Graduate Theses & Dissertations


Mfsd8 regulates growth and multicellular development in Dictyostelium discoideum
The neuronal ceroid lipofuscinoses (NCLs), commonly known as Batten disease, are a family of inherited neurodegenerative lysosomal storage disorders. CLN7 disease is a subtype of NCL that is caused by mutations in the MFSD8 gene. MFSD8 encodes a lysosomal transmembrane protein that is predicted to play a role in transporting small substrates across membranes. However, little is known about its role and substrate specificity. Previous work identified an ortholog of human MFSD8 in the social amoeba Dictyostelium discoideum and reported its localization to endocytic compartments. In this study, the effects of mfsd8 loss during Dictyostelium growth and multicellular development were further characterized. Dictyostelium mfsd8- cells displayed increased rates of proliferation and pinocytosis in liquid media. During growth, loss of mfsd8 altered lysosomal enzymatic activities and reduced the intracellular and extracellular levels of autocrine proliferation repressor A. mfsd8- cells grown on a lawn of bacteria formed plaques in a shorter period of time compared to WT cells, providing additional support for the enhanced growth of mfsd8- cells. Upon starvation, the aggregation of mfsd8- cells was delayed, and mfsd8- cells formed more mounds that were smaller in size, which may be attributed to the reduced cell-substrate adhesion and altered lysosomal enzymatic activities observed for mfsd8- cells. Following aggregation, tipped mound formation was delayed, however, loss of mfsd8 did not affect the timing of slug/finger and fruiting body formation. Additionally, slug migration was reduced in mfsd8- cells. These aberrant phenotypes, excluding fruiting body formation, were effectively or partially rescued when Mfsd8-GFP was introduced into mfsd8- cells. Overall, these results show that Mfsd8 plays a role in regulating growth and developmental processes in Dictyostelium via lysosomal-associated functions. Author Keywords: CLN7, Dictyostelium discoideum, Lysosomes, MFSD8, Neuronal Ceroid Lipofuscinoses
mycobiome and skin chemistry of bat wings in relation to white-nose syndrome
White-nose syndrome (WNS) is a skin disease of bats caused by the fungus Pseudogymnoascus destructans (Pd) that damages flight membranes during hibernation and can lead to death. The disease causes mortality of multiple bat species in eastern North America and is spreading into western North America. Future impacts of WNS on naïve bat populations are unknown. Variation in host susceptibility occurs among and within species, but mechanisms driving this variation are unclear. Multiple studies have characterized immunological responses to WNS, but skin physiology as a barrier to pathogens is understudied. The unique ability of Pd to actively penetrate the normal, intact skin of its mammalian host makes WNS an interesting study system to understand skin defenses. Aspects of the mammalian skin environment that can influence disease susceptibility include pH, sebaceous lipids, and microbiomes. I found skin mycobiomes of WNS-susceptible species had significantly lower alpha diversity and abundance compared to bat species resistant to Pd infection. Using these data, I predicted that most naïve bat species in western North America will be susceptible to WNS based on the low diversity of their skin mycobiomes. Some fungi isolated from bat wings inhibited Pd growth in vitro, but only under specific salinity and pH conditions, suggesting the microenvironment on wings can influence microbial interactions and potentially WNS-susceptibility. I measured the wing-skin pH of bats in eastern Canada and found that Eptesicus fuscus (WNS-tolerant) had more acidic skin than M. lucifugus (WNS-susceptible). Differences in sebum quantity and composition among and within mammalian species may help explain variation in skin disease susceptibility and the composition of skin microbiomes. This is due to the antimicrobial properties of sebum and the use of sebum as a nutrition source by microbes. Outcomes of this work further our understanding of inter- and intra-specific differences among bat species and individuals in skin mycobiomes and physiology, which may contribute to variation in WNS-susceptibility. Future research should focus on characterizing the physical and chemical landscape of skin as this is essential for understanding mechanisms structuring skin microbial assemblages and skin disease susceptibility in wildlife. Author Keywords: bat, fungi, microbiome, mycology, physiology, white-nose syndrome
Impacts of Cover Crops on Soil Health, Soil Nitrogen Dynamics, and Cytokinin Profiles
In Ontario, the dominant cash crop rotations consist of soybean (SB), which is a leguminous crop grown in rotation with maize (MZ) and winter wheat (WW). In addition to these crops, some farmers integrate cover crops (CC) into crop rotation, especially during the fallow period and winter seasons, to reduce nitrogen (N) losses via nitrate (NO3-) leaching and emission of N2 and the greenhouse gas nitrous oxide (N2O). This thesis focused on understanding the impact of crop phases in a MZ-(SB-WW)-CC rotation on the abundance of N-cycling bacterial communities that mediate nitrification and denitrification pathways. In addition, the influence of CCs on soil cytokinin (CK) profiles, which are plant growth-promoting hormones, were studied in a greenhouse trial to assess their potential impacts when integrating CCs into crop rotations. In particular, the relationship between traditional soil health parameters and the soil