Graduate Theses & Dissertations

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Phytohormone-enhanced heavy metal responses in Euglena gracilis
Phytohormones, Cytokinin (CK) and Abscisic acid (ABA), are best known for controlling plant growth and stress responses; but they also mediate various developmental perspectives in alga. Yet, their mode of action in algal adaptive strategies to heavy metal responses, their involvement in orchestration of phytohormone crosstalk remain largely unknown and a molecular framework of phytohormone-controlled heavy metal uptake is absent. I found that three metals known globally to contaminate aquatic ecosystems, nickel (Ni), lead (Pb), and cadmium (Cd), cause changes in the levels of endogenous CKs, ABA, auxins, and gibberellins (GAs) in the green alga Euglena gracilis. Exogenous ABA or CK (trans-zeatin) alleviated metal toxicity through improved metal uptake efficiency and the regulation of the endogenous CKs activity profiles and GAs activity. This new evidence suggests that E. gracilis possesses functional phytohormone signals and metabolic pathways that are under metal stress response. Exogenously applied ABA or CK provoked the coordinated activation of metal uptake, likely via enhanced accumulation of metal binding compounds (i.e., proline, glycine, cysteine containing peptides), which are effective for metal sequestration. Using untargeted metabolomics analysis and functional annotation, this thesis further established that, CK and ABA modified pathways and metabolites, which were mainly involved in metal acclimation and resistance. These modified metabolites that were under the influence of phytohormones in algal cells growing under metal stress conditions were associated with: lipid pathways, riboflavin metabolism, biosynthesis of cofactors/vitamin, and carbohydrate metabolism. Bioactive secondary compounds (e.g., terpenoids, alkaloids, flavonoids, carotenoids) were also modified in algal cells treated with phytohormones. The present study highlights that ABA and CKs are important regulators of algal metal accumulation/acclimation strategies based on increased metal uptake, enhanced CK metabolism, regulation of hormonal crosstalk and regulation of some core cellular metabolism pathways, all of which improve metal uptake efficiency. Finally, our results suggest that ABA and CK form a novel strategy for metal bioremediation techniques and for sourcing microalgal value-added metabolites. Author Keywords: abscisic acid, cadmium, cytokinin, Euglena gracilis, lead, nickel
Landscape fitness
Variation in habitat quality and disturbance levels can strongly influence a species’ distribution, leading to spatial variation in population density and influencing population dynamics. It is therefore critical to understand how density can lead to variability in demographic responses for effective conservation and recovery of species. My dissertation illustrates how density and spatial familial networks can be integrated together to gain a better understanding of the influence of density on population dynamics of boreal caribou. First, I created an analytical framework to assess results from empirical studies to inform spatially-explicit capture-recapture sampling design, using both simulated and empirical data from noninvasive genetic sampling of several boreal caribou populations in Alberta, Canada, which varied in range size and estimated population density. Analysis of the empirical data indicated that reduced sampling intensity had a greater impact on density estimates in smaller ranges, and the best sampling designs did not differ with estimated population density but differed between large and small population ranges. Secondly, I used parent-offspring relationships to construct familial networks of boreal caribou in Saskatchewan, Canada to inform recovery efforts. Using network measures, I assessed the contribution of individual caribou to the population with several centrality measures and then determined which measures were best suited to inform on the population demographic structure. I found substantial differences in the centrality of individuals in different local areas, highlighting the importance of analyzing familial networks at different spatial scales. The network revealed that boreal caribou in Saskatchewan form a complex, interconnected familial network. These results identified individuals presenting different fitness levels, short- and long-distance dispersing ability across the range, and can be used in support of population monitoring and recovery efforts. Finally, I used a spatial capture-recapture analytical framework with covariates to estimate spatial density of boreal woodland caribou across the Saskatchewan Boreal Plains, and then reconstructed parent-offspring relationships to create a familial network of caribou and determined whether spatial density influenced sex-specific network centrality, dispersal distance, individual reproductive success, and the pregnancy status of females. I show that caribou densitygreatly varied across the landscape and was primarily affected by landscape composition and fragmentation, and density had sex-specific influences on dispersal distance, reproductive success, and network centrality. The high density areas reflected good-quality caribou habitat, and the decreased dispersal rates and female reproductive output suggest that these remnant patches of habitat may be influencing demographic responses of caribou. Author Keywords: boreal caribou, density, familial networks, population dynamics, rangifer tarandus caribou, spatial capture-recapture
