Effects of metformin on wild fathead minnows (Pimephales promelas) using in-lake mesocosms in a boreal lake ecosystem.

Due to its widespread use for the treatment of Type-2 diabetes, metformin is routinely detected in surface waters globally. Laboratory studies have shown that environmentally relevant concentrations of metformin can adversely affect the health of adult fish, with effects observed more frequently in males. However, the potential risk to wild fish populations has yet to be fully elucidated and remains a topic of debate. To explore whether environmentally relevant metformin exposure poses a risk to wild fish populations, the present study exposed wild fathead minnows (Pimephales promelas) to 5 or 50 µg/L metformin via 2 m diameter in-lake mesocosms deployed in a natural boreal lake in Northern Ontario at the International Institute for Sustainable Development - Experimental Lakes Area (IISD-ELA). Environmental monitoring was performed at regular intervals for 8-weeks, with fish length, weight (body, liver and gonad), condition factor, gonadosomatic index, liver-somatic index, body composition (water and biomolecules) and hematocrit levels evaluated at test termination. Metabolic endpoints were also evaluated using liver, brain and muscle tissue, and gonads were evaluated histologically. Results indicate that current environmental exposure scenarios may be sufficient to adversely impact the health of wild fish populations. Adult male fish exposed to metformin had significantly reduced whole body weight and condition factor and several male fish from the high-dose metformin had oocytes in their testes. Metformin-exposed fish had altered moisture and lipid (decrease) content in their tissues. Further, brain (increase) and liver (decrease) glycogen were altered in fish exposed to high-dose metformin. To our knowledge, this study constitutes the first effort to understand metformin's effects on a wild small-bodied fish population under environmentally relevant field exposure conditions.

The acute toxicity of bitumen-influenced groundwaters from the oil sands region to aquatic organisms.

The extraction of surface mined bitumen from oil sands deposits in northern Alberta, Canada produces large quantities of liquid tailings waste, termed oil sands process-affected water (OSPW), which are stored in large tailings ponds. OSPW-derived chemicals from several tailings ponds migrating past containment structures and through groundwater systems pose a concern for surface water contamination. The present study investigated the toxicity of groundwater from near-field sites adjacent to a tailings pond with OPSW influence and far-field sites with only natural oil sands bitumen influence. The acute toxicity of unfractionated groundwater and isolated organic fractions was assessed using a suite of aquatic organisms (Pimephales promelas, Oryzias latipes, Daphnia magna, Hyalella azteca, Lampsilis spp., Ceriodaphnia dubia, Hexagenia spp., and Vibrio fischeri). Assessment of unfractionated groundwater demonstrated toxicity towards all invertebrates in at least one far-field sample, with both near-field and far-field samples with bitumen influence toxic towards P. promelas, while no toxicity was observed for O. latipes. When assessing the unfractionated groundwater and isolated organic fractions from near-field and far-field groundwater sites, P. promelas and H. azteca were the most sensitive to organic components, while D. magna and L. cardium were most sensitive to the inorganic components. Groundwater containing appreciable amounts of dissolved organics exhibited similar toxicities to sensitive species regardless of an OSPW or natural bitumen source. The lack of a clear distinction in relative acute toxicities between near-field and far-field samples indicates that the water-soluble chemicals associated with bitumen are acutely toxic to several aquatic organisms. This result, combined with the similarities in chemical profiles between bitumen-influenced groundwater originating from OSPW and/or natural sources, suggests that the industrial bitumen extraction processes corresponding to the tailings pond in this study are not contributing unique toxic substances to groundwater, relative to natural bitumen compounds present in groundwater flow systems.

Comparative Effects of Embryonic Metformin Exposure on Wild and Laboratory-Spawned Fathead Minnow (Pimephales promelas) Populations.

