Graduate Theses & Dissertations


Expression of Giardia intestinalis flavoenzyme GiOR-1 and characterization of its electron transfer properties
Giardia intestinalis possesses four isotypes of cytochrome b5 (gCYTB-I-IV) that differ from their mammalian counterparts, suggesting different functions in this protozoan parasite. Although the recently discovered Giardia flavoenzyme, GiOR-1, reduces these cytochromes, its properties have not been thoroughly studied, owing to the difficulty in its expression. Here I describe successful conditions for expression of GiOR-1 using autoinduction. GiOR-1 is obtained with flavins bound as indicated by its UV-visible spectrum. Its ability to catalyze electron transfer from donors (NADH, NADPH) to acceptors (oxygen, ferricyanide, cytochrome c, gCYTB5-III) were studied in spectrophotometric rate assays. NADPH is the preferred electron donor, while cytochromes are the preferred electron acceptors. Interestingly, the His-tag used to purify gCYTB5-III decreases its reaction rate with GiOR-1, as an untagged version has slightly faster rates. These findings establish the appropriate conditions for further studies on GiOR-1, including the identification of endogenous electron acceptors. Author Keywords: Autoinduction, Cytochrome b5, Cytochrome P450 oxidoreductase, Giardia intestinalis, GiOR-1, Polyhistidine tag
Prey abundance and habitat during the breeding season for Piping Plovers in the Ontario Great Lakes region
Similar to other shorebird trends around the world, the Piping Plover population (Charadrius melodus circumcinctus) is projected to decline if concerted conservation efforts are relaxed. To date, there is insufficient understanding of the connection between habitat type, prey abundance, and chick behaviour of the Piping Plover breeding population in Ontario. The aim of my thesis was to gain knowledge about prey abundance at recent and historic breeding locations, understanding how habitat influences prey abundance and chick behaviour across the Piping Plover breeding range in Ontario’s Great Lakes. The objective of my first study was to understand prey abundance across the breeding region Great Lakes of Ontario from 2018-2019, including occupied and unoccupied sites, and to quantify variation among habitats and periods of reproduction. To evaluate resources, I sampled 17 locations to compare prey abundance using invertebrate traps (n= 3,507). Sampling took place over the reproductive periods of nest initiation, post-hatch, and fledging and in four habitat types of shoreline, wrack, berm, and back dune. Occupied breeding sites had higher prey abundances, and different assemblages of invertebrate prey than unoccupied sites. Additionally, breeding sites had higher prey abundance during nest initiation and supported higher amounts of prey in shoreline and wrack habitat. The objective of my second study was to understand how habitat types influence chick behaviour. To evaluate behaviour-habitat trends, instantaneous chick observations were recorded at the four nest sites from the post-hatch to fledging stages. In total there were 23 fledged chicks that we observed across the two years. Chicks in this study spent 60.9% of their time foraging, 11.9% of the time displaying alert behaviour, 21.4% of their time resting or being brooded, and 5.9% of their time preening. Chicks spent a large proportion of time foraging in the shoreline, resting in the back dune, and alert in berm habitat. The frequency of these alert, defensive behaviours differed among sites, with Sauble Beach chicks spending more time in defensive behaviours compared to other sites. I concluded that in both nesting and brood-rearing periods, habitat is selected non-randomly by adult and young Piping Plovers to maximize access to invertebrate prey for growth and survival. Access by chicks to the most productive habitats should be considered in local management decisions. Author Keywords: chick behaviour, endangered, Great Lakes Region, habitat, Piping Plovers, prey abundance
Sensitivity of Forest Soils to Acidic Deposition Downwind of an Aluminum Smelter, Kitimat, B.C.
