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Expression optimization and NMR spectroscopy of Giardia intestinalis cytochrome b5 isotype III
The parasitic protist Giardia intestinalis does not synthesize heme and lacks many common eukaryotic heme proteins, yet it expresses four cytochrome b5 (gCYTB5) isotypes of unknown function. These have low reduction potentials and distinct subcellular locations that are consistent with structural features and biological functions that differ from their mammalian counterparts. Isotype III (gCYTB5-III) is particularly fascinating for its unusual location in the nuclei of Giardia. This thesis reports the optimization of recombinant gCYTB5-III overexpression for structural studies by NMR spectroscopy. Vital optimization factors for isotope labelling were first identified, finding that auto-induction promotes the optimization of many other conditions, such as colony selection, starter cultures, media components, temperature, pH and aeration. Optimized conditions were then applied to the expression and NMR spectroscopy of isotope-labelled gCYTB5-III and bovine cytochrome b5 as a control. These results can be extended to other heme proteins and will expand our biochemical knowledge of Giardia.
Author Keywords: Auto-induction, Cytochrome b5, Giardia intestinalis, Isotope Labelling, Nuclear Magnetic Resonance Spectroscopy, Recombinant Protein
Paleolandscape Reconstruction of Burleigh Bay, Ontario 12,600 cal BP to Present: Modeling Archaeological Site Potential for the Late Paleoindian and Early Archaic Period in a Lacustrine Shield Environment
This thesis presents a palaeotopographic reconstruction of the Burleigh Bay region of Stony Lake (Kawartha Lakes Region, Ontario) from 12,600 cal BP to present. The paleotopographic reconstructions are used to model paleoshoreline locations and archaeological site potential for the Late Paleoindian and early Archaic periods. Isostatic rebound following the end of the last glacial period has altered the topography in the region and water levels are now artificially managed by dams constructed in the 1830s. I completed a high-resolution bathymetric survey using a kayak equiped with a GPS coupled single-beam sonar. Utilizing GIS technology and isostatic rebound response surface models, I created paleotopographic reconstructions for 12,600 cal BP, 11,500 cal BP, 7,000 cal BP, 5,700 cal BP, and present. Results show that water levels in Burleigh Bay have been regressing over time until dam construction. Early site potential is centered in northern inland areas. Site potential following 7,000 cal BP is concentrated in northern areas flooded by the dam. Based on the reconstructions, surveys in lacustrine granite shield regions that follow the Ontario Standards and Guidelines for Consultant Archaeologists risk missing areas of high archaeological potential for early sites in these environments. Paleolandscape reconstructions would alleviate this issue by modeling paleoshoreline changes over time.
Author Keywords: Canadian Shield, Early Archaic, Isostatic Rebound, Kawartha Lakes, Late Paleoindian, Paleolandscape
Shorebird Habitat Use and Foraging Ecology on Bulls Island, South Carolina During the Non-Breeding Season
Recent declines in North American shorebird populations could be linked to habitat loss on the non-breeding grounds. Sea-level rise and increased frequency of coastal storms are causing significant erosion of barrier islands, thereby threatening shorebirds who rely on shoreline habitats for foraging. I conducted shorebird surveys on Bulls Island, South Carolina in the winters of 2018 and 2019 and examined habitat selection and foraging behaviour in Dunlin (Calidris alpina), Sanderling (Calidris alba), Semipalmated Plovers (Charadrius semipalmatus), and Piping Plovers (Charadrius melodus). Area, tidal stage, and invertebrate prey availability were important determinants of shorebird abundance, behaviour, and distribution. My study highlights the importance of Bulls Island's habitat heterogeneity to supporting a diverse community of non-breeding shorebirds. Considering both the high rate of erosion and the increased frequency of disturbance along the shoreline of the island, intertidal habitats should be monitored to predict negative effects of changes in habitat composition and area on non-breeding shorebirds.
Author Keywords: foraging behaviour, habitat loss, habitat selection, invertebrate prey, non-breeding, shorebirds
Discriminating grey wolf (Canis lupus) predation events in a multi-prey system in central Saskatchewan
I investigated if spatio-temporal behaviour of grey wolves (Canis lupus) determined via GPS collar locations could be used to discriminate predation events generally, and among prey species, in Prince Albert National Park during winter, 2013-2017. I used characteristics of spatio-temporal GPS clusters to develop a predictive mixed-effect logistic regression model of which spatial clusters of locations were wolf kill sites. The model suffered a 60 % omission error when tested with reserved data due to the prevalence of deer kills with correspondingly low handling time. Next, I found a multivariate difference in the percentage of habitat classes used by wolves in the 2 hours preceding predation events of different prey species, suggesting that wolf habitat use reflects prey selection at a fine-scale. My results highlight the difficulty and future potential for remoting discriminating wolf predation events via GPS collar locations in multi-prey ecosystems.