Metformin is routinely detected in aquatic ecosystems because of its widespread use as a treatment for Type 2 diabetes. Laboratory studies have shown that exposure to environmentally relevant concentrations of metformin can alter metabolic pathways and impact the growth of early life stage (ELS) fish; however, it is unknown whether these effects occur in wild populations. Herein, we evaluate whether findings from laboratory studies are representative and describe the relative sensitivities of both populations. Duplicate exposures (0, 5, or 50 µg/L metformin) were conducted using wild- and lab-spawned fathead minnow (Pimephales promelas) embryos. Apart from the water source, exposure conditions remained constant. Wild embryos were exposed to previously dosed lake water to account for changes in bioavailability, while reconstituted freshwater was used for the laboratory study. Developmental metformin exposure differentially impacted the growth and morphology of both cohorts, with energy dyshomeostasis and visual effects indicated. The fitness of wild-spawned larvae was impacted to a greater extent relative to lab-spawned fish. Moreover, baseline data reveal important morphological differences between wild- and lab-spawned ELS fatheads that may diminish representativeness of lab studies. Findings also confirm the bioavailability of metformin in naturally occurring systems and suggest current exposure scenarios may be sufficient to negatively impact developing fish.

Temperature modulates the impacts of wastewater exposure on the physiology and behaviour of fathead minnow.

Municipal wastewater treatment plant (WWTP) effluent is a substantial source of pollution in aquatic habitats that can impact organisms across multiple levels of biological organization. Even though wastewater effluent is discharged continuously all year long, its impacts across seasons, specifically during winter, have largely been neglected in ecotoxicological research. Seasonal differences are of particular interest, as temperature-driven metabolic changes in aquatic organisms can significantly alter their ability to respond to chemical stressors. In this study, we examined the effects of multiple levels of wastewater effluent exposure (0, 25, or 50% treated effluent) on the physiological and behavioural responses of adult fathead minnow (Pimephales promelas) at temperatures simulating either summer (20 °C) or winter (4 °C) conditions. At 20 °C, wastewater exposure posed a metabolic cost to fish, demonstrated by higher standard metabolic rate and was associated with increased haematocrit and a reduction in boldness. In contrast, fish exposed to wastewater at 4 °C experienced no change in metabolic rate but performed fewer social interactions with their conspecifics. Taken together, our results demonstrate that wastewater exposure can lead to metabolic and behavioural disruptions, and such disruptions vary in magnitude and direction depending on temperature. Our findings highlight the importance of studying the interactions between stressors, while also underscoring the importance of research during colder periods of the year to broaden and deepen our understanding of the impacts of wastewater contamination in aquatic ecosystems.

Transformation of bitumen-derived naphthenic acid fraction compounds across surface waters of wetlands in the Athabasca Oil Sands region.

Bitumen is extracted from oil sands in the Athabasca Oil Sands region (AOSR) of Alberta, Canada. Much of the bitumen-derived toxicity in mine waste is attributable to naphthenic acid fraction compounds (NAFCs). Mines in the AOSR are required to be returned to a natural state after closure; thus, cost-effective strategies are needed to reduce toxicity from NAFCs. Previous studies have demonstrated the capability of constructed wetlands to attenuate NAFCs. However, the capacity of wetlands in the natural environment to degrade and transform NAFCs to less toxic components is poorly understood. To better understand the spatial distribution and fate of NAFCs in natural wetlands, samples were collected across the surfaces of two mature opportunistic wetlands near active oil sands mines. The first wetland has a well-defined surface flow pathway and inflows affected by overburden containing lean bitumen ore. The second wetland, in contrast, is a stagnant water body with raw bitumen visible along its edges. For the wetland with a well defined flow path, NAFCs decreased in concentration down gradient, while oxidized NAFCs constituted a greater proportion of NAFCs with increase in flow path. Likewise there was a decrease in the molecular weights of NAFCs, similar to trends observed in constructed wetland treatment systems. In comparison, NAFCs were more uniformly distributed across the relatively stagnant wetland. Overall, these data provide new evidence that mature opportunistic wetlands in the AOSR can promote the degradation and oxidation of bitumen-derived naphthenic acids into less toxic compounds.

