Graduate Theses & Dissertations

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Using Fluorescent Carbon Dots for Biosensing Applications of Amino Acids
Amino acids make up proteins, which are the building blocks of life. A balance of amino acids is needed to maintain a healthy state. Tyrosine (Tyr) is synthesized from the metabolism of phenylalanine, which is an essential amino acid, meaning it can only be obtained from the diet. It is related to many metabolic and neurodegenerative diseases. Tyr can undergo post-translational modifications such as phosphorylation and nitration, which are implicated in cancer and nitrative stress, respectively. Although there are many methods to detect Tyr and its analogues, phosphotyrosine (pTyr) and nitrotyrosine (nTyr), these methods are time-consuming, involve expensive instruments and involve tedious process. This research proposes a new type of nanomaterials, carbon dots (CDs), to detect these amnio acids. Data indicate that CDs can be used to detect nTyr with a limit of detection of 34 μM in the linear range of 20 - 105 μM. The amenability of CD-nTyr assay was also tested in various biological matrices and biological molecules and was shown to be sensitive to nTyr. Nitration of Tyr was carried out in the presence of sodium nitrite and hydrogen peroxide catalyzed by either Cu(II) or Fe(III) to mimic biological reactions and CDs were tested as both inhibitors and indicators of Tyr nitration. Although CDs did not inhibit the nitration reaction of Tyr, they did not serve as indicators of Tyr nitration due to the quenching of CDs by the nitrating agents. This shows the importance of using CDs to detect nTyr and further use it for biological applications to detect diseased states. Author Keywords: amino acids, carbon dots, nanomaterials, sensor, spectroscopy, tyrosine
Using DNA Barcoding to Investigate the Diet and Food Supply of a Declining Aerial Insectivote, the Barn Swallow (Hirundo rustica)
Barn Swallow (Hirundo rustica) populations have declined in North America over the past 40 years and they are listed as Threatened in Ontario, Canada. Changes in the food supply have been hypothesized as a potential cause of this population decline. I used DNA barcoding to investigate the diet and food supply of Barn Swallows and to determine if the food supply affects their reproductive performance. In two breeding seasons, I monitored nests, collected fecal samples, and monitored prey availability by collecting insects from the habitat surrounding breeding sites using Malaise traps. I used DNA barcoding to identify insect specimens collected from the habitat and to identify prey items from Barn Swallow nestling fecal samples. I found that Barn Swallow nestlings were fed a very broad range of prey items but were fed larger prey items more frequently. Prey availability was not related to the timing of reproduction, the number of nests at a breeding site, or the reproductive output of individual nests. This study provides information on the diet composition of Barn Swallows in North America and suggests that food limitation during the breeding season may not be a major factor in their population decline. Author Keywords: aerial insectivore, diet, DNA barcoding, Hirundo rustica, metabarcoding, reproductive success
Understanding the establishment of Typha spp. in North America using population genetics and common garden studies
There are three cattail (Typha) taxa in Canada: T. latifolia (native), T. angustifolia (introduced), and their hybrid T. x glauca. The latter is invasive in regions around the Laurentian Great Lakes, and I investigated the potential role that commercial suppliers may be playing in the introduction of non-native Typha by comparing genotypes of North American, European, and commercially available plants. I found that Ontario garden centres are importing both hybrids and non-native lineages of T. angustifolia into Canada, but was unable to identify the provenance of T. latifolia. I also investigated the possibility that the hybrid cattail leaf litter shade and leachate influences germination and early growth of the parental species of the hybrids. Using three common garden experiments, I found that T. x glauca leaf litter suppresses germination rates of the three taxa. In the early seedling growth experiment, plant performance varied by taxa, and for the competition experiment there were no intra- or interspecific competition or treatment effects on the performance of plants. Overall, my research identified a potential mechanism allowing T. x glauca to dominate wetlands, and also shows that non-native lineages are being introduced into Canada through commercial trade Author Keywords: Competition, Germination, Non-native lineages, Plant nurseries, Seedling Growth, Typha spp.
