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Aquatic receiving environments have long been used to dilute municipal wastewater effluents (MWWE) which are the largest discharge by volume into the aquatic environment in Canada. These treated effluents are a complex mixture of environmental contaminants that includes natural and synthetic hormones, pharmaceuticals, industrial chemicals, nutrients and ammonia. Discharge of MWWE may lead to serious problems in aquatic environments such as eutrophication, hypoxia as well as increased occurrence of disease and toxicity in resident aquatic biota. Reproductive impairment in fish has also been widely reported in association with exposure to wastewaters. Recently, concerns have been raised about the potential for municipal effluents to cause endocrine disruption in fish and other organisms. The effects of MWWE on fish and fish populations in Canada are currently poorly understood. The overall objective of this thesis is to contrast the impact of MWWE discharged into two Canadian rivers on sentinel fish species across levels of biological organization ranging from biochemical responses to changes at the fish community level. Results from these studies support the development of robust effects-based biological monitoring approaches to assess the effectiveness of regulations and remedial actions for minimizing the effects of MWWE. Understanding the temporal changes in physiological and reproductive parameters across the annual cycle of a sentinel species is necessary to optimize biomonitoring programs. The annual variability in terms of survival, reproduction and energy storage in the Greenside Darter (Etheostoma blennioides), a potential sentinel species for the Grand River, Ontario, was documented at a reference site across two years. Variation in energy storage and reproductive development indicated by somatic indices (i.e., relative organ size) and steroid production suggest that biomonitoring can be optimized for this species by sampling in late fall or early spring (pre-spawning). With this new knowledge, field studies conducted with small bodied species, including Rainbow Darter (E. caeruleum), Brook Stickleback (Culaea inconstans) and Fathead Minnow (Pimephales promelas) have demonstrated that, when sampled during the appropriate season (e.g., pre-spawning), they can be used as effective biomonitoring tools to detect fish responses associated with exposure to MWWEs. Two sentinel fish species, Rainbow Darter and Greenside Darter, were then used to examine the impact of two MWWE discharges on fish in the Grand River, Ontario, relative to reference sites in two seasons (fall and early spring). Fish responses, in terms of energy storage (condition factor, liver size), energy utilization (gonadosomatic indices) and reproduction (in vitro sex steroid production, cellular development and intersex) were assessed at each site. Both sentinel species were longer and heavier downstream of the wastewater outfalls. However, these larger fish did not demonstrate consistent increases in condition and liver somatic indices. MWWE-exposed male Rainbow and Greenside Darters had impaired capacity to produce androgens in vitro, lower gonadosomatic indices and altered sperm cell staging. Exposed female fish also had impaired capacity to produce estrogens in vitro, however, they did not demonstrate differences in oocyte development. Male Rainbow and Greenside Darters collected downstream of both MWWE discharges showed increased incidence of intersex (33 - 100%) in contrast to very low occurrences of this condition in upstream agricultural and urban reference sites. This increased incidence of intersex coincided with reductions in gonadosomatic indices and capacity to produce steroids, demonstrating the ability of MWWE to alter the reproductive systems of these fish. The fish communities downstream of the the MWWE outfalls demonstrated differences in abundance, diversity, and species composition when compared to reference sites. MWWE exposed sites had few of the darter species that dominate the fish community at reference sites. More mobile fish species such as suckers (Catostomidae spp.) and sunfish (Centrarchidae spp.) were more common downstream of the outfalls, with occurances becoming more pronounced downstream of the second sewage discharge. Wascana Creek, Saskatchewan, downstream of the wastewater treatment plant for the City of Regina can be up to 100% treated municipal wastewater. Brook Stickleback and Fathead Minnow exhibited delayed spawning and altered gonadal development downstream of the wastewater outfall. Exposed male Fathead Minnows were feminized, having lower expression of secondary sexual characteristics (i.e., loss of nuptial tubercles, dorsal pad, and dorsal fin dot) and induction of the female egg-yolk precursor protein, vitellogenin. Fathead Minnows also showed cellular damage to the gills and kidneys. These responses indicate exposure to a variety of environmental contaminants in the effluent such as ammonia as well as endocrine disruptors. The potential effect of MWWE discharges in these two Canadian watersheds on fish responses was demonstrated across various levels of biological organization including reduced sex steroid production, altered gonadal development, reduction in gonadosomatic indices, delayed spawning, and changes in fish assemblages. An effects-based monitoring approach using sentinel species can be successfully applied to detect changes associated with MWWE outfalls, as long as sampling of sentinel species is conducted during optimal time periods (i.e., when somatic indices are maximized and variability among individuals is minimized). MWWE can impair the reproductive potential of fish beyond a threshold where impacts are expressed at higher levels of organization such as populations or communities. It is essential to make mechanistic linkages between responses at different levels to determine the overall potential impact of effluents on fish. The collection of responses across multiple levels of biological organization can complement and support development of biomonitoring approaches that are focused at the population and community levels such as those being proposed for MWWE in Canada.