A Deep Dive into the Complex Chemical Mixture and Toxicity of Tire Wear Particle Leachate in Fathead Minnow.

The ecological impact of tire wear particles in aquatic ecosystems is a growing environmental concern. We combined toxicity testing, using fathead minnow (Pimephales promelas) embryos, with nontarget high-resolution liquid chromatography Orbitrap mass spectrometry to characterize the toxicity and chemical mixture of organic chemicals associated with tire particle leachates. We assessed: 1) exposure to tire particle leachates after leaching for 1-, 3-, and 10-d; and 2) the effect of the presence and absence of small tire particulates in the leachates. We observed a decrease in embryonic heart rates, hatching success, and lengths, as well as an increase in the number of embryos with severe deformities and diminished eye and body pigmentation, after exposure to the leachates. Overall, there was a pattern whereby we observed more toxicity in the 10-d leachates, and greater toxicity in unfiltered leachates. Redundancy analysis showed that several benzothiazoles and aryl-amines were correlated with the toxic effects observed in the embryos. These included benzothiazole, 2-aminobenzothiazole, 2-mercaptobenzothiazole, N,N'-diphenylguanidine, and N,N'-diphenylurea. However, many other chemicals characterized as unknowns are likely to also play a key role in the adverse effects observed. Our study provides insight into the types of chemicals likely to be important toxicological drivers in tire leachates, and improves our understanding of the ecotoxicological impacts of tire wear particles. Environ Toxicol Chem 2022;41:1144-1153. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Chronic Embryo-Larval Exposure of Fathead Minnows to the Pharmaceutical Drug Metformin: Survival, Growth, and Microbiome Responses.

Metformin is a glucose-lowering drug commonly found in municipal wastewater effluents (MWWEs). The present study investigated the chronic effects of metformin in early-life stages of the fathead minnow (Pimephales promelas). Endpoints assessed were growth, survival, and deformities. The larval gut microbiome was also examined using 16 S ribosomal RNA gene amplicon sequencing to determine microbial community composition and alpha and beta diversity. Eggs and larvae were exposed to metformin measured concentrations (mean [standard deviation]) of 0.020 (0.017) µg/L (for controls) and 3.44 (0.23), 33.6 (1.6), and 269 (11) µg/L in a daily static-renewal setup, with 20 embryos per beaker. The low and middle metformin exposure concentrations represent river and MWWE concentrations of metformin. To detect small changes in growth, we used 18 replicate beakers for controls and 9 replicates for each metformin treatment. Over the 21-d exposure (5 d as embryos and 16 d posthatch [dph]), metformin did not affect survival or growth of larval fish. Hatch success, time to hatch, deformities in hatched fry, and survival were similar across all treatments. Growth (wet wt, length, and condition factor) assessed at 9 and 16 dph was also unaffected by metformin. Assessment of the microbiome showed that the larvae microbiome was dominant in Proteobacteria and Firmicutes, with small increases in Proteobacteria and decreases in Firmicutes with increasing exposure to metformin. No treatment effects were found for microbiome diversity measures. Control fish euthanized with the anesthetic tricaine methane sulfonate had decreased alpha diversity compared to those sampled by spinal severance. This experiment demonstrates that metformin at environmentally relevant concentrations (3.44 and 33.6 µg/L) and at 10 times MWWE concentrations (269 µg/L) does not adversely affect larval growth or gut microbiome in this ubiquitous freshwater fish species. Environ Toxicol Chem 2022;41:635-647. © 2021 Her Majesty the Queen in Right of Canada. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. Reproduced with the permission of the Minister of Environment and Climate Change Canada.

Polycyclic aromatic compounds in the Canadian Environment: Aquatic and terrestrial environments.