Maximum permitted SO2 emissions from an aluminum smelter in Kitimat, B.C., increased after modernization in 2015. An increase in acidic deposition can potentially acidify forest soils. Monitoring was conducted at two long-term soil monitoring plots at near (7 km) and far (41 km) sites downwind from the smelter. Change in soil properties was assessed between 2015 and 2018: for the near plot, there was significant decrease in pH and exchange acidity; far plot soils exhibited significant decrease of base cations and exchange acidity in the 0–5 cm layer only. The average total SO42- deposition at near and far plots were estimated to be between 8.2–12.1 and 6.7–7.4 kg/ha/yr, respectively. It was concluded no soil acidification was detected. Observed changes were attributed to measured differences in organic matter, likely influenced by sampling difficulty and measurement process discrepancies. Estimated SO42- deposition levels pose no risk to soil base cation depletion. Author Keywords: acid forest soils, acidic deposition, aluminum smelter, exchangeable base cations, long-term monitoring, minimum detectable change
Assessing effects and fate of environmental contaminants in invasive, native, and endangered macrophytes
Macrophytes play an important role in aquatic ecosystems, and thus are integral to ecological risk assessments of environmental contaminants. In this dissertation, I address gaps in the assessments of contaminant fate and effects in macrophytes, with focus on glyphosate herbicide use for invasive plant control. First, I evaluated the suitability of Typha as future standard test species to represent emergent macrophytes in risk assessments. I concluded that Typha is ecologically relevant, straight-forward to grow, and its sensitivity can be assessed with various morphological and physiological endpoints. Second, I assessed effects from glyphosate (Roundup WeatherMAX® formulation) spray drift exposure on emergent non-target macrophytes. I performed toxicity tests with five taxa, Phragmites australis, Typha × glauca, Typha latifolia, Ammannia robusta, and Sida hermaphrodita, which in Canada collectively represent invasive, native, and endangered species. I found significant differences in glyphosate sensitivity among genera, and all species’ growth was adversely affected at concentrations as low as 0.1% (0.54 g/L), much below the currently used rate (5%, 27 g/L). Third, I assessed the potential for glyphosate accumulation in and release from treated plant tissues. I found that P. australis and T. × glauca accumulate glyphosate following spray treatment, and that accumulated glyphosate can leach out of treated plant tissues upon their submergence in water. Finally, I assessed effects of released glyphosate on non-target macrophytes. I found that P. australis and T. × glauca leachate containing glyphosate residues can stimulate the germination and seedling growth of T. latifolia, but can exert an inhibiting effect on A. robusta, although leachate without glyphosate caused similar responses in both plants. Additionally, I found no negative effects in A. robusta when exposed to glyphosate residues in surface water, or when grown with rhizosphere contact to an invasive plant that was wicked (touched) with glyphosate. My results show that non-target macrophytes can be at risk from glyphosate spray for invasive plant control, but risks can be mitigated through informed ecosystem management activities, such as targeted wick-applications or removing plant litter. Integrating contaminant fate and effect assessments with emergent macrophytes into ecological risk assessments can support the protection of diverse macrophyte communities. Author Keywords: Ecosystem management, Ecotoxicology, Glyphosate, Herbicide, Invasive plant, Species at risk
Robust assessment of changes in wild mammal occupancy and activity relative to livestock and human disturbance
Anthropogenic activities such as human activity and livestock grazing are responsible for the global rise in disturbance impacts on wildlife and may underlie regional changes in biodiversity and ecosystem dynamics. Few studies have tried to disentangle the effects of different anthropogenic activities on wildlife behaviour, leaving a major gap in our understanding of conservation and management needs in disturbed areas. Human activity and livestock grazing are increasing in spread and intensity worldwide, thereby imposing pressure on both wildlife and natural areas. In this thesis, I used a camera trapping and occupancy modeling framework to assess whether human presence and livestock grazing had different impacts on site occupancy and activity of 10 wild mammal species, and how responses differed across taxa. Specifically, I predicted that all species would be sensitive to disturbance, but the type and intensity of the response would depend on disturbance type. I detected different responses to each disturbance type across species, but response type (displacement, activity change, crepuscularity) was not associated with species characteristics such as body. Importantly, disturbance intensity had a strong effect on wildlife activity levels, with many species exhibiting marked reductions in activity at high human or livestock disturbance intensity. It remains unclear whether all species’ responses are a direct consequence of disturbance versus indirect outcomes of shifts in behaviour of other species in the wildlife community (i.e., disturbance-related changes in prey activity may affect predator activity). Although on the whole disturbance intensity and effect sizes tended to be relatively low in this study, responses were exhibited across all species, implying that disturbance responses may be universal in wild mammals and largely underestimated. Ultimately, my work offers a template for the robust assessment of disturbance impacts on wildlife and provides new avenues for future research to deepen our understanding of wildlife sensitivity to anthropogenic activities. Author Keywords: activity, anthropogenic disturbance, human activity, livestock, occupancy, wildlife
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
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


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