Graduate Theses & Dissertations

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Responses of Primary Producers and Grazers to Silver Nanoparticle Exposure
The increasing production and use of silver nanoparticles (AgNPs) raise concerns on environmental exposure and impact. A large scale in situ enclosure study was conducted at the Experimental Lakes Area to determine the effect of AgNPs on natural phytoplankton and zooplankton communities. This study investigated AgNPs of varying concentrations (4, 16 and 64 μg/L), dosing regimens (chronic vs. pulse), and capping agents (poly-vinyl pyrrolidone vs. citrate). Phytoplankton communities were influenced only by the natural limnological properties of the system signifying tolerance to AgNPs. Zooplankton community structure significantly changed with AgNP concentration and dosing regimen indicating AgNP sensitivity. A microcosm study investigating the effect of AgNPs and phosphorus-dosed periphyton before and after grazing by two benthic invertebrate species (snails and caddisfly larvae) showed reduced periphyton stoichiometry with AgNP exposure. Grazers foraged less on silver dosed periphyton indicating a preferential choice in food quality. Phosphorus reduced the detrimental effects of AgNPs across all conditions. These studies verify the need for in situ experimental designs to fully investigate the effects of AgNPs and their interaction with environmental factors, multiple species assemblages, and across trophic levels. Author Keywords: benthic invertebrate, Experimental Lakes Area, periphyton, phytoplankton, silver nanoparticles, zooplankton
Evaulating the American Woodcock Singing-Ground Survey Protocol in Ontario using Acoustic Monitoring Devices
The breeding phenology of American Woodcocks (Scolopax minor) was evaluated in Ontario, Canada to determine if changes in dates of courtship activity have introduced negative bias into the American Woodcock Singing-ground Survey (SGS). Long-term woodcock phenology and climate data for Ontario were analysed using linear regression to determine if woodcock breeding phenology has changed between 1968 and 2014. There was no significant trend in woodcock arrival date, but arrival date was correlated with mean high temperature in March. In 2011-2013, programmable audio-recording devices (song meters) were deployed at known woodcock singing-grounds to determine if peaks in courtship activity coincided with survey dates used by the SGS. Spectrogram interpretation of recordings and data analyses using mixed-effects models indicated the SGS survey dates were still appropriate, except during the exceptionally early spring in 2012 when courtship displays were waning in one region during the survey window. The methods for interpretation of song meter recordings were validated by conducting point counts adjacent to song meters deployed at singing-grounds, and at randomly selected locations in woodcock habitat. Recommendations for the SGS protocol are included. Author Keywords: detectability, phenology, Scolopax minor, Singing-ground Survey, song meter
Detection of four at-risk freshwater pearly mussel species (Bivalvia
Environmental DNA (eDNA) detection uses species-specific markers to screen DNA from bulk samples, such as water, to infer species presence. This study involved the development and testing of species-specific markers for four freshwater pearly mussels (Unionidae). The markers were applied to water samples from intensively sampled mussel monitoring sites to compare species detections from eDNA with established sampling method detections. Target species were detected using eDNA at all sites where they had previously been detected by quadrat sampling. This paired design demonstrated that eDNA detection was at least as sensitive as quadrat sampling and that high species specificity can be achieved even when designing against many sympatric unionids. Detection failures can impede species conservation efforts and occupancy estimates; eDNA sampling could improve our knowledge of species distributions and site occupancy through increased sampling sensitivity and coverage. Author Keywords: conservation genetics, cytochrome oxidase subunit I (COI), environmental DNA (eDNA), quantitative PCR (qPCR), species at risk (SAR)
Chemical characterization of dissolved organic matter in relation with hydrography in the Arctic Ocean
