Graduate Theses & Dissertations

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Population Genetics and Gut Microbiome Composition Reveal Subdivisions and Space Use in a Generalist and Specialist Ungulate
Natural populations are often difficult and costly to study, due to the plethora of confounding processes and variables present. This is of particular importance when dealing with managed species. Ungulates, for example, act as both consumers and prey sources; they also provide economic benefit through harvest, and as such, are of high ecological and economic value. I addressed conservation and management concerns by quantifying subdivision in wild populations and combined movement with non-invasive sampling to provide novel insight on the physiological drivers of space use in multiple species. This thesis explored biological patterns in ungulates using two distinct approaches: the first used molecular genetics to quantify gene flow, while the second examined the relationship between movement and the gut microbiome using high-throughput sequencing and GPS tracking. The goal of the first chapter was to quantify gene flow and assess the population structure of mountain goats (Oreamnos americanus) in northern British Columbia (BC) to inform management. I used microsatellites to generate genotype data and used a landscape genetics framework to evaluate the possible drivers behind genetic differentiation. The same analyses were performed at both a broad and fine scale, assessing genetic differentiation between populations in all of northern BC and in a case management study area northeast of Smithers BC. The results indicated panmixia among mountain goats regardless of scale, suggesting distance and landscape resistance were minimally inhibiting gene flow. Therefore, management at local scales can continue with little need for genetically informed boundaries, but regulations should be tailored to specific regions incorporating data on local access and harvest pressure. My second chapter aimed to determine the extent to which the gut microbiome drives space-use patterns in a specialist (mountain goat) and generalist (white-tailed deer, Odocoileus virginianus) ungulate. Using fecal samples, we generated genomic data using 16S rRNA high-throughput sequencing to evaluate gut diversity and gut microbiome characteristics. Additionally, individuals were fitted with GPS collars so that we could gain insight into movement patterns. Gut microbiome metrics were stronger predictors of space use and movement patterns with respect to home range size, whereas they were weaker predictors of habitat use. Notably, factors of both the gut microbiome and age of a given species were correlated with changes in space use and habitat use. Ultimately, this research linked high-throughput sequencing and GPS data to better understand ecological processes in wild ungulates. Author Keywords: gene flow, genomics, gut microbiome, home range, population genetic structure, ungulates
Assessing Brook Trout (Salvelinus fontinalis) Seasonal Occupancy in Haliburton County, ON Using Environmental DNA
Brook trout (Salvelinus fontinalis) are declining across Ontario in both numbers and distribution, prompting concern for their future. Here, conventional, emerging, and predictive tools were combined to document brook trout occupation across seasons using streams in Haliburton County, ON as model systems. By using the Ontario Ministry of Natural Resources and Forestry’s (OMNRFs) Aquatic Ecosystem Classification (AEC) system variables with environmental DNA (eDNA) sampling and backpack electrofishing, my research supports the development of species occupancy models (SOMs) and eDNA as tools to document brook trout occurrence. To do this, eDNA sampling was validated in Canadian Shield stream environments by comparison with single-pass backpack electrofishing before seasonally sampling two river systems across their main channel and tributaries to assess occupancy. Streams were classified as potential high, moderate, and low-quality brook trout habitats using indicator variables within the AEC and sampled seasonally with eDNA to quantify occupancy and relate it to habitat potential at the county scale. Results showed eDNA to be an effective tool for monitoring fish across Canadian Shield landscapes and that brook trout occupancy varied seasonally within and across watersheds, suggesting that habitat and fish management strategies need to consider seasonal movement and spatial connectivity. Using these tools will enable biologists to efficiently predict and document brook trout occurrences and habitat use across the landscape. Author Keywords: Aquatic Ecosystem Classification, brook trout, Canadian Shield, connectivity, environmental DNA, seasonal occupation