Aquatic receiving environments have long been used to dilute municipal wastewater effluents (MWWE) which are the largest discharge by volume into the aquatic environment in Canada. These treated effluents are a complex mixture of environmental contaminants that includes natural and synthetic hormones, pharmaceuticals, industrial chemicals, nutrients and ammonia. Discharge of MWWE may lead to serious problems in aquatic environments such as eutrophication, hypoxia as well as increased occurrence of disease and toxicity in resident aquatic biota. Reproductive impairment in fish has also been widely reported in association with exposure to wastewaters. Recently, concerns have been raised about the potential for municipal effluents to cause endocrine disruption in fish and other organisms. The effects of MWWE on fish and fish populations in Canada are currently poorly understood. The overall objective of this thesis is to contrast the impact of MWWE discharged into two Canadian rivers on sentinel fish species across levels of biological organization ranging from biochemical responses to changes at the fish community level. Results from these studies support the development of robust effects-based biological monitoring approaches to assess the effectiveness of regulations and remedial actions for minimizing the effects of MWWE. Understanding the temporal changes in physiological and reproductive parameters across the annual cycle of a sentinel species is necessary to optimize biomonitoring programs. The annual variability in terms of survival, reproduction and energy storage in the Greenside Darter (Etheostoma blennioides), a potential sentinel species for the Grand River, Ontario, was documented at a reference site across two years. Variation in energy storage and reproductive development indicated by somatic indices (i.e., relative organ size) and steroid production suggest that biomonitoring can be optimized for this species by sampling in late fall or early spring (pre-spawning). With this new knowledge, field studies conducted with small bodied species, including Rainbow Darter (E. caeruleum), Brook Stickleback (Culaea inconstans) and Fathead Minnow (Pimephales promelas) have demonstrated that, when sampled during the appropriate season (e.g., pre-spawning), they can be used as effective biomonitoring tools to detect fish responses associated with exposure to MWWEs. Two sentinel fish species, Rainbow Darter and Greenside Darter, were then used to examine the impact of two MWWE discharges on fish in the Grand River, Ontario, relative to reference sites in two seasons (fall and early spring). Fish responses, in terms of energy storage (condition factor, liver size), energy utilization (gonadosomatic indices) and reproduction (in vitro sex steroid production, cellular development and intersex) were assessed at each site. Both sentinel species were longer and heavier downstream of the wastewater outfalls. However, these larger fish did not demonstrate consistent increases in condition and liver somatic indices. MWWE-exposed male Rainbow and Greenside Darters had impaired capacity to produce androgens in vitro, lower gonadosomatic indices and altered sperm cell staging. Exposed female fish also had impaired capacity to produce estrogens in vitro, however, they did not demonstrate differences in oocyte development. Male Rainbow and Greenside Darters collected downstream of both MWWE discharges showed increased incidence of intersex (33 - 100%) in contrast to very low occurrences of this condition in upstream agricultural and urban reference sites. This increased incidence of intersex coincided with reductions in gonadosomatic indices and capacity to produce steroids, demonstrating the ability of MWWE to alter the reproductive systems of these fish. The fish communities downstream of the the MWWE outfalls demonstrated differences in abundance, diversity, and species composition when compared to reference sites. MWWE exposed sites had few of the darter species that dominate the fish community at reference sites. More mobile fish species such as suckers (Catostomidae spp.) and sunfish (Centrarchidae spp.) were more common downstream of the outfalls, with occurances becoming more pronounced downstream of the second sewage discharge. Wascana Creek, Saskatchewan, downstream of the wastewater treatment plant for the City of Regina can be up to 100% treated municipal wastewater. Brook Stickleback and Fathead Minnow exhibited delayed spawning and altered gonadal development downstream of the wastewater outfall. Exposed male Fathead Minnows were feminized, having lower expression of secondary sexual characteristics (i.e., loss of nuptial tubercles, dorsal pad, and dorsal fin dot) and induction