Graduate Theses & Dissertations

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Evaluating Environmental DNA (eDNA) Detection of Invasive Water Soldier (Stratiotes Aloides)
In 2008, the first North American water soldier (Stratiotes aloides) population was discovered in the Trent River, Ontario. Water soldier is an invasive aquatic plant with sharp, serrated leaves that has the potential to spread rapidly through dispersed vegetative fragments. Although it is too late to prevent water soldier establishment in the Trent River, its local distribution remains limited. In this study, environmental DNA (eDNA) was explored as a potential tool for early detection of water soldier. Species-specific markers were designed from chloroplast DNA regions matK and rbcL, and a qPCR assay with rbcL primers yielded the most sensitive detection of water soldier eDNA. Positive detections were obtained from six of 40 sampling locations, of which five were collected in Seymour Lake, an area with large patches of water soldier. As water soldier plants were known to be present at these sites, high eDNA concentrations were expected. The sixth positive detection from Trent Lock 5 (50 km downstream of Lake Seymour) was unexpected as it was obtained at a site with no water soldier sightings. This is one of the first studies to demonstrate the effectiveness of eDNA detection from aquatic plants. Author Keywords: aquatic plant, eDNA, environmental DNA, invasive species, Stratiotes aloides, water soldier
Functional Genetic Diversity in American Mink (Neovison vison)
The release of domestic organisms to the wild is considered a threat to biodiversity because the introduction of domestic genes through interbreeding can negatively impact wild conspecifics via outbreeding and local extinction. In North America, captive American mink (Neovison vison) are frequently escaping into the wild, yet the impact of these events on the functional genetic diversity of wild mink populations is unclear. I characterized domestic and wild mink in Ontario at 17 trinucleotide microsatellites located in functional genes thought to be associated with traits affected by domestication. I found low functional genetic diversity, as only 4 of 17 genes were variable and of those four there was little evidence of allele frequency differences between domestic and wild mink. Using redundancy analysis and a spatial analysis of principal components on the four variable loci (AR, ATN1, IGF-1, and TOB1) I found no evidence to suggest domestic release events are affecting functional genetic diversity of free-ranging mink at the set of markers assessed. Author Keywords: American mink, domestication, functional gene, introgression, Neovison vison
Yearly variation in fall movements of adult female American black bears (Ursus americanus) in central Ontario, Canada
I investigated site fidelity and habitat selection of American black bears (Ursus americanus) from 15 GPS-collared adult females in central Ontario, Canada over nine years. I used generalized linear mixed models to determine the factors affecting between-year variation in fall fidelity and the habitat selection in movement paths. I assessed second and third-order habitat preference by female bears moving between seasonal home ranges. I found that 66% of bears returned to the same fall area between years, expressed as range overlap, influenced negatively by whether they had cubs. When moving between seasonal ranges, bears selected for mixedwood, hardwood and wetlands cover but selected ridge tops over other habitat features at both scales. With increases in climatic uncertainty and habitat fragmentation, these results emphasize the need for wildlife management to consider annual variation in seasonal movements and habitat use by wide-ranging, opportunistic animals. Author Keywords: American black bear, Habitat Selection, Logistic Regression, Site Fidelity
Pathogen vs. Predator
Stressors are often an inescapable part of an organism’s life. While the effects of many stressors have been well studied individually, potential interactions between stressors exist that may result in greater than additive negative effects. Stressors may be linked by conflicting demands on energy budgets, interfering with important physiological pathways, or necessitating incompatible adaptive responses. Using Ranavirus (FV3) and larval dragonfly predators (Anax spp.) in a 2x2 factorial experiment on green frog (Lithobates clamitans) tadpoles, I investigate the interactions in behaviour, morphology, and metabolism when both stressors were applied in concert. I demonstrate that activity and feeding are reduced additively by both stressors, and tadpoles increase distance between conspecifics in FV3-exposed tanks, but only in the absence of predators. I also note decreases in mass, and a non-significant marginal increase in metabolic rate of tadpoles exposed to FV3. Interestingly, I provide evidence that FV3 can compromise morphometric responses through antagonistic interactions with perceived predation risk exposure, which may result in significantly elevated mortality even when either stressor is present in sub-lethal quantities. Thus, I conclude that sub-lethal exposure to stressors can nonetheless have substantial impacts on organisms and a more integrative approach to examining the impacts of stressors on individual physiology and fitness is necessary. Author Keywords: Behaviour, Interaction, Morphology, Predation Risk, Ranavirus, Tadpoles
