Environmental and Life Sciences

In recent decades, declining calcium concentrations have been reported throughout lakes across the southern edge of the Canadian Shield. This raises concern as Daphnia populations have shown to be decreasing as they require calcium not only for survival but to mitigate predation risks. Therefore, the purpose of my thesis was to study the adaptability of Daphnia under calcium limitation and predation risk from Chaoborus. Firstly, I examined the effects of calcium limitation and Chaobours kairomones on daphniid life-history and population growth. I found that low calcium concentrations and Chaoborus kairomones affected Daphnia calcium content, life-history traits, and survival. Next, I focused on how calcium concentrations and Chaoborus abundance affected the calcium content and abundance of daphniids. During this study, I also examined the relationship between the abundance of Daphnia and a competitor Holopedium. I found that calcium concentrations and the abundance of Chaoborus affects daphniid abundance. Overall, results from this study show the importance of considering both predation risk and calcium declines to better determine daphniid losses.

Author Keywords: anti-predator responses, Chaoborus, competition , Life-History traits, predator cues, Zooplankton

Maximum permitted SO2 emissions from an aluminum smelter in Kitimat, B.C., increased after modernization in 2015. An increase in acidic deposition can potentially acidify forest soils. Monitoring was conducted at two long-term soil monitoring plots at near (7 km) and far (41 km) sites downwind from the smelter. Change in soil properties was assessed between 2015 and 2018: for the near plot, there was significant decrease in pH and exchange acidity; far plot soils exhibited significant decrease of base cations and exchange acidity in the 0–5 cm layer only. The average total SO42- deposition at near and far plots were estimated to be between 8.2–12.1 and 6.7–7.4 kg/ha/yr, respectively. It was concluded no soil acidification was detected. Observed changes were attributed to measured differences in organic matter, likely influenced by sampling difficulty and measurement process discrepancies. Estimated SO42- deposition levels pose no risk to soil base cation depletion.

Author Keywords: acid forest soils, acidic deposition, aluminum smelter, exchangeable base cations, long-term monitoring, minimum detectable change

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

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

Evaluating the impact of climate change is arguably one of the main goals of conservation biology, which can be addressed in part by studying the demographic history of species in the region of interest. In North America, landscape and species composition during the most recent Pleistocene epoch was primarily influenced by glaciation cycles. Glacial advance and retreat caused species ranges to shift as well, leaving signatures of past population bottlenecks in the genetic code of most species. Genomic tools have shown to be important tools for understanding these demographic events to enhance conservation biology measures in several species. In my thesis I first reviewed the state of ungulate genomics, with a focus on how such data sets can be used in understand demography, adaptation, and inform conservation and management. Importantly, the review introduces key analyses like the pairwise sequentially Markovian coalescent and features like variation in antlers and horns and selection pressures that are used throughout subsequent chapters. Using the North American mountain goat as a model species, I then explored the genomic and phenotypic variation in this alpine specialist mammal. Starting with the generation of the first genome assembly for the mountain goat, I identified genes unique to the mountain goat and modeled demographic history going back millions of years using a pairwise sequentially Markovian coalescent approach. Species' effective population size generally paralleled climatic trends over the past one hundred thousand years and severely declined to under a thousand individuals during the last glacial maximum. Given the biological importance of horns in mountain goats and the recent scientific interest in genetic basis of headgear, I analyzed over 23,000 horn records from goats harvested in British Columbia, Alaska and Northwest Territories from 1980 to 2017. Overall, variation in horn size over space and time was low; goats harvested further North had shorter horn lengths and smaller horn circumferences in one year old and 4 years and older age classes and 4 years and older age class, respectively. Proximity of roads, which was used as an indicator of artificial selection, had a small effect on horn size, with larger horns being harvested closer to major roads. Finally, I used two range-wide genomic data sets sequenced with a whole genome re-sequencing and reduced representation approaches to provide estimates of genetic diversity, contemporary effective population sizes and population structure. These insights can help inform management and will potentially make an impact in preserving the mountain goat.

Author Keywords: genome assembly, horn size, Oreamnos americanus, population demography, reduced representation sequencing, whole genome resequencing

Recent agricultural land use change in east-central Ontario, including the expansion of intensive agriculture (corn and soybean crops) and tile drainage (TD) infrastructure, may alter the fluxes of both phosphorus (P) and nitrogen (N) to the Lower Great Lakes. Through intensive monitoring of several sub-watersheds that encompassed a range of row crop and TD extents, this study examined differences in stream N and P concentrations both seasonally and during variable flow conditions, to better understand land use/land cover (LULC) relationships with nutrient export. There was no clear relationship between total P (TP; concentration or export) and agricultural LULC, and instead, TP delivery was highly sensitive to flow events, and TP concentrations (especially particulate P; PP) were significantly higher during event flow compared with baseflow. In contrast, the dissolved fraction of P (total dissolved P; TDP) and dissolved nitrogen as nitrate (NO3-N) were not sensitive to flow, but were instead positively related to row crop agriculture, and associations between NO3-N concentrations/export and tile-drained row crop area were particularly strong (concentration r2 = 0.93; export r2 = 0.88). Regression relationships showed that with every 10% increase in row crop area in watersheds, NO3-N and TDP flow-weighted concentrations increased by 0.34 mg/L and 1.5 µg/L, respectively. As well, the same 10% increase in row crop agriculture translated to an increase of NO3-N export of as much as 130 kg/km2. Geospatial records of TD are incomplete in east-central Ontario, which presents challenges for evaluating the contribution of TD to nutrient export. Understanding the response of nutrients to changes in agriculture and agricultural practices is an integral part of watershed management as rapid changes in both urban and agricultural LULC continue to put pressure on water quality in the Lower Great Lakes.

Author Keywords: East-Central Ontario, Nutrient Export, Row crops, Streams, Tile Drainage, Water Quality

Selenium is an essential trace nutrient to many organisms, yet in high concentrations it is toxic. Organic selenium is more bioavailable to aquatic biota than inorganic selenium, but is usually found in much lower concentrations. Algae are known to biotransform inorganic selenium into several organo-selenium compounds, but it is unknown whether any of these bioaccumulate in the food chain. In this study, selenium was incorporated into the methionine residues of an algal photosynthetic protein, c-phycocyanin from Spirulina spp. The extent of selenium incorporation was quantified by inductively coupled plasma-mass spectrometry (ICP-MS), and the protein was identified using electrospray mass spectrometry (ES-MS).

C-phycocyanin was isolated and purified from Spirulina with a final recovery of 20-30 % of the total c-phycocyanin present. Selenomethionine replaced 92.8% ± 1.22 of the methionine residues in c-phycocyanin when grown in 2.5 ppm sodium selenite. ES-MS was used to obtain protein spectra, and pure c-phycocyanin was identified. Data of full scans provided estimated masses of both protein subunits--α-chain measured at 18,036 Da; β-chain measured at 19,250 Da--close to the theoretical masses. Protein fragmentation by collision-induced dissociation and electron capture dissociation provided approximately 52 % amino acid sequence match with c-phycocyanin from Spirulina platensis. This study demonstrates the incorporation of selenium into an algal protein, and the identification of c-phycocyanin using electrospray ionization-mass spectrometry.

Nitrogen is a major constituent of agricultural fertilizers, and nitrogen inputs to stream water via runoff and groundwater lead to a variety of negative environmental impacts. In order to quantify the movement of nitrogen through aquatic food webs, fourteen streams with varying land uses across South-Central Ontario were sampled for two species of fish, freshwater mussels, and water for measurement of isotope ratios of δ15N and δ13C. I found that nitrogen isotopes in fish, water, and mussels were related to the percentage of riparian monoculture, and that carbon isotopes were unrelated to monoculture. Though all species were enriched as monoculture increased, the rate of δ15N enrichment as monoculture increased did not vary between species. This study has improved our understanding of how monoculture affects nutrient enrichment in stream food webs, and assesses the validity of using nitrogen isotopes to measure trophic positions of aquatic organisms across an environmental gradient.

Author Keywords: agriculture, fish, food webs, nitrogen, stable isotopes, streams

Stormwater ponds (SWPs) are a common feature in new urban developments where they are designed to minimize runoff peaks from impervious surfaces and retain particulate matter. As a consequence, SWPs can be efficient at retaining particle-bound nutrients, but may be less efficient at retaining nutrients that are present primarily in the dissolved form, like nitrogen (N). However, the forms of nutrients (e.g. particulate vs. dissolved) likely differ with hydrologic and seasonal conditions and few studies have examined year-round differences in nutrient forms and concentrations at urban SWPs. In order to contrast total suspended solids (TSS), phosphorus (P) and nitrogen (N) levels between low and high flow conditions, sampling was conducted at an urban SWP in Peterborough, ON between November 2012 and October 2013. Only an increase in TSS levels at the outflow between low and high flow conditions was observed, as well as a decrease in TSS levels at the outflow compared to Inflow 1 under low flow conditions. Nitrate-N (NO3-N) was the dominant form of N entering the pond under all flow conditions, whereas the fraction of total-P (TP) that was particulate increased under high flow conditions. Nevertheless, the dissolved fraction of TP was consistently high in these urban inlets. Only NO3-N was significantly greater in the inflows than outflow and only under low flow conditions. Increases in the proportions of organic-N and ammonium-N

in the outlet suggest that biological processing is important for N retention.

Author Keywords: nitrogen, Ontario, phosphorus, stormwater ponds, total suspended solids

Ustilago maydis (DC) Corda is the causal agent of 'common smut of corn'. Completion of the U. maydis lifecycle is dependent on development inside its host, Zea mays. Symptoms of U. maydis infection include chlorosis and the formation of tumours on all aerial corn tissues. Within the tumours, thick-walled diploid teliospores form; these are the reproductive and dispersal agent for the fungus. U. maydis is the model to study basidiomycete biotrophic plant-pathogen interactions. It holds this status in part because of the completely sequenced 20.5 Mb genome; however, thorough genome annotation is required to fully realize the value of this resource. The research presented here improved U. maydis genome annotation through the analysis of cDNA library sequences and comparative genomics. These analyses identified and characterized pathogenesis-related genes, and identified putative meiosis genes. This enabled the use of U. maydis as a model for investigating 'host-induced' meiosis. Further, the cDNA library analyses identified non-coding RNAs (ncRNAs) and natural antisense transcripts (NATs). NATs are endogenous RNA molecules with regions complementary to a protein-coding transcript. Although NATs have been identified in a wide variety of mammals, plants, and fungi, very few have been functionally characterized. Over 200 U. maydis NATs were annotated by analyzing full-length cDNA sequences. NAT structural features were characterized. Strand-specific RT-PCR was used to detect NATs in U. maydis and in a related smut fungus, U. hordei. The data supported a common role for NATs in smut teliospore development, independent of the RNA interference pathway. Analysis of the expression of one U. maydis NAT, as-um02151, in haploid cells, led to a model for NAT function in U. maydis during teliospore dormancy. This model proposed NATs facilitate the maintenance of stored mRNAs through the formation of double-stranded RNA. In testing this model, it was determined that the deletion of two separate upstream regulatory regions, one of which contained a ncRNA (ncRNA1), altered NAT levels and decreased pathogenesis. These studies strengthened U. maydis as a model organism, and began the functional investigation of NATs in U. maydis, which identified a new class of fungal pathogenesis genes.

Author Keywords: cDNA library analysis, genome annotation, mRNA stability, natural antisense transcripts, pathogenesis, Ustilago maydis