Understanding Historical and Contemporary Gene Flow Patterns of Ontario Black Bears
Consequences of habitat loss and fragmentation include smaller effective population sizes and decreased genetic diversity, factors that can undermine the long-term viability of large carnivores that were historically continuously distributed. I evaluated the historical and contemporary genetic structure and diversity of American black bears (Ursus americanus) in Ontario, where bear habitat is largely contiguous, except for southern regions that experience strong anthropogenic pressures. My objectives were to understand gene flow patterns in a natural system still largely reflective of pre-European settlement to provide context for the extent of genetic diversity loss in southern populations fragmented by anthropogenic influences. Phylogeographic analyses suggested that Ontario black bears belong to a widespread "continental" genetic group that further divides into 2 subgroups, likely reflecting separate recolonization routes around the Great Lakes following the Last Glacial Maximum. Population genetic analyses based on individual genotypes showed that Ontario black bears are structured into 3 contemporary genetic clusters. Two clusters, located in the Northwest (NW) and Southeast (SE), are geographically vast and genetically diverse. The third cluster is less diverse, and spatially restricted to the Bruce Peninsula (BP). Microsatellite analyses revealed that the NW and SE clusters are weakly differentiated from each other relative to mitochondrial DNA findings, suggesting male-biased dispersal and isolation by distance across the province. I also conducted simulations to assess competing hypotheses that could explain the reduced genetic diversity on the BP, which supported a combination of low migration and recent demographic bottlenecks. I showed that management actions to increase genetic variation in BP black bears could include restoring landscape connectivity between BP and SE; however, the irreversible human footprint in the area makes regular translocations from SE individuals a more practical alternative. Overall, my work suggests that: 1) historical genetic processes in Ontario black bears were likely predominated by isolation by distance, 2) large mammalian carnivores such as black bears can become isolated and experience reduced diversity in only a few generations, and 3) maintaining connectivity in regions under increased anthropogenic pressures could prevent populations from becoming small and geographically and genetically isolated, and should be a priority for conserving healthy populations. Author Keywords: American black bear, carnivore, conservation genetics, Ontario, phylogeography, population genetics
Understanding Angler Dynamics in a Recreational Lake Trout (Salvelinus namaycush) Fishery in Algonquin Provincial Park Using Long-Term Access Creel Data
In order to effectively manage recreational fisheries, it is important to understand how the resource is being used. In this thesis, long-term creel census data, collected on Lake Opeongo in Algonquin Provincial Park, Ontario, Canada was used to assess fine-scale angler dynamics within a recreational Lake Trout (Salvelinus namaycush) fishery. The spatial distribution of angler reported catch locations of Lake Trout within the lake, was assessed using an Optimized Hotspot Analysis in ArcGIS. Areas of significant clustering of catch locations were revealed during all time periods and varied in size and location both seasonally and temporally. Cormack-Jolly-Seber models were used to evaluate the probability of individual angling boats persisting in the regional fishery and being detected on Lake Opeongo through time, as well as to examine the effect of angler travel distance and gas prices on participation parameters. Time-varying models revealed that the probability of an angler persisting in the fishery varied, while detection estimates remained stable. Travel distance had a negative effect on both parameters while increased gas prices only had a slight negative effect on detection estimates. Additionally, among Lake Opeongo anglers, angling avidity varied as did lake specific fishing experience. Average CUE was found to be higher among angling parties who visited the lake more often than fishing parties who visited relatively few times. Author Keywords:
USE OF SALIVARY CORTISOL TO EVALUATE THE INFLUENCE OF RIDES ON THE STRESS PHYSIOLOGY OF DROMEDARY CAMELS (CAMELUS DROMEDARIUS)