Automated Separation and Preconcentration of Ultra-Trace Levels of Radionuclides in Complex Matrices by Online Ion Exchange Chromatography Coupled with Inductively Coupled Plasma Mass Spectrometry (ICP-MS)
Radionuclides occur in the environment both naturally and artificially. Along with weapons testing and nuclear reactor operations, activities such as mining, fuel fabrication and fuel reprocessing are also major contributors to nuclear waste in the environment. In terms of nuclear safety, the concentration of radionuclides in nuclear waste must be monitored and reported before storage and/or discharge. Similarly, radionuclide waste from mining activities also contains radionuclides that need to be monitored. In addition, a knowledge of ongoing radionuclide concentrations is often required under certain ‘special’ conditions, for example in the area surrounding nuclear and mining operations, or when nuclear and other accidents occur. Thus, there is a huge demand for new methods that are suitable for continuously monitoring and rapidly analyzing radionuclide levels, especially in emergency situations. In this study, new automated analytical methods were successfully developed to measure ultra trace levels of single or multiple radionuclides in various environmental samples with the goal of faster analysis times and less analyst involvement while achieving detection limits suitable for typical environmental concentrations. Author Keywords: automation, ICP-MS, ion exchange, radionuclide
Enhanced weathering and carbonation of kimberlite residues from South African diamond mines
Mafic and ultramafic mine wastes have the potential to sequester atmospheric carbon dioxide (CO2) through enhanced weathering and CO2 mineralization. In this study, kimberlite residues from South African diamond mines were investigated to understand how weathering of these wastes leads to the formation of secondary carbonate minerals, a stable sink for CO2. Residues from Venetia Diamond Mine were fine-grained with high surface areas, and contained major abundances of lizardite, diopside, and clinochlore providing a maximum CO2 sequestration capacity of 3–6% of the mines emissions. Experiments utilized flux chambers to measure CO2 drawdown within residues and unweathered kimberlite exhibited greater negative fluxes (-790 g CO2/m2/year) compared to residues previously exposed to process waters (-190 g CO2/m2/year). Long-term weathering of kimberlite residues was explored using automated wet-dry cycles (4/day) over one year. Increases in the δ13C and δ18O values of carbonate minerals and unchanged amount of inorganic carbon indicate CO2 cycling as opposed to a net increase in carbon. Kimberlite collected at Voorspoed Diamond Mine contained twice as much carbonate in yellow ground (weathered) compared to blue ground, demonstrating the ability of kimberlite to store CO2 through prolonged weathering. This research is contributing towards the utilization of kimberlite residues and waste rock for CO2 sequestration. Author Keywords: CO2 fluxes, CO2 mineralization, CO2 sequestration, Enhanced weathering, Kimberlite, Passive carbonation
Fungal pathogen emergence
The emergence of fungal hybrid pathogens threatens sustainable crop production worldwide. To investigate hybridization, the related smut fungi, Ustilago maydis and Sporisorium reilianum, were selected because they infect a common host (Zea mays), can hybridize, and tools are available for their analysis. Hybrid dikaryons exhibited filamentous growth on plates but reduced virulence and limited colonization in Z. mays. Select virulence genes in the hybrid had similar transcript levels on plates and altered levels during infection of Z. mays relative to each parental dikaryon. Virulence genes were constitutively expressed in the hybrid to determine if its pathogenic development could be influenced. Little impact was observed in hybrids with increased expression of effectors known to modify host response and metabolism. However, increased expression of transcriptional regulators of stage specific pathogenic development increased the hybrid’s capacity to induce symptoms. These results establish a base for investigating molecular aspects of fungal hybrid pathogen emergence. Author Keywords: effectors, hybrid pathogenesis assays, Sporisorium reilianum, transcription factors, Ustilago maydis, virulence factors
Surface temperature regulation during stress exposure
