Graduate Theses & Dissertations

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Effects of Opioids on the Development and Reproductive Capacity of Japanese Medaka, Oryzias latipes
Opioid drugs are among the microcontaminants that are discharged with domestic wastewater into the aquatic environment with the potential to affect the development and reproductive capacity of aquatic organisms. To study the effects of exposure of fish to opioid drugs, Japanese medaka (Oryzias latipes) were exposed over a full life cycle to environmentally relevant (i.e. ng/L) concentrations of two opioids, codeine and fentanyl. Acute exposure of early life stage (ELS) of medaka to codeine resulted in slightly extended times to hatch, but no increase in embryo mortalities or reduced hatching success. Chronic exposure to codeine at a nominal concentration of 25,000 ng/L over a full life cycle interfered with growth, reducing the weight of the fish at maturity. Life cycle treatments with codeine at all test concentrations resulted in a significant reduction in the number of eggs produced in reproductive trials, as well as a reduction in the numbers of mature oocytes in adult females. High treatments with codeine also increased the percentage of immature sperm cells in adult males. Life cycle exposures to codeine also resulted in reduced whole-body concentrations of several hypothalamic-pituitary-gonadal (HPG) axis hormones, including reduced levels of luteinizing hormone in male and female fish and a reduction in 11-ketotestosterone in males and 17β-estradiol in females. Fentanyl did not affect reproduction or HPG hormones at the exposure concentrations tested, but high fentanyl exposures increased the mortality rate among ELS’s of the F1 generation offspring from life cycle exposed adults. The results of this thesis contribute to the literature on the environmental impacts of microcontaminants of wastewater origin and, the potential for effects in fish exposed to opioids.Keywords: Opioids, microcontaminants, fentanyl, codeine, sexual development, reproduction. Author Keywords: codeine, fentanyl, microcontaminants, opioids, reproduction, sexual development
Effects of Recycled Media on Culture Growth and Hormone Profiles in Heterotrophic Euglena gracilis
The rapid expansion of the worldwide population has caused an urgent need for the development of new, more environment-conscious, food sources. In this context, algae, such as Euglena, are of interest thanks to their capacity to naturally produce essential nutrients such as proteins and oils commonly found in animals and plant sources. While these processes are currently being investigated, underlying measures affecting growth of Euglena gracilis like hormonal influences and growth stress like nutrient deprivation are poorly understood. From this vantage point, this thesis seeks to understand the role of phytohormones cytokinin (CKs) and abscisic acid (ABA) in complex mechanisms underlying heterotrophic growth of Euglena gracilis under recycled, organic media conditions with no supplementation. Hormone profiles were quantified by HPLC-ESI-MS/MS and compared to culture growth dynamics of pH, weight accumulation, glucose content, cell count and morphology. It was expected that ABA acted as an inhibitory hormone and this was confirmed by its higher levels when CKs where low and vice versa. Contrastingly, it was expected that CKs stimulated growth, in which this was shown not to be the case. Interestingly, it was revealed that both hormone groups increase with increasing recycling. Other key findings include: E. gracilis synthesizes CKs via the tRNA-degradation pathway and is cZ and iP dominated, recycling E. gracilis medium is viable for growth, however, the percentage (25% or less) is crucial to cell viability and markedly no ABA was detected in E. gracilis pellet fractions from recycled media. Therefore, this data revealed that recycled media has a striking influence on physiological aspects of growth and illustrated unique changes in hormone profiles of which could be manipulated to help the food industry. Author Keywords: cytokinin, endogenous hormones, Euglena gracilis, heterotrophic, large scale microalgae cultivation, recycled medium
Effects of Silver Nanoparticles on Lake Bacterioplankton