Many facilities attempt to alleviate the risk of chronic stress in captivity by providing environmental enrichment shown to minimize behavioural disorders and stress in several species. One potential form of enrichment used in zoos is training animals to perform rides for guests, however, the effect of this activity on the welfare of individual animals has never been examined. I validated the use of saliva for assessing stress in dromedary camels (Camelus dromedarius) an animal commonly used for rides. I then measured variation in salivary cortisol in four male camels during animal rides for guests at the Toronto Zoo. The camels were sampled during the ride season (from June to August) using four treatments: 1) in their pasture, 2) at the ride area not performing rides, 3) performing a low number of rides (n=50/day) and 4) performing a high number of rides (n=150/day). Furthermore, samples were taken before and after the ride season for comparison. There was a significant difference between the post-ride season treatment and the three treatments involving guest presence during the ride season (ride area, low rides, high rides. This indicates that performing rides is not a stressful experience based on the stress metrics I used, and suggests that rides may be a form of enrichment for dromedary camels. Author Keywords: ACTH challenge, animal welfare, camels, environmental enrichment, salivary cortisol, stress
Tracking Mercury and Mercury Stable Isotopes Throughout the Wabigoon/English River System
In the Wabigoon/English River system, mercury concentrations downstream from Dryden, ON, where there was a former chlor-alkali plant, remain elevated in sediments and biota. Understanding the current extent and severity of mercury contamination downstream from the former chlor-alkali plant is of great interest in furthering the clean-up of mercury within the traditional territory of Asubpeeschoseewagong Netum (Grassy Narrows) First Nation. The objective of this study was to evaluate the current level and extent of mercury contamination within sediments, crayfish, Hexagenia mayflies, yellow perch, spottail shiner and walleye in the Wabigoon/English River system. An additional objective was to use mercury stable isotope analysis to distinguish between legacy mercury from the former chlor-alkali plant and mercury from geogenic sources. Mercury contamination within surface sediments and biota at locations as far as 178 kms downstream of the historical source of mercury contamination are elevated relative to the reference lake, Wabigoon Lake. Isotope ratios in young of the year fish and sediments collected from within the system were distinct from fish from the reference lake, Wabigoon Lake, indicating that anthropogenic mercury contamination is distinguishable from geogenic mercury. Author Keywords:
Trace Metal Geochemistry in Peatlands
Peatlands can be found widely across all latitudes and play a significant role in global cycles within the earth’s biosphere. The anoxic conditions in peatlands promotes the accumulation of organic matter through decreased rates of decomposition and the storage of certain elements, which have received contaminant loading over the course of human existence, with significant increases occurring during the period of industrialization. We assessed global patterns of metal enrichment in peatlands in 439 cores distributed across 5 continents and 21 countries and measured 35 elements by depth increments and by peatland type. Global patterns in enrichment factors (EF’s) were determined for all metals with the majority of metals being found to have a median EF < 2 indicating relatively minor enrichment. Principal component analysis indicated EF’s of 6 metals (Cd, Co, Cu, Ni, Pb, and Zn), 2 metalloids (As and Sb) and Se in the upper peat horizon had similar spatial patterns among peatlands and these elements had generally the highest EF’s with many cores exceeding EF >10 and some having EF values >100. Significant differences in EF’s were found for these 9 “pollution” elements by peatland type and to a greater extent by geographic region, with higher EF values typically occurring in Europe and North America. Enrichment factors for most elements exhibited weak but significant positive correlations with modelled [1850 – present] S deposition. Estimated pools for the “pollution metals” within the 0 - 40 cm depth varied considerably, with median global pools in peat ranging from 12.9 mg m-2 (Sb) to 439 mg m-2 (Zn) for these 9 metals. Climate changes presents a significant risk to global peatland geochemistry due to expected changes in hydrologic regimes, resulting in potentially increased metal mobility though drought-induced peatland acidification, with historic areas previously impacted by industrial activities presenting the greatest risk of metal release to downstream receiving environments. Using a case study, we examined the impact of simulated 30-day drought on pore water chemistry at six sites in a peatland complex in Elliot Lake Ontario that were historically impacted by uranium (U) mining activities. All sites responded similarly to simulated drought with pore water pH significantly declining. The decline in pore water pH was likely due to increasing sulphate (SO42-) concentrations, which accompanied large increases in