impact of selection harvesting on soil properties and understory vegetation in canopy gaps and skid roads in central Ontario
Tree harvesting alters nutrient cycling and removes nutrients held in biomass, and as a result nutrient availability may be reduced, particularly in naturally oligotrophic ecosystems. Selection harvesting is a silvicultural technique limited to tolerant hardwood forests where individual or small groups of trees are removed creating a “gap” in the forest canopy. In order for harvesting machinery to gain access to these individual trees, trees are felled to create pathways, known as skid roads. The objective of this study was to characterize differences in soil chemical and physical properties in gaps, skid roads and uncut areas following selection harvesting in central Ontario as well as documenting differences in the understory vegetation community and sugar maple (Acer saccharum) seedlings chemical composition post harvest. First year seedlings were collected for elemental analysis from unharvested areas, canopy gaps, and skid roads in 2014, eight months after harvesting. In 2015, first and second year sugar maple seedlings were collected. Soil bulk density and water infiltration were measured in the three areas of the catchment as well as soil moisture, organic matter content, exchangeable base cations, and net nitrification. Seedlings in the disturbed sites had lower concentrations of Mg, K, P, and N compared with unharvested sites and soil nitrification was significantly lower in the skid roads. Water infiltration rates in the gap and skid roads were slower than the control and concentrations of metals (e.g. Fe, Al, Ca) and litter mass increased in litter bags deployed over 335 days, likely reflecting an increase in soil erosion in the skid roads. Understory vegetation was markedly different amongst sites, particularly the dominance of Carex spp. in the skid roads. The sustainability of industrial logging is dependent on successful tree regeneration, however, increased soil compaction, establishment and growth of grasses and shrubs, as well as low nutrient concentrations in seedlings may ultimately restrict forest succession. Author Keywords: Canadian Shield, nitrification, selection harvesting, soil compaction, sugar maple seedling, understory vegetation
Assessing Canada Lynx Dispersal Across an Elevation Barrier
Mountain ranges are often thought to restrict movement of wildlife, yet previous studies evaluating the role of the Rocky Mountains as a dispersal barrier for Canada lynx (Lynx canadensis) have been contradictory. Our study uses neutral microsatellite loci to evaluate the role of the Rocky Mountains as a barrier to gene flow for lynx. Although lynx exhibited low genetic differentiation, we detected a limited effect of the mountains. Furthermore, we inferred the role played by landscape variables in gene flow (genetic differentiation predicted by landscape resistance). Limited gene flow most strongly related to resistance from physical factors (low snow cover and elevation), rather than other topographic and ecological factors (high terrain roughness, low forest cover, low habitat suitability, and geographic distance). Structural connectivity was a relatively poor predictor of functional connectivity. Overall, the Rockies represent an area of reasonably high functional connectivity for lynx, with limited resistance to gene flow. Author Keywords: Canada lynx, connectivity, gene flow, genetic structure, landscape genetics, Rocky mountains
Habitat Preferences and Feeding Ecology of Blackfin Cisco (Coregonus nigripinnis) in Northern Algonquin Provincial Park
Blackfin Cisco (Coregonus nigripinnis), a deepwater cisco species once endemic to the Laurentian Great Lakes, was discovered in Algonquin Provincial Park in four lakes situated within a drainage outflow of glacial Lake Algonquin. Blackfin habitat preference was examined by analyzing which covariates best described their depth distribution using hurdle models in a multi-model approach. Although depth best described their distribution, the nearly isothermal hypolimnion in which Blackfin reside indicated a preference for cold-water habitat. Feeding structure differentiation separated Blackfin from other coregonines, with Blackfin possessing the most numerous (50-66) gill rakers, and, via allometric regression, the longest gill rakers and lower gill arches. Selection for feeding efficiency may be a result of Mysis diluviana affecting planktonic size structure in lakes containing Blackfin Cisco, an effect also discovered in Lake Whitefish (Coregonus clupeaformis). This thesis provides insight into the habitat preferences and feeding ecology of Blackfin and provides a basis for future study. Author Keywords: allometric regression, blackfin cisco, habitat, hurdle models, lake whitefish, mysis