Using genomic and phenotypic data to explore the evolution and ecology of the North American mountain goat
Evaluating the impact of climate change is arguably one of the main goals of conservation biology, which can be addressed in part by studying the demographic history of species in the region of interest. In North America, landscape and species composition during the most recent Pleistocene epoch was primarily influenced by glaciation cycles. Glacial advance and retreat caused species ranges to shift as well, leaving signatures of past population bottlenecks in the genetic code of most species. Genomic tools have shown to be important tools for understanding these demographic events to enhance conservation biology measures in several species. In my thesis I first reviewed the state of ungulate genomics, with a focus on how such data sets can be used in understand demography, adaptation, and inform conservation and management. Importantly, the review introduces key analyses like the pairwise sequentially Markovian coalescent and features like variation in antlers and horns and selection pressures that are used throughout subsequent chapters. Using the North American mountain goat as a model species, I then explored the genomic and phenotypic variation in this alpine specialist mammal. Starting with the generation of the first genome assembly for the mountain goat, I identified genes unique to the mountain goat and modeled demographic history going back millions of years using a pairwise sequentially Markovian coalescent approach. Species’ effective population size generally paralleled climatic trends over the past one hundred thousand years and severely declined to under a thousand individuals during the last glacial maximum. Given the biological importance of horns in mountain goats and the recent scientific interest in genetic basis of headgear, I analyzed over 23,000 horn records from goats harvested in British Columbia, Alaska and Northwest Territories from 1980 to 2017. Overall, variation in horn size over space and time was low; goats harvested further North had shorter horn lengths and smaller horn circumferences in one year old and 4 years and older age classes and 4 years and older age class, respectively. Proximity of roads, which was used as an indicator of artificial selection, had a small effect on horn size, with larger horns being harvested closer to major roads. Finally, I used two range-wide genomic data sets sequenced with a whole genome re-sequencing and reduced representation approaches to provide estimates of genetic diversity, contemporary effective population sizes and population structure. These insights can help inform management and will potentially make an impact in preserving the mountain goat. Author Keywords: genome assembly, horn size, Oreamnos americanus, population demography, reduced representation sequencing, whole genome resequencing
Effects of Intensive Agriculture on Stream Nutrient Export in East-Central Ontario, Canada
Recent agricultural land use change in east-central Ontario, including the expansion of intensive agriculture (corn and soybean crops) and tile drainage (TD) infrastructure, may alter the fluxes of both phosphorus (P) and nitrogen (N) to the Lower Great Lakes. Through intensive monitoring of several sub-watersheds that encompassed a range of row crop and TD extents, this study examined differences in stream N and P concentrations both seasonally and during variable flow conditions, to better understand land use/land cover (LULC) relationships with nutrient export. There was no clear relationship between total P (TP; concentration or export) and agricultural LULC, and instead, TP delivery was highly sensitive to flow events, and TP concentrations (especially particulate P; PP) were significantly higher during event flow compared with baseflow. In contrast, the dissolved fraction of P (total dissolved P; TDP) and dissolved nitrogen as nitrate (NO3-N) were not sensitive to flow, but were instead positively related to row crop agriculture, and associations between NO3-N concentrations/export and tile-drained row crop area were particularly strong (concentration r2 = 0.93; export r2 = 0.88). Regression relationships showed that with every 10% increase in row crop area in watersheds, NO3-N and TDP flow-weighted concentrations increased by 0.34 mg/L and 1.5 µg/L, respectively. As well, the same 10% increase in row crop agriculture translated to an increase of NO3-N export of as much as 130 kg/km2. Geospatial records of TD are incomplete in east-central Ontario, which presents challenges for evaluating the contribution of TD to nutrient export. Understanding the response of nutrients to changes in agriculture and agricultural practices is an integral part of watershed management as rapid changes in both urban and agricultural LULC continue to put pressure on water quality in the Lower Great Lakes. Author Keywords: East-Central Ontario, Nutrient Export, Row crops, Streams, Tile Drainage, Water Quality
