Metcalfe, Chris D

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

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

Road runoff is a vector for the transport of potentially toxic chemicals into receiving waters. In this study, selected tire-derived chemicals were monitored in surface waters of rivers adjacent to two high traffic highways in the Greater Toronto Area in Ontario, Canada. Composite samples were collected from the Don River and Highland Creek in the GTA during 5 hydrological events that occurred in the period between early October 2019 and late March 2020, as well as an event in August 2020. Grab samples were collected from these rivers during a period of low flow in August 2020, as well as during a storm event in July of 2020. Analysis was performed using ultra-high pressure liquid chromatography with high resolution mass spectrometric detection (UHPLC-HRMS). Hexamethoxymethylmelamine (HMMM), a cross-linker of tire material, was detected at elevated concentrations (> 1 µg/L) during rain events in the fall and winter of 2019-20 and during a period of rapid snow melt in early March of 2020. These samples were also analyzed for the tire additive, 6PPD, and its oxidation by-product, 6PPD-quinone, as well as 1,3-diphenylguanidine (DPG). In many samples collected from the Don River and Highland Creek during storm events, the estimated concentrations of 6PPD-quinone exceeded the reported LC50 of 0.8 µg/L for Coho salmon exposed to this compound. Temporal samples collected at 3-hour intervals throughout rain event the October 2020 showed that there was a delay of several hours after the start of the event before these compounds reached their peak concentrations. In addition, 26 candidate transformation products and precursor compounds of HMMM were monitored; 15 of these compounds were detected in surface waters in the GTA. The maximum total concentration of this class of methoxymethylmelamine compounds in surface water samples was estimated to be 18 µg/L. There is limited knowledge about the properties of HMMM, its precursor contaminants, and its transformation compounds, as well as their fate in the environment. COSMO-RS solvation theory was used to estimate the physico-chemical properties of HMMM and its derivatives. Using the estimated values for these properties (e.g., solubility, vapour pressure, log Kow) as inputs to the Equilibrium Criterion (EQC) fugacity-based multimedia model, the compounds were predicted to readily partition into aqueous media, with mobility in water increasing with the extent of loss of methoxymethyl groups from HMMM. Overall, this study contributes to the growing literature indicating that potentially toxic tire-wear compounds are transported via road runoff into urban surface waters. In addition, this study provides insight into the environmental behaviour of HMMM and its transformation products.

Author Keywords: 6PPD-quinone, COSMOtherm, Fugacity, Hexamethoxymethylmelamine, Road runoff, Tire wear

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

Micropollutants are discharged into the aquatic environment with industrial and domestic wastewater and these compounds may cause toxic effects in aquatic organisms. In this study, the toxic effects to fish of micropollutants extracted from ozonated and nonozonated municipal wastewater effluent (MWWE) were measured in order to assess the effectiveness of ozonation in reducing toxicity. Juvenile rainbow trout (Oncorhynchus mykiss) injected with extracts prepared from ozonated MWWE had significantly reduced induction of plasma vitellogenin (VTG), significantly reduced hepatic total glutathione (tGSH) levels and an elevated oxidized-to-total glutathione (GSSG-to-tGSH) ratio. Exposure of Japanese medaka (Oryzias latipes) embryos to extracts prepared from both ozonated and non-ozonated MWWE resulted in elevated developmental toxicity in both treatment groups. These results indicate that wastewater treatment by ozonation reduces the estrogenicity of wastewater, but treatment may induce oxidative stress and embryonic developmental toxicity due to the production of toxic by-products.

Author Keywords: Estrogenicity, Micropollutants, Oxidative stress, Ozonation, Toxic by-products, Wastewater