In this thesis, water mass distribution of dissolved organic matter (DOM) characteristics (i.e. molecular weight, fluorescent components, thiols and humic substances concentration) was observed in the Arctic Ocean. For the first time, DOM molecular weight (MW) in Beaufort Sea was assessed using asymmetrical flow field-flow fractionation, as well as the first monitoring of thiols and humic substances (HS) using cathodic stripping voltammetry (CSV) in the Arctic Ocean. Based on fluorescence property, DOM characterization was carried out using parallel factor analysis – excitation-emission matrices. Pacific winter waters in the Canada Basin showed higher MW DOM associated with higher fluorescence intensity. High HS was associated with the Arctic outflow waters in top 300 m of the Canadian Arctic Archipelago. Interestingly, maximum thiol concentration was associated with the subsurface chlorophyll-a maximum at most sites, but not universal along the study area. Comparable distributions of CSV-based HS and humic-like fluorescent components suggest similar sources/ processes in the Arctic Ocean. The findings in this thesis suggested DOM characteristics could be used as fingerprints in tracing water masses in the Arctic Ocean. Author Keywords: Asymmetrical flow field-flow fractionation, Cathodic stripping voltammetry, DOM, Metal-binding ligands, Molecular weight, PARAFAC-EEMs
Investigating wheat rust virulence evolution through transcriptome analysis of a recently emerged race of Puccinia triticina
Puccinia triticina, wheat leaf rust (WLR), is the most economically damaging fungal rust of wheat on a global scale. This study identified transcriptome changes in a recently emerged race of WLR in Ontario with a new virulence type relative to a possible ancestor race. Also, this study focused on detecting variation in candidate virulence genes and uncovering novel insight into WLR virulence evolution. Various race-by-variety interactions were evaluated using RNA-seq experiments. A list of genes with statistically significant expression changes in each comparison was prepared and predicted effectors were retained for further analysis. Proteins with nonsynonymous substitutions were run through BLASTx to identify potential orthologs. Over 100 candidate effectors with a 2-fold or higher change in transcript level were identified. Seven of these candidate effector genes were recognized to contain single nucleotide polymorphisms (SNPs) which altered the amino acid sequence of the resulting protein. The information gained may aid in targeted breeding programs to combat new WLR races as well as provide the basis for functional analysis of WLR using potential orthologs in a model basidiomycete. Author Keywords: effectors, RNA-seq, rust fungi, SNPs, transcriptome, wheat leaf rust
Nunavik Inuit Knowledge of Beluga
Socio-ecological systems are inherently complex and marine mammals are fundamentally challenging to study. In the Arctic, marine mammals occupy a central ecological role, as nutrient cyclers and as a source of food and culture for Indigenous peoples. Inuit have developed a rich knowledge system, which has not been fully actualized in application in most Arctic research. Considering the need for the best available information in marine mammal ecology, the research question guiding this dissertation was: How can multiple methods and approaches be used to more effectively gather, understand, and represent Inuit Knowledge for an improved understanding of marine mammal ecology? The dissertation investigates this question using a case study of beluga in Nunavik (Arctic Quebec) drawing on the expertise of hunters and Elders to better understand complex questions in marine mammal ecology. The thesis uses a transdisciplinary approach to address the dissertation objective and is comprised of a general introduction, followed by four chapters formatted as journal manuscripts, and closes with an integrated discussion and conclusion. The first manuscript examines the contributions of Traditional Ecological Knowledge (TEK) of beluga to marine mammal literature. The second manuscript uses a sub-set of data gathered through participant mapping to apply a mapping method to explore how the spatial aspects of TEK could be better documented, analyzed, and represented. The third and fourth papers are based on the knowledge shared by hunters and Elders. The third explores the questions ‘why do beluga migrate?’ and ‘what factors influence beluga movement?’. The fourth investigates aspects of beluga foraging ecology. This dissertation makes methodological contributions through the application of kernel density estimators to participant maps as a method for transforming multiple mapped narratives into a quantitative database. The understandings shared by hunters and Elders make significant ecological contributions, particularly to foraging (e.g. diet composition and seasonal energy intake), and movement ecology (e.g. potential drivers of migration). Broadly these findings contribute to our collective understanding of beluga ecology and have implications for wildlife management. Author Keywords: Arctic, Beluga biology, foraging ecology, Inuit Knowledge, migration, transdisciplinary