Polycyclic aromatic compounds (PACs) are ubiquitous across environmental media in Canada, including surface water, soil, sediment and snowpack. Information is presented according to pan-Canadian sources, and key geographical areas including the Great Lakes, the Alberta Oil Sands Region (AOSR) and the Canadian Arctic. Significant PAC releases result from exploitation of fossil fuels containing naturally-derived PACs, with anthropogenic sources related to production, upgrading and transport which also release alkylated PACs. Continued expansion of the oil and gas industry indicates contamination by PACs may increase. Monitoring networks should be expanded, and include petrogenic PACs in their analytical schema, particularly near fuel transportation routes. National-scale roll-ups of emission budgets may not expose important details for localized areas, and on local scales emissions can be substantial without significantly contributing to total Canadian emissions. Burning organic matter produces mainly parent or pyrogenic PACs, with forest fires and coal combustion to produce iron and steel being major sources of pyrogenic PACs in Canada. Another major source is the use of carbon electrodes at aluminum smelters in British Columbia and Quebec. Temporal trends in PAC levels across the Great Lakes basin have remained relatively consistent over the past four decades. Management actions to reduce PAC loadings have been countered by increased urbanization, vehicular emissions and areas of impervious surfaces. Major cities within the Great Lakes watershed act as diffuse sources of PACs, and result in coronas of contamination emanating from urban centres, highlighting the need for non-point source controls to reduce loadings.

Assessing natural recovery from contaminants in a river using sediment chemistry and toxicity from different depth ranges.

To determine whether natural recovery was occurring in a depositional area of the St. Marys River (Ontario, Canada) known as East Bellevue Marine Park (EBMP), sediment was collected from two depth ranges, 0-5 cm and 0-10 cm, and subjected to a series of laboratory toxicity tests and chemical analysis. Toxicological responses (survival, growth, reproduction, development) of four benthic invertebrates and the fathead minnow were compared at test vs. reference sites using univariate and multivariate (ordination) techniques. Temporal trends in sediment chemistry and invertebrate toxicity were examined with time series data from 2008 through to 2018. Polycyclic aromatic hydrocarbons (PAHs; ≤ 37 mg/kg) and petroleum hydrocarbons (PHCs; ≤ 6266 mg/kg) were elevated in EBMP compared to reference sites (PAHs, ≤ 1.6 mg/kg; PHCs ≤ 180 mg/kg). Comparatively, the 0-5 cm sediment layer had lower concentrations of all contaminants than the 0-10 cm layer at three of four test sites. Over time, contaminant concentrations have mostly remained stable or have decreased. There were no significant differences in survival, growth, or development of the larval fish in EBMP compared to the upstream reference sites, and no differences between sampling depths. However, most EBMP sediments were toxic to invertebrates, driven by reduced reproduction by the worm Tubifex and reduced survival by the amphipod Hyalella. Among habitat variables, a combination of different classes of compounds based on ordination scores (PHCs, oil and grease, metals) was most strongly correlated to toxicological response. There was little to no difference in toxicity between sampling depths based on integrated endpoint response; however, individual endpoints showed mostly greater toxicity from exposure to the 0-10 cm layer. Over time, toxicity has mostly remained stable or showed improvement. These results provided some positive indications that gradual natural recovery is occurring in EBMP.

Characterization of naphthenic acid fraction compounds in water from Athabasca oil sands wetlands by Orbitrap high-resolution mass spectrometry.

