Graduate Theses & Dissertations

Comparing Biological Responses to Contaminants in Darters (Etheostoma spp.) Collected from Rural and Urban Regions of the Grand River Watershed, Ontario
Urban and agricultural activities may introduce chemical stressors, including contaminants of emerging concern (CECs) and current use pesticides (CUPs) into riverine systems. The objective of this study was to determine if fish collected from sites in a river show biomarkers of exposure to these classes of contaminants, and if the biomarker patterns vary in fish collected from urbanized and agricultural sites. The watershed selected for this study was the Grand River in southern Ontario, which transitions from areas dominated by agricultural land use in the north to highly urbanized locations in the southern part of the watershed. Rainbow darters (Etheostoma caerluem) and fantail darters (Etheostoma flabellare) were collected from the Grand River in June, 2014 for biomarker analysis from two urbanized sites and three agricultural sites (n=20 per site). Over the same period of time, Polar Organic Chemical Integrative Samplers (POCIS) were deployed for 2 weeks at each site to monitor for the presence of CUPs and CECs. The amounts of the target compounds accumulated on POCIS, determined using LC-MS/MS were used to estimate the time weighted average concentrations of the contaminants at each site. Data on the liver somatic index for darters indicate site-specific differences in this condition factor (p<0.05). Significant differences in the concentrations of thiobarbituric acid reactive substances (TBARS) in gill tissue (p<0.05) indicate differences in oxidative stress in fish collected from the various sites. Measured concentrations of ethoxyresorufin-O-deethylase (EROD) in liver tissue were significantly different between sites (p<0.05), indicating differences in CYP1A metabolic activity. Finally, acetylcholinesterase (AChE) activity in brain tissue was significantly different between fish from rural and urban sites (p<0.05). The analysis of these biomarkers indicates that fish may be experiencing different levels of biological stress related to different land uses. These data may be useful in developing mitigation strategies to reduce impacts on fish and other aquatic organisms in the watershed. Author Keywords: AChE, Biomarker, Darter, EROD, POCIS, TBARS
Effect of the neonicotinoid imidacloprid on embryogenesis and anuran survivorship in frog virus 3 infected tadpoles
Exposure of pre-metamorphic amphibians to neonicotinoid insecticides may be contributing to the global decline in amphibian populations. In this study, anuran embryos and tadpoles of the African clawed frog (Xenopus laevis) and the North American leopard frog (Lithobates pipiens) were used to determine the effects of embryonic exposure to neonicotinoids. In addition, Xenopus was used to determine if prolonged exposure to neonicotinoids influenced tadpole sensitivity to frog virus 3 (FV3). Exposure of anuran embryos to concentrations of the neonicotinoid insecticide, imidacloprid, ranging from 1 -20 ppm induced a concentration dependent increase in malformations of the retina in Xenopus embryos. However, similar responses were not observed with embryos of leopard frogs. Exposure of Xenopus tadpoles to 500 ppb concentration of imidacloprid followed by challenge with FV3 showed that pesticide exposure unexpectedly decreased the rates of mortality, although total mortalities by the end of the experiment were not significantly different from controls. This unexpected observation may be attributed to a reduced inflammatory response induced by exposure to imidacloprid. Despite the low acute toxicity of neonicotinoid insecticides to vertebrates, these studies indicate that exposure to this class of insecticides causes sublethal effects in anuran species during early life stages. Author Keywords: embryogenesis, Lithobates pipiens, neonicotinoid, ranavirus, tadpole, Xenopus laevis
Effects of Opioids on the Development and Reproductive Capacity of Japanese Medaka, Oryzias latipes
Opioid drugs are among the microcontaminants that are discharged with domestic wastewater into the aquatic environment with the potential to affect the development and reproductive capacity of aquatic organisms. To study the effects of exposure of fish to opioid drugs, Japanese medaka (Oryzias latipes) were exposed over a full life cycle to environmentally relevant (i.e. ng/L) concentrations of two opioids, codeine and fentanyl. Acute exposure of early life stage (ELS) of medaka to codeine resulted in slightly extended times to hatch, but no increase in embryo mortalities or reduced hatching success. Chronic exposure to codeine at a nominal concentration of 25,000 ng/L over a full life cycle interfered with growth, reducing the weight of the fish at maturity. Life cycle treatments with codeine at all test concentrations resulted in a significant reduction in the number of eggs produced in reproductive trials, as well as a reduction in the numbers of mature oocytes in adult females. High treatments with codeine also increased the percentage of immature sperm cells in adult males. Life