of the female egg-yolk precursor protein, vitellogenin. Fathead Minnows also showed cellular damage to the gills and kidneys. These responses indicate exposure to a variety of environmental contaminants in the effluent such as ammonia as well as endocrine disruptors. The potential effect of MWWE discharges in these two Canadian watersheds on fish responses was demonstrated across various levels of biological organization including reduced sex steroid production, altered gonadal development, reduction in gonadosomatic indices, delayed spawning, and changes in fish assemblages. An effects-based monitoring approach using sentinel species can be successfully applied to detect changes associated with MWWE outfalls, as long as sampling of sentinel species is conducted during optimal time periods (i.e., when somatic indices are maximized and variability among individuals is minimized). MWWE can impair the reproductive potential of fish beyond a threshold where impacts are expressed at higher levels of organization such as populations or communities. It is essential to make mechanistic linkages between responses at different levels to determine the overall potential impact of effluents on fish. The collection of responses across multiple levels of biological organization can complement and support development of biomonitoring approaches that are focused at the population and community levels such as those being proposed for MWWE in Canada.

A thesis submitted to the School of Graduate Studies in partial fulfilment of the requirements for the degree of Master of Science in the graduate academic unit of Biology / Microparticles enter aquatic environments through many sources, including wastewater treatment plants (WWTPs), but their uptake by aquatic organisms is poorly understood. Freshwater bivalves accumulate multiple contaminants, making them potential bioindicators for MP pollution. This study aims to understand the abundance and characteristics of microparticles that accumulate in wild bivalves. Samples were collected from 5 locations along the Grand River (Ontario, Canada) in 2021-2022, including 3 municipal WWTPs where both an upstream and downstream site were sampled. At each site, fingernail clams (Sphaeriidae, n=5 composite samples), flutedshell mussels (Lasmigona costata, n=10), and surface water (n=3) were sampled. Within the mussels, the gill, digestive gland, and hemolymph tissues were targeted and compared. Microparticles were isolated and quantified via stereomicroscopy but have not yet been confirmed as plastic; as such, they will be referred to herein as microparticles. Fibers were the dominant morphology and clear, blue, and black were the most common colours, but there were some differences among sites and sample types. Most microparticles were between 80 μm and 1 mm in length. Fingernail clams contained the highest microparticle counts per mass of tissue at 35.5 ± 29.4 microparticles/g, mussel tissues ranged from 4.3 ± 4.2 microparticles/mL to 6.5 ± 8.1 microparticles/g, and water samples contained the lowest counts at 0.0055 ± 0.0028 microparticles/mL. Elevated microparticle counts at downstream sites were only seen in mussel gills and not other bivalve tissues. Surface water samples did not show elevated counts downstream of the WWTPs and microparticle exposures were similar across sites. This study provides baseline data for future monitoring and informs toxicity studies to fully assess the risk of microparticles to vulnerable freshwater bivalves and other aquatic organisms. It also suggests microparticles in freshwater bivalves are coming from sources in addition to WWTPs and are ubiquitous in the Grand River. / Thesis / Master of Science (MSc) / Microplastics are found in nearly every environment, especially freshwater ecosystems. These plastics come from a variety of sources, and this study focuses on assessing the characteristics of microparticles in freshwater clams and mussels (bivalves) that have been exposed to municipal wastewaters. Bivalves and water samples were collected from 5 locations along the Grand River (Ontario, Canada) in 2021-2022, and microparticles were extracted and analyzed from each sample. Fibers were the most abundant type of microparticle, with colours consisting mostly of clear, blue, and black. Clams had the highest number of microparticles per mass of tissue collected and the lowest counts were found in water samples. Higher microparticle counts were only seen in one (mussel gill) of the four tissues from bivalves collected downstream of wastewater outfalls. This study provides baseline data on microparticle characteristics in freshwater bivalves and will guide future studies on the toxicity of microparticles to these animals.