Classical naphthenic acids (NAs) are known to be primary aquatic toxicants of concern in the Athabasca oil sands region (AOSR), and are a component of naphthenic acid fraction compounds (NAFCs). Recent studies conducted in the AOSR have examined metals and polycyclic aromatic hydrocarbons in regional wetlands. However, few studies have described NAs and/or NAFCs in AOSR wetlands. To address this gap, we examined NAFC profiles in the water of different wetlands in the AOSR, including naturalized borrow pits (i.e., abandoned pits created by excavation of road-building materials), and opportunistically-formed wetlands associated with reclamation activities. For comparison, NAFC profiles from these wetlands were compared to an opportunistic wetland formed from tailings pond dyke seepage. Samples were prepared using solid-phase extraction and analyzed using negative-ion high-resolution Orbitrap mass spectrometry. Principal component analyses (PCA) revealed patterns to the NAFC profiles in the wetlands. The first distinct grouping of wetlands included water bodies associated with reclamation activities that are located on and/or adjacent to mining overburden. One other wetland, HATS5w, separated from all other wetlands during PCA, and had a unique NAFC profile; detailed examination of NAFCs revealed HATS5w contained the heaviest (i.e., high m/z components) and most unsaturated NAFCs among study locations, demonstrating the usefulness of high-resolution mass spectrometry for characterizing individual wetlands. The NAFCs of HATS5w are also substantially different from bitumen-derived inputs in overburden-adjacent opportunistic wetlands. Collectively, the NAFC profiles presented provide new information on background levels of polar bitumen-derived organics in AOSR wetlands.

Treatment of oil sands process affected waters by constructed wetlands: Evaluation of designs and plant types.

Constructed wetland treatment systems (CWTS)s can be used to treat various wastewaters. The main constituent in oil sands process-affected water (OSPW) with uncertain treatment by CWTS are naphthenic acid fraction compounds (NAFC)s. The NAFCs are also among the primary contributors of toxicity to aquatic organisms. While there is preliminary evidence that some CWTSs are capable of treating OSPW for future potential discharge, there is little information comparing the effectiveness and efficiencies of different CWTS designs. Obtaining large volumes of OSPW for testing can be difficult, and while it is known that synthetic NAFCs are simpler and have different toxicity than OSPW-NAFCs, it is unknown whether they could serve as a proxy for optimization of CWTS design and operation. This study presents a comprehensive comparison of CWTS performance operated with both synthetic OSPW and OSPW for four CWTS designs differing in plant type, aeration, flow path, water depth, and substrate type. This study evaluated the potential biodegradation of NAFCs including: (1) decrease in total NAFC concentration, (2) shifts in Ox-NAFC fractions from O2- to O3-, O4-, and O5-NAFC, (3) decrease in carbon number, (4) decrease of the double bond equivalencies (DBE), and (5) change in toxicity of the waters to test organisms. CWTS planted with Sedge achieved the greatest extent of NAFC treatment and detoxification regardless of design. Although CWTSs planted with Cattail and Bulrush also degraded NAFCs and decreased toxicity, a greater hydraulic retention time was required, and the total extent of treatment was less than the CWTSs planted with Sedge. While synthetic OSPW was more toxic and experienced faster degradation rates, it showed similar trends to OSPW in terms of CWTS design efficiencies and function. Although synthetic OSPW would not be appropriate for modelling or scaling of CWTSs, it can be useful for testing designs and operating conditions.

Lethal and sublethal toxicity of perfluorooctanoic acid (PFOA) in chronic tests with Hyalella azteca (amphipod) and early-life stage tests with Pimephales promelas (fathead minnow).

Perfluoroalkyl substances (PFAS), including perfluorooctanoic acid (PFOA), are industrial chemicals that are of concern due to their environmental presence, persistence, bioaccumulative potential, toxicity, and capacity for long-range transport. Despite a large body of research on environmental exposure, insufficient chronic aquatic toxicity data exist to develop water quality targets for clean-up of federal contaminated sites in Canada. Thus, our objective was to assess the aqueous toxicity of PFOA in chronic tests with Hyalella azteca (amphipod) and early-life stage tests with Pimephales promelas (fathead minnow). Toxicity data were analyzed based on measured PFOA concentrations. Amphipod exposures were 42 d (0.84-97 mg/L) and examined survival, growth, and reproduction. Fathead minnow exposures were 21 d (0.010-76 mg/L), which encompassed hatching (5 d) and larval stages until 16 d post-hatch; endpoints included hatching success, deformities at hatch, and larval survival and growth. Amphipod survival was significantly reduced at 97 mg/L (42-d LC50 = 51 mg/L), but growth and reproduction were more sensitive endpoints (42-d EC50 for both endpoints = 2.3 mg/L). Fathead minnows were less sensitive than amphipods, exhibiting no significant effects in all endpoints with the exception of uninflated swim bladder, which was significantly higher at 76 mg/L (15%) than controls (0%). Maximum concentrations of PFOA are generally in the ng/L range in global surface waters, but can reach the µg/L range in close proximity to major source inputs; therefore, environmental concentrations are well below those that caused toxicity in the current study. Our data will provide valuable information with which to assess the risk of PFOA at contaminated sites, and to set a target for site remediation.