effects of Dissolved Organic Matter (DOM) sources on Pb2+, Zn2+ and Cd2+ binding
Metal binding to dissolved organic matter (DOM) determines metal speciation and strongly influences potential toxicity. The understanding of this process, however, is challenged by DOM source variation, which is not always considered by most existing metal speciation models. Source determines the molecular structure of DOM, including metal binding functional groups. This study has experimentally showed that the allochthonous-dominant DOM (i.e. more aromatic and humic) consistently has higher level of Pb binding than the autochthonous-dominant DOM (i.e. more aliphatic and proteinaceous) by more than two orders of magnitude. This source-discrimination, however, is less noticeable for Zn and Cd, although variation still exceeds a factor of four for both metals. The results indicate that metal binding is source-dependent, but the dependency is metal-specific. Accordingly, metal speciation models, such as the Windermere Humic Aqueous Model (WHAM), needs to consider DOM source variations. The WHAM input of active fraction of DOM participating in metal binding (f) is sensitive to DOM source. The commonly-used f = 0.65 substantially overestimated the Pb and Zn binding to autochthonous-dominant DOM, indicating f needs to be adjusted specifically. The optimal f value (fopt) linearly correlates with optical indexes, showing a potential to estimate fopt using simple absorbance and/or fluorescence measurements. Other DOM properties not optically-characterized may be also important to determine fopt, such as thiol, which shows strong affinity to most toxic metals and whose concentrations are appreciably high in natural waters (< 0.1 to 400 nmol L-1). Other analytical techniques rather than Cathodic Stripping Voltammetry (CSV) are required to accurately quantify thiol concentration for DOM with concentration > 1 mg L-1. To better explain the DOM-source effects, the conditional affinity spectrum (CAS) was calculated using a Fully Optimized ContinUous Spectrum (FOCUS) method. This method not only provides satisfactory goodness-of-fit, but also unique CAS solution. The allochthonous-dominant DOM consistently shows higher Pb affinity than autochthonous-dominant DOM. This source-discrimination is not clearly observed for Zn and Cd. Neither the variability of affinity nor capacity can be fully explained by the variability of individual DOM properties, indicating multiple properties may involve simultaneously. Together, the results help improve WHAM prediction of metal speciation, and consequently, benefit geochemical modelling of metal speciation, such as Biotic Ligand Model for predicting metal toxicity. Author Keywords: Dissolved organic matter, Metal binding, Source, Windermere Humic Aqueous Model
effects of in-stream woody debris from selective timber harvest on nutrient pools and dynamics within Precambrian Shield streams
Timber harvest can influence the rate of transfer of organic matter from the terrestrial catchment to streams, which may have both direct and indirect effects on in-stream nutrient pools and dynamics. In the interest of developing sustainable forestry practices, the continued study of the effects of forestry on nutrient dynamics in aquatic systems is paramount, particularly in sensitive nutrient-poor oligotrophic systems. The goal of this study was to investigate the impacts of harvest-related woody debris on stream nutrient status in streams located in the Canadian Shield region of south-central Ontario. Surveys showed greater large (> 10 cm) and small (< 10 cm) woody debris dry masses and associated nutrient pools in streams located in recently (2013) selectively harvested catchments, when compared with catchments not harvested for at least 20 years. Experimental releases of flagging tape underlined the importance of woody debris as a mechanism of coarse particulate organic matter (CPOM) retention. Sediment surveys showed a significant exponential decline in both OM content and nutrients associated with coarse sediment with distance upstream from debris dams. Laboratory leaching experiments suggest that fresh woody debris may be an important short-term source of water-soluble nutrients, particularly phosphorus and potassium. This study suggests that woody debris from timber harvest is both a direct and indirect source of nutrients, as trapped wood and leaves that accumulate behind debris dams can augment stream nutrient export over long time periods. Author Keywords: nutrient leaching, nutrient pools, organic matter retention, selection harvest, southern Ontario, woody debris