Bank Swallow (Riparia riparia) Breeding in Aggregate Pits and Natural Habitats
I examined Bank Swallow (Riparia riparia) colony persistence and occupancy, in lakeshore, river and man-made aggregate pit habitat. Habitat persistence was highest on the lakeshore and lowest in aggregate pits, likely due to annual removal and relocation of aggregate resources. Bank Swallow colonies in aggregate pit sites were more likely to persist if a colony was larger or if burrows were located higher on the nesting face. I also compared nest productivity and health factors of Bank Swallows in lakeshore and aggregate pit habitats. While clutch size was the same in both habitat types, the number of fledglings from successfully hatched nests was significantly higher in aggregate pit sites than from lakeshore sites. Mass of fledgling Bank Swallows did not differ significantly between habitat types, however mass of adults from aggregate pits decreased significantly over the nesting season. Parasite loads on fledgling Bank Swallows were significantly lower in aggregate pits than in lakeshore sites. According to these indicators, aggregate pits appear to provide equivalent or higher quality habitat for Bank Swallows than the natural lakeshore sites, making them adequate and potentially key for this species’ recovery. Aggregate pit operators can manage for swallows by (1) creating longer, taller faces to attract birds and decrease predation, and (2) supplementing their habitat with water sources to encourage food availability. Author Keywords: Aerial insectivore, aggregate pits, Bank Swallow, colony persistence, ectoparasites, substitute habitat
Biology and Management of Stratiotes Aloides in the Trent River, Ontario
Invasive aquatic plants can create negative ecological, economic and social impacts when they displace local vegetation, interfere with shipping and navigation and inhibit water-based recreational activities. In 2008, the first North American occurrence of the invasive plant Stratiotes aloides (Water soldier) was identified in the Trent River, Ontario. This research measured offset photosynthesis and turion germination to determine the light compensation point (5.2-5.4m) and maximum depth of colonization (4-6m) for S. aloides propagules using in situ incubations and controlled growth experiments. The effects of spring and fall chemical (Diquat) and physical (hand raking) treatments on S. aloides biomass, local macrophyte recovery and community dynamics in the Trent River were also measured. The target of a 75% minimum reduction in S. aloides biomass was not attained using any of the treatment methods and no perceivable recovery of the local plant community was observed. Significant S. aloides regrowth was recorded for both treatment methods regardless of application timing. Author Keywords: herbicide, invasive species, macrophyte, photosynthesis, propagule
Interactome Study of Giardia Intestinalis Cytochromes B5
Giardia intestinalis is an anaerobic protozoan that lacks common eukaryotic heme-dependent respiratory complexes and does not encode any proteins involved in heme biosynthesis. Nevertheless, the parasite encodes several hemeproteins, including three members of the Type II cytochrome b5 sub-group of electron transport proteins found in anaerobic protist and amitochondriate organisms. Unlike the more well-characterized cytochrome b5s of animals, no function has been ascribed to any of the Type II proteins. To explore the functions of these Giardia cytochromes (gCYTB5s), I used bioinformatics, immunofluorescence microscopy (IFM) and co-immunoprecipitation assays. The protein-protein interaction in silico prediction tool, STRING, failed to identify relevant interacting partners for any of the Type II cytochromes b5 from Giardia or other organisms. Differential cellular localization of the gCYTB5s was detected by IFM: gCYTB5-I in the perinuclear space; gCYTB5-II in the cytoplasm with a staining pattern similar to peripheral vacuole-associated protein; and gCYTB5-III in the nucleus. Co-immunoprecipitation with the gCYTB5s as bait identified potential interacting proteins for each isotype. The most promising candidate is the uncharacterized protein GL50803_9861, which was identified in the immunoprecipitate of both gCYTB5-I and II, and which co-localizes with both. Structural analysis of GL50803_9861 using Swiss Model, Phyre2, I-TASSER and RaptorX predicts the presence of a nucleotide-binding domain, which is consistent with a potential redox role involving nicotinamide or flavin-containing cofactors. Finally, the protein GL50803_7204 which contains a RNA/DNA binding domain was identified a potential partner of gCYTB5-III. These findings represent the first steps in the discovery of the roles played by these proteins in Giardia. Author Keywords: Cytochrome b5, Giardia intestinalis, Heme, Interactome, Protein structure prediction