The influence of stress exposure on the body temperature of vertebrates has been known for nearly two-thousand years. While the proximate mechanisms supporting this phenomenon are well described, the ultimate mechanisms remain enigmatic. In this thesis, I propose a novel hypothesis which states that changes in body surface temperature (henceforth "surface temperature") following stress exposure occur to reduce energetic expenditure toward thermoregulation, thus freeing energy for use in the stress response (hereafter, the "Thermoprotective Hypothesis"). Using a paired experimental design, I first show that black-capped chickadees (Poecile atricapillus, Linnaeus, 1766) exposed to repeated stressors decrease their surface temperatures at low ambient temperatures, and increase their surface temperatures at high ambient temperatures relative to unstressed controls. These changes in surface temperature contribute to a relative reduction in heat loss in the cold, and a relative increase in heat dissipation in the warmth among stress-exposed individuals, thus reducing their energetic demands toward more costly thermoregulatory strategies. Next, I show that stress-induced changes in surface temperature are most pronounced in chickadees that experience naturally-occurring resource-restrictions, suggesting that this response occurs to balance allocation of energy among the stress response and thermoregulation (i.e. a true energetic trade-off ). Third, I show that the magnitudes of chronic, stress-induced changes in surface temperature are highly variable among, and highly consistent within, chickadees, therefore suggesting that this response could hold adaptive significance if such variation among individuals is heritable. Finally, using domestic pigeons (Columba livia domestica, Gmelin, 1789) as a model species, I show that stress-induced changes in surface temperature are highly pronounced at bare tissues with a critical role in thermoregulation for some avian species (the bill), when compared with responses at surrounding bare tissues (the eye region). Together, these findings strongly support the Thermoprotective Hypothesis and suggest that endotherms may trade energetic investment toward thermoregulation with that toward the stress response in resource-limiting environments. Author Keywords: heat transfer, infrared thermography, stress physiology, surface temperature, thermoregulation, trade-off
influence of tree species litterfall on soil chemistry and implications for modelling soil recovery from acidification
Decades of acidic deposition have depleted base cation pools in soil over large parts of eastern north America, including the Muskoka-Haliburton region of central Ontario. This region has also experienced a shift in forest species composition over the past 200 years, favouring sugar maple (Acer saccharum Marsh.) at the expense of species such as white pine (Pinus strobus L.) and eastern hemlock (Tsuga canadensis (L.) Carr.). This shift in species composition may have changed soil chemistry over time due to differences in nutrient and metal inputs in litterfall. An analysis of litterfall and soil chemistry was conducted for five tree species commonly found across central Ontario. Stands were established in the Haliburton Forest & Wild Life Reserve and were dominated by one of balsam fir (Abies balsamea (L.) Mill.), eastern hemlock, white pine, sugar maple, or yellow birch (Betula alleghaniensis Britt.). Analysis of mineral soil oxides suggested that these stands were established on similar parent material. Deciduous dominated stands (maple and birch) had greater litterfall mass compared with conifer stands (fir, hemlock, and pine), generally leading to greater macronutrient inputs to the soil. Elemental cycling through the organic horizons was more rapid in deciduous stands, with base cations having the shortest residence times. This suggests that a change from greater conifer dominance to mixed hardwood forests may lead to more rapid elemental cycling and alter the distribution of elements in soil. Forests in the region are typically mixed and the resulting differences in soil chemistry may influence model predictions of soil recovery from acidification. Laboratory leaching tests indicated that both stand type and the acidity of simulated rainwater inputs influenced soil solution chemistry, with deciduous stands generally having a greater buffering capacity than sites dominated by coniferous species. Changes in soil chemistry were examined for each stand type using the Very Simple Dynamic (VSD) biogeochemical model. Simulations showed that soil base saturation began to increase following lows reached around the year 2000, and similar patterns were observed for all stands. When sulphur (S) and nitrogen (N) deposition were held constant at present rates, soil base saturation recovery (toward pre-1900 levels) was marginal by 2100. With additional deposition reductions, further increases in base saturation were minor at all sites. In conjunction with additional deposition reductions, the elimination of future forest harvesting allowed for the greatest potential for recovery in all stands. Overall, these results suggest that changes in forest cover may influence soil chemistry over time, most notably in the organic soil horizons. However, forecasted recovery from acidification is expected to follow similar patterns among stands, since differences in soil chemistry were less significant in the mineral soil horizons which compose a greater proportion of the soil profile. Author Keywords: base cation decline, forest harvesting, litterfall, mineral weathering, soil acidification, VSD model
Short photoperiod lowers visceral adiposity and shifts the molecular clock in Peromyscus leucopus
The length of photoperiod can alter circadian rhythms and change fat depot mass whencombined with environmental temperatures below thermoneutral. To isolate photoperiod effects, we compared the effects of long and short photoperiod exposure at thermoneutrality in photoperiod sensitive, F1 generation adult male white-footed mice (P. leucopus). Mice were housed in long-day or short-day photoperiod conditions at thermoneutrality for 4 weeks. Short photoperiod decreased vWAT mass without changing body weight, scWAT or iBAT mass, or calorie consumption. Short photoperiod increased Adrβ3 and Lpl mRNA expression in vWAT with no change in Ucp1, Pgc1a or Hsl. vWAT Per1, Per2 and Nr1d1 mRNA expression were aligned to the onset of dark and food intake, while Bmal1 and Clock were misaligned. These findings suggest that short photoperiod per se can decrease visceral fat accumulation, without activating thermogenesis, reinforcing that environmental photoperiod should be considered when researching cause and prevention of obesity. Author Keywords: adiposity, circadian rhythm, clock genes, obesity, Peromyscus, photoperiod
Differential expression of cytochrome b5s in Giardia intestinalis during nitrosative stress and encystation
The waterborne protozoan Giardia intestinalis cycles between the environmentally-resistant and infectious cyst and the metabolically-active trophozoite that adheres to the epithelial lining of the small intestine. Adhesion can trigger the innate immune response in epithelial cells, including the synthesis of the free radical nitric oxide (NO) that inhibits cell proliferation and encystation of trophozoites. In this work changes in protein expression of three Giardia isotypes of the redox heme protein cytochrome b5 (gCYTb5 I, II and III) were studied in response to either nitrosative stress or induction of encystation. Two nitrosative stressors, sodium nitrite and the NO donor DETA-NONOate, were used at sub-lethal concentrations (0.5 mM and 0.05 mM, respectively) that do not affect cell proliferation until later time points so that subtle changes in protein expression could be observed in the absence of other confounding factors. Nucleolar gCYTb5-I and nucleoplasmic gCYTb5-III expression patterns were similar in trophozoites exposed to either stressor, showing gradual increases in expression with peaks between 4 and 12 hours, which indicates these cytochromes respond to nitrosative stress and possibly to potential DNA damage in Giardia. In contrast, gCYTb5-II of the peripheral vacuoles, which are part of the endocytic pathway of Giardia, showed little change in expression in response to either stressor. However, changes in gCYTb5-II expression were observed in encysting trophozoites, with a 1.4-fold increase in protein levels at seven hours after induction of encystation, followed by a gradual decrease in expression. These changes are consistent with previous mRNA analysis done in our laboratory and suggest a role for gCYTb5-II in the increase in nutrient uptake during early encystation. Author Keywords: cytochrome, encystation, Giardia, heme, nitrosative, parasite
Assessment of Potential Threats to Eastern Flowering Dogwood (Cornus florida) in Southern Ontario
In Canada, eastern flowering dogwood (Cornus florida L.) is an endangered tree that occurs only in the Carolinian forest of southern Ontario. Threats to this species include habitat fragmentation and the fungal pathogen dogwood anthracnose (Discula destructiva). I conducted a population genetic analysis using seven nuclear microsatellite markers to determine if fragmented populations are genetically isolated from one another and have low levels of genetic diversity. Genetic comparisons suggest on-going dispersal among sites and relatively high genetic diversity within most sites; however, smaller populations and younger trees were less genetically diverse. I also used linear mixed effects models to assess potential relationships between several ecological variables and the prevalence of dogwood anthracnose. Disease severity was higher in trees on shallow slopes and in larger trees; the latter also had higher likelihood of infection. Insights from this study will be important to incorporate into future management strategies. Author Keywords: Cornus florida, Discula destructiva, dogwood anthracnose, Eastern flowering dogwood, endangered, population genetics