Time-consuming and expensive benthic surveys are currently the most common means of determining impacts on invertebrates and fish habitat in lotic systems. We propose using the rate of chitobiase production by benthic invertebrates as a complement for determining impacts on freshwater systems. We successfully modified the existing assay to a microplate approach for high throughput analysis of chitobiase activity. Next we conducted two case studies in: 1) the Dead Horse Creek, Manitoba, to determine if changes in chitobiase could detect impacts on the benthic community from wastewater effluent and; 2) in Snake and Kinch Creeks, Manitoba to see if chitobiase could be used to assess fish habitat quality. In both cases, we observed no strong relationships between chitobiase and traditional metrics (e.g., abundance, biomass, diversity). We recommend further studies concerning the timing of chitobiase release in lotic systems and assessments of its use in mesocosm and microcosm toxicity studies / May 2016

A avaliação da presença de compostos orgânicos voláteis nos efluentes líquidos é importante para atendimento à legislação vigente e proteção do sistema de esgotamento sanitário e da saúde ocupacional. Objetivos: o uso de técnica rápida de identificação e quantificação de compostos orgânicos voláteis do grupo BTEX presentes em amostras de efluentes líquidos através das emissões gasosas permite verificar quais delas não atendem à legislação vigente, através de faixas de concentração de interesse. A possibilidade de efetuar em campo essa avaliação promove maior agilidade nas ações de controle da qualidade do efluente bem como da exposição ocupacional. Também permite racionalizar os custos de monitoramento através da redução das amostras enviadas aos laboratórios. Materiais e métodos: os compostos avaliados causam agravos à saúde e, por serem altamente voláteis, possuem altos percentuais de distribuição no ar. O benzeno é comprovadamente cancerígeno, não existindo limites seguros para exposição a esse composto. A legislação brasileira fornece os subsídios necessários para o controle da produção, usos e disposição de BTEX. Há ações em andamento para um melhor controle e monitoramento da exposição ocupacional ao benzeno, em especial, e também ações para o monitoramento das populações expostas a ele. Com o objetivo de efetuar leituras no campo que identifiquem e quantifiquem os compostos do grupo BTEX, optou-se pela utilização de espectrofotômetro de infravermelho portátil. Resultados: os resultados das leituras das emissões desses compostos foram comparados com os resultados das análises de BTEX efetuadas nos efluentes líquidos. Para tanto, foram considerados os parâmetros existentes na legislação relativos à exposição ocupacional e à legislação de lançamento de efluentes. Conclusão: a avaliação dos resultados permite dizer que é factível a utilização da leitura das emissões gasosas para identificar a presença de BTEX em amostras de efluentes líquidos e que essas leituras podem quantificar valores que ultrapassem as faixas de concentração de interesse, através dos modelos de regressão identificados nessa avaliação. Para a construção de modelos de regressão mais confiáveis entre as concentrações das emissões gasosas e das concentrações no efluente líquido, são necessários estudos complementares. / The evaluation of the presence of volatile organic compounds in wastewater is important for compliance with current regulations, protection of the wastewater sewer systems and occupational health. Objectives: Identification and quantification of volatile organic compounds of BTEX group present in samples of liquid effluents through their gaseous emissions, allows checking which of them do not meet the current regulation, through concentration ranges of interest. The possibility of performing this evaluation in the field promotes greater flexibility in efforts to control the quality of the effluent as well as occupational exposure. It also allows streamlining the monitoring costs through reduction of liquid samples to be sent to laboratories. Materials and methods: The compounds evaluated cause health problems and for being highly volatile have high percentages of distribution in the air. Benzene is a proven carcinogen and there are no safe limits for exposure to this compound. Brazilian regulations provide the necessary subsidies for production control, uses and disposal of BTEX subsidies needed to control the production, uses and disposal of BTEX. There are ongoing activities to better control and monitoring of occupational exposure and also actions for monitoring populations exposed to benzene. An infrared spectrophotometer was used in air analyses to identify BTEX. Results: The results of the compounds emissions scans were compared with the results of BTEX analyzes in wastewater. For both parameters were considered in the regulation relating to occupational exposure and the rules of effluent discharge. Conclusion: The evaluation of the results allows saying that it is feasible to use gaseous emissions scans to identify the presence of BTEX in wastewater samples and they can indicate scans that exceed the concentration ranges of interest, through mathematical models this assessment. For mathematical models that provide precise curves of the correlation between the concentrations of gaseous emissions and liquid effluent concentrations, complementary studies are needed.

A avaliação da presença de compostos orgânicos voláteis nos efluentes líquidos é importante para atendimento à legislação vigente e proteção do sistema de esgotamento sanitário e da saúde ocupacional. Objetivos: o uso de técnica rápida de identificação e quantificação de compostos orgânicos voláteis do grupo BTEX presentes em amostras de efluentes líquidos através das emissões gasosas permite verificar quais delas não atendem à legislação vigente, através de faixas de concentração de interesse. A possibilidade de efetuar em campo essa avaliação promove maior agilidade nas ações de controle da qualidade do efluente bem como da exposição ocupacional. Também permite racionalizar os custos de monitoramento através da redução das amostras enviadas aos laboratórios. Materiais e métodos: os compostos avaliados causam agravos à saúde e, por serem altamente voláteis, possuem altos percentuais de distribuição no ar. O benzeno é comprovadamente cancerígeno, não existindo limites seguros para exposição a esse composto. A legislação brasileira fornece os subsídios necessários para o controle da produção, usos e disposição de BTEX. Há ações em andamento para um melhor controle e monitoramento da exposição ocupacional ao benzeno, em especial, e também ações para o monitoramento das populações expostas a ele. Com o objetivo de efetuar leituras no campo que identifiquem e quantifiquem os compostos do grupo BTEX, optou-se pela utilização de espectrofotômetro de infravermelho portátil. Resultados: os resultados das leituras das emissões desses compostos foram comparados com os resultados das análises de BTEX efetuadas nos efluentes líquidos. Para tanto, foram considerados os parâmetros existentes na legislação relativos à exposição ocupacional e à legislação de lançamento de efluentes. Conclusão: a avaliação dos resultados permite dizer que é factível a utilização da leitura das emissões gasosas para identificar a presença de BTEX em amostras de efluentes líquidos e que essas leituras podem quantificar valores que ultrapassem as faixas de concentração de interesse, através dos modelos de regressão identificados nessa avaliação. Para a construção de modelos de regressão mais confiáveis entre as concentrações das emissões gasosas e das concentrações no efluente líquido, são necessários estudos complementares. / The evaluation of the presence of volatile organic compounds in wastewater is important for compliance with current regulations, protection of the wastewater sewer systems and occupational health. Objectives: Identification and quantification of volatile organic compounds of BTEX group present in samples of liquid effluents through their gaseous emissions, allows checking which of them do not meet the current regulation, through concentration ranges of interest. The possibility of performing this evaluation in the field promotes greater flexibility in efforts to control the quality of the effluent as well as occupational exposure. It also allows streamlining the monitoring costs through reduction of liquid samples to be sent to laboratories. Materials and methods: The compounds evaluated cause health problems and for being highly volatile have high percentages of distribution in the air. Benzene is a proven carcinogen and there are no safe limits for exposure to this compound. Brazilian regulations provide the necessary subsidies for production control, uses and disposal of BTEX subsidies needed to control the production, uses and disposal of BTEX. There are ongoing activities to better control and monitoring of occupational exposure and also actions for monitoring populations exposed to benzene. An infrared spectrophotometer was used in air analyses to identify BTEX. Results: The results of the compounds emissions scans were compared with the results of BTEX analyzes in wastewater. For both parameters were considered in the regulation relating to occupational exposure and the rules of effluent discharge. Conclusion: The evaluation of the results allows saying that it is feasible to use gaseous emissions scans to identify the presence of BTEX in wastewater samples and they can indicate scans that exceed the concentration ranges of interest, through mathematical models this assessment. For mathematical models that provide precise curves of the correlation between the concentrations of gaseous emissions and liquid effluent concentrations, complementary studies are needed.