Finding Community
This thesis explores the history of the Indigenous child welfare system in Manitoba and the effects of the Millennium Scoop on children in care. My research question is: what was the experience of children in care in Manitoba from 1990 to 2015? A related question is: how do survivors find healing? The thesis begins with a discussion of the history of acts, policies, and practices that began with the Indigenous child welfare system during the running of Residential schools. Then the acts, reviews, and policies that have shaped the child welfare system in Manitoba are discussed more thoroughly. The focus of the thesis is on the stories of Phoenix Sinclair, Tina Fontaine, and Natasha Reimer. I share their stories and provide an analysis of how the child welfare system has affected their lives. The negative effects of being a child in care are numerous. Being a child in care leaves behind grief, loss of identity, and loss of security. The systemic issues of the child welfare system include inadequate funding, overloaded case workers, staff burnout, and a lack of transparency. These overarching failures translate into the failure of children in care: details are overlooked, wrong decisions are made, and children are left to fend for themselves. Or they fall into the cracks and do not receive adequate care. This then translates into the deaths of children in care, or they are left to navigate life on their own and forced to create their circle of supports. Despite all the complications and negative impacts, some children can succeed while in care. Natasha’s story is a perfect example of such resilience. Author Keywords: child welfare, indigenous studies, millennium scoop, sixties scoop
Combining Line Transect Sampling and Photographic-Identification Surveys to Investigate the Abundance and Distribution of Cetaceans
Line transect sampling and photographic-identification (photo-ID) are common survey techniques for estimating the abundance and distribution of cetaceans. Combining these approaches in the field (‘combined LTPI’ surveys) and using data from both components has the potential for generating comprehensive ecological knowledge that can be far more valuable than when these techniques and their data are used independently. In this thesis, I evaluated the results and conclusions from these two methods, used singly and in tandem, by investigating the population dynamics of two humpback dolphin (Sousa chinensis spp.) populations: the large and widely distributed Chinese white dolphin (S. c. chinensis) of the Pearl River estuary (PRE), and the small and geographically isolated subspecies of Taiwanese white dolphin (S. c. taiwanensis) in the eastern Taiwan Strait. Data from combined LTPI surveys in Hong Kong waters, at the eastern edge of the PRE, revealed a shift in space use with individuals spending less time in these waters than at the start of surveys. Data from combined LTPI surveys in Taiwan provided further support for a subspecies restricted to the central western waters, and identified a commonly used area at the northern part of their limited range. These two case studies demonstrated an overall efficacy of combined LTPI surveys in ecological studies of cetaceans. However, a multi-criteria analysis revealed that combined LTPI surveys with a line transect focus (e.g., Hong Kong) performed better than a LTPI survey with a photo-ID focus (e.g., Taiwan) when considering ecological aspects of the study populations, labour and data requirements, and ecological output. Even so, the photo-ID focus of Taiwan’s monitoring program led to better assessments of individual space use patterns, likely helped by the Taiwanese white dolphin population’s smaller size and intensive photographic effort. In both cases, the ecological output of combined LTPI surveys could be improved by expanding the study area or extending the field season or frequency of surveys. Overall, I showed that by following a set of general guidelines, different iterations of the combined LTPI approach (i.e., photo-ID focus or LT focus) can serve as powerful tools for uncovering multi-dimensional ecological information on cetaceans. Author Keywords: abundance, cetacean, distribution, line transect sampling, multi-criteria analysis, Photo-ID
Biodiversity patterns along a forest time series in a remediated industrial landscape
Sudbury, Ontario is an epicenter of research on industrially degraded landscapes. Regreening efforts over the past 40 years have changed the landscape, leading to an increase in forest cover in the “barrens”, that once covered more than 100,000 ha. This study characterized changes in plant and insect composition using a space for time approach in the pine plantations. A total of 25 treated sites were sampled and soil characteristics, understory plants and insect communities were assessed. All sites were contaminated with copper and nickel, but the metals had little influence on biodiversity. Vegetation diversity metrics were more strongly correlated with the pH of the organic soil horizons, while the insect community shows little response to site characteristics, and rather vegetation cover. Plant composition changes are similar to those in pine stands undergoing natural recovery and as liming effects fade there may be a decline in insect community richness. Author Keywords: Biodiversity, Heavy Metals, Mining, Remediation