Ecosystem Response to Above Canopy Nitrogen Addition in a Jack Pine (Pinus banksiana) Forest in the Athabasca Bituminous Sands Region of Northeastern Alberta, Canada
In this study we seek to better understand the potential effects of short-term (5-year) N fertilization on jack pine forest biogeochemistry, vascular plant community composition and to project a temporal endpoint of nitrogen leaching below the major rooting zone. Aqueous ammonium nitrate (NH4NO3) was applied above the forest canopy across five treatment plots (20 x 80 m) four times annually. The experimental deposition gradient followed those known for localized areas around the major open pit operations at 0, 5, 10, 15, 20 and 25 kg N ha-1 yr-1 over a five-year period (2011 – 2015). Nitrate recovery in throughfall was significantly higher than NH4+ (p < 0.05), indicating canopy NH4+ immobilization. There was a strong treatment effect (p < 0.05) of N on the epiphytic lichen thalli concentrations of Hypogymnia physodes and Evernia mesomorpha after five years. The canopy appeared to approach saturation at the highest deposition load (25 kg N ha-1 yr-1) during the fifth year of N additions as most N added above the canopy was accounted for in throughfall and stemflow. The non-vascular (lichen and moss) vegetation pool above the forest floor was the largest receptor of N as cryptogam foliar and thalli N concentrations showed a significant treatment effect (p < 0.05). Nitrogen in decomposing litter (25 kg N ha-1 yr-1) remained immobilized after five years, while treatments ≤ 20 kg N ha-1 yr-1 started to mobilize. Understory vascular plant cover expansion was muted when deposition was ≥ 10 kg N ha-1 yr-1. Finally, modeling suggests the forest may not leach N below the rooting zone until around 50 years after chronic addition begin (25 kg N ha-1 yr-1). The modeling results are consistent with empirical data from a high exposure (~20 - 25 kg N ha-1 yr-1) jack pine site approximately 12 km west of the experimental site that has not yet experienced N leaching. Author Keywords: Biogeochemistry, Canopy, Deposition, Jack Pine, Nitrogen, Understory
Phosphorus delivery in the Rainy-River Lake of the Woods Watershed
Lake of the Woods (LOW) is a large international waterbody which suffers from frequent and widespread algae blooms. Previous studies have highlighted the importance of the lake's largest tributary, the Rainy River (RR) and its significance in total phosphorus (TP) delivery to the LOW. Unfortunately, little is known about TP contributions from the RR and its tributaries within the Canadian portion of the watershed. This thesis examines patterns and sources of TP from four tributaries on the Canadian side of the lower RR region, two of which are predominantly natural, and two that are predominantly agricultural. Relationships between water quality parameters, land use and geologic characteristics were observed over a complete hydrologic year (Oct 1, 2018 - Sept 31, 2019), and through an intensive sampling campaign using a nested watershed approach during the spring high flow and summer low flow periods. Results revealed that TP and total suspended sediment (TSS) concentrations (>100 µg/L and >20 mg/L respectively), and loads (>20 kg/km2 and >3500 kg/km2, respectively), were greater at agricultural sites compared with natural sites (<65 µg/L TP and <15 mg/L TSS concentration, and <20 kg/km2 TP and <4000 kg/km2 TSS export). Total P, TSS, Fe, and Al were significantly positively correlated (R2= 0.26-0.59; p<0.05) and intensive sampling revealed that these relationships were strongest during the spring and at the agricultural sites (R2= 0.73-0.98; p<0.05). In contrast, the summer intensive sampling revealed that TP and redox sensitive Fe were significantly correlated (R2= 0.72; p<0.005), whereas redox insensitive Al and TSS were not, suggesting TP may be sourced via redox processes in the summer due to favourable hydrologic conditions. This was observed not only at sites with high wetland influence, but also at sites with more agricultural presence suggesting that redox sourced TP may also originate from mineral stream bed sediment during low flow periods. This research suggested two primary TP sources in the lower RR region: erosion in the spring, and redox processes (internal release) in the summer. It is recommended that intensive monitoring continue in Canada, and further research be conducted to fully understand the significance of internal P release in the tributaries. Author Keywords: erosion, land use, nutrients, particulates, redox, water quality

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