Stress Axis Function and Regulation in New World Flying Squirrels
Across vertebrate taxa, the hypothalamic-pituitary-adrenal axis (or the stress axis) is highly conserved, and is central to vertebrate survival because it allows appropriate responses to psychological stressors. Habitat shapes successful physiological and ecological strategies, and to appreciate how individual species respond to stressors in their environment, it is essential to have a thorough knowledge of the basic stress physiology of each species. In this dissertation, I studied the functioning and evolution of the stress physiology of New World flying squirrels. I showed that baseline, circulating cortisol levels in northern (Glaucomys sabrinus) and southern (G. volans) flying squirrels are some of the highest ever reported for mammals, indicating that their stress axes operate at a higher set point than most other species. I also assessed other aspects of their acute stress response, including free fatty acid and blood glucose levels, and indices of immune function, and showed that the flying squirrels’ physiological reaction to stressors may differ from that of other mammals. Using immunoblotting, I found that corticosteroid-binding globulin (CBG) expression levels in flying squirrels appeared to be higher than previously reported using alternative methods. I also concluded however, that these levels did not appear to be high enough to provide their tissues with the protective CBG-bound buffer from their high circulating cortisol concentrations experienced by the majority of vertebrates. Thus, this arm of cortisol regulation within the flying squirrel stress axes may be weak or non-existent. Following this, I focused on southern flying squirrels and showed evidence that the second arm of cortisol regulation — the negative feedback mechanism at the level of the brain — functions effectively, but that this species is glucocorticoid resistant. Their tissue receptors appear to have a reduced affinity for cortisol, and this affinity may change seasonally to allow for the onset of other biological processes required for survival and reproduction. Due to their distinctive stress physiology, northern and southern flying squirrels may provide comparative physiologists with model systems for further probing of the function and evolution of the stress axis among vertebrates. Author Keywords: corticosteroid-binding globulin, flying squirrel, Glaucomys, glucocorticoids, physiological ecology, stress physiology
Reproductive Fitness of Smallmouth Bass (Micropterus dolomieu) Under Heterogeneous Environmental Conditions
Identifying the biotic and abiotic factors that influence individual reproductive fitness under natural conditions is essential for understanding important aspects of a species’ evolutionary biology and ecology, population dynamics, and life-history evolution. Using next generation sequencing technology, I developed five microsatellite multiplex reactions suitable for conducting large scale parentage analysis of smallmouth bass, Micropterus dolomieu, and used molecular pedigree reconstruction techniques to characterize the genetic mating system and mate selection in adult smallmouth bass nesting in Lake Opeongo, Ontario, Canada. I used multivariate spatial autocorrelation analysis to indirectly infer the occurrence and extent of natal philopatry among spawning adults, to assess the strength and direction of sex-bias in natal dispersal patterns, and to evaluate the degree of nest site fidelity and breeding dispersal of spawning adults. I also evaluated how differences in littoral zone water temperature caused by wind-induced seiche events influence the relative reproductive success of spawning adults. Lastly, I provide a synopsis of potential future research aimed at further exploring factors that influence the reproductive fitness of smallmouth bass in Lake Opeongo. This information will contribute to our understanding of the factors regulating smallmouth bass populations, and provide insight into the factors controlling the variance in individual reproductive success and thus recruitment dynamics in this species. Author Keywords: Dispersal, Fitness, Mate selection, Mating systems, Philopatry

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