Graduate Theses & Dissertations

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Stress Axis Function and Regulation in New World Flying Squirrels
Across vertebrate taxa, the hypothalamic-pituitary-adrenal axis (or the stress axis) is highly conserved, and is central to vertebrate survival because it allows appropriate responses to psychological stressors. Habitat shapes successful physiological and ecological strategies, and to appreciate how individual species respond to stressors in their environment, it is essential to have a thorough knowledge of the basic stress physiology of each species. In this dissertation, I studied the functioning and evolution of the stress physiology of New World flying squirrels. I showed that baseline, circulating cortisol levels in northern (Glaucomys sabrinus) and southern (G. volans) flying squirrels are some of the highest ever reported for mammals, indicating that their stress axes operate at a higher set point than most other species. I also assessed other aspects of their acute stress response, including free fatty acid and blood glucose levels, and indices of immune function, and showed that the flying squirrels’ physiological reaction to stressors may differ from that of other mammals. Using immunoblotting, I found that corticosteroid-binding globulin (CBG) expression levels in flying squirrels appeared to be higher than previously reported using alternative methods. I also concluded however, that these levels did not appear to be high enough to provide their tissues with the protective CBG-bound buffer from their high circulating cortisol concentrations experienced by the majority of vertebrates. Thus, this arm of cortisol regulation within the flying squirrel stress axes may be weak or non-existent. Following this, I focused on southern flying squirrels and showed evidence that the second arm of cortisol regulation — the negative feedback mechanism at the level of the brain — functions effectively, but that this species is glucocorticoid resistant. Their tissue receptors appear to have a reduced affinity for cortisol, and this affinity may change seasonally to allow for the onset of other biological processes required for survival and reproduction. Due to their distinctive stress physiology, northern and southern flying squirrels may provide comparative physiologists with model systems for further probing of the function and evolution of the stress axis among vertebrates. Author Keywords: corticosteroid-binding globulin, flying squirrel, Glaucomys, glucocorticoids, physiological ecology, stress physiology
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
Study of Aerosols for use in Water Remediation of Pharmaceutical Pollutants
In this thesis, aerosolization was studied as a possible means of water remediation for several environmentally relevant pharmaceutical pollutants, known for their persistence in wastewater effluent and potable water sources. Seven different pharmaceutical compounds and a well-known plasticizer were all shown to decrease considerably in concentration in aerosol that was produced and subsequently collected within a short time span. Strong evidence is presented that an enhanced rate of partitioning into the gas phase at the air-water interface of water droplets exists for every compound tested relative to that occurring in bulk solution. UV photolysis in aerosols was also explored and shown for sulfamethoxazole to be at least an order of magnitude faster in aerosols then in bulk solution. The implications towards both the environmental fate, and removal of these compounds from water sources is discussed. Author Keywords: Aerosols, Air-water partitioning, Pharmaceuticals, Photolysis, Sulfamethoxazole
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
Syrphidae (Diptera) of northern Ontario and Akimiski Island, Nunavut
Syrphids, also known as hover flies (Diptera: Syrphidae) are a diverse and widespread family of flies. Here, I report on their distributions from a previously understudied region, the far north of Ontario, as well as Akimiski Island, Nunavut. I used samples collected through a variety of projects to update known range and provincial records for over a hundred species, bringing into clearer focus the distribution of syrphids throughout this region. I also analysed a previously un-tested trap type for collecting syrphids (Nzi trap), and report on results of DNA analysis for a handful of individuals, which yielded a potential new species. Author Keywords: Diptera, Ontario, range extension, Syrphidae
THE EFFECTS OF ROTATIONAL GRAZING AND HAY MANAGEMENT ON THE REPRODUCTIVE SUCCESS OF BOBOLINK AND EASTERN MEADOWLARK IN EASTERN ONTARIO