Polycyclic aromatic compounds (PACs) in the Canadian environment: A review of sampling techniques, strategies and instrumentation.

A wide variety of sampling techniques and strategies are needed to analyze polycyclic aromatic compounds (PACs) and interpret their distributions in various environmental media (i.e., air, water, snow, soils, sediments, peat and biological material). In this review, we provide a summary of commonly employed sampling methods and strategies, as well as a discussion of routine and innovative approaches used to quantify and characterize PACs in frequently targeted environmental samples, with specific examples and applications in Canadian investigations. The pros and cons of different analytical techniques, including gas chromatography - flame ionization detection (GC-FID), GC low-resolution mass spectrometry (GC-LRMS), high performance liquid chromatography (HPLC) with ultraviolet, fluorescence or MS detection, GC high-resolution MS (GC-HRMS) and compound-specific stable (δ13C, δ2H) and radiocarbon (Δ14C) isotope analysis are considered. Using as an example research carried out in Canada's Athabasca oil sands region (AOSR), where alkylated polycyclic aromatic hydrocarbons and sulfur-containing dibenzothiophenes are frequently targeted, the need to move beyond the standard list of sixteen EPA priority PAHs and for adoption of an AOSR bitumen PAC reference standard are highlighted.

Increased Temperature and Turbulence Alter the Effects of Leachates from Tire Particles on Fathead Minnow (Pimephales promelas).

Tire particles are of concern as a stressor due to the combination of their chemical constituents, high emission rates, and global distribution. Once in the environment, they will interact with physical parameters (e.g., UV, temperature). The interaction of chemical pollution with changing physical environmental parameters is often underestimated in ecotoxicology. Here, we investigate the role of temperature, mechanical stress (i.e., turbulence), UV, and CO2 on the effects of tire leachates on fish. Two samples of tire particles were exposed to four different levels of each physical stressor. A toxicological assessment was performed with fathead minnow embryos assessing five end points (hatching success, time to hatch, length, deformities, and heart rate). Results showed that variations of temperature and mechanical stress affect the toxicological impact of tire leachates. Zn and/or polycyclic aromatic hydrocarbons (pyrene, phenanthrene, chrysene, benzo[a]pyrene, anthracene, naphthalene, fluoranthene, and benzo[ghi]perylene) were identified in the leachate and tire samples by Raman/surface-enhanced Raman spectroscopy and gas chromatography with mass spectroscopy, respectively.

Fish Performance Indicators Adjacent to Oil Sands Activity: Response in Performance Indicators of Slimy Sculpin in the Steepbank River, Alberta, Adjacent to Oil Sands Mining Activity.