Silver nanoparticles (AgNP) released into aquatic environments could threaten natural bacterial communities and ecosystem services they provide. We examined natural lake bacterioplankton communities' responses to different exposures (pulse vs chronic) and types (citrate and PVP) of AgNPs at realistic environmental conditions using a mesocosm study at the Experimental Lakes Area. An in situ bioassay examined interactions between AgNPs and phosphorus loading. Bacterial communities exposed to high AgNP concentrations regardless of exposure or capping agent type accumulated silver. We observed increases in community production during additions of polyvinylpyrrolidone (PVP) -capped AgNPs and that site and nutrient-specific conditions are important to AgNPs toxicology in aquatic systems. Toxicological effects of AgNP are attenuated in natural conditions and differ from results from laboratory studies of AgNP toxicity. Our results demonstrate more studies are needed to fully assess the risk posed by these novel chemicals to the environment. This work could be useful in forming risk assessment policies which are largely based on lab studies and typically demonstrate strong toxic effects. Author Keywords: bacterial production, bacterioplankton communities, ecological stoichiometry, Experimental Lakes Area, mesocosms, silver nanoparticles
Effects of Silver Nanoparticles on Lower Trophic Levels in Aquatic Ecosystems
Due to their effective antibacterial and antifungal properties, silver nanoparticles (AgNPs) have quickly become the most commonly used nanomaterial, with applications in industry, medicine and consumer products. This increased use of AgNPs over the past decade will inevitably result in an elevated release of nanoparticles into the environment, highlighting the importance of assessing the environmental impacts of these nanomaterials on aquatic ecosystems. Although numerous laboratory studies have already reported on the negative effects of AgNPs to freshwater organisms, only a handful of studies have investigated the impacts of environmentally relevant levels of AgNPs on whole communities under natural conditions. This thesis examines the effects of chronic AgNP exposure on natural freshwater littoral microcrustacean, benthic macroinvertebrate and pelagic zooplankton communities. To assess the responses of these communities to AgNPs, I focused on a solely field-based approach, combining a six-week mesocosm study with a three-year whole lake experiment at the IISD – Experimental Lakes Area (Ontario, Canada). Our mesocosm study tested the effects of AgNP concentration (low, medium and high dose), surface coating (citrate- and polyvinylpyrrolidone [PVP]-coated AgNPs), and type of exposure (chronic and pulsed addition) on benthic macroinvertebrates in fine and stony sediments. Relative abundances of metal-tolerant Chironomidae in fine sediments were highest in high dose PVP-AgNP treatments; however, no negative effects of AgNP exposure were seen on biodiversity metrics or overall community structure throughout the study. I observed similar results within the whole lake study that incorporated a long-term addition of low levels of AgNPs to an experimental lake. Mixed-effects models and multivariate methods revealed a decline in all species of the littoral microcrustacean family Chydoridae in the final year of the study within our experimental lake, suggesting that this taxon may be sensitive to AgNP exposure; however, these effects were fairly subtle and were not reflected in the overall composition of littoral communities. No other negative effects of AgNPs were observed on the pelagic zooplankton or benthic macroinvertebrate communities. My results demonstrate that environmentally relevant levels of AgNPs have little impact on natural freshwater microcrustacean and benthic macroinvertebrate communities. Instead, biodiversity metrics and community structure are primarily influenced by seasonal dynamics and nutrient concentrations across both lakes. This thesis highlights the importance of incorporating environmental conditions and the natural variability of communities when examining the potential risks posed by the release of AgNPs into the environment, as simplistic laboratory bioassays may not provide an adequate assessment of the long-term impacts of AgNPs on freshwater systems. Author Keywords: Benthic macroinvertebrates, IISD - Experimental Lakes Area, Littoral microcrustaceans, Silver nanoparticles, Whole lake experiment, Zooplankton
Effects of biodiversity and lake environment on the decomposition rates of aquatic macrophytes in the Kawartha Lakes, Ontario
Decomposition of aquatic macrophytes has an important role in defining lake carbon (C) storage and nutrient dynamics. To test how diversity impacts decomposition dynamics and site-quality effects, I first examined whether the decomposition rate of aquatic macrophytes varies with species richness. Generally, I found neutral effects of mixing, with initial stoichiometry of component species driving decomposition rates. Additionally, external lake conditions can also influence decomposition dynamics. Therefore, I assessed how the decomposition rate of a submersed macrophyte varies across a nutrient gradient in nine lakes. I found decomposition rates varied among lakes. Across all lakes, I found Myriophyllum decomposition rates and changes in stoichiometry to be related to both nutrients and water chemistry. During the incubation changes in detrital stoichiometry were related to lake P and decomposition rates. Aquatic plant community composition and stoichiometry could alter decomposition dynamics in moderately nutrient enriched lakes. Author Keywords: Aquatic Plants, Decomposition, Diversity, Littoral, Macrophytes, Nutrients