Author Keywords: Canis lupus, GPS clusters, GPS collars, grey wolf, habitat use, predation
A multi-faceted approach to evaluating the detection probability of an elusive snake (Sistrurus catenatus)
Many rare and elusive species have low detection probabilities, thereby imposing unique challenges to monitoring and conservation. Here, we assess the detection probability of the Eastern Massasauga (Sistrurus catenatus) in contrast to a more common and conspicuous species, the Eastern Gartersnake (Thamnophis sirtalis). We found that patterns of detection probability differed between species, wherein S. catenatus was detected less often and under a more specific set of sampling conditions. Correspondingly, detection trials with S. catenatus found a high non-detection rate, while detection trials with artificial models suggest that regional differences in detection probability are driven by variation in population density and habitat use. Our results suggest that current monitoring efforts are not sufficient, and that S. catenatus is frequently undetected. Accordingly, we highlight the importance of species-specific monitoring protocols when monitoring rare and elusive species, and recommend a multi-faceted approach that estimates detection probability and identifies species-specific challenges to monitoring.
Author Keywords: detection probability, elusive species, monitoring programs, non-detection, S. catenatus, snakes
Academic Procrastination, Self-Regulation, Anxiety and Personality (ASAP): 'Some' Day is not a Day of the Week
Academic procrastination is analogous to a common cold: pervasive and with no established cure. Students experience repercussions that are not inclusive to academic performance, but rather, are experienced across mental, physical, social and emotional domains as well. While this necessitates treatment, much of the behaviour has yet to be explained. In the current study, a state approach was integrated to explore procrastination in terms of the Yerkes-Dodson paradigm. Procrastination was hypothesized to arise for one of two reasons: students are either too low or too high in anxiety. A sample of 847 Ontario undergraduate students completed measures of procrastination, state-anxiety, personality, and self-regulation. Results suggest 88% of students procrastinate regularly, and contrary to what was hypothesized, procrastination and anxiety were linearly related. Independent t-tests establish that relative to non-procrastinators, procrastinators are significantly more emotionally and biologically dysregulated and less socially integrated. Factor analysis demonstrated three central reasons contributing to procrastination, which were used as the basis to establish types of procrastination: social (n=61), self-doubt (n=70) and low-energy (n=76) procrastinators. Results indicate divergences across procrastination type with respect to personality as well as biological, affective and social domains. Findings also inform that the expression of procrastination, varies across student. Overall, findings suggest that academic interventions should acknowledge and address the variability among procrastinators.
Author Keywords: ACADEMIC PROCRASTINATION, ANXIETY, PERSONALITY, SELF-REGULATION
Advanced broadband CARS microscopy based on a supercontinuum-generating photonic crystal fiber
I have developed and improved a coherent anti-Stokes Raman scattering (CARS) microscope based on the spectral focusing (SF) technique. The CARS microscope uses an 800 nm oscillator and a photonic crystal fibre module to generate the supercontinuum Stokes. The photonic crystal fibre was originally designed to generate light beyond 945 nm which is useful for CARS microscopy in the CH/OH frequencies but essentially prevents access to the important fingerprint region at lower frequencies. With expert and nontraditional approaches to generating supercontinuum with sufficient power at wavelengths below 945 nm, I substantially extend the usefulness of the module for SF-CARS microscopy deep into the fingerprint region. Moreover, with the invention of a dynamic supercontinuum generation scheme we call "spectral surfing," I improve both the brightness of the CARS signal and extend the accessible CARS frequency range to frequencies as low as 350 cm$^{-1}$ and as high as 3500 cm$^{-1}$---all in a single scan-window. I demonstrate the capabilities of our broadband SF-CARS system through CARS and four-wave mixing hyperspectroscopy on samples such as astaxanthin, lily pollen and glass; liquid chemicals such as benzonitrile, nitrobenzene and dimethyl sulfoxide; and on pharmaceutical samples such as acetaminophen, ibuprofen, and cetirizine. Furthermore, In search of more useful Stokes supercontinuum sources, I compare the performance of two commercial photonic crystal fibre modules for use in SF-CARS applications, ultimately finding that one module provides better spectral characteristics for static supercontinuum use, while the other provides improved characteristics when spectral surfing is implemented.