I investigated the impact of beef-cattle farm management on the reproductive success of Bobolink (Dolichonyx oryzivorus) and Eastern Meadowlark (Sturnella magna) within Eastern Ontario. I monitored rotational grazing management regimes and hay cut dates while assessing breeding phenology and reproductive success of Bobolinks and Eastern Meadowlarks. In pasture paddocks the major factor determining Bobolink reproductive success was the date that cattle entered a paddock to graze, with earlier entries resulting in lower reproductive success. On a landscape scale, within a series of paddocks grazed by a single herd, as the number of paddocks grazed during the nesting season increased, the number of Bobolinks that reproduced successfully decreased. Experimental quantification of trampling showed that cattle exposure to clay pigeon targets, regardless of stocking rates, resulted in the majority of targets being trampled. In hayfields associated with beef- cattle operations, grassland birds had a higher likelihood of success when cutting occurred after 4 July. The best method to improve the reproductive success of Bobolinks and Eastern Meadowlarks is to leave some hayfields and pasture paddocks undisturbed until nesting is complete. Author Keywords: Bobolink, Dolichonyx oryzivorus, farm management, hayfield, pasture, rotational grazing
THE LIFE HISTORY STRATEGY, GROWTH, BODY CONDITION, AND DIET OF STOCKED AMERICAN EEL (Anguilla rostrata) IN THE UPPER ST. LAWRENCE RIVER AND LAKE ONTARIO
My study was primarily focused on the comparison of life history traits between stocked American eel and their naturally recruited conspecifics in Lake Ontario and the upper St. Lawrence River (USLR/LO). I found that stocked eels experienced faster annual growth than their naturally recruited conspecifics and were comprised of a greater proportion of males. These findings indicate that the life history patterns of American eel may be genetically predisposed. Additionally, my study served to characterize the diets of stocked American eel and examine possible associations between eel and prey size. The eels consumed a number of macroinvertebrate prey orders as well as fishes and macrocrustaceans, with the latter prey items being more prevalent in the diets of larger eel specimens. A disparity in eel growth and body condition was observed between two primary stocking locations and were likely attributable to differences in available forage and habitat. Lastly, growth, body condition, and stocked eel diet were compared between lentic and lotic habitats. Eels from lotic streams experienced slower annual growth and had reduced body condition, and their diets were comprised of smaller prey items. The results of this study suggest that the current stocking methods employed in the USLR/LO are not suitable to restore the natural recruitment of individuals that will exhibit desired life history traits. Author Keywords:
Tabanidae and Culicidae in the Northern Boreal Region of Ontario
I studied the abundance, distribution and diversity of horse fly and deer fly species (Diptera: Tabanidae) and mosquito species (Diptera: Culicidae) in the boreal forest region of northern Ontario in 2011 and 2012. I collected 19 mosquito species, including one species new for Ontario, Aedes pullatus (Coquillett). I documented 11 northern and one southern range extension. I also collected a total of 30 species of horse and deer flies, including one new species of horse fly for Ontario, Hybomitra osburni (Hine). Results were inconsistent with a hypothesis of colonization of dipteran species from west to east. I examined the trapping biases of Malaise and sweep sampling for horse and deer flies and found that Malaise traps collected fewer individuals than sweep netting (850 versus 1318) but more species (28 versus 22). Consequently, I determined that surveys of diversity benefit from the use of multiple trapping methods. I also examined how blood-feeding (anautogeny) requirements affect the distribution patterns of Tabanidae. Ultimately, there are likely multiple factors that affect the expression of anautogeny in Tabanidae. Author Keywords: Autogeny, Culicidae, Diversity, Hudson Bay Lowlands, Northern Ontario, Tabanidae
Temperature effects on the routine metabolic rates of brook trout (Salvelinus fontinalis) eggs, alevin and fry
Early developmental stages of cold-adapted ectotherms such as brook trout (Salvelinus fontinalis) are at risk of mortality with increasing water temperatures because of their sensitivity to changes in their environment. I studied the mass and routine metabolic rate (RMR) of wild-origin brook trout eggs, alevin and young fry reared at normal (5°C) and elevated (9°C) temperatures for the duration of the study or at mismatched temperatures. This setup determined if preconditioning acclimation for one temperature benefits or hinders the organism later in life. Three levels of biological organization (ancestry, population, family) were studied using Akaike’s Information Criterion (AIC) to identify models that best accounted for variation in the data. Family, mass and temperature were most important in predicting body mass and mass-adjusted RMR, although population and ancestral-level differences were also detected at some life stages. Strong variation in body mass and mass-adjusted RMR among families may indicate adaptive potential within brook trout populations to respond to increases in water temperature with climate change. Author Keywords: Acclimation, AIC, Brook trout (Salvelinus fontinalis), Environmental matching, Routine metabolic rate, Temperature
Temporal Variability of Coloured Dissolved Organic Matter in the Canada Basin, Arctic Ocean (2007-2017)
This thesis investigated coloured and fluorescent dissolved organic matter in the Canada Basin, Arctic Ocean from 2007 to 2017. The first interannual time-series of its kind in the Canada Basin incorporated the use of EEM-PARAFAC to validate a seven-component model. Statistical temporal tests revealed (1) an increasing protein-like intensity in the upper polar mixed layer (UPML); (2) increasing intensities of humic-like components in the halocline due to increasing freshwater content; and (3) no change in DOM composition in deeper Atlantic waters (AW) congruent with the long residence time of the water mass (> 30 years). The significant decline in sea ice concentration was related to a decrease in humic-like FDOM due to enhanced photodegradation and an increase in protein-like FDOM, likely the results of increased biological activities in surface layers. This research provides evidence that the changes in physical and biological environment in the Arctic regions have already profound impacts on the composition and distribution of FDOM. Author Keywords: absorbance, Arctic Ocean, dissolved organic matter, fluorescence, parallel factor analysis, time-series
Temporal variation of dissolved organic matter and diffusive gradient in thin films-labile mercury in the Quesnel river, BC, and the Goose Creek tributary of Churchill river, MB
This study examined dissolved organic matter (DOM) and labile Mercury (from diffusive gradient in thin films (DGT)) in the Quesnel river, British and the Goose creek tributary of the Churchill river, Manitoba. DOM properties were measured with optical measurements of absorption coefficient (a254), spectral slopes (S275-295, SR) and fluorescence indices (HIX, BIX, FI). The DOC proxy measurements (a254) were almost 10 times higher at the Churchill site (Mean a254 116.77 cm-1) compared with the Quesnel river site (Mean a254 12.06 cm-1) during the study periods. While DGT labile Hg concentrations at the Quesnel site (2.17 to 98.97ppt) were almost 10 times more than the levels reported at the Churchill site (0.03 to 9.06 ppt). Fluorescence indices and the rise of labile Hg concentrations in spring indicated mostly terrestrial sources of DOM at both the sites. Spectral slopes and fluorescence indices substantiated that Churchill site had high molecular weight, complex and more humic DOM compared with Quesnel. DOM at both the sites was prone to temporal variation and affected by environmental conditions. Correlation between DGT labile-Hg and DOM parameters suggested that DGT collected Hg-organic complexes along with inorganic labile-Hg complexes. Author Keywords: Churchill, Diffusive gradient in thin films, Dissolved organic matter, Labile Hg, Mercury, Quesnel
Temporo-spatial patterns of occupation and density by an invasive fish in streams
Since its introduction to North America in the 1990s, the Round Goby has spread throughout the Great Lakes, inland through rivers and is now moving into small tributary streams, a new environment for this species in both its native and invaded ranges. I explored density and temporal occupation of Round Gobies in four small streams in two systems in south-central Ontario, Canada in order to determine what habitat variables are the best predictors of goby density. Two streams are tributaries of Lake Ontario and two are tributaries of the Otonabee River, and all of these streams have barriers preventing upstream migration. I found that occupation and density differed between the systems. In the Otonabee River system, Round Gobies occupy the streams year round and the most important factor determining adult density is distance from a barrier to upstream movement, with the entire stream occupied but density highest next to the barriers. In the Lake Ontario system, density is highest at mid-stream and Round Gobies appear to occupy these streams mainly from spring to fall. Adult density in Lake Ontario tributaries is highest in sites with a high percentage of cobble/boulder and low percentage of gravel substrate, while substrate is less important in Otonabee River tributaries. Occupation and density patterns may differ due to contrasting environmental conditions in the source environments and distance to the first barrier preventing upstream movement. This study shows diversity in invasion strategies, and provides insight into the occurrence and movement patterns of this species in small, tributary streams. Author Keywords: biological invasion, Generalised Additive Mixed Model, habitat, Neogobius melanostomus, Round Goby, stream

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