With the ever increasing demand for potable water due to the continued increase in population coupled with the threat of California’s current drought, water will remain a limited resource that must be managed responsibly. In order to strategically plan and manage water use in the most beneficial manner, water providers must take into account all sources of water, including recycled water and their applications. Recycled water as a source for supplementing high quality potable water is a sustainable strategy that will prove to be an essential tactic in any water management plan. The purpose of this project is to emphasize the importance of supplementing potable water in the City of Banning by discussing the characteristics of California’s current water drought; evaluating the City of Banning’s available water supplies and current water demand; discussing the Soil Aquifer Treatment process; summarizing California’s regulations related to recycled water; and discussing the quality of recycled water available at NP-1, an unequipped City owned water well, by examining water quality testing on water samples taken from NP-1. Analysis of water available at NP-1 showed that with additional disinfection, the water pumped from NP-1 could meet the recycled water requirements in order to be used on a local golf course. The local golf course is currently being irrigated with potable water, which would be supplemented with the recycled water from NP-1.

Municipal wastewater effluent (MWWE) is an important source of pollution in the aquatic environment impacting fish. MWWE is a complex mixture of chemicals including pharmaceuticals, personal care products, industrial chemicals and pesticides. A link between reproductive endocrine disruption and MWWE exposure has been established in fish, but less is known about the effects of MWWE on non-reproductive endocrine disruption. The overall objective of this thesis was to examine the impacts of MWWE exposure on the stress response and intermediary metabolism in rainbow trout (Oncorhynchus mykiss). In fish, the primary adaptive organismal stress response involves the activation of hypothalamic-sympathetic-chromaffin axis to produce catecholamines, predominantly epinephrine, and the hypothalamic-pituitary-interrenal (HPI) axis to produce cortisol. Both of these hormones play a key role in elevating plasma glucose levels that is essential to fuel the increased energy demand associated with stress. Along with the organismal stress response, the cellular stress response, involving the synthesis of a suite of heat shock proteins (hsps), also plays an important role in protecting cellular protein homeostasis in response to stressors, including toxicants. The impact of MWWE on stress-related pathways were identified using a low-density trout cDNA microarray enriched with genes encoding for proteins involved in endocrine-, stress- and metabolism-related processes. This was further confirmed by assessing plasma hormone and metabolite levels and stress-related targeted genes and proteins expression and enzyme activities in select tissues in rainbow trout. Studies were carried out in controlled field (caging) and laboratory experiments to examine the impacts of MWWE on stress and tissue-specific metabolic responses in rainbow trout. Further in vitro studies using rainbow trout hepatocytes in primary cultures were carried out to investigate the mechanism of action of two pharmaceuticals, atenolol and venlafaxine, found in relatively high concentrations in MWWE in impacting the stress-mediated glucose response. In caged fish, MWWE exposure significantly elevated plasma cortisol and glucose concentrations, and altered the mRNA abundance of a number of stress-related genes, hormone receptors, glucose transporter 2 and genes related to immune function. When fish were exposed to an acute handling stress following a 14 d exposure to MWWE, the cortisol response was abolished and the glucose response was attenuated. The effects on cortisol did not correlate with changes in the expression of genes involved in cortisol biosynthesis, but were associated with an increase in hepatic glucocorticoid receptor (GR) protein expression. Upon further investigation in controlled laboratory studies, MWWE exposure elevated constitutive hsp 70 and hsp90 expression after 8 d exposure, which correlated with a decrease in glycogen levels in the liver in fish exposed to a high concentration of MWWE compared to control fish, pointing to a MWWE-induced increase in liver energy demand. By 14 d, glycogen stores were replenished, and this was commensurate with increases in liver gluconeogenic capacity, including increases in the activities of phosphoenolpyruvate carboxykinase (PEPCK) and alanine aminotransferase (AlaAT), along with a decrease in liver GR expression. In the heart, GR protein expression increased in treated fish, and the activity of pyruvate kinase