Gueguen, Celine

This study assesses the ability of a unicellular protist, Euglena gracilis, to remove Cu and Ni from solution in mono- and bi-metallic systems. Living Euglena cells and non-living Euglena biomass were examined for their capacity to sorb metal ions. Adsorption isotherms were used in batch systems to describe the kinetic and equilibrium characteristics of metal removal. In living systems results indicate that the sorption reaction occurs quickly (<30 min) in both Cu (II) and Ni (II) mono-metallic systems and adsorption follows a pseudo-second order kinetics model for both metals. Sorption capacity and intensity was greater for Cu than Ni (p < 0.05) and were described by the Freundlich model. In bi-metallic systems sorption of both metals appears equivalent. In non-living systems sorption occurred quickly (10-30 min) and both Cu and Ni equilibrium uptake increased with a concurrent increase of initial metal concentrations. The pseudo-first-order model was applied to the kinetic data and the Langmuir and Freundlich models effectively described single-metal systems. The biosorption capacity of Cu (II) and) was 3x times greater than that of Ni (II). Sorption of one metal in the presence of relatively high concentrations of the other metal was supressed. Generally, it was found that living Euglena remove Cu and Ni more efficiently than non-living Euglena biomass in both mono- and bi-metallic systems. It is anticipated that this work should contribute to the identification of baseline uptake parameters and capacities for Cu and Ni by Euglena as well as to the increasing amount of research investigating sustainable bioremediation.

Author Keywords: accumulation, biosorption, Cu, Euglena gracilis, kinetics, Ni

As part of a mandate to control the spread of Stratiotes aloides (WS; water soldier) in the Trent Severn Waterway, the Ministry of Natural Resources (MNR) created a management plan to eradicate WS. However, one of the biggest challenges in eradicating WS or any invasive aquatic plant is the ability to estimate the extent of its spread and detect new populations. While current detection methods can provide acceptable detection, these methods often require extensive time and effort. The purpose of this thesis was to assess the use optical properties of WS and WS exudates for detection, in order to improve on current detection methods. The optical properties of WS were sampled at three different sites during three different seasons (spring, summer, and fall) by a) randomly sampling tissue from WS and the local plant community at each site, and recording the reflectance properties in a laboratory setting b) collecting dissolved organic matter (DOM) samples from plant incubations and river water in the field. Significant differences in the reflectance properties of WS were observed among samples from different sites and different sampling times; however, changes in fluorescence properties were only seasonal. Despite spatial differences in WS reflectance; WS was detectable using both hyperspectral and multispectral reflectance. When hyperspectral reflectance was used, significant differences between WS and the local plant community were found in June using two bands (i.e. bands 525 and 535, R 2 = 0.46 and 0.48, respectively). Whereas multispectral reflectance was significant different in October using the coastal and blue band. While WS produced a unique signal using both reflectance types, multispectral reflectance had a greater potential for detection. Its greater potential for detection was due to the reduced noise produced by background optical properties in October in comparison to June. DOM derived from WS was also characterized and compared with whole-river DOM samples in order to find unique markers for WS exudates in river samples. While similarities in DOM concentrations of WS exudates to Trent River water limited the ability to detect WS using compositional data, the ratio of C4/C5 components were compared in order to find components that were proportionally similar. Based on the results of this study multispectral and fluorescence techniques are better suited for the detection of a unique WS signature. The results derived from this work are intended to have practical applications in plant management and monitoring, DOM tracing, as well as remote sensing.

Author Keywords: Dissolved organic matter, Hyperspectral reflectance, Invasive species management, Multispectral reflectance, PARAFAC, Stratiotes aloides

Dissolved organic matter (DOM) is a mixture of molecules with dynamic structure and composition that are ubiquitous in aquatic systems. DOM has several important functions in both natural and engineered systems, such as supporting microorganisms, governing the toxicity of metals and other pollutants, and controlling the fate of dissolved carbon. The structure and composition of DOM determine its reactivity, and hence its effectiveness in these ecosystem functions.

While the structure, composition, and reactivity of riverine and marine DOM have been previously investigated, those of allochthonous DOM collected prior to exposure to microbes and sunlight have received scant attention. The following dissertation constitutes the first in-depth study of the structure, composition, and reactivity of allochthonous DOM at its point of origin (i.e. leaf leachates, LLDOM), as detected by measuring its size and optical properties. Concomitantly, novel chemometric methods were developed to interpret size-resolved data obtained using asymmetrical flow field-flow fractionation, including spectral deconvolution and the application of machine learning algorithms such as self-organizing maps to fluorescence data using a dataset of more than 1000 fluorescence excitation-emission matrices.

