Graduate Theses & Dissertations

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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
Clonal structure and mating patterns in a natural population of Sagittaria latifolia
Increased plant size is expected to have negative consequences for mating by increasing pollen transfer among the same plant. However, recent theoretical studies have demonstrated that this may not be true for clonal plants. Instead, clonal expansion could enhance outcrossing opportunities without increasing selfing by reducing distances to potential mates. I investigated how the spatial structure of clones influences patterns of pollen dispersal, selfing rates and siring success in a natural population of Sagittaria latifolia. I found that pollen dispersal distances typically exceeded the spatial extent of clones and there was a positive association between clone size and the likelihood that clones were intermingled. Together, this resulted in a weak positive association between clone size and selfing rates, and a strong positive association between clone size and outcross siring success. This is the first empirical support for the theoretical expectation that any negative effects of selfing in large clones might be offset by increased siring success. Author Keywords: clonal growth, fitness gain curve, geitonogamy, plant mating, plant reproductive ecology, sex allocation theory
Molecular Architectures for Improved Biomaterials Derived from Vegetable Oils – Application to Energy Storage and Lubricants
The replacement of petroleum with renewable feedstock for energy and materials has become a priority because of concerns over the environment and finite nature of petroleum. The structures of the available natural biomass feedstocks fall short in delivering key functionality required in materials such as lubricants and phase change energy storage materials (PCMs). The approach taken in this thesis was to combine select functional groups with vegetable oil derivatives to create novel PCMs and lubricantswhich deliver desired functionality. One series of diester PCMs were prepared with terephthalic acid and fatty alcohols to address known shortcomings of esters. The second class of PCMs are sulfones prepared from oxidation of fatty sulfides to improve thermal energy storage. Overall, the new PCMs presented narrow phase change temperature ranges, high transition temperature (between 67 to 110℃), high transition enthalpy (210 to 266J/g), minimal supercooling and congruent phase transitions unaffected by cooling rates. They also demonstrated higher thermal degradation stability with onset of degradation from 290 to 310℃. The series of lubricants studied consists of sulfide and sulfonyl functional groups attached to the unsaturation sites of oleyl oleate as pendant groups to improve the thermal and flow properties. The new lubricants present subzero crystallization temperatures, very low crystallization enthalpy and dynamic viscosity as high as 180mPas. Furthermore, they also presented high onset of degradation (up to 322℃) and oxidation (up to 298℃). The PCMs and lubricants of the present thesis demonstrate that select functional groups can be used with common structural elements of vegetable oil such as fatty acids, ester groups and unsaturation sites to make a variety of molecular structures capable of delivering desired properties Author Keywords: Crystal Structure, Lubricant, Phase Change Material, Renewable, Structure-Property Relationships, Vegetable Oil
Differences and similarities in exploration and risk-taking behaviours of two Myotis bat species.
AbstractDifferences and similarities in exploration and risk-taking behaviours of two Myotis bat species. Laura Michele Scott Behaviours that are repeatable across circumstances and time determine an individual’s personality. Personality and behavioural variation are subject to selective pressures, including risks related to the use of different habitat types. I explored the ecological and evolutionary consequences of habitat selection by comparing the behaviour of two sympatric bat species, Myotis leibii and M. lucifugus. These species display overlap in roosting preferences, however, M. leibii tend to roost in crevices on the ground, while M. lucifugus tend to roost in crevices or cavities that are raised off the ground. I hypothesized that the habitat selection patterns of these two species create behavioural reaction norms at the species level. I predicted that ground roosting behaviour favours bolder personality and more exploratory and active traits when compared with bats that do not ground roost. I examined inter- and intra-specific variation in behaviour using a modified, three-dimensional open-field test and quantified the frequency and duration of behaviours such as flying, landing, and crawling. Bats were continuously video-recorded over 1-hour nocturnal and diurnal trials. I used a priori mixed models with combinations of individual characteristics and life-history traits to select the models that best describe each species. We found that M. leibii (n = 15) displayed more exploratory and bolder behaviours than M. lucifugus while on the ground (n = 21) and higher overall activity during the trial. I also found that M. leibii displayed crawling behaviours and movements consistent with foraging while on the ground which is a rare behaviour in bats and only observed in a few species (Desmodus rotundus and Mystacina tuberculate to my knowledge). Future research should explore biomechanical adaptations associated with ground-foraging in M. leibii. Author Keywords: Bats, Behaviour, Exploration, Myotis leibii, Myotis lucifugus, Roosting
Contributions of Mayflies (EPHEMEROPTERA
Walleye (Sander vitreus) are an ecologically and economically significant fish harvestedby recreational and commercial fisheries across Ontario. Adult Walleye are piscivores, but anecdotal evidence from anglers suggests that Walleye often target aquatic insects such as mayfly larvae (Ephemeroptera). My research examined the diet of Walleye caught from May to September in Lake St. Joseph in northern Ontario. I examined the stomach contents of angle harvested Walleye to identify the prey over two summers. Through morphological analysis of stomach contents, mayflies were found to be a significant prey source for Walleye, during larval emergence events in early summer, and to a lesser extent throughout the rest of the summer season. These findings are important for long term management of Walleye populations and associated resources. I also assess the potential and problems of Walleye management and research from my experiences of having worked with industry, government, and university partners on this project. Author Keywords: alternative prey, Food web interactions, invertebrate, piscivore, Predator prey interactions
Effect of Attending a Virtual Oncology Camp on Childhood Cancer Patient's Pyshcosocial Functioning and Parental Stress - A Pilot Study
Objectives/purpose: The current study examined whether attending a 1-month virtual oncology camp (VOC) improved resilience and hope in childhood cancer patients and parental/caregiver stress. Methods:Childhood cancer patients/survivors and their parent/caregivers enrolled for VOC, participated in an online anonymous survey: before, after and 3-months after VOC. The survey included the Child and Youth Resilience Measure (CYRM) and the Snyder’s Children’s Hope Scale (CHS) for the childhood cancer patients/survivors and the Pediatric Inventory for Parents (PIP) for parent/caregivers. Results:CYRM scores increased from T1 to T2 (d=0.86). Compared to T1, at T2 CHS scores also increased (d=1.33). Both CHS and CYRM scores remained higher at T3 compared with T1 (d=1.34; d=0.86). There were no changes in PIP scores between any time points. Conclusion and significance: Our study demonstrated that participation in a VOC improved children’s resilience and hope but did not change parental stress. Highlighting the clinical significance of these VOCs and the impacts they have on childhood cancer patients/survivors. Author Keywords: cancer, children, hope, parental stress, resilience, virtual oncology camp
Linking large scale monitoring and spatially explicit capture–recapture models to identify factors shaping large carnivore densities
Understanding the spatial ecology of large carnivores in increasingly complex, multi-use landscapes is critical for effective conservation and management. Complementary to this need are robust monitoring and statistical techniques to understand the effect of bottom-up and top-down processes on wildlife population densities. However, for wide-ranging species, such knowledge is often hindered by difficulties in conducting studies over large spatial extents to fully capture the range of processes influencing populations. This thesis addresses research gaps in the above themes in the context of the American black bear (Ursus americanus) in the multi-use landscape of Ontario, Canada. First, I assess the performance of a widely adopted statistical modelling technique – spatially explicit capture-recapture (SECR) – for estimating densities of large carnivores (Chapter 2). Using simulations, I demonstrate that while SECR models are generally robust to unmodeled spatial and sex-based variation in populations, ignoring high levels of this variation can lead to bias with consequences for management and conservation. In Chapter 3, I investigate fine-scale drivers of black bear population density within study areas and forest regions by applying SECR models to a large-scale, multi-year black bear spatial capture-recapture dataset. To identify more generalizable patterns, in Chapter 4 I then assess patterns of black bear density across the province and within forest regions as a function of coarse landscape-level factors using the same datasets and assess the trade-offs between three different modeling techniques. Environmental variables were important drivers of black bear density across the province, while anthropogenic variables were more important in structuring finer-scale space use within study areas. Within forest regions these variables acted as both bottom-up and top-down processes that were consistent with ecological influences on black bear foods and intensity of human influences on the species’ avoidance of developed habitats. Collectively, this thesis highlights the opportunities and challenges of working across multiple scales and over expansive landscapes within a SECR framework. Specifically, the multi-scale approach of this thesis allows for robust inference of the mechanisms structuring fine and broad scale patterns in black bear densities and offers insight to the relative influence of top-down and bottom-up forces in driving these patterns. Taken together, this thesis provides an approach for monitoring large carnivore population dynamics that can be leveraged for the species conservation and management in increasingly human-modified landscapes. Author Keywords: animal abundance, black bear, capture-recapture, density estimation, statistical ecology, wildlife management
SARS-CoV-2 Protein-based Detection Using Localized Surface Plasmon Resonance
During the COVID-19 pandemic, nucleic acid and antibody-based testing methods were heavily relied upon, but can be costly, time-consuming and exhibit high false -negative and -positive rates. Thus, alternative strategies are needed. Viral antigens such as the SARS-CoV-2 spike (S) glycoprotein are critical in the function of the virus and useful as diagnostic biomarkers for viral infections. For biosensing applications, aptamers are suitable high-affinity and cost-effective binding partners for their specific targets. Using localized surface plasmon resonance (LSPR), real-time, rapid acquisition of results can be achieved, essential for improving the efficacy of a sensor. Herein, LSPR aptamer sensors were fabricated for the detection of the SARS-CoV-2 protein. Data indicate that the best performing aptasensor was the streptavidin-biotin sensor, while the current gold aptasensor exhibited lower sensitivity and the fabrication of the carboxyl aptasensor was unsuccessful. The S1 aptamer selectively bound the S1 protein with high binding affinity. Excellent shelf-life stability, reusability, and high recovery in complex matrices was also maintained. Additionally, a receptor binding domain (RBD) functionalized sensor was fabricated to examine the interactions with angiotensin converting enzyme 2 (ACE2), for future assessment of inhibitors used in drug therapies. Overall, LSPR has been demonstrated as a viable tool for measuring SARS-CoV-2 related aptamer-protein and protein-protein interactions, and this strategy may be applied to other viral or non-viral antigen targets. Author Keywords: Antigen-based Detection, Coronavirus, COVID-19, Inhibition, Localized Surface Plasmon Resonance, SARS-CoV-2
Exploring the Role of Natural Antisense Transcripts in the Stress Response of Ustilago maydis
Fungal pathogens adapt to environmental changes faster than their hosts, due in part to their adaptive mechanisms exhibited in response to stress. Ustilago maydis was used to investigate potential natural antisense transcript (NAT) RNA-mediated mechanisms that enhance fungal adaptation to stress. Of the 349 NATs conserved amongst U. maydis and two related smut fungi, five NATs were identified as having altered transcript levels in response to multiple stress conditions. Subsequently, antisense transcript expression vectors were created for select NATs and transformed into U. maydis haploid cells. When exposed to stress conditions, two antisense expressing mutant strains exhibited alterations in growth. RT-qPCR analysis of mRNA complementary to expressed NATs revealed no significant change in mRNA levels, which suggests NAT expression may influence stress response through dsRNA formation or other RNA mediated mechanisms. These results establish a basis for further investigations into the connection between NATs and the stress response of fungi. Author Keywords: natural antisense transcripts, non-coding RNAs, stress response, Ustilago maydis
Gene flow directionality and functional genetic variation among Ontario, Canada Ursus americanus populations.
Rapidly changing landscapes introduce challenges for wildlife management, particularly for large mammal populations with long generation times and extensive spatial requirements. Understanding how these populations interact with heterogeneous landscapes aids in predicting responses to further environmental change. In this thesis, I profile American black bears using microsatellite loci and pooled whole-genome sequencing. These data characterize gene flow directionality and functional genetic variation to understand patterns of dispersal and local adaptation; processes key to understanding vulnerability to environmental change. I show dispersal is positively density-dependent, male biased, and influenced by food productivity gradients suggestive of source-sink dynamics. Genomic comparison of bears inhabiting different climate and forest zones identified variation in genes related to the cellular response to starvation and cold. My thesis demonstrates source-sink dynamics and local adaption in black bears. Population management must balance dispersal to sustain declining populations against the risk of maladaptation under future scenarios of environmental change. Author Keywords: American black bear, Dispersal, Functional Genetic Variation, Gene Flow Directionality, Genomics, Local Adaptation
Interseeded Cover Crops in Ontario Grain Corn Systems
Ontario grain corn is highly valuable, accounting for 60% of Canada’s total corn output. Grain producers are increasingly interested in including cover crops (CCs) in their cropping systems, but they have concerns regarding successful CC establishment and potential adverse competitive effects on corn yield and nutrient status. One option to improve the success of CC establishment is the interseeding in corn at the V4 -V6 stages. Interseeding improves the chances of good CC establishment, with potential benefits for soil health, weed control, and plant productivity. This thesis research was conducted to evaluate the short-term effectiveness of interseeding annual ryegrass (AR), red clover (RC), and their mixture (MIX) in grain corn at three locations in central and southwestern Ontario. Cover crop and corn yields, and their nitrogen (N) uptake, residual soil N, soil biological parameters, weed biomass, and residue decomposition rates were measured. CC biomass was highly variable (range: 0 - 1.6 Mg ha-1), influenced by climatic conditions, location, and CC type. Total carbon (C) and N contributions from CCs were similarly influenced by site-year and CC type. Regression analyses showed significant influence of corn biomass on CC establishment. Red clover had a significantly lower C/N ratio (11.8) than AR (18.2) and MIX (15.6). Strikingly, the amount of CC biomass accumulated in early spring reduced weeds by 50%. Moreover, CCs did not reduce corn grain or stover yield, nor N uptake, and soil mineral N in either fall or spring. Soil metabolic activity measured by BIOLOG Ecoplates was significantly greater in plots with AR than RC, MIX or NOCC. Soil biological parameters showed no CC effect. Results of residue decomposition i.e., C and N mineralization showed negligible CC residue effects on corn stover decomposition or N immobilization. The findings from this research suggest the need for assessing a more diverse range of CCs over longer durations to establish more specific CC niches for improving soil health in Ontario corn systems. Author Keywords: CLPP, cover crops, grain corn, nitrogen uptake, residue decomposition, soil health

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Format: 2024/03/28