Environmental and Life Sciences

Cytokinins (CKs) are hormones that promote cell division. During the formation of tumors in the Ustilago maydis-Zea mays pathosystem, the levels of CKs are elevated. Although CK levels are increased, the origins of these CKs have not been determined and it is unclear as to whether they promote the formation of tumors. To determine this, we measured the CK levels, identified CK biosynthetic genes as well as CK signaling genes and measured the transcript levels during pathogenesis. By correlating the transcript levels to the CK levels, our results suggest that increased biosynthesis and signaling of CKs occur in both organisms. The increase in CK biosynthesis by the pathosystem could lead to an increase in CK signaling via CK translocation and promote tumor formation. Taken together, these suggest that CK biosynthesis, signaling and translocation play a significant role during the formation of tumors in the Ustilago maydis-Zea mays pathosystem.

Author Keywords: Biosynthesis, Cytokinins, Signaling, Translocation, Ustilago maydis, Zea mays

Leaf litter decomposition represents a major pathway for nutrient cycling and carbon flow in aquatic ecosystems, and macroinvertebrates play an important role in the processing of this material. To assess the causes of variable leaf breakdown and nutrient fluxes, I measured decomposition rates and the nutrient release ratios of decomposing leaf material across a broad latitudinal gradient in Ontario boreal lakes which varied in nutrients, temperature, and pH. I examined the effects of macroinvertebrates using inclusion and exclusion bags. Generally, leaves decomposed faster in nutrient-rich, warmer lakes. Macroinvertebrates increased decomposition rates but their effects were relatively small compared to regional effects of nutrients and temperature. In addition, we found differential effects of nutrients and temperature on nutrient release ratios, which were partially determined by the release and retention of N and P. These results indicate that changes in these important environmental lake variables could alter decomposition dynamics in Ontario lakes, with implications for nutrient cycling and the storage of this important external carbon source.

I studied the biogeography of predaceous diving beetles (Coleoptera: Dytiscidae) in two remote and understudied regions: the Far North of Ontario, and Akimiski Island, Nunavut. I identified 35 species from northern Ontario, including three first provincial records for Ontario, Acilius athabascae Larson (1975), Hygrotus unguicularis (Crotch 1874), and Nebrioporus depressus (Fabricius 1775). I also documented three significant range extensions and six gap-infills for this region. I collected and identified 16 species from Akimiski Island, Nunavut, which include several first time reports for these species for the Nunavut territory. My collections also extend the known ranges of five species into the Hudson Plains Ecozone. This work provides important baseline information on the distribution of diving beetles for these regions.

Author Keywords: biodiversity, Boreal Shield, decomposition, Dytiscidae, ecological stoichiometry, macroinvertebrates

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

Range contractions and expansions are important ecological concepts for species management decisions. These decisions relate not only to rare and endangered species but to common and invasive species as well. The development of the broad spatiotemporal extent models that are helpful in examining range fluctuations can be challenging given the lack of data expansive enough to cover the time periods and geographic extents needed to fit the models. Archival records such as museum databases and harvest data can provide the spatiotemporal extent needed but present statistical challenges given they represent presence-only location information. In this thesis, I used maximum entropy and Bayesian hierarchical occupancy algorithms fitted with archival presence-only records to develop spatiotemporal models covering broad spatial and temporal extents for snowshoe hare and Canada lynx. These two algorithm types are well suited for presence-only data records and can be adapted to include biological and physical processes, thus improving the ecological realism of the models. Using these modelling methods, I found the extent of occurrence (EOO) and area of occupancy (AOO) varied greatly over time and space for both snowshoe hare and Canada lynx, suggesting that management decisions for these species should include consideration of these variations. While the presence-only data were appropriate for model development and understanding changing values in EOO and AOO, it sometimes lacked the locational accuracy and precision needed to create fine scale ecological analyses, thus resulting in somewhat coarse but potentially relevant conclusions.

Author Keywords: Area of occupancy, Bayesian hierarchical models, Canada lynx, Extent of occurrence, Presence-only data, Snowshoe hare

Environmental DNA (eDNA) metabarcoding targets sequences with interspecific

variation that can be amplified using universal primers allowing simultaneous detection

of multiple species from environmental samples. I developed novel primers for three

barcodes commonly used to identify plant species, and compared amplification success

for aquatic plant DNA against pre-existing primers. Control eDNA samples of 45 plant

species showed that species-level identification was highest for novel matK and preexisting

ITS2 primers (42% each); remaining primers each identified between 24% and

33% of species. Novel matK, rbcL, and pre-existing ITS2 primers combined identified

88% of aquatic species. The novel matK primers identified the largest number of species

from eDNA collected from the Black River, Ontario; 21 aquatic plant species were

identified using all primers. This study showed that eDNA metabarcoding allows for

simultaneous detection of aquatic plants including invasive species and species-at-risk,

thereby providing a biodiversity assessment tool with a variety of applications.