The size and fluorescence properties of LLDOM are highly distinct. Indeed, LLDOM was correctly classified as one of 13 species/sources with 92.5% accuracy based on its fluorescence composition, and LLDOM was distinguished from riverine DOM sampled from eight different rivers with 98.3% accuracy. Additionally, both fluorescence and size properties were effective conservative tracers of DOC contribution in pH-controlled mixtures of leaf leachates and riverine DOM over two weeks. However, the structure of LLDOM responded differently to pH changes for leaves/needles from different tree species, and for older needles. Structural changes were non-reversible.

Copper-binding strength (log K) differed for the different fluorescent components of DOM in a single allochthonous source by more than an order of magnitude (4.73 compared to 6.11). Biotransformation preferentially removed protein/polyphenol-like fluorescence and altered copper-binding parameters: log K increased from 4.7 to 5.5 for one fluorescent component measured by fluorescence quenching, but decreased from 7.2 to 5.8 for the overall DOM, as measured using voltammetry. The complexing capacity of DOM increased in response to biotransformation for both fluorescent and total DOM. The relationship between fluorescence and size properties was consistent for fresh allochthonous DOM, but differed in aged material.

Since the size and fluorescence properties of LLDOM are strikingly different from those of riverine DOM, deeper investigation into transformative pathways and mixing processes is required to elucidate the contribution of riparian plant species to DOM signatures in rivers.

Author Keywords: Analytical chemistry, Chemometrics, Dissolved organic matter (DOM), Field-flow fractionation, Fluorescence spectroscopy, Parallel factor analysis (PARAFAC)

This study assesses the bioavailable metal (Cu, Ni, Zn, Pb) species in the Athabasca-Mackenzie watersheds using diffusive gradient in thin films (DGT) devices. Metal toxicity is not only based on the concentration of metal in natural waters, but also on the nature of metal species. Four main forms in aquatic systems are: free ion, inorganic species, DOM bound (humic) species and metal colloidal species. The free ion and inorganic species and very small humic species are known as DGT-labile species and, are considered to be more bioavailable to micro-organisms due to the size and thus may be toxic to microorganisms. In this study, DGT devices were applied to (1) monitor the DGT-labile metal species in the lower Athabasca River and the Mackenzie River watershed and (2) assess the DGT-labile metal concentrations on temporal and spatial scales. In the lower Athabasca River, comparison between the DGT results and the Windermere Humic Acid Model (WHAM) calculation indicated good agreements for all metals when the precipitated iron(III) hydroxide was assumed as an active binding surface. No significant variations in labile species were found over 2003-2012 (RAMP database) despite the development of oil sands. In the Mackenzie River, no significant difference in DGT-labile metal concentrations and DOC concentrations was found in yearly basis 2012-2014. Only DOC was lower in August (6.98 and 3.85 ppm, respectively; p< 0.05) due to dilution from heavy rain events. Spatially, DGT-labile Cu and Ni in the downstream Mackenzie River were higher than upstream (1.79 and 0.58 ppb for Cu, 1.68 and 0.77 ppb for Ni, 4.06 and 6.91 ppm for DOC; p < 0.05). Overall the in situ measurements of metals constitute a benchmark for future studies in water quality and be helpful in environmental management in Alberta and the Northwest Territories in Canada.

Author Keywords: Athabasca River, DGT, Mackenzie River, Speciation, Trace Metal, WHAM

A bedrock fracture hosting arsenic (As) contaminated groundwater was suspected to be transported to Ramsey Lake, a drinking water resource for more than 50,000 residents of Sudbury, Ontario. A high resolution, spatial, water quality mapping technique using an underwater towed vehicle (UTV) was used to identify sources of upwelling groundwater into lake water and localize the upwelling As contaminated groundwater vent site. The top 7 cm of lake sediments (in-situ) at this vent site were observed to adsorb 93 % of the dissolved As, thus inhibiting lake water quality degradation from this contaminant source. Sediment samples from this location were used in laboratory experiments to assess the potential for this system to become a source of As contamination to Ramsey Lake water quality and elucidate As(III) fractionation, transformation and redistribution rates and processes during aging. Arsenic speciation is important because As(III) has been shown to be more toxic than As(V). To accomplish this a sequential extraction procedure (SEP) that maintains As(III) and As(V) speciation in (sub)oxic sediments and soils was validated for the operationally defined fractions: easily exchangeable, strongly sorbed, amorphous Fe oxide bound, crystalline Fe oxide bound, and the residual fraction for total As because the characteristics of the reagents required to extract the final fraction do not maintain As species.