Effects of flooding on nutrient budgets and ecosystem services
Increases in flooding due to anthropogenic influences such as climate change and reservoir creation will undoubtedly impact aquatic ecosystems, affecting physical, chemical, and biological processes. We used two approaches to study these impacts: a whole-ecosystem reservoir flooding experiment and a systematic literature review. In the whole-ecosystem experiment, we analyzed the impact of flooding on nutrient release from stored organic matter in an upland forest. We found that flooded organic matter produced N (nitrogen) and P (phosphorus), but that more N was released relative to P, increasing the N:P ratio over time. In the systematic literature review, we linked small (<10 year recurrence interval) and extreme (>100 year recurrence interval) floods to changes in 10 aquatic ecosystem services. Generally, extreme floods negatively impacted aquatic ecosystem service provisioning, while small floods contributed positively. Overall, we found that flood impacts vary depending on ecosystem properties (organic matter content) and flood characteristics (magnitude). Author Keywords: ecosystem services, flooding, nutrients, reservoirs, rivers
Effects of hydrologic seasonality on dissolved organic matter composition, export, and biodegradability in two contrasting streams
Environmental and seasonal processes are important watershed drivers controlling the amount, composition, and fate of dissolved organic matter (DOM) in aquatic ecosystems. We used ten months of water samples and eight months of bioassay incubations from two contrasting catchments (agriculture and natural, forested) to assess the effects of seasonal variability on the composition, export, and biodegradability of DOM. As expected, the DOM composition and exports were more allochthonous-like and autochthonous-like in the forest and agriculture streams, respectively. However, we found no relationship between DOM composition and biodegradability in our study, suggesting that broad environmental factors play a large part in determining bioavailability of DOM. We found that both differences between the catchments and seasonal variability in hydrology and water temperature cause shifts in DOM composition that can affect exports and potentially affect its susceptibility to microbial activity. More research is needed to fully understand the impact of land use and temporal variability on bioavailability and delivery to downstream ecosystems. Author Keywords: Bioavailable dissolved organic carbon, Biodegradability, Dissolved organic matter, Export, Seasonality, Streams
Effects of road salt sodium on soil
While previous studies have focused on how road salt affects water quality and vegetation, limited research has characterized road salt distribution through soil and the resulting impacts. The potential for sodium (Na+) to be retained and impact soil physical and chemical properties is likely to vary depending on the soil’s parent material, and more specifically on the extent of base saturation on the cation exchange complex. This thesis contrasted Na+ retention, impacts, and mobility in roadside soils in two different parent materials within southern Ontario. Soils were sampled (pits and deep cores) during fall 2013 and spring 2014 from two sites along highways within base-poor, Precambrian Shield soil and base-rich soil, respectively. Batch experiments were subsequently performed to investigate the influence of parent material and the effect of co-applied Ca2+-enriched grit on the longevity of Na+ retention in soils. Less Na+ is adsorbed upon the co-application of Ca2+, suggesting grit has a protective effect on soil by increasing cation exchange competition. Positive correlations between Na+ and pH, and negative correlations between Na+ and soil organic matter, % clay and base cations within Shield soils suggest that they are more vulnerable to Na+ impacts than calcareous soils due to less cation exchange competition. However, Na+ is more readily released from calcareous roadside soils, suggesting there is greater potential for Na+ transfer to waterways in regions dominated by calcareous soils. Author Keywords: cation exchange, parent material, road salt, sodium retention, urban soil
Effects of wood ash addition on soil chemical properties and sugar maple (Acer saccharum, Marsh.) seedling growth in two northern hardwood forest sites in central Ontario