Author Keywords: coherent anti-Stokes Raman scattering, nonlinear microscopy, scanning microscopy, spectroscopy, supercontinuum generation, vibrational spectroscopy
The Lives of Young Deliquents: Relationships of Power in the Royal Philanthropic Society
The Royal Philanthropic Society (RPS) was the first institution in England to care for young offenders. While historians have demonstrated the legal importance of this institution, none have examined the experience of the youths it attempted to reform. The admission registers of the RPS reveal the importance of adults and peers in the experiences of the inmates of the institution, as both could be sources of conflict and support. Youths could express power over their own lives by resisting the authority of adults, but also by conforming to the rules of the RPS. Inmates could restrict the choices of their peers by working with the RPS, but also through peer pressure or violence. Networks of collaboration and youth culture could also exert a positive impact on peers. Because this thesis represents male youths as actors, it makes a significant addition to recent histories emphasizing the impact of the subaltern groups on eighteenth-century reform movements.
Author Keywords: Conformity, Juvenile Delinquency, Penal Reform, Power Relationships, Royal Philanthropic Society, Youth Culutre
Sustainable Development and Environmental Security in the Canadian Arctic: Inuvik and Tuktoyaktuk
This study identifies and examines interlinkages between climate change and sustainable development, environmental security, and adaptive capacity through a case study of two communities in the Inuvialuit Settlement Region: Inuvik and Tuktoyaktuk. It seeks to understand how these two communities perceive climate change and define sustainable development, particularly in relation to oil and gas development. This thesis discusses the constraints that may be arising for local communities to benefit from emerging opportunities due to competing notions of sustainable development and environmental security. The findings indicate how notions of environmental security and sustainable development act upon multiple levels to impact the adaptive capacity of communities in the Arctic. The general findings also suggest that regionally specific understandings of sustainable development, sustainability, and environmental security need to be acknowledged in order to develop successful governance coordination and cooperation strategies and paradigms related to economic, social, infrastructure and environmental issues in the NWT and the Canadian Arctic.
Author Keywords: Arctic, Beaufort Sea, climate change, Environmental Security, Inuvialuit, Sustainable Development
Effects of Invasive Wetland Macrophytes on Habitat Selection by Turtles
Invasive species that alter habitats can have significant impacts on wildlife. The invasive graminoids Phragmites australis (Cav.) Trin. ex Steud, hereafter Phragmites, and Typha × glauca Godr. are rapidly spreading into North American wetlands, replacing native vegetation. Invasive Phragmites is considered a potential threat to several species-at-risk (SAR), including some turtle species. My study wetland contained large stands of Phragmites, as well as Typha spp. (including invasive T. × glauca) that have similar structural traits to Phragmites. To explore the hypothesis that Phragmites and Typha spp. do not provide suitable habitat for turtles, I tested the prediction that turtles avoid Phragmites- and Typha-dominated habitats. I used VHF-GPS transmitters to follow Blanding's turtles (Emydoidea blandingii, n = 14) and spotted turtles (Clemmys guttata, n = 12). I found that both turtle species did not avoid Phragmites- or Typha-dominated habitats when choosing a home range, or while moving within their home range. I also tested whether the microhabitat selection of Blanding's turtles and spotted turtles is affected by shoot density of Phragmites, Typha spp., or both. I compared shoot densities of Phragmites and Typha spp. in 4 m2 plots, from locations used by tracked turtles with paired, random locations in these turtles' home ranges. For both turtle species, the densities of Phragmites and Typha shoots were comparable between used and random locations within the home ranges (generalized linear mixed model; p > 0.05). The use of Phragmites- and Typha-dominated habitats by Blanding's turtles and spotted turtles suggests that these habitats do not automatically constitute "unsuitable habitats" for turtles. Phragmites and Typha spp. (especially T. × glauca) can replace preferred habitats of some turtle species, and the control of these invasive macrophytes can help to preserve habitat heterogeneity. However, the presence of SAR turtles in Phragmites and Typha spp. stands should inform risk-assessments for invasive plant species control methods that include mechanical rolling of stands, where heavy machinery might encounter turtles.
Author Keywords: Blanding's turtles, compositional analysis, habitat selection, Phragmites australis, spotted turtles, Typha x glauca