Since 2009, the Canadian and Alberta governments have been developing monitoring plans for surface water quality and quantity of the lower Athabasca River and its tributaries (2010-2013). The objectives of the present study to the fish monitoring program were to 1) assess the current status of fish in a tributary of the lower Athabasca River, 2) identify existing differences between upstream reference and within the oil sands deposit exposure sites, and 3) identify trends/changes in fish performance indicators relative to historical studies. The present study examines the fish performance indicators in slimy sculpin (Cottus cognatus) in the Steepbank River, Alberta, in terms of growth, gonad size, condition, and hepatic 7-ethoxyresorufin-O-deethylase (EROD) activity as an indicator of exposure to oil-sands-related compounds. The sampling program followed historical sampling methods (1999-2000) to provide comparable data over time with an additional upstream site (n = 2) added as development progressed. Consistent changes were documented in sculpin collected from downstream sections of the Steepbank River within the oil sands deposit (n = 2) in 2010 through 2013. Sculpin demonstrated increased liver size with corresponding induction of EROD activity consistent with historical data and reductions in energy investment relative to reproductive development and gonadal steroid production capacity. There was no consistent evidence of changes in fish performance indicators with increased surface mining development, particularly adjacent to the Steepbank River Mid site. Although physical development in the Steepbank watershed has increased over the last 15 yr, these results are consistent with historical data suggesting that the magnitude of the response in the aquatic environment adjacent to the development has not changed. Environ Toxicol Chem 2020;39:396-409. © 2019 SETAC.

Development of environmental effects monitoring protocol in Brazil: a fish guide study of three river estuaries.

In Brazil, there are no unified and effective environmental monitoring models for bodies of water. Thus, several methodologies are used that result in information that is often difficult to compare, especially for stakeholders involved in regional water management. Studies in some countries such as Australia, Chile, the USA, and Sweden use the monitoring model implemented in Canada that was developed in the early 1990s. This model was designed to evaluate whether the current environmental regulations are sufficiently protective for pulp and paper effluents and for metal mining effluents. In this study, the Canadian Environmental Effects Monitoring methodologies were applied to three different Brazilian river basins, with the goal of constructing a framework for monitoring environmental effects. Pilot studies were carried out in the estuarine regions of the Benevente, Jucu, and Santa Maria da Vitória river basins, which are important rivers in the state of Espírito Santo. Evaluations included fish health, bioaccumulation studies, benthic invertebrate survey, and physical-chemical analyses of water and sediment. The quality of the environments was evaluated by means of seasonal samplings and comparisons between discharge, upstream, and downstream areas. This study made it possible to identify appropriate fish species to be used in environmental effects monitoring and the environmental quality of the rivers themselves as well as knowledge and policy gaps to implement such monitoring programs in Brazil. The study raises questions about the adequacy of Brazilian environmental legislation concerning tidal rivers.

Transcriptome Profiling in Larval Fathead Minnow Exposed to Commercial Naphthenic Acids and Extracts from Fresh and Aged Oil Sands Process-Affected Water.

Surface mining and extraction of oil sands results in the generation of and need for storage of large volumes of oil sands process-affected water (OSPW). More structurally complex than classical naphthenic acids (NAs), naphthenic acid fraction components (NAFCs) are key toxic constituents of OSPW, and changes in the NAFC profile in OSPW over time have been linked to mitigation of OSPW toxicity. Molecular studies targeting individual genes have indicated that NAFC toxicity is likely mediated via oxidative stress, altered cell cycles, ontogenetic differentiation, endocrine disruption, and immunotoxicity. However, the individual-gene approach results in a limited picture of molecular responses. This study shows that NAFCs, from aged or fresh OSPW, have a unique effect on the larval fathead minnow transcriptome and provides initial data to construct adverse outcome pathways for skeletal deformities. All three types of processed NAs (fresh, aged, and commercial) affected the immunome of developing fish. These gene networks included immunity, inflammatory response, B-cell response, platelet adhesion, and T-helper lymphocyte activity. Larvae exposed to both NAFCs and commercial NA developed cardiovascular and bone deformities, and transcriptomic networks reflected these developmental abnormalities. Gene networks found only in NAFC-exposed fish suggest NAFCs may alter fish cardiovascular health through altered calcium ion regulation. This study improves understanding regarding the molecular perturbations underlying developmental deformities following exposure to NAFCs.

The Alberta oil sands eroded bitumen/sediment transitional journey: Influence on sediment transport dynamics, PAH signatures and toxicological effect.

This paper investigated the sediment/contaminant continuum of bitumen containing sediment, from eroded exposed natural bitumen outcrops to river depositional zