Environmental and Life Sciences

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

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

Exposure of pre-metamorphic amphibians to neonicotinoid insecticides may be contributing to the global decline in amphibian populations. In this study, anuran embryos and tadpoles of the African clawed frog (Xenopus laevis) and the North American leopard frog (Lithobates pipiens) were used to determine the effects of embryonic exposure to neonicotinoids. In addition, Xenopus was used to determine if prolonged exposure to neonicotinoids influenced tadpole sensitivity to frog virus 3 (FV3). Exposure of anuran embryos to concentrations of the neonicotinoid insecticide, imidacloprid, ranging from 1 -20 ppm induced a concentration dependent increase in malformations of the retina in Xenopus embryos. However, similar responses were not observed with embryos of leopard frogs. Exposure of Xenopus tadpoles to 500 ppb concentration of imidacloprid followed by challenge with FV3 showed that pesticide exposure unexpectedly decreased the rates of mortality, although total mortalities by the end of the experiment were not significantly different from controls. This unexpected observation may be attributed to a reduced inflammatory response induced by exposure to imidacloprid. Despite the low acute toxicity of neonicotinoid insecticides to vertebrates, these studies indicate that exposure to this class of insecticides causes sublethal effects in anuran species during early life stages.

Author Keywords: embryogenesis, Lithobates pipiens, neonicotinoid, ranavirus, tadpole, Xenopus laevis

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

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

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

Researchers have shown increasing interest in biological invasions for the associated ecological and economic impacts as well as for the opportunities they offer to study the mechanisms that induce range expansion in novel environments. I investigated the strategies exhibited by invasive species that facilitate range expansion. Invasive populations exhibit shifts in life-history strategy that may enable appropriate responses to novel biotic and abiotic factors encountered during range expansion. The spatio-temporal scales at which these shifts occur are largely unexplored. Furthermore, it is not known whether the observed dynamic shifts represent a consistent biological response of a given species to range shifts, or whether the shifts are affected by the abiotic characteristics of the new systems. I examined the life-history responses of female round gobies Neogobius melanastomus across fine and coarser spatial scales behind the expansion front and investigated whether invasive populations encountering different environmental conditions (Ontario vs France) exhibited similar life-history shifts. In both study systems, I found an increase in reproductive investment at invasion fronts compared to longer established areas at coarse and fine scales. The results suggest a similar response to range shifts, or a common invasion strategy independent of environmental conditions experienced, and highlight the dynamic nature of an invasive population's life history behind the invasion front.

The second part of my research focused on the development of an appropriate eDNA method for detecting invasive species at early stages of invasion to enable early detection and rapid management response. I developed a simple, inexpensive device for collecting water samples at selected depths for eDNA analysis, including near the substrate where eDNA concentration of benthic species is likely elevated. I also developed a protocol to optimise DNA extraction from water samples that contain elevated concentration of inhibiters, in particular near-bottom samples. Paired testing of eDNA and conventional surveys was used to monitor round goby expansion along its invasion pathway. Round gobies were detected in more sites with eDNA, permitting earlier, more accurate, upstream detection of the expansion front. My study demonstrated the accuracy and the power of using eDNA survey method to locate invasion fronts.

Author Keywords: Age-specific reproductive investment, DNA extraction, Energy allocation, Fecundity, Invasion front, Range expansion

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

The Oak Ridges Moraine (ORM) is a key hydrogeologic feature in southern Ontario. Previous work has emphasized the importance of depression-focused recharge (DFR) for the timing and location of groundwater recharge to the ORM's aquifers. However, the significance of DFR has not been empirically demonstrated and the relative control of land cover, topography, and surficial geology on DFR is unclear. The potential for DFR was examined for topographic depressions under forested and open, agricultural land covers with similar soils and surficial geology. Recharge (R) was estimated at the crest and base of each depression during the 2012-13 and 2013-14 winter-spring periods (~December – May) using both a 1-dimensional water balance approach and a surface-applied Br- tracer. At each depression, air temperatures, precipitation, snow depth and water equivalent, soil water contents, soil freezing, and depression surface-water levels were monitored and soil properties (texture, bulk density, porosity, and hydraulic conductivity) were measured. Both forested and agricultural land covers experienced soil freezing; however, concrete frost did not develop in the more porous and conductive forest soils. Concrete frost in agricultural depressions resulted in overland flow, episodic ponding and drainage of rain-on-snow and snowmelt inputs. Recharge was an order-of-magnitude greater at the base of open depressions. Observations of ponding (as evidence of DFR) were made at an additional 14 depressions with varying land cover, geometry, and soil type during the 2014 snowmelt period and measurements of pond depth, pond volume, land cover (i.e., percentage of agricultural vs. forested cover), depression geometry (i.e., contributing area, average slope, relief ratio) and soil texture were made. Ponding was restricted to depressions under mostly agricultural cover and a positive, non-linear relationship between pond volume and average slope was shown for sites with similar land cover and soil texture, but neither pond depth nor volume were related to any other depression characteristics. Results suggest that DFR is a significant hydrologic process during winter and spring under agricultural land cover on the ORM. Topographic depressions under agricultural land cover on the ORM crest may serve as critical recharge "hot spots" during winter and spring, and the ability of the unsaturated zone beneath these depressions to modify the chemistry of recharging water deserves further attention.

Author Keywords: Concrete frost, Depression-focused groundwater recharge, Oak Ridges Moraine, Ponding, Topographic depressions, Water balance

As an emerging contaminant, the antimicrobial agent silver nanoparticles (AgNPs) have been receiving considerable attention to determine their potential effects to aquatic ecosystems. However, estimates of aquatic consumer survivorship and other toxicological endpoints vary considerably among experiments, largely due to the environment in which the test takes place. Throughout this thesis I aim to understand which natural environmental variables impact toxicity to the common aquatic consumer Daphnia. I focus on the effects of particulate matter as it may play a role in animal nutrition as well as interact with AgNPs. I explore particulate matter's effect on survival in the complex matrices including other natural variables that could impact toxicity. I conduct a series of complimentary field and laboratory studies to understand how particles impact AgNP toxicity and how those interactions vary within whole lake ecosystems. Using laboratory studies, I establish that algal particles mitigate the toxic effects of AgNPs on Daphnia survival through removing Ag from the water column and that phosphorus increases this effect. Using wild Daphnia and lake water, I demonstrate the ability of particulate matter to mitigate toxicity in complex natural settings. It was also one of the major predictors of AgNP toxicity to Daphnia along with dissolved organic carbon and daphnid seasonal health. Finally, using a whole lake AgNP addition experiment, I demonstrate that particles and AgNPs interact variably in the lake. Silver from AgNPs binds to particles and is removed to the sediments through the actions of settling particles without impacting the dynamics of living communities. Overall, I am able to demonstrate that the natural components of lake ecosystems, especially particulate matter, are able to mitigate the effects of AgNPs in lake ecosystems to a point where they likely will be never pose a threat to the survivorship of aquatic consumers such as Daphnia.

Author Keywords: Daphnia, ecotoxicity, particulate matter, Silver nanoparticles, whole lake experiment