Investigation of fugitive dust emissions from nepheline syenite mine tailings near Nephton, Ontario
A set of experiments was designed to investigate the factors—atmospheric and surficial—controlling fugitive dust emissions from the tailings ponds of UNIMIN Canada, a mining company that extracts and produces nepheline syenite (feldspar) at two adjacent sites (Nephton and Blue Mountain) located north of Havelock, Ontario. Using wind tunnel measurements, the combined influence of relative humidity and temperature (represented by the absolute matric potential, |Ψ|) on dust emission was quantified and modelled. About 300 experimental runs were conducted under various conditions of wind speed (4.5-6.25 ms-1), temperature (0-30oC) and relative humidity (10-70%). Generally, dust flux decreased as a logarithmic function of matric potential, with dust emission strongly suppressed for RH > 60% or |Ψ|<70 MPa. Field measurements also confirmed the role of relative humidity in suppressing dust emission. Irrigation, which is widely used by mines to control dust emissions, reduced ambient dust concentration at the study site only about 60% of the time, with the highest mitigation efficiencies (average of 90%) occurring when the total depth of water applied intermittently over a few hours was greater than 10 mm. In the absence of emergent vegetation, the terrestrial laser scanning (TLS) technique proved to be a promising method for detecting and estimating both spatial and temporal moisture content changes in the field environment, particularly for the very thin surface layer, which is the most important layer for dust emission. It is hoped that the results from this study will help mines to optimize their dust management programs for the range of climate and topographic conditions found at their sites, and also serve as a source of useful information and input data for atmospheric dispersion models, such as AERMOD and CALPUFF, whose accuracy depends on the quality of the input data such as the emission rate. Author Keywords: dust mitigation efficiency, Fugitive dust, nepheline syenite, relative humidity, tailings, terrestrial laser scanning
effects of particulate matter on the fate and toxicity of silver nanoparticles
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
significance of topographically-focused groundwater recharge during winter and spring on the Oak Ridges Moraine, southern Ontario
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
Studies of the Giardia intestinalis trophozoite cell cycle
To study the Giardia intestinalis cell cycle, counterflow centrifugal elutriation (CCE) was used to separate an asynchronous trophozoite culture into fractions enriched for cells at the different stages of the cell cycle. For my first objective, I characterized the appearance of a third peak (Peak iii) in our flow cytometry analysis of the CCE fractions that initially suggested the presence of 16N cells that are either cysts or the result of endoreplication of Giardia trophozoites. I determined that this third peak consists of doublets of the 8N trophozoites at the G2 stage of the cell cycle that were not removed effectively by gating parameters used in the analysis of the flow cytometry data. In the second objective, I tested the use of a spike with RNA from the GS isolate of Giardia as an external normalizer in RT-qPCR on RNA from CCE fractions and encystation cultures of Giardia from the WB isolate. My results showed that the GS RNA spike is as effective as the use of previously characterized internal normalizer genes for these studies. For the third objective, I prepared two sets of elutriation samples for RNA seq analysis to determine the transcriptome of the Giardia trophozoite cell cycle. I confirmed the results of the cell cycle specific expression of several genes we had previously tested by RT-qPCR. Furthermore, our RNA-seq identified many genes in common with those identified from a microarray analysis of the Giardia cell cycle conducted by a collaborator. Finally, I observed an overall <4 fold change in differentially expressed genes during the G1/S and G2/M phase of the cell cycle. This is a modest change in gene expression compared to 10 - 30 fold changes for orthologous genes in mammalian cell cycles. Author Keywords: Cell cycle, Counterflow Centrifugal Elutriation, Flow Cytometry, RNA-sequencing, RT-qPCR

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