Author Keywords: aquatic plants, biodiversity, bioinformatics, environmental DNA (eDNA), high-throughput sequencing, metabarcoding

Soluble uranium (U) has been observed continuously in the porewaters of Bentley Lake,

a lake with semi-permanent anoxic sediments, despite the fact that reduced U(IV) is known to be

insoluble. To be able to predict the fate and mobility of U that has been deposited in lake

sediments, it is very important to understand the factors that determine soluble uranium in anoxic

environments. Understanding soluble U species is crucial for predicting its behavior in natural

systems as well as for the development of U remediation schemes.

To explore the factors affecting soluble U in natural environments, anoxic lake sediments

and porewaters were tested using two analytic methods, ICP-MS and ESI-HR-MS. Reduced

uranium (U(IV)) can be precipitated as U(IV)-NdF3. Using this method revealed that most of the

uranium in porewater is not able to be co-precipitated with NdF3. In addition, UO2+ was found

using ESI-HR-MS, showing uranyl ions exist in reduced porewater. However, the UO2+ might be

attached to some organic groups rather than present as free ions.

Seasonal variation and air exposure experiments on the mobility of U between sediments

and porewater were observed to test for changes of the redox state of U as a function of sample

collection and storage. The results of this study will contribute to better remediation strategies for

U tailings and will help U mining operations in the future.

This thesis looks at the genus Pollenia: historically where they were first introduced into Canada and spatially, where they are found now. This project involved me identifying 2211 files, sorted from the 3 years of field specimens obtained in 2011, 2012, 2013. P. pediculata was the most abundant and widespread, yielding 1272 specimens out of 2211, and it was found in all provinces sampled. The previous understanding of all Pollenia specimens as being P. rudis appears to be incorrect both in terms of actual number of species – which is known – and how prevalent it is. P. rudis comprised only 20% of the entire collection. The least common was P. griseotomentosa, occurring as 45 of 2211, or 2%.

I found new eight first provincial records: four species in Alberta (P. angustigena, P. labialis, P. rudis, P. vagabunda) , one species for Saskatchewan (P. pediculata), two for New Brunswick (P. griseotomentosa, P. labialis), and one for Nova Scotia (P. labialis). P. labialis was new to three provinces, the other species to one province each.

Author Keywords: Calliphoridae, Canada, Cluster Fly, Distribution, Pollenia, Provincial Records

Neuroblastoma (NB) has one of the highest mortality rates in pediatric oncology due to relapsed and refractory disease. Current aggressive multi-modal treatments are inhibited by dose-limiting toxicities and are associated with late-effects and secondary malignancies, emphasizing the necessity for novel therapeutics. Uniquely, most NB cells highly express disialoganglioside (GD2) a cell surface glycolipid that can provide a target for tumour-specific delivery. This study demonstrates a comprehensive evaluation of silver nanoparticles (AgNPs) and the first preliminary evaluation of anti-GD2-AgNP antibody-drug conjugates (ADCs) against NB in vitro. This present study validates the potential for AgNPs as an anti-cancer agent against NB as AgNPs demonstrated preferential toxicity towards NB cells through metabolic inhibition and indicative morphological alterations, while a less tumorigenic cell line demonstrated resistance to AgNP treatment. Therefore, this work identified an AgNP cell-type-dependent cytotoxicity effect. Low conjugation efficiency of the anti-GD2 monoclonal antibody, 14.G2a, to NHS-activated AgNPs failed to exert greater toxicity than the AgNPs alone. Collectively, this thesis provides novel information regarding the anti-cancer effects of AgNPs against NB with recommendations for anti-GD2-AgNP ADCs.

Author Keywords: ADC, Chemotherapy, GD2, Neuroblastoma, Silver nanoparticles

Salamanders are capable of tissue regeneration throughout all life-stages, which requires the dedifferentiation of mature cells to regrow lost tissues. Dedifferentiation is promoted by degradation of the extracellular matrix by matrix metalloproteases, as well as lysosomal degradation of intracellular and cell-surface proteins that mark cells as part of a mature lineage. Salamanders are also capable of developing cannibalistic phenotypes, plastic traits that are elicited by environmental stressors that result in elevated circulating glucocorticoid (e.g., corticosterone) levels that underlie many fundamental adaptive changes in morphology. Interestingly, the direct effect of corticosterone on regeneration and the cannibalistic phenotype have yet to be examined. In the present thesis, axolotls (Ambystoma mexicanum) were exposed to exogenous corticosterone and 50% of the distal tail tissue was removed. The effects of high corticosterone levels on matrix metalloprotease (MMP-2, MMP-9) and lysosomal acid phosphatase (LAP) activity were assessed; these are two classes of enzymes which are markers of extracellular matrix and intracellular remodeling during regeneration, respectively. We found that elevated corticosterone levels inhibited tissue regeneration, by prolonging the dedifferentiation phase as indicated by increased LAP and reduced MMP-2 and MMP-9 activity. Elevated corticosterone levels also promoted the cannibalistic morphology and this effect was strongest among smaller individuals.

Author Keywords: amphibian, cannibalistic morphology, corticosterone, dedifferentiation, regeneration, stress

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