Environmental and Life Sciences

The purpose of this study was to quantify the variation of baseline carbon and nitrogen stable isotope signatures in the Lake Simcoe watershed and relate that variation to various physicochemical parameters. Particulate organic matter samples from 2009 and 2011 were used as representatives of baseline isotopic values. Temporal data from two offshore lake stations revealed that δ15N of POM was lowest mid-summer and highest after the fall turnover. POM δ13C was variable throughout the summer before declining after fall turnover. Spatial data from the lake and the tributaries revealed that POM stable isotope signatures were highly variable. Various physicochemical parameters indicative of phytoplankton biomass were significantly positively correlated with POM δ15N and significantly negatively correlated with POM δ13C. The correlations were mostly significant in the tributaries, not the lake. Moreover, many of the correlations involving δ15N of POM were driven by extreme values in Cook's Bay and its tributaries. In general, it's likely that different processes or combination of processes were affecting the δ15N and δ13C POM in the Lake Simcoe watershed as physicochemical parameters alone could not explain the variability. Measuring the δ15N of ammonium and nitrate, as well as the δ13C of DIC would help discern the dominant nitrogen and inorganic carbon cycling processes occurring in the Lake Simcoe watershed.

Author Keywords: δ13C, δ15N, isotopic baseline, particulate organic matter, spatial variation, stable isotopes

Effective population size (Ne) is a key concept in population genetics, evolutionary biology and conservation biology that describes an important facet of genetic diversity and the capacity of populations to respond to future evolutionary pressures. The importance of Ne in management and conservation of wild populations encouraged the development of numerous genetic estimators which rely on a variety of methods. Despite the number and diversity of available Ne methods, however, tests of estimator performance have largely relied on simulations, with relatively few tests based on empirical data. I used well-studied wild populations of brook trout (Salvelinus fontinalis) in Algonquin Park, Ontario as a model system to assess the comparative performance of multiple Ne estimation methods and programs, comparing the resultant Ne estimates against demographic population size estimates.

As a first step, the genetic diversity and ancestry of wild brook trout populations was determined using 14 microsatellite loci. Genetic structure of brook trout populations showed variable contributions from historical supplemental stocking and also identified localized gene pools within and between watersheds, reflecting variable levels of connectivity and gene flow. Once the genetic ancestry and connectivity of populations had been resolved, single sample (point) and two samples (temporal) genetic estimators were used to estimate Ne of populations with pure native ancestry. Values obtained from genetic estimators utilizing both methods were variable within as well as among populations. Single sample (point) estimators were variable within individual populations, but substantially less than was observed among the temporal methods. The ratios of Ne to the estimated demographic population size (N) in small populations were substantially higher than in larger populations. Variation among estimates obtained from the different methods reflects varying assumptions that underlay the estimation algorithms. This research further investigated the effect of sampling effort and number of microsatellite loci used on Ne values obtained using the linkage disequilibrium (LD) estimation method. Ne estimates varied substantially among values generated from subsets of loci and genotyped individuals, highlighting the necessity for proper sampling design for efforts aiming to measure Ne. Despite the variation observed among and within estimation methods, the Ne concept is a valuable for the conservation and management of both exploited and endangered species.

Author Keywords: Brook Trout, Effective population size, Genetic Diversity, Genetic Structure

ABSTRACT

Calving site selection and fidelity in a restored elk (Cervus elaphus) herd in Bancroft

Ontario, Canada.

Michael R. Allan

Parturition site selection by ungulates is believed to be influenced by forage abundance and concealment from predators. In 2011 and 2012, I used vaginal implant transmitters and movements to identify calving sites for 23 GPS collared elk (Cervus elaphus) from a restored herd. I tested the hypothesis that maternal elk used sites with higher forage and denser concealment compared to pre-calving sites at micro and macrohabitat levels. I detected no significant microhabitat differences from direct measurements of vegetation. At the macrohabitat scale, based on proximity of landcover classes, mean distances to hardwood forests was significantly less for calving (153 m) than pre-calving sites (198 m).

Site fidelity is hypothesized to offer security in terms of familiarity to an area. I tested the hypothesis that females demonstrated fidelity to their previous year's location during pre-partum, parturition, post partum, breeding and winter periods. Elk were more philopatric during parturition and post partum than during breeding. Compared to winter elk were more philopatric during pre-partum, parturition and post-partum periods. Expressed as distance between consecutive-year calving locations, site fidelity varied with 27% of females exhibiting high (<1 km), 18% moderate and 55% (>2.9 km) low fidelity.

I measured nearest-neighbour distances at calving time, exploring the hypothesis that females distance themselves from conspecifics. Elk increased the average distances

to collared conspecifics during parturition; however, sample sizes were small. This strategy might influence calving site selection.

Rapid movement prior to parturition, low site fidelity and spacing-out of females during parturition appear to be strategies to minimize predator risk and detection. Little evidence of selection for vegetation structure suggests this may not be limiting to these elk.

Author Keywords: calving, elk, fidelity, movement, parturition, selection

This study investigated the role of the JNK signaling pathway in cadmium-treated mouse embryo forelimb bud cells in vitro. Primary cultures of forelimb bud cells harvested at day 11 of gestation were pre-treated with JNK inhibitor SP600125, and incubated with or without CdCl2 for 15, 30, 60, 120 minutes and 24, 48 hours or 5 days. Endpoints of toxicity were measured through cell differentiation by Alcian Blue Assay and phosphorylation of JNK proteins by Western blot. The results demonstrated that, in the cell differentiation assay, inhibiting JNK activation by 20 μM SP600125 causes an enhanced toxic effect in limb cells and inhibits cell differentiation, whereas 2 μM decreases differentiated nodule numbers under both cadmium stress and normal conditions. In conclusion, the JNK pathway has an essential role in the differentiation processes of limb bud cells in normal growth conditions.

Author Keywords: Cadmium, Cell Signaling, JNK, Limbs, Mouse Embryo, Teratology

Models of partitioning, uptake, and toxicity of neutral organic chemicals in fish

Alena Kathryn Davidson Celsie

A novel dynamic fugacity model is developed that simulates the uptake of chemicals in fish by respiration as applies in aquatic toxicity tests. A physiologically based toxicokinetic model was developed which calculates the time-course of chemical distribution in four tissue compartments in fish, including metabolic biotransformation in the liver. Toxic endpoints are defined by fugacity reaching a 50% mortality value. The model is tested against empirical data for the uptake of pentachloroethane in rainbow trout and from naphthalene and trichlorobenzene in fathead minnows. The model was able to predict bioconcentration and toxicity within a factor of 2 of empirical data. The sensitivity to partition coefficients of computed whole-body concentration was also investigated. In addition to this model development three methods for predicting partition coefficients were evaluated: lipid-fraction, COSMOtherm estimation, and using Abraham parameters. The lipid fraction method produced accurate tissue-water partitioning values consistently for all tissues tested and is recommended for estimating these values. Results also suggest that quantum chemical methods hold promise for predicting the aquatic toxicity of chemicals based only on molecular structure.

Author Keywords: COSMOtherm, fish model, fugacity, Partition coefficient, tissue-water, toxicokinetics

Benthic macroinvertebrtes (BMI) are functionally important in aquatic ecosystems; as such, knowledge of their community structure and function is critical for understanding these systems. BMI were sampled from ten lakes in each of two regions of south-central Ontario to investigate which chemical and physical variables could be shaping their community structure and function. Ten Precambrian Shield lakes in the Muskoka-Haliburton region, and ten St Lawrence lowland lakes in the Kawartha lakes region were sampled. These lakes are geologically and chemically distinct, creating natural chemical and physical gradients within and between both regions. Community function was assessed using stable isotope analysis to elucidate carbon transfer dynamics (δ13C) and food web interactions (δ15N). It was predicted that the BMI from Shield lakes would have a δ13C signature indicative of allochthonous carbon subsidies, whereas the lowland lake BMI signatures would reflect autochthonous production. Additionally, it was predicted that the food web length (measured in δ15N units) would be different in Shield and lowland lakes. Both of these predictions were supported; however, the data indicate that δ13C signatures are more likely influenced by catchment geology (represented by bicarbonate concentration) than the extent of allochthony. The best predictor of food web length was found to be region. To assess BMI community structure, taxonomic richness, %EPT (% Ephemeroptera, Plecoptera, Trichoptera; a water quality index), and distribution of functional feeding groups were examined. Based on chemistry it was expected that the Shield lakes would be more speciose, and of greater water quality (relatively lower nutrient levels). These predictions were rejected; since there were no significant regional differences in taxonomic richness or biologically inferred water quality (%EPT). However, sediment size was found to best explain the variability in both metrics, with greater richness and %EPT found at sites with medium and small substrates than those with large substrates. Significant regional differences were found in the distribution of functional feeding groups. Most notably, there were significantly greater proportions of scrapers and shredders in the lowland and Shield lakes, respectively. Based on the feeding mechanisms of these invertebrates it can be inferred that allochthonous subsidies are likely of greater importance to Shield lake BMI communities than those of the lowland lakes; supporting the carbon transfer prediction. These findings provide insight about the structure and function of BMI communities from two dominant lake types in Ontario, and could be useful when determining how future chemical and physical changes will impact these communities.

Author Keywords: benthic macroinvertebrates, community function, community structure, Precambrian Shield, stable isotopes, St. Lawrence lowlands

Eastern wolves (Canis lycaon) have hybridized extensively with coyotes (C. latrans) and gray wolves (C. lupus) and are listed as a `species of special concern' in Canada. Previous studies have not linked genetic analysis with field data to investigate the mechanisms underlying Canis hybridization. Accordingly, I studied genetics, morphology, mortality, and behavior of wolves, coyotes, and hybrids in and adjacent to Algonquin Provincial Park (APP), Ontario. I documented 3 genetically distinct Canis types within the APP region that also differed morphologically, corresponding to putative gray wolves, eastern wolves, and coyotes. I also documented a substantial number of hybrids (36%) that exhibited intermediate morphology relative to parental types. I found that individuals with greater wolf ancestry occupied areas of higher moose density and fewer roads. Next, I studied intrinsic and extrinsic factors influencing survival and cause-specific mortality of canids in the hybrid zone. I found that survival was poor and harvest mortality was high for eastern wolves in areas adjacent to APP compared with other sympatric Canis types outside of APP and eastern wolves within APP. Contrary to previous studies of wolves and coyotes elsewhere, I hypothesized that all Canis types exhibit a high degree of spatial segregation in the Ontario hybrid zone. My hypothesis was supported as home range overlap and shared space use between neighboring Canis packs of all ancestry classes were low. Territoriality among Canis may increase the likelihood of eastern wolves joining coyote and hybrid packs and exacerbate hybridization. Canids outside APP modified their use of roads between night and day strongly at high road densities (selecting roads more at night), whereas they responded weakly at lower road densities (generally no selection). Individuals that survived exhibited a highly significant relationship between the difference in their night and day selection of roads and availability of roads, whereas those that died showed a weaker, non-significant response. My results suggest that canids in the unprotected landscape outside APP must balance trade-offs between exploiting benefits associated with secondary roads while mitigating risk of human-caused mortality. Overall, my results suggest that the distinct eastern wolf population of APP is unlikely to expand numerically and/or geographically under current environmental conditions and management regulations. If expansion of the APP eastern wolf population (numerically and in terms of its geographic distribution) is a conservation priority for Canada and Ontario, additional harvest protection in areas outside of APP may be required. If additional harvest protection is enacted, a detailed study within the new areas of protection would be important to document specific effects on eastern wolf population growth.

Author Keywords: Canis, coyotes, eastern wolves, hybridization, resource selection, survival

Silver nanoparticles (AgNP) released into aquatic environments could threaten natural bacterial communities and ecosystem services they provide. We examined natural lake bacterioplankton communities' responses to different exposures (pulse vs chronic) and types (citrate and PVP) of AgNPs at realistic environmental conditions using a mesocosm study at the Experimental Lakes Area. An in situ bioassay examined interactions between AgNPs and phosphorus loading. Bacterial communities exposed to high AgNP concentrations regardless of exposure or capping agent type accumulated silver. We observed increases in community production during additions of polyvinylpyrrolidone (PVP) -capped AgNPs and that site and nutrient-specific conditions are important to AgNPs toxicology in aquatic systems. Toxicological effects of AgNP are attenuated in natural conditions and differ from results from laboratory studies of AgNP toxicity. Our results demonstrate more studies are needed to fully assess the risk posed by these novel chemicals to the environment. This work could be useful in forming risk assessment policies which are largely based on lab studies and typically demonstrate strong toxic effects.

Author Keywords: bacterial production, bacterioplankton communities, ecological stoichiometry, Experimental Lakes Area, mesocosms, silver nanoparticles

In this thesis, aerosolization was studied as a possible means of water remediation for several environmentally relevant pharmaceutical pollutants, known for their persistence in wastewater effluent and potable water sources. Seven different pharmaceutical compounds and a well-known plasticizer were all shown to decrease considerably in concentration in aerosol that was produced and subsequently collected within a short time span. Strong evidence is presented that an enhanced rate of partitioning into the gas phase at the air-water interface of water droplets exists for every compound tested relative to that occurring in bulk solution. UV photolysis in aerosols was also explored and shown for sulfamethoxazole to be at least an order of magnitude faster in aerosols then in bulk solution. The implications towards both the environmental fate, and removal of these compounds from water sources is discussed.

Author Keywords: Aerosols, Air-water partitioning, Pharmaceuticals, Photolysis, Sulfamethoxazole

Microfibres, which are threadlike particles < 5 mm, are the most common type of microplastic reported in the environment. However, few studies have focused on their abundance in background natural environments. This study assessed the abundance of microfibres in rainfall samples (from four precipitation monitoring stations) and across three headwater lake catchments that were in remote, undeveloped areas, away from anthropogenic disturbance and anthropogenic emission sources (i.e., sites were background natural environments). Anthropogenic microfibres were observed in all samples using visual identification methods, with Raman spectroscopy confirming the presence of polyester film and synthetic pigments, e.g., indigo and hostasol green. The estimated annual average atmospheric deposition of microfibres was ~28,800 mf m-2. Meteorological variables, e.g., rain, wind direction, and relative humidity were correlated with the abundance of microfibres. The average abundance of microfibres in headwater lake catchments was 24 mf g-1 in moss, 0.70 mf m-3 in surface trawl, 9,690 mf m-3 in subsurface, 910 mf kg-1 in lake sediment and 576 mf kg-1 in lakeshore sediment.

Author Keywords: Atmospheric Deposition, Background Environments, Headwater Lake Catchments, Microfibres, Microplastics, Rainfall