cycle exposures to codeine also resulted in reduced whole-body concentrations of several hypothalamic-pituitary-gonadal (HPG) axis hormones, including reduced levels of luteinizing hormone in male and female fish and a reduction in 11-ketotestosterone in males and 17β-estradiol in females. Fentanyl did not affect reproduction or HPG hormones at the exposure concentrations tested, but high fentanyl exposures increased the mortality rate among ELS’s of the F1 generation offspring from life cycle exposed adults. The results of this thesis contribute to the literature on the environmental impacts of microcontaminants of wastewater origin and, the potential for effects in fish exposed to opioids.Keywords: Opioids, microcontaminants, fentanyl, codeine, sexual development, reproduction. Author Keywords: codeine, fentanyl, microcontaminants, opioids, reproduction, sexual development
Effects of Silver Nanoparticles on Lake Bacterioplankton
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
Fate and Effects of Silver Nanoparticle Addition in a Lake Ecosystem
The potential release of nanoparticles into aquatic environments is raising global concerns. As antimicrobials, silver nanoparticles (AgNPs) are among the most prominent form in use. Despite this, their fate, long-term toxicity, and ecological relevance have yet to be investigated largely under natural settings with seasonality and environmental complexity. To better understand the environmental significance, we released AgNPs into Lake 222 at the Experimental Lakes Area over two years. AgNPs remained suspended in the water column and were detected throughout the lake and in the lower food web. Total Ag concentrations ranged from below 0.07 to 18.9 μg L-1 in lake water, and were highly dynamic seasonally both in the epilimnion and hypolimnion depending on the physical, chemical and biological conditions of the lake. Approximately 60% of the measured Ag mass in October was present in the sediment in 2014 and 50% in 2015 demonstrating relatively high sedimentation and removal from the water column. During winter months, Ag was largely absent in the water column under the ice. After ice melt and before summer stratification, Ag concentrations increased in the lake suggesting AgNPs may not be tightly bound to the sediment and are able re-enter the water column during spring mixing events. Despite temporal variation, total Ag was highly synchronous across spatial locations for both years, indicating rapid dispersal upon lake entry. When investigating AgNP sizes using spICPMS, size distributions were similar across spatial locations, with the 40-60 nm size class constituting approximately 60% of all particles identified. Large aggregates (>100 nm) and dissolved Ag were infrequently detected within the lake. Ag accumulated in the lower food web ranging from 0.27-16.82 μg Ag mg C-1 in the bacterioplankton and 0.17-6.45 μg Ag mg C-1 in algae (particulate fraction). Partial least squares models revealed the highest predictors of Ag accumulation were dissolved nutrients including DOC, TDN, TDP in bacterioplankton. Major predictors for particulate Ag included temperature, dissolved oxygen, and sampling date. The diversity of predictors among biological compartments emphasizes the importance of understanding the role of environmental complexity within the lower food web. Despite Ag accumulation we did not detect strong negative effects on the lake food web. An increase in particulate and bacterioplankton chlorophyll-a occurred after addition in contrast to reference lakes, which may indicate a hormetic response to low dose AgNP concentrations. Our findings provide the first whole-lake perspective regarding Ag fate and toxicity, suggesting small scale experiments may overestimate environmental responses. Author Keywords: Ecotoxicity, Fate, Lower food web, Silver Nanoparticles, Whole-lake addition
Molecular Composition of Dissolved Organic Matter Controls Metal Speciation and Microbial Uptake
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
Responses of Primary Producers and Grazers to Silver Nanoparticle Exposure
The increasing production and use of silver nanoparticles (AgNPs) raise concerns on environmental exposure and impact. A large scale in situ enclosure study was conducted at the Experimental Lakes Area to determine the effect of AgNPs on natural phytoplankton and zooplankton communities. This study investigated AgNPs of varying concentrations (4, 16 and 64 μg/L), dosing regimens (chronic vs. pulse), and capping agents (poly-vinyl pyrrolidone vs. citrate). Phytoplankton communities were influenced only by the natural limnological properties of the system signifying tolerance to AgNPs. Zooplankton community structure significantly changed with AgNP concentration and dosing regimen indicating AgNP sensitivity. A microcosm study investigating the effect of AgNPs and phosphorus-dosed periphyton before and after grazing by two benthic invertebrate species (snails and caddisfly larvae) showed reduced periphyton stoichiometry with AgNP exposure. Grazers foraged less on silver dosed periphyton indicating a preferential choice in food quality. Phosphorus reduced the detrimental effects of AgNPs across all conditions. These studies verify the need for in situ experimental designs to fully investigate the effects of AgNPs and their interaction with environmental factors, multiple species assemblages, and across trophic levels. Author Keywords: benthic invertebrate, Experimental Lakes Area, periphyton, phytoplankton, silver nanoparticles, zooplankton