Batch reaction experiments using sediment spiked with As(III) or As(V) and aged for up to 32 d were sequentially extracted and analysed for As(III) and As(V). Consecutive reaction models illustrate As(III) is first adsorbed to the sediment then oxidized to As(V). Fractionation analyses show As(III) most rapidly adsorbs to the easily exchangeable fraction where it is oxidized and redistributes to the strongly sorbed and amorphous Fe oxide bound fractions. Oxidation of As(III) adsorbed to the amorphous and crystalline Fe oxide bound fractions is less efficient and possibly inhibited. Select samples amended with goethite provide evidence supporting Mn(II) oxidation is catalyzed by the goethite surface, thus increasing As(III) oxidation by Mn(III/IV) complexed with the strongly sorbed fraction. Although As immobilization through groundwater sediment interactions may be inhibited by increased ion activity, particularly phosphate or lake eutrophication, this threat in Ramsey Lake is likely low.

Author Keywords: arsenic, fractionation, modelling, redistribution, speciation, water quality mapping

Owen Sound Bay, which is located within the traditional territory of the Saugeen Ojibway Nation (SON), is contaminated as a result of historical industrial and shipping activity. Gross contamination of the sediments in the inner part of the Bay (i.e., Owen Sound Harbour) includes high concentrations of polycyclic aromatic hydrocarbons (PAHs) and other organic compounds, as well as metals that may pose a risk to the SON fishery for lake whitefish (Coregonus clupeaformis). However, evaluating the environmental risks posed by contaminated sediments is a challenge, as these risks are dependent upon several factors and require multiple lines of evidence. Including Indigenous communities in environmental risk assessment and the management of those risks is vital for sustaining ecosystem integrity, as well as respecting Treaty Rights. In this study, a risk assessment framework was developed that included several risk assessment tools used in Western science and also encompassed the concerns and values of the SON, including the application of SON-ecological knowledge. Methods for risk evaluation included gathering lines of evidence though community workshops, as well as field sampling in the Bay to determine the concentrations of PAHs and other organic contaminants in sediments and in the water column. Laboratory studies of toxicity to early life stages of lake whitefish and Japanese medaka (Oryzias latipes) and sediment disturbance simulations to evaluate biological responses in juvenile lake whitefish were also completed as lines of evidence. The results indicate that leaving the harbour "as is" without a thorough analysis of remediation options fails to address the concerns of the people within the SON communities. Overall, this research demonstrated a successful process for developing a collaborative risk assessment framework that recognizes the sovereignty of Indigenous peoples and promotes Nation-to-Nation decision making.

Author Keywords: biomarkers, Coregonus clupeaformis, Indigenous knowledge, polycyclic aromatic hydrocarbons, risk assessment, source tracking

Microalgae are a promising source of valuable compounds relevant to biofuels, biomaterials, nutraceuticals as well as animal and human nutriment. Unfortunately, low cell density and slow growth result in reduced economic feasibility. Heterotrophic cell culturing using an organic carbon source in lieu of light has proven to be an effective alternative to photobioreactors; however, further improvement may be possible with the addition of growth promoting phytohormones. In this thesis, growth and endogenous hormone profiles in heterotrophic cultures of Euglena gracilis were evaluated using glucose and ethanol as carbon sources. Cytokinin (CK) and abscisic acid (ABA) were quantified by HPLC-ESI-MS/MS and compared to culture growth dynamics. Exogenous phytohormones treatments were also conducted to determine if they may mitigate nutrient reduction and improve growth. Phytohormones CK and ABA were purified and analyzed at seven points along the growth curve in small scale (250 mL flasks, 100 mL working volume) cultures. Among the key findings was that ethanol cultures undergoing exponential growth, primarily synthesize freebase cytokinins (FBCKs) and methylthiol-cytokinins (MeSCKs), while not producing detectable levels of ABA. In exogenous studies, dry biomass was positively influenced with the addition of exogenous ABA; however, the most notable result revealed the ability of transZ to alleviate nutrient reduction. These findings suggest a communication network in algal culture using FBCKs and MeSCKs, as well as the potential for exogenous hormone supplementation to increase growth rates and overall biomass productivity.

Author Keywords: abscisic acid, cytokinin, Euglena gracilis, heterotrophy, phytohormones