CK profiles was studied to understand how CKs might reflect biotic interactions and soil vitality. Results indicate N fertilizer application mono ammonium phosphate (MAP) and starter N:P: K (24:6:24) during WW planting in fall largely supported nitrifying bacterial communities (amoA) and potentially contributed to NO3- leaching. Management of MZ, which included spring-applied MAP resulted in larger denitrifying (nirK) bacterial communities, increasing the potential risk of N-loss via emission of dinitrogen gas (N2) and greenhouse gas N2O. However, CC soils had significantly lower nirK than MZ, reflecting the importance of strong and deep root systems of CCs, which have a higher ability to scavenge the substrates for denitrifying communities (NO3-). This highlights the importance of growing CCs in reducing the potential risk for N-loss via leaching and denitrification. Additionally, in the greenhouse trial, the ability of CCs to affect CK was detected, highlighting the importance of integrating CC in crop rotations. This is particularly noteworthy, given that total CK profiles showed strong associations with traditional soil health parameters such as labile or active carbon and soil microbial community diversity. It was concluded that total soil CK can be used as a novel and dynamic soil health measure. Future research on quantifying N2O fluxes and levels of NO3- in leachates would provide a more precise understanding of the impact of different crop rotation phases on N-dynamics in these fields. Further studies on single or combined measures of soil CKs are warranted to develop its potential as a practical and effective soil health parameter. Author Keywords: Cover crops, Crop rotations, Cytokinin hormone, Nitrogen Cycle, qPCR, Soil health
Relationships between bird densities and distance to mines in Northern Canada
Increased mining activity in the Canadian Arctic has resulted in significant changes to the environment that may be influencing some tundra-nesting bird populations. In this thesis I examine the direct and indirect effects of mining on birds nesting in the Canadian Arctic. I first perform a literature review of the effects that mining in the Arctic has on northern environments and wildlife and outline several ways in which mines affect Arctic-breeding birds. By using the Program for Regional and International Shorebird Monitoring (PRISM) Arctic plot-based bird survey data from across the Canadian Arctic, collected from 1995 to 2018, I identify the effects of distance to mining operations on the occupancy patterns of Arctic-breeding bird species. Six species’ densities were significantly impacted by mine proximity (Canada/Cackling Goose, Long-tailed Duck, Long-tailed Jaeger, Pectoral Sandpiper, Savannah Sparrow, and Rock Ptarmigan) across five major mine sites. Each species has its own unique relationship to distance from mining activity. Author Keywords: Bird populations, Canadian Arctic, Mining, Mining activities, PRISM, Tundra-nesting birds
Larval lake whitefish (Coregonus clupeaformis) recruitment dynamics in Lake Huron
Lake whitefish (Coregonus clupeaformis) recruitment has declined substantially in several regions of the Laurentian Great Lakes since the establishment of non-native dreissenid mussels in the early 1990’s. In Lake Huron, the reasons for the observed recruitment declines are currently unknown and there is limited knowledge about larval life stage. In our study, we determined whether larval hatching and growth rates have changed before and after dreissenid mussel invasion, and the role of several key environmental variables in influencing annual variation in larval densities. Larval fish were collected in the Fishing Islands spawning shoal during two time periods: a historical period (1976-1986) and a contemporary period (2017-2019). Larval densities and growth were lower in recent years, suggesting that recruitment is being limited at the larval life stage and that reduced food availability may be further limiting the growth during the larval stage. Annual variation of larval densities were influenced by spawning stock biomass, water levels, and dreissenid mussel presence, with higher water levels and the presence of dreissenid mussels being associated with higher larval densities. The direction of the effect of spawning stock biomass was either negative or positive depending on the model. We also found that larval density was a significant predictor of age 4 recruitment, indicating that year-class strength may be partly established at the larval life stage. Author Keywords: Coregonus clupeaformis, Great Lakes, Lake Huron, Lake whitefish, Larval, Recruitment
Behavioural ecology and population dynamics of freshwater turtles in a semi-urban landscape at their northern range limit
Species are faced with a variety of challenges in the environment, including natural challenges, such as variability in ambient temperature, and anthropogenic threats, such as habitat transformation associated with urbanisation. Understanding how animals respond to these kinds of challenges can advance the field of behavioural ecology and guide management decisions for wild species. Yet, we still have limited understanding of the extent of natural and human-caused impacts on animal behaviour and population dynamics, and lack robust assessment of behaviour in free-ranging animals. Using novel miniaturised biologging technologies, I characterised and validated behaviour in two freshwater turtle species: Blanding’s turtles (Emydoidea blandingii) and Painted turtles (Chrysemys picta). Further, I investigated how these two ectothermic species navigate a thermally heterogeneous landscape near their northern range limit, by comparing selected and available ambient temperatures. I showed that turtles preferred locations that were, on average, warmer and less variable in temperature than the available environment, and that this thermal sensitivity was greatest early in the year, and at fine spatial scales that likely matched the species' perception of the environment. Lastly, I assessed whether urban development was compatible with long-term viability of a Blanding’s turtle population, by monitoring habitat change and turtle survival over one decade of ongoing residential and road development. I found that Blanding’s turtle habitat quantity and connectivity declined in the area, which coincided with high road mortality and severe declines in turtle survival and population size, especially in adult females. I concluded that urban development and current road mortality rates are incompatible with the long-term viability of this at-risk turtle population. Overall, my findings demonstrate the importance of variation in the thermal environment and anthropogenic impacts on habitat in shaping the behaviour and population dynamics of this species-at-risk. Author Keywords: animal behaviour, biologging, ectotherms, habitat selection, temperature, urbanisation
Phytohormone-enhanced heavy metal responses in Euglena gracilis
Phytohormones, Cytokinin (CK) and Abscisic acid (ABA), are best known for controlling plant growth and stress responses; but they also mediate various developmental perspectives in alga. Yet, their mode of action in algal adaptive strategies to heavy metal responses, their involvement in orchestration of phytohormone crosstalk remain largely unknown and a molecular framework of phytohormone-controlled heavy metal uptake is absent. I found that three metals known globally to contaminate aquatic ecosystems, nickel (Ni), lead (Pb), and cadmium (Cd), cause changes in the levels of endogenous CKs, ABA, auxins, and gibberellins (GAs) in the green alga Euglena gracilis. Exogenous ABA or CK (trans-zeatin) alleviated metal toxicity through improved metal uptake efficiency and the regulation of the endogenous CKs activity profiles and GAs activity. This new evidence suggests that E. gracilis possesses functional phytohormone signals and metabolic pathways that are under metal stress response. Exogenously applied ABA or CK provoked the coordinated activation of metal uptake, likely via enhanced accumulation of metal binding compounds (i.e., proline, glycine, cysteine containing peptides), which are effective for metal sequestration. Using untargeted metabolomics analysis and functional annotation, this thesis further established that, CK and ABA modified pathways and metabolites, which were mainly involved in metal acclimation and resistance. These modified metabolites that were under the influence of phytohormones in algal cells growing under metal stress conditions were associated with: lipid pathways, riboflavin metabolism, biosynthesis of cofactors/vitamin, and carbohydrate metabolism. Bioactive secondary compounds (e.g., terpenoids, alkaloids, flavonoids, carotenoids) were also modified in algal cells treated with phytohormones. The present study highlights that ABA and CKs are important regulators of algal metal accumulation/acclimation strategies based on increased metal uptake, enhanced CK metabolism, regulation of hormonal crosstalk and regulation of some core cellular metabolism pathways, all of which improve metal uptake efficiency. Finally, our results suggest that ABA and CK form a novel strategy for metal bioremediation techniques and for sourcing microalgal value-added metabolites. Author Keywords: abscisic acid, cadmium, cytokinin, Euglena gracilis, lead, nickel
Assessing and Mitigation the Impacts of Mining-induced Flooding on Arctic-nesting Birds
Mining and resource development are growing industries in the Arctic, resulting in increased conflict with wildlife. Best practices for mitigation require an understanding of the potential ecological effects. One such effect concerns the flooding of terrestrial bird habitat from dewatering of lakes during mining pit development. I first assessed the efficacy of bird deterrents to mitigate impacts of mining-induced flooding on arctic-nesting birds at a gold mine in Nunavut. I used a Before-After Control Impact (BACI) design to determine changes in male territory densities, between year and treatment types (Control, High and Low Deterrent Intensity). Additionally, I assessed whether deterrents impacted daily survival rates of two passerine species, and the incubation behaviour of female Lapland Longspur. Finally, I quantified nest losses during the breeding season due to direct flooding of the tundra nesting habitat caused by mining operations. Deterrents did not affect male territory densities and neither deterrent treatment nor year affected the daily survival rate of nesting passerines. Female Lapland Longspurs exposed to deterrents exhibited more incubation off-bouts than control females. I documented six flooded nests. Deterrents used in this study appear to be ineffective in mitigating nesting in potential zones of impact. Incidental take accounted for about 1.2% of all nests found in the 0.48 km2 Whale Tail Lake study area. Author Keywords: arctic-nesting birds, audio deterrents, incidental take, mining and resource development, nest incubation, visual deterrents
Assessing the environmental correlates of a lethal amphibian pathogen, Batrachochytrium dendrobatidis, in Ontario wetlands
Many emerging infectious diseases are caused by pathogens that possess free-living life stages, in which they interact with the environment directly rather than through the mediation of a host. These diseases represent major impediments to wildlife conservation; however, the dynamics of their interaction with the environment are poorly studied, often due to the difficulty of detecting these microscopic pathogens in environmental samples. One of these pathogens is Batrachochytrium dendrobatidis (Bd), a fungus that has been linked to declines in many amphibian species. In this thesis I use an emerging technique, environmental DNA detection (eDNA), to detect and quantify Bd in the water of southern Ontario (Canada) wetlands and examine its correlation with a variety of aspects of water quality, surrounding habitat, and seasonal timing. My purpose was to inform not only on potential environment-pathogen dynamics for Bd in northern environments, but to provide insight into the use of eDNA as a disease surveillance tool. I found that not only was there high geographic variation in Bd detection and intensity, but also high temporal variation within the same site on time scales as low as two weeks. While Bd prevalence was not strongly correlated with any of the variables tested, intensity showed strong correlation with canopy cover, with greater canopy cover over a waterbody correlating to lower Bd intensity. My results present several promising avenues for further examination of Bd in northern ecosystems, and indicate that, while caution is warranted in its implementation, eDNA may become an important tool in amphibian pathogen surveillance. Author Keywords: amphibian disease, Batrachochytrium dendrobatidis, disease monitoring, environment-pathogen dynamics, environmental DNA, wildlife disease
Comparative efficacy of eDNA and conventional methods for monitoring wetland anuran communities
Identifying population declines and mitigating biodiversity loss require reliable monitoring techniques, but complex life histories and cryptic characteristics of anuran species render conventional monitoring challenging and ineffective. Environmental DNA (eDNA) detection is a highly sensitive and minimally invasive alternative to conventional anuran monitoring. In this study, I conducted a field experiment in 30 natural wetlands to compare efficacy of eDNA detection via qPCR to three conventional methods (visual encounter, breeding call, and larval dipnet surveys) for nine anuran species. eDNA and visual encounter surveys detected the greatest species richness, with eDNA methods requiring the fewest sampling events. However, community composition results differed among methods, indicating that even top performing methods missed species detections. Overall, the most effective detection method varied by species, with some species requiring two to three methods to make all possible detections. Further, eDNA detection rates varied by sampling season for two species (A. americanus and H. versicolor), suggesting that species-specific ecology such as breeding and larval periods play an important role in eDNA presence. These findings suggest that optimized monitoring of complex anuran communities may require two or more monitoring methods selected based on the physiology and biology of all target species. Author Keywords: amphibian, anuran, conventional monitoring, eDNA, environmental DNA, species richness
Assessing the Cost of Reproduction between Male and Female Sex Functions in Hermaphroditic Plants
The cost of reproduction refers to the use of resources for the production of offspring that decreases the availability of resources for future reproductive events and other biological processes. Models of sex-allocation provide insights into optimal patterns of resource investment in male and female sex functions and have been extended to include other components of the life history, enabling assessment of the costs of reproduction. These models have shown that, in general, costs of reproduction through female function should usually exceed costs through male function. However, those previous models only considered allocations from a single pool of shared resources. Recent studies have indicated that the type of resource currency can differ for female and male sex functions, and that this might affect costs of reproduction via effects on other components of the life history. Using multiple invasibility analysis, this study examined resource allocation to male and female sex functions, while simultaneously considering allocations to survival and growth. Allocation patterns were modelled using both shared and separate resource pools. Under shared resources, allocation patterns to male and female sex function followed the results of earlier models. When resource pools were separate, however, allocations to male function often exceeded allocations to female function, even if fitness gains increased less strongly with investment in male function than with investment in female function. These results demonstrate that the costs of reproduction are affected by (1) the types of resources needed for reproduction via female or male function and (2) via trade-offs with other components of the life history. Future studies of the costs of reproduction should examine whether allocations to reproduction via female versus male function usually entail the use of different types of resources. Author Keywords: Cost of Reproduction, Gain Curve, Life History, Resource Allocation Patterns, Resource Currencies
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


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Format: 2023/06/07