Wastewater Impacts on Freshwater Mussels and Water Quality in a Tributary of the Lower Grand River in Southwestern Ontario, Canada
The main goal of this thesis was to assess the potential impacts of discharges of treated effluent from a small facultative sewage lagoon serving approximately 300 residents of the Mississaugas of the Credit First Nation to freshwater mussel populations in Boston Creek, a small tributary of the lower Grand River. The current resident mussel populations inhabiting Boston Creek were assessed using semi-qualitative visual surveying methods. In addition to various population level observations, other possible point and non-point influences on water quality in Boston Creek were identified. Following this, Lasmigona costata mussels were deployed as biomonitoring organisms alongside passive samplers during the October 2017 lagoon discharge period. Time weighted average (TWA) concentrations of select Contaminants of Emerging Concern (CECs) and Polycyclic Aromatic Hydrocarbons (PAHs) were estimated from levels of these compounds accumulated on passive samplers to understand the influence of wastewater on water quality in Boston Creek. Finally, mussel tissues were analyzed for various biomarkers of exposure to contaminants. Population surveys indicated that Boston Creek supports a plentiful and diverse community of freshwater mussels and may be a refuge for the Species of Special Concern, Villosa iris. Passive sampling revealed that most PAHs measured were present at concentrations below detection limits, while CECs were typically detected at relatively low concentrations (ng/L) directly downstream of the lagoon discharge. Biomarker responses detected in Lasmigona costata generally could not be attributed to exposure to the lagoon effluent but these data may indicate response to other point and non-point sources of pollution that could be affecting resident freshwater mussel populations in Boston Creek. The mussels surveyed in Boston Creek may be displaying community level effects of exposure to other sources of pollution in the area. The results of this thesis will help in establishing water quality guidelines in the lower Grand River watershed that will assist in the recovery strategy for freshwater mussel species at risk in Ontario. Author Keywords: Biomarkers, Biomonitoring, CECs, First Nations, Freshwater Mussels, SAR
collaborative ecotoxicological risk assessment of in-place pollutants in Owen Sound Bay, Lake Huron within the Saugeen Ojibway Nation Territory
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
effects of particulate matter on the fate and toxicity of silver nanoparticles
As an emerging contaminant, the antimicrobial agent silver nanoparticles (AgNPs) have been receiving considerable attention to determine their potential effects to aquatic ecosystems. However, estimates of aquatic consumer survivorship and other toxicological endpoints vary considerably among experiments, largely due to the environment in which the test takes place. Throughout this thesis I aim to understand which natural environmental variables impact toxicity to the common aquatic consumer Daphnia. I focus on the effects of particulate matter as it may play a role in animal nutrition as well as interact with AgNPs. I explore particulate matter’s effect on survival in the complex matrices including other natural variables that could impact toxicity. I conduct a series of complimentary field and laboratory studies to understand how particles impact AgNP toxicity and how those interactions vary within whole lake ecosystems. Using laboratory studies, I establish that algal particles mitigate the toxic effects of AgNPs on Daphnia survival through removing Ag from the water column and that phosphorus increases this effect. Using wild Daphnia and lake water, I demonstrate the ability of particulate matter to mitigate toxicity in complex natural settings. It was also one of the major predictors of AgNP toxicity to Daphnia along with dissolved organic carbon and daphnid seasonal health. Finally, using a whole lake AgNP addition experiment, I demonstrate that particles and AgNPs interact variably in the lake. Silver from AgNPs binds to particles and is removed to the sediments through the actions of settling particles without impacting the dynamics of living communities. Overall, I am able to demonstrate that the natural components of lake ecosystems, especially particulate matter, are able to mitigate the effects of AgNPs in lake ecosystems to a point where they likely will be never pose a threat to the survivorship of aquatic consumers such as Daphnia. Author Keywords: Daphnia, ecotoxicity, particulate matter, Silver nanoparticles, whole lake experiment

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