Al, Ni, Cu, Pb, Zn, and U. Dissolved organic carbon (DOC) increased, which may further enhance Al, Cu, and U mobility as these metals are strongly complexed by organic acids. Metal partitioning (Kd) values could be significantly predicted by pH and DOC although the strength of the relationship varied considerably among sites. Multiple linear regression and the inclusion of SO4-2 improved predictions, indicating that declines in pH as a result of SO4-2 and H+ production primarily governs metals and U partitioning in peatland soils. The results from both studies show that metal enrichment in global peatlands is highly variable, with northern peatlands in industrialized areas presenting the greatest risk of metal release to downstream surface waters based on expected hydrologic impacts from climate change due to historical and on-going metal and S deposition. Author Keywords: Acidification, Climate Change, Drought, Enrichment Factors, Global, Peatlands
Time-dependent effects of predation risk on stressor reactivity and growth in developing larval anurans (Lithobates pipiens)
The predator vs. prey dynamic is an omnipresent factor in ecological systems that may drive changes in life history patterns in prey animals through behavioural, morphological, and physiological changes. Predation risk can have profound effects on the life history events of an animal, and is influenced by the neuroendocrine stress response. Activation of the hypothalamic-pituitary-adrenal/interrenal axis, and the induction of stress hormones (e.g., corticosterone (CORT)) have been shown to mediate the onset of inducible anti-predator defensive traits including increased tail-depth, and reduced activity. The predator-prey relationship between dragonfly nymphs and tadpoles can be a powerful model system for understanding mechanisms that facilitate changes in the stress response in accordance with altered severity of risk. It has been well demonstrated early in tadpole ontogeny that increased corticosterone (CORT) levels, observed within three weeks of predator exposure, are correlated with increased tail depth morphology. However, the reactivity of the stress response in relation to the growth modulation in developing prey has yet to be fully explored. Accordingly, this thesis assessed the stress and growth response processes in tadpoles that were continuously exposed to perceived predation risk later in ontogeny. Continuous exposure of prey to predation risk for three weeks significantly increased CORT levels, and tail depth. However, tadpoles exposed to six weeks of predation risk acclimated to the presence of the predator, which was observed as a significant reduction of stressor-induced CORT levels. In addition, although increased tail depth has been attributed to predator defense, predator-naïve tadpoles began to display similar tail depth morphology as treated tadpoles at the six week time point. Thus, this thesis suggests that the stress response in lower vertebrate systems (e.g., tadpoles) may operate in a similarly complex manner to that observed in higher vertebrates (e.g., rats), for which severity of risk associated with the stressor aids in defining activity of the stress response. Moreover, the lack of morphological difference between treatments among tadpoles exposed later in ontogeny suggests that the mechanisms for inducing defenses are normal morphological traits in the development of the animal. This thesis paves the way for future research to elucidate the relationship between the neuroendocrine stress response and hormonal pathways involved in growth modulation in the presence of environmental pressures. Author Keywords: Acclimation, Corticosterone, Growth Modulation, Predation Risk, R. pipiens, Tadpole
Time to adapt
To better understand species’ resilience to climate change and implement solutions, we must conserve environments that maintain standing adaptive genetic variation and the potential generation of new beneficial alleles. Coding trinucleotide repeats (cTNRs) providing high-pace adaptive capabilities via high rates of mutation are ideal targets for mitigating the decline of species at risk by characterizing adaptively significant populations. Ultimately, adaptive genetic information will inform the protection of biological diversity below the species level (i.e., “Evolutionarily Significant Units” or “ESUs”). This dissertation investigates cTNRs within candidate genes to determine their prevalence and influence under selection in North American mammals. First, I evaluated the potential for somatic mosaicism in Canada lynx (Lynx canadensis), and found that tissue-specific mosaicism does not confound cTNR genotyping success in lynx. Second, I assessed a selection of clock gene cTNRs across characterized mammals and found that these repeats are abundant and highly variable in length and purity. I also identified preliminary signatures of selection in 