Environmental and Life Sciences

Range expansion by the bobcat (Lynx rufus) may be contributing to range contraction by the Canada lynx (Lynx canadensis), but interactions between them are not well understood. To investigate the potential for competition, I conducted a literature review of hierarchical habitat selection by these two species. I determined that the lynx and the bobcat select different resources at the first and second orders, and that the fourth order is under-studied compared to higher orders. I therefore conducted a snow-tracking study of fine-scale habitat selection by lynx and bobcat in an area of sympatry in northern Ontario. I found that the two species selected similar resources at the fourth order, but appeared to be allopatric at the level of the home range. These results suggest that competition is unlikely to occur between lynx and bobcat, and other factors should be considered as more probable causes of the lynx range contraction.

Author Keywords: Bobcat, Canada lynx, Competition, Habitat selection, Scale, Snow tracking

Environmental monitoring and investigation of metal biogeochemical cycling has been

carried out in the Pearl River Estuary (PRE), an important and complex system in Southern

China. In this study, rare earth element (REE) patterns as well as isotope ratios (i.e., Zn and Cd)

were evaluated as tools to identify contamination sources in environmental compartments (i.e.,

water and suspended particles (SP)) as well as in oysters collected from estuarine sites. Results

show elevated concentrations (also called anomalies) of Pr, Nd, Dy and Ho, relative to other

REE elements, in water samples, potentially from REE recycling and other industrialized

activities in this area. Unlike water samples, no REE anomalies were found in SP or oysters,

suggesting that the dominate REE uptake pathway in oysters is from particles. Secondly, site to

site variations in Zn isotope ratios were found in water and SP, showing the complexity of the

source inputs in this area. Also, in estuarine locations, larger spatially differences in Zn isotope

ratios were found in water collected in wet season than those in dry season, which may due to

mixing of different source inputs under the water circulations in different seasons.

A series of laboratory experiments were conducted during which changes in Zn isotope

ratios were measured during uptake under varying salinity and Zn concentrations and during

depuration. Neither in vivo Zn transportation among the various tissues within the oysters nor

water exposure conditions (i.e., different salinities or Zn concentrations) caused Zn isotopic

fractionation in the oysters.

Cd and Zn isotope ratios were also determined in oysters obtained from the PRE. Large variations in Cd and Zn isotope ratios suggest that oysters were receiving contaminants from different input sources within the PRE. A consistent difference (approximately 0.67‰) was observed for Zn isotope ratios in oysters collected from the east side of the PRE compared to those from sampling locations on the western side of the PRE, suggesting different Zn sources in these two areas. Ultimately, by combining biogeochemistry with physiology, this study represents a first attempt to assess pollution status, monitor contaminants using oysters and model/identify contamination sources using both REEs and metal isotope ratios.

Contemporary evolution has the potential to help limit the biological impact of rapidly changing climates, however it remains unclear whether wild populations can respond quickly enough for such adaptations to be effective. In this thesis, I used the introduction of native North American Pumpkinseed (Lepomis gibbosus) into the milder climate of Europe over 140 years ago, as a 'natural' experiment to test for contemporary evolution to a change in climate in wild populations. In 2008, four outdoor pond colonies were established in central Ontario using adult Pumpkinseed from two native Canadian populations, and two non-native populations from northeastern Spain. By raising native and non-native Pumpkinseed within a common environment, this design minimized the impact of phenotypic plasticity on differential trait expression, and allowed me to interpret differences in the phenotype among pond-reared Pumpkinseed as evidence of genetic differences among populations. I demonstrated that Canadian and Spanish Pumpkinseed have similar thermal physiology except when acclimated to seasonally warm temperatures; trait differences are consistent with Spanish Pumpkinseed being better adapted to a warmer climate. Populations also had similar overwintering ecology, however some differences, such as higher survival under starvation conditions and greater energetic benefits associated with winter feeding, indicated that Canadian populations are better adapted to harsh winter conditions typical of the native range. Finally, I determined that the relatively fast life history expressed in wild European Pumpkinseed is largely driven by plastic responses to the local environment; however, the higher reproductive investment by European populations has a genetic basis. Most climate change research considers taxa that are expected to be negatively impacted by warming: my research demonstrates that even warm-tolerant taxa that are unlikely to experience strong climatic selective forces can respond to a warming environment through evolutionary changes. The potential for adaptive contemporary evolution in warm-tolerant taxa should be taken into account when predicting future ecosystem effects of climate change, and when planning management strategies for species introduced into novel climates.

Author Keywords: climate change, contemporary evolution, fish, non-native species, thermal biology, winter ecology

Aquatic contaminant mobility and biological availability is strongly governed by the complexation of organic and inorganic ligands. Dissolved organic matter (DOM) is a complex, heterogeneous mixture of organic acids, amino acids, lipids, carbohydrates and polyphenols that vary in composition and can complex to dissolved metals thereby altering their fate in aquatic systems. The research conducted in this doctoral dissertation addresses 1) how DOM composition differs between phytoplankton taxa and 2) how DOM composition affects metal speciation and its subsequent microbial bioavailability in laboratory and field conditions. To accomplish this, a series of analytical methods were developed and applied to quantify thiols, sulphur containing DOM moieties, and the molecular composition of DOM. The works presented in this thesis represents one of the first comprehensive and multipronged analyses of the impact of phytoplankton metabolite exudates on microbial metal bioavailability. This dissertation demonstrated the analytical versatility of high-resolution mass spectrometry as a tool for compound specific information, as well as having the capabilities to obtain speciation information of organometallic complexes. The work presented in this PhD strengthens the understanding compositional differences of both autochthonous and allochthonous DOM and their effects on metal biogeochemistry.