One possible solution to acidification and losses of base cations in central Ontario forest soils may be the application of wood ash. Wood ash is generally high in pH and contains large amounts of calcium (Ca) and other nutrients essential for ecosystem health, however it also contains trace metals. Understanding the chemistry of soils following ash application to forests is crucial for future policy recommendations and remediation efforts. In this study, soil and soil water chemistry was measured at two acidic forest sites in central Ontario. Sugar maple (Acer saccharum, Marsh.) seedling growth and chemistry, as well as understory vegetation composition, were also measured. At site one, plots (2 m x 2 m) were established with sugar maple, white pine (Pinus strobus L.) and yellow birch (Betula alleghaniensis Britt.) residential wood ash treatments and applied at rates of 0 and 6 Mg ha-1. The effects of residential wood ash on soil and understory vegetation were measured three- and 12-months following ash addition. At site two, plots (5 m x 5 m) were established with both fly and bottom industrial grade bark ash treatments of 0, 4 and 8 Mg ha-1 (n=4), and tension lysimeters were positioned in each plot at 30, 50, and 100 cm depths. The effects of industrial grade wood ash on soil, soil water and understory vegetation were measured four years following ash addition. Metal concentrations in the ashes were generally low but were higher in the fly ash and yellow birch ash types. At site one, significant increase in soil pH, and Ca and magnesium (Mg) concentrations were observed after three months, however changes varied by treatment. Some metal concentrations increased in the upper organic horizons, but metals were likely immobilized in the soil due to increases in soil pH, electrical conductivity (EC) and high organic matter content of the soil. After one year, changes to metal concentrations in soils could be seen in mineral horizons, and a few metals (aluminum (Al), zinc (Zn), copper (Cu), chromium (Cr), strontium (Sr)) increased in treatment plots. At site two, the effects of industrial-grade bark ashes on soil pH could still be seen after four years and soil water metal concentrations were not elevated relative to controls. Changes to understory vegetation composition following ash application were observed, but ash addition had no significant effect on sugar maple seedling growth (root:shoot ratio) and did not lead to significant increases in foliar metal concentrations. There were significant differences in root chemistry, suggesting metal translocation and uptake could be restricted. Mass balance estimates indicate that the organic horizon is a sink for all metals and simulated drought in this horizon led to a decrease in soil pH and increase in soil water metal concentration, but this occurred in all treatments including control. These results suggest that application of industrial and residential wood ash in moderate doses with trace metal concentrations below or near regulatory limits will increase soil pH and base cation concentrations, as well as increase seedling tissue nutrient concentrations in northern hardwood forest soils. However, depending on the parent material of the ash, increased metal availability can also occur. Author Keywords: Acer saccharum, calcium decline, forest soil amendment, Haliburton Forest and Wildlife Reserve, heavy metal, wood ash
Electrochemical Characterization of Giardia Intestinalis Cytochromes b5
Giardia intestinalis is a protozoan parasite that causes waterborne diarrheal disease in animals and humans. It is an unusual eukaryote as it lacks the capacity for heme biosynthesis; nonetheless it encodes heme proteins, including three cytochrome b5 isotypes (gCYTB5s) of similar size. Homology modelling of their structures predicts increased heme pocket polarity compared to mammalian isotypes, which would favour the oxidized state and lower their reduction potentials (E°’). This was confirmed by spectroelectrochemical experiments, which measured E°’ of -171 mV, -140 mV and -157 mV for gCYTB5-I, II, III respectively, compared to +7 mV for bovine microsomal cytochrome b5. To explore the influence of heme pocket polarity in more detail, five gCYTB5-I mutants in which polar residues were replaced by nonpolar residues at one of three positions were investigated. While these substitutions all increased the reduction potential, replacement of a conserved tyrosine residue at position-61 with phenylalanine had the most significant effect, raising E°’ by 106 mV. This tyrosine residue occurs in all gCYTB5s and is likely the greatest contributor to their low reduction potentials. Finally, complementary substitutions were made into a bovine microsomal cytochrome b5 triple mutant to lower its reduction potential. These not only lowered the E°’ by more than 140 mV but also weakened the interaction of heme with the protein. The lower reduction potentials of the gCYTB5s may indicate that these proteins have different roles from their more well-known mammalian counterparts. Author Keywords:
Elemental Variation in Daphnia
Environmental variation can affect consumer trait expression and alter ecological and evolutionary dynamics in natural populations. However, although dietary nutrient content can vary by an order of magnitude in natural ecosystems, intra-specific differences in consumer responses to food quality have not been thoroughly investigated. Therefore, the purpose of my dissertation was to examine the influence of dietary nutrition and other environmental factors on consumer phenotypic variation using the freshwater cladoceran Daphnia. I conducted a series of complementary laboratory and field studies where I examined the effects of dietary phosphorus (P) content and additional biological/environmental variables (multi-elemental limitation, genetic variation, and temperature) on daphnid life-history, biochemistry, body elemental composition, and population growth. In general, phenotypic expression within a species varied significantly in response to all experimental variables, but the relative influence of each was highly context dependent. In my first chapter, I found that dietary P content and environmental calcium (Ca) concentrations both altered Daphnia body Ca:P ratios and growth rates of individuals and affected intrinsic rates of increase at the population level. However, food quality appeared to have a much larger effect on trait expression, and body Ca:P ratios were highly sensitive to other forms of dietary nutrient limitation. Next, I documented significant quantitative genetic variation and phenotypic plasticity in daphnid P content, growth, and P use efficiency of field collected animals grown across dietary P gradients. Trait expression was also influenced by genotype X diet interactions suggesting that consumer responses to dietary nutrient limitation can be heritable and may be adaptive in different nutrient environments. Finally, I found that temperature appeared to override food quality effects and decouple P metabolism in natural Daphnia populations, but total biomass production was affected by both dietary P content and temperature, depending on the nutrient content of the lake. Overall, my dissertation shows that consumer responses to nutrient limitation can vary significantly within a species and that changes in trait expression may be modified by other environmental variables. These results should be incorporated into existing stoichiometric models and used to investigate the eco-evolutionary consequences of consumer phenotypic variation in response to nutritional stress. Author Keywords: ecological stoichiometry, evolution, life-history, nutrient limitation, nutrient metabolism, zooplankton
Enduring Attack
Numerous prey taxa employ defensive postures for protection against attack by predators. Defensive postures mitigate predation risk at various stages of the predator-prey sequence, including through crypsis, mimicry, thanatosis, aposematism, and deflection. In terrestrial salamanders, defensive postures may be aposematic, or deflect attacks away from vital body parts and towards the tail, however the extent to which these strategies act exclusively or synergistically remains poorly understood. Herein I demonstrate a novel approach to study the function of salamander defensive postures through experimental manipulation of predator response to antipredator behaviour in a natural field setting. I deployed 1600 clay salamander prey on Pelee Island, Ontario, manipulating prey size (small, large) and posture (resting, defensive) and documented attack rates across three predator types to further assess the effect of prey body size and predator type on antipredator efficacy. My research suggests that irrespective of prey body size, defensive posture does not function through aposematism, but rather acts to deflect predator attacks to the tail, which is commonly noxious and expendable in terrestrial salamanders. An intriguing possibility is that this behaviour facilitates taste-rejection by predators. Overall, my research should further contribute to our understanding of the importance and potential evolutionary significance of defensive posturing in Ambystoma salamanders, and more broadly, on the determinants of prey vulnerability to predation. I also briefly discuss the implications of my results to the conservation of Ambystoma populations on Pelee Island. Author Keywords: Anti-predator behaviour, Aposematism, Attack deflection, Predator avoidance, Small-mouthed salamander, Taste-rejection

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Format: 2021/10/17