Assessing and Mitigation the Impacts of Mining-induced Flooding on Arctic-nesting Birds
Mining and resource development are growing industries in the Arctic, resulting in increased conflict with wildlife. Best practices for mitigation require an understanding of the potential ecological effects. One such effect concerns the flooding of terrestrial bird habitat from dewatering of lakes during mining pit development. I first assessed the efficacy of bird deterrents to mitigate impacts of mining-induced flooding on arctic-nesting birds at a gold mine in Nunavut. I used a Before-After Control Impact (BACI) design to determine changes in male territory densities, between year and treatment types (Control, High and Low Deterrent Intensity). Additionally, I assessed whether deterrents impacted daily survival rates of two passerine species, and the incubation behaviour of female Lapland Longspur. Finally, I quantified nest losses during the breeding season due to direct flooding of the tundra nesting habitat caused by mining operations. Deterrents did not affect male territory densities and neither deterrent treatment nor year affected the daily survival rate of nesting passerines. Female Lapland Longspurs exposed to deterrents exhibited more incubation off-bouts than control females. I documented six flooded nests. Deterrents used in this study appear to be ineffective in mitigating nesting in potential zones of impact. Incidental take accounted for about 1.2% of all nests found in the 0.48 km2 Whale Tail Lake study area. Author Keywords: arctic-nesting birds, audio deterrents, incidental take, mining and resource development, nest incubation, visual deterrents
Immunotherapies Targeting the Amyotrophic Lateral Sclerosis-Associated Protein TDP-43
Transactive response (TAR) DNA-binding protein of 43 kDa (TDP-43) pathology, including fibrillar aggregates and mutations, develops in amyotrophic lateral sclerosis (ALS) and is characterized by hyperphosphorylation and aggregation patterns, a mechanism largely understudied. In addition, ALS remains without a cure. Herein, in vitro aggregation of phosphorylated TDP-43 was explored, and the anti-TDP-43 antibodies tested for their inhibitor efficacies. Additionally, in vitro phosphorylation of TDP-43 by protein kinases was conducted to identify which protein kinases catalyze phosphorylation. The aggregation of phosphorylated and unphosphorylated full-length TDP-43 protein (pS410) was monitored by transmission electron microscopy (TEM), turbidity absorbance, and thioflavin (ThT) fluorescence spectroscopy. The protein aggregates were largely insoluble, ThT-positive and characterized with heterogeneous morphologies. Antibodies specific to epitopes within the RNA-recognition motifs and the C-terminal domains reduced the formation of β-sheets and insoluble aggregates, with outcomes highly dependent on the type of antibodies, indicating dual functionality. The only protein kinase able to phosphorylate TDP-43 at S410 was MARK4, indicating its role in the onset of PTMs in the protein. Thus, targeting TDP-43 epitopes for inhibition of aggregation and in vitro phosphorylation represent viable biochemical assays for screening protein kinase inhibitors as potential drugs against ALS. Author Keywords: aggregation, ALS, antibody-based inhibition, phosphorylation, protein kinase, TDP-43
Demographic history and conservation genomics of caribou (Rangifer tarandus) in Québec
Genetic variation is the raw material and basis for evolutionary changes in nature. The loss of genetic diversity is a challenge many species are facing, with genomics being a potential tool to inform and prioritize decision making. Whole genome analysis can be an asset to conservation biology and the management of species through the generation of more precise and novel metrics. This thesis uses whole genome re-sequencing to characterize the demographic history and quantify genomic metrics relevant to conservation of caribou (Rangifer tarandus) in Québec, Canada. We calculated the ancestral and contemporary patterns of genomic diversity of five representative caribou populations and applied a comparative population genomics framework to assess the interplay between demographic events and genomic diversity. When compared to the census size, NC, the endangered Gaspésie Mountain caribou population had the highest ancestral Ne:NC ratio which is consistent with recent work suggesting high ancestral Ne:NC is of conservation concern. These ratios were highly correlated with genomic signatures (i.e. Tajima’s D) of recent population declines and explicit demographic model parameters. Values of contemporary Ne, estimated from linkage-disequilibrium showed Gaspêsie having among the highest contemporary Ne:NC ratio. Importantly, classic conservation genetics theory would predict this population to be of less concern based off this metric alone. Inbreeding measures suggested nuanced patterns of inbreeding and correlated to the demographic models. This study suggests that while the Québec populations are all under decline, they harbour enough ancestral genetic variation to replenish any lost diversity, if conservation decisions are made in favour of these populations, specifically supporting NC. Author Keywords:
Assessing the environmental correlates of a lethal amphibian pathogen, Batrachochytrium dendrobatidis, in Ontario wetlands
Many emerging infectious diseases are caused by pathogens that possess free-living life stages, in which they interact with the environment directly rather than through the mediation of a host. These diseases represent major impediments to wildlife conservation; however, the dynamics of their interaction with the environment are poorly studied, often due to the difficulty of detecting these microscopic pathogens in environmental samples. One of these pathogens is Batrachochytrium dendrobatidis (Bd), a fungus that has been linked to declines in many amphibian species. In this thesis I use an emerging technique, environmental DNA detection (eDNA), to detect and quantify Bd in the water of southern Ontario (Canada) wetlands and examine its correlation with a variety of aspects of water quality, surrounding habitat, and seasonal timing. My purpose was to inform not only on potential environment-pathogen dynamics for Bd in northern environments, but to provide insight into the use of eDNA as a disease surveillance tool. I found that not only was there high geographic variation in Bd detection and intensity, but also high temporal variation within the same site on time scales as low as two weeks. While Bd prevalence was not strongly correlated with any of the variables tested, intensity showed strong correlation with canopy cover, with greater canopy cover over a waterbody correlating to lower Bd intensity. My results present several promising avenues for further examination of Bd in northern ecosystems, and indicate that, while caution is warranted in its implementation, eDNA may become an important tool in amphibian pathogen surveillance. Author Keywords: amphibian disease, Batrachochytrium dendrobatidis, disease monitoring, environment-pathogen dynamics, environmental DNA, wildlife disease
Frequency-time and polarization considerations in spectral-focusing-based CARS microscopy
Spectral-focusing-based coherent anti-Stokes Raman scattering (SF-CARS) microscopy is a powerful imaging technique that involves temporally and spectrally stretching ultrashort laser pulses and controlling their frequency-time characteristics. However, a broader and more detailed understanding of the frequency-time characteristics of the laser pulses and signals involved, how they are related, and how they influence important aspects such as the spectral resolution is needed to understand the full potential of SF-CARS systems. In this work, I elucidate these relationships and discuss how they can be exploited to optimize SF-CARS microscopy setups. I present a theoretical analysis of the relationships between the spectral resolution, the degree of chirp-matching, and pulse bandwidth in SF-CARS. I find that, despite allowing better ultimate spectral resolution when chirp-matching is attained, the use of the broadest bandwidth pulses can significantly worsen the spectral resolution if the pulses are not chirp-matched. I demonstrate that the bandwidth of the detected anti-Stokes signal is minimized when the pump is twice as chirped as the Stokes, meaning that (perhaps counter-intuitively) a narrow anti-Stokes bandwidth does not imply good spectral resolution. I present approximate expressions that relate the bandwidths of the pump, Stokes, and anti-Stokes pulses to the degree of chirp-matching and outline how these could be used to estimate the amount of glass needed to attain chirp-matching. I develop a spectral-focusing-based polarization-resolved (SFP-CARS) setup, by modifying our existing system, to explore the merits of integrating polarization-dependent detection as an add-on to existing SF-CARS setups. By using the system to study polarization-dependent features in the CARS spectrum of benzonitrile, I assess its capabilities and demonstrate its ability to accurately determine Raman depolarization ratios. Ultimately, the detected anti-Stokes signals are more elliptically polarized than desired, hindering a complete suppression of the non-resonant background. Nevertheless, I find that the SFP-CARS setup is a useful tool for studying polarization-dependent features in the CARS spectra of various samples and is worthy of further investigation. This work clarifies several technical aspects of SF-CARS microscopy and provides researchers with valuable information to consider when working with SF-CARS microscopy systems. Author Keywords: coherent anti-Stokes Raman scattering, nonlinear microscopy, polarization, spectral focusing, spectroscopy

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Format: 2024/03/28