3 clock gene cTNRs in 3 pairs of congeneric North American mammal species, highlighting the importance of cTNRs for understanding the evolution and adaptation of wild populations. I further evaluated the influence of selection on the NR1D1 cTNR within Canada lynx sampled across Canada using environmental correlation, where I estimated the variation in NR1D1 cTNR alleles explained by environmental and spatial variables after removing the effects of neutral population structure. Although most variation was explained by neutral structure, environment and spatial patterns in eastern lynx populations significantly explained some of the variation in NR1D1 alleles. To examine the role of island populations in the generation and distribution of adaptive genetic variation, I used 14 neutral microsatellites and a dinucleotide repeat within a gene linked to mammalian body size, IGF-1, and found that both genetic drift and natural selection influence the observed genetic diversity of insular lynx. Finally, I estimated the divergence dates of peripheral lynx populations and made recommendations towards the conservation of Canada lynx; high levels of genetic differentiation coupled with post-glacial colonization histories and patterns of divergence at cTNR loci suggest at least 4 ESUs for Canada lynx across their range. Author Keywords: adaptation, Canada lynx, candidate genes, coding trinucleotide repeats, evolution, natural selection
Tests of the Invasional Meltdown Hypothesis in invasive herbaceous plant species in southern Ontario
According to the Invasional Meltdown Hypothesis (IMH), invasive species may interact in their introduced range and facilitate future invasions. This study investigated the possibility that Alliaria petiolata, an invasive allelopathic herbaceous plant in Ontario, is facilitating invasions by additional alien species. Two allelopathic focal species were chosen for this study: the native Solidago canadensis and the invasive A. petiolata. Field surveys in southern Ontario that quantified plant biodiversity in plots that included one or both focal species revealed no support for the IMH, although fewer species co-existed with A. petiolata than with S. canadensis. A year-long recruitment experiment in Peterborough, Ontario, also produced results inconsistent with the IMH, although did provide some evidence that A. petiolata limited recruitment of other species. These results collectively show negative impacts on regional biodiversity by A. petiolata, even in the absence of an invasional meltdown. Author Keywords: allelopathy, Alliaria petiolata, co-occurrence surveys, invasional meltdown hypothesis, invasive species, Solidago canadensis
Testing for Interspecific Hybridization and a Latitudinal Cline Within the Clock Gene Per1 of the Deer Mouse (Peromyscus maniculatus) and the White-Footed Mouse (Peromyscus leucopus)
The recent northward expansion of the white-footed mouse (Peromyscus leucopus) in response to climatic changes provides a natural experiment to explore potential adaptive genetic variation within the clock gene Per1 in Peromyscus undergoing latitudinal shifts, as well as, the possibility of hybridization and introgression related to novel secondary contact with its sister species the deer mouse (Peromyscus maniculatus). Because clock genes influence the timing of behaviors critical for survival, variations in genotype may reflect an organism’s ability to persist in different environments. Hybridization followed by introgression may increase the adaptive potential of a species by quickly generating adaptive variation through novel genetic recombination or by the transfer of species-specific alleles that have evolved in response to certain environments. In chapter 2, I used microsatellite and mtDNA markers to test for hybridization and introgression between P. maniculatus and P. leucopus and found that interbreeding is occurring at a low frequency (<1%). In chapter 3, I tested for a latitudinal cline in a polyglycine repeat located within the Per1 gene of Peromyscus and discovered a putative cline in the Per1-142 and Per1-157 allele of P. leucopus and P. maniculatus, respectively. Chapter 4, further expands upon these findings, limitations, and the lack of evidence supporting introgression at the Per1 locus. Despite this lack of evidence, it is possible that novel hybridization has or could lead to adaptive introgression of other genes, allowing for the exchange of adaptive alleles or traits that could be advantageous for range expansion and adaption to future environmental changes. Author Keywords: Clock genes, Hybridization, Latitudinal gradient, Per1, Peromyscus, Range Expansion

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Graduate student theses produced at Trent University. You may also browse theses and dissertations by year, program, or author.

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