in ligand stabilization, as it had only a single Fe-CO stretching mode. This stretching mode was higher in energy than the open conformer of the wild type, indicating a residual hydrogen bonding interaction, likely provided by the E7 glutamine (Q54). In contrast the L58A mutant had no effect on the configurational nature of the enzyme. This was unexpected, as the side chain of L58 sits atop the heme and is thought to regulate the access of distal residues to the heme-bound ligand. The similar spectroscopic properties of wild type and L58A suggest that any such regulation would involve rapid conformational dynamics within the heme pocket. Author Keywords: B10 Tyrosine, Catalytic Globin, E11 Leucine, Flavohemoglobin, gFlHb, Giardia intestinalis
Electrochemical Characterization of Giardia Intestinalis Cytochromes b5
Giardia intestinalis is a protozoan parasite that causes waterborne diarrheal disease in animals and humans. It is an unusual eukaryote as it lacks the capacity for heme biosynthesis; nonetheless it encodes heme proteins, including three cytochrome b5 isotypes (gCYTB5s) of similar size. Homology modelling of their structures predicts increased heme pocket polarity compared to mammalian isotypes, which would favour the oxidized state and lower their reduction potentials (E°’). This was confirmed by spectroelectrochemical experiments, which measured E°’ of -171 mV, -140 mV and -157 mV for gCYTB5-I, II, III respectively, compared to +7 mV for bovine microsomal cytochrome b5. To explore the influence of heme pocket polarity in more detail, five gCYTB5-I mutants in which polar residues were replaced by nonpolar residues at one of three positions were investigated. While these substitutions all increased the reduction potential, replacement of a conserved tyrosine residue at position-61 with phenylalanine had the most significant effect, raising E°’ by 106 mV. This tyrosine residue occurs in all gCYTB5s and is likely the greatest contributor to their low reduction potentials. Finally, complementary substitutions were made into a bovine microsomal cytochrome b5 triple mutant to lower its reduction potential. These not only lowered the E°’ by more than 140 mV but also weakened the interaction of heme with the protein. The lower reduction potentials of the gCYTB5s may indicate that these proteins have different roles from their more well-known mammalian counterparts. Author Keywords:
Comparison of the Optical Properties of Stratiotes aloids and the Local Plant Community
As part of a mandate to control the spread of Stratiotes aloides (WS; water soldier) in the Trent Severn Waterway, the Ministry of Natural Resources (MNR) created a management plan to eradicate WS. However, one of the biggest challenges in eradicating WS or any invasive aquatic plant is the ability to estimate the extent of its spread and detect new populations. While current detection methods can provide acceptable detection, these methods often require extensive time and effort. The purpose of this thesis was to assess the use optical properties of WS and WS exudates for detection, in order to improve on current detection methods. The optical properties of WS were sampled at three different sites during three different seasons (spring, summer, and fall) by a) randomly sampling tissue from WS and the local plant community at each site, and recording the reflectance properties in a laboratory setting b) collecting dissolved organic matter (DOM) samples from plant incubations and river water in the field. Significant differences in the reflectance properties of WS were observed among samples from different sites and different sampling times; however, changes in fluorescence properties were only seasonal. Despite spatial differences in WS reflectance; WS was detectable using both hyperspectral and multispectral reflectance. When hyperspectral reflectance was used, significant differences between WS and the local plant community were found in June using two bands (i.e. bands 525 and 535, R 2 = 0.46 and 0.48, respectively). Whereas multispectral reflectance was significant different in October using the coastal and blue band. While WS produced a unique signal using both reflectance types, multispectral reflectance had a greater potential for detection. Its greater potential for detection was due to the reduced noise produced by background optical properties in October in comparison to June. DOM derived from WS was also characterized and compared with whole-river DOM samples in order to find unique markers for WS exudates in river samples. While similarities in DOM concentrations of WS exudates to Trent River water limited the ability to detect WS using compositional data, the ratio of C4/C5 components were compared in order to find components that were proportionally similar. Based on the results of this study multispectral and fluorescence techniques are better suited for the detection of a unique WS signature. The results derived from this work are intended to have practical applications in plant management and monitoring, DOM tracing, as well as remote sensing. Author Keywords: Dissolved organic matter, Hyperspectral reflectance, Invasive species management, Multispectral reflectance, PARAFAC, Stratiotes aloides
Nutrient Metabolism of an Aquatic Invertebrate and its Importance to Ecology
Aquatic consumers frequently face nutritional limitation, caused in part, by imbalances between the nutrients supplied by primary producers and the metabolic demands of the consumers. These nutritional imbalances alter many ecological processes including consumer life-history traits, population dynamics, and food web properties. Given the important ecological role of organismal nutrition, there is a need to have precise and specific indicators of nutritional stress in animals. Despite this need, current methods used to study nutrition are unable to distinguish between different types of nutritional limitation. Here I studied nutritional metabolism in the freshwater zooplankter, Daphnia. A greater understanding of nutritional metabolism would allow for the development of dietary bio-indicators that could improve the study of the nutritional ecology of animal consumers. Specifically, I addressed the question: What affects the biochemical composition of a generalist aquatic consumer? My overall hypothesis was that the quantity and quality of the diet affects the biochemical composition in a nutrient specific manner. To test this hypothesis, I examined various response variables involved in nutrient metabolism such as alkaline phosphatase activity, whole metabolome, and free amino acid composition. For each response variable, I grew Daphnia under various nutritional stressors and determined if responses are nutrient specific or are a general stress response. I found the current method of measuring alkaline phosphatase was not a phosphorus specific indicator, as activity increased in all nutrient stressed treatments. Analyzing the whole metabolome resulted in nutritional stressors being separated in multivariate space, with many identified metabolites being significantly different from nutrient rich Daphnia. Upon further examination the daphnids free amino acids profiles are caused by differences between the supply of amino acids from the algae and the demand within the Daphnia. These differences in supply and demand resulted in the ability to classify the nutritional status of Daphnia with the use of discriminant analysis, a classification multivariate model. In addition to a deeper understanding and advanced knowledge of the physiological changes caused by nutrient limitation, this research has provided strong evidence for the application of nutritional biomarkers/profiles to identified the nutritional status of Daphnia. Author Keywords: Bio-indictor, Ecological stoichiometry, Metabolism, Nutritional limitation, Nutritional status
Legume species, nitrogen rate and arbuscular mycorrhizal fungi inoculation effects on crop biomass and nitrogen requirement in a corn-legume system
Interseeding legume cover crops in grain corn may improve the environmental sustainability of corn production system in Southern Ontario. This study aimed to assess the effects of legume species, nitrogen (N) fertilizer rate and arbuscular mycorrhizal fungi (AMF) inoculation on biomass and N requirement in a corn-legume system. Corn was grown with red clover (RCl), microclover (MCl), hairy vetch (HV), or beans at 10 and 80 kg N ha-1 rates with and without AMF inoculation in a greenhouse for 7 weeks. Corn dry matter (DM) and N uptake were reduced by beans and HV (average 35%) compared with control; however, the DM for beans and HV was 7 and 3 times higher than RCl and MCl, respectively. The N2 fixation ability was similar among legume species and no significant N transfer from legume was detected. Overall, species collection was critical to the success of incorporating legumes into grain corn production. Author Keywords: Arbuscular mycorrhizal fungi, corn, legume cover crop, nitrogen

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