increased, indicating an increase in glycolytic capacity. Subjecting the MWWE exposed fish to a secondary handling disturbance (acute stress) led to an attenuated plasma cortisol and glucose response compared to the control group. This corresponded with a reduced liver gluconeogenic capacity and a lower liver and heart glycolytic capacities, reflecting a disturbance in the energy substrate repartitioning that is essential to cope with stress. While it is difficult to establish causative agents from a complex mixture such as MWWE, the two pharmaceutical that were tested impacted glucose production. Specifically, atenolol and venlafaxine disrupted the epinephrine-induced glucose production, but did not modify cortisol-mediated glucose production in trout hepatocytes. The suppression of epinephrine-mediated glucose production by atenolol and venlafaxine was abolished by cAMP analogue (8-bromo cAMP) or glucagon (a metabolic hormone that increases glucose production). This suggests that both drugs disrupt β-adrenoceptor signaling, while it remains to be determined if the response is receptor isoform-specific. Altogether MWWE exposure disrupts the organismal and cellular stress responses in trout. Key targets for MWWE impact leading to the impaired cortisol and metabolic responses to stress include liver and heart GR expression, liver gluconeogenic capacity, and liver, heart and gill glycolytic capacities. Most significantly, MWWE impairs the ability to metabolically adjust to a secondary acute stressor, which is an important adaptive process that is integral to successful stress performance. From an environmental stand-point, long-term exposure to MWWE will lead to reduced fitness and will compromise the capacity of fish to cope with additional stressor, including escape from predators.

Fast paced population growth in urban areas of southern Ontario is putting increased pressure on the surrounding aquatic environment. The City of Guelph uses the Speed River to assimilate its municipal wastewater effluent. With a projected 57% population increase in the watershed by 2031, the assimilative capacity of the river may be challenged in the coming years. The Guelph Wastewater Treatment Plant uses tertiary treatment methods greatly reducing ammonia, suspended solids and phosphate concentrations in the effluent. However there are still impacts detectable related to excessive nutrients released into this relatively small river (6th order) which promotes algae and aquatic macrophyte growth. There is also concern about a variety of emerging contaminants that may enter the river and impact the health of the ecosystem. The research in this thesis examined the seasonal and spatial variability and extent of the impacts of the wastewater effluent on the riffle fish communities in the Speed River. Stable isotope signatures (δ13C and δ15N) were used to understand the changes in the dominant benthic fish species, Rainbow Darters (Etheostoma caeruleum) and Greenside Darters (E. blennioides), relative to changes in invertebrate signatures and their abundance. Rainbow Darters were extremely abundant relative to Greenside Darters at the site immediately downstream of the effluent outfall, particularly in August. The benthic invertebrate community was distinctly different downstream of the effluent outfall, especially in the summer, with a reduced abundance of Elmidae beetle larvae and increased abundance of isopods (Caecidotea intermedius) compared to upstream. δ13C and δ15N of the two darters species were similar at all sites in May and July, but in August and October Rainbow Darter signatures were more enriched in the two heavier isotopes at sites downstream of the effluent outfall. The vast majority of invertebrate taxa sampled were also enriched at the downstream sites. An analysis of Rainbow and Greenside Darter stomach contents revealed that Rainbow Darters incorporated more isopods and other invertebrates in their diet, especially at the immediate downstream sites suggesting that they are more adaptable to the altered downstream environment. The feeding habits of Greenside Darters appear to change between July and August in response to changes in habitat and food availability. They are potentially consuming food organisms with less enriched isotopic signatures, which results in their isotopic signatures not rising during these months like most of the invertebrates and other fish. Alternatively, the Greenside Darters may move across the stream to feed on invertebrates that remain unexposed to the wastewater effluent. These impacts, although subtle, may be a reflection of the Speed River ecosystem being compromised by nutrient inputs from the wastewater effluent. With the impending increase in demand on the treatment plant (e.g., population growth), ongoing treatment and infrastructure improvements may be needed in the future to maintain the current ecosystem structure.