Author Keywords: Dissolved Organic Matter, Mercury, Metal Accumulation, Phytoplankton, Spring Melts, Thiol

Ophidiomycosis (snake fungal disease) is caused by the pathogen Ophidiomyces ophiodiicola. Infected snakes exhibit dermal lesions, occasional systemic infections, and, in some cases, mortality. We studied snakes at Rondeau Provincial Park, Ontario, Canada, to explore whether ophidiomycosis develops during brumation or year-round. Throughout their active season, we quantified the prevalence of clinical signs of the disease on snakes and conducted qPCR of skin swabs to determine the prevalence of O. ophiodiicola on snakes. Prevalence of O. ophiodiicola and disease symptoms were highest on eastern foxsnakes (Pantherophis vulpinus) and very rare on other snake species. In P. vulpinus, pathogen and clinical sign prevalence was highest, directly after emergence from overwintering, with the majority of P. vulpinus being able to resolve clinical signs of ophidiomycosis by the return of winter. When we analyzed the survivorship of P. vulpinus we determined that the likelihood of a snake dying with ophidiomycosis is similar to a snake dying without ophidiomycosis. Given that P. vulpinus were the most affected species at our study site, ophidiomycosis does not appear to pose an imminent threat to our study population of P. vulpinus under current conditions.

Author Keywords: Eastern Foxsnake, Fitness, Ophidiomycosis, Pantherophis vulpinus, Seasonal trends, Snake fungal disease

One possible solution to acidification and losses of base cations in central Ontario forest soils may be the application of wood ash. Wood ash is generally high in pH and contains large amounts of calcium (Ca) and other nutrients essential for ecosystem health, however it also contains trace metals. Understanding the chemistry of soils following ash application to forests is crucial for future policy recommendations and remediation efforts. In this study, soil and soil water chemistry was measured at two acidic forest sites in central Ontario. Sugar maple (Acer saccharum, Marsh.) seedling growth and chemistry, as well as understory vegetation composition, were also measured. At site one, plots (2 m x 2 m) were established with sugar maple, white pine (Pinus strobus L.) and yellow birch (Betula alleghaniensis Britt.) residential wood ash treatments and applied at rates of 0 and 6 Mg ha-1. The effects of residential wood ash on soil and understory vegetation were measured three- and 12-months following ash addition. At site two, plots (5 m x 5 m) were established with both fly and bottom industrial grade bark ash treatments of 0, 4 and 8 Mg ha-1 (n=4), and tension lysimeters were positioned in each plot at 30, 50, and 100 cm depths. The effects of industrial grade wood ash on soil, soil water and understory vegetation were measured four years following ash addition. Metal concentrations in the ashes were generally low but were higher in the fly ash and yellow birch ash types. At site one, significant increase in soil pH, and Ca and magnesium (Mg) concentrations were observed after three months, however changes varied by treatment. Some metal concentrations increased in the upper organic horizons, but metals were likely immobilized in the soil due to increases in soil pH, electrical conductivity (EC) and high organic matter content of the soil. After one year, changes to metal concentrations in soils could be seen in mineral horizons, and a few metals (aluminum (Al), zinc (Zn), copper (Cu), chromium (Cr), strontium (Sr)) increased in treatment plots. At site two, the effects of industrial-grade bark ashes on soil pH could still be seen after four years and soil water metal concentrations were not elevated relative to controls. Changes to understory vegetation composition following ash application were observed, but ash addition had no significant effect on sugar maple seedling growth (root:shoot ratio) and did not lead to significant increases in foliar metal concentrations. There were significant differences in root chemistry, suggesting metal translocation and uptake could be restricted. Mass balance estimates indicate that the organic horizon is a sink for all metals and simulated drought in this horizon led to a decrease in soil pH and increase in soil water metal concentration, but this occurred in all treatments including control. These results suggest that application of industrial and residential wood ash in moderate doses with trace metal concentrations below or near regulatory limits will increase soil pH and base cation concentrations, as well as increase seedling tissue nutrient concentrations in northern hardwood forest soils. However, depending on the parent material of the ash, increased metal availability can also occur.

Author Keywords: Acer saccharum, calcium decline, forest soil amendment, Haliburton Forest and Wildlife Reserve, heavy metal, wood ash

Water storage is a fundamental component of drainage basins, controlling the synchronization between precipitation input and streamflow output. The ability of a drainage basin to store water and regulate streamflow may mediate sensitivity to climate and land cover change. There is currently no agreement on the best way to quantify basin storage. This study compares results of a combined hydrometric and isotopic approach for characterizing inter-basin differences in storage across the Oak Ridges Moraine (ORM) in southern Ontario. The ratio of the standard deviation of the stable isotope signature of streamflow relative to that of precipitation has been shown to be inversely proportional to mean water transit times, with smaller ratios indicating longer water transit times and implying greater storage. Stable isotope standard deviation ratios were inversely related to baseflow index values. Basins demonstrating longer transit times were associated with hydrological characteristics that promote infiltration and recharge of storage.

Author Keywords: baseflow, basin storage, climate change, mean transit time, Oak Ridges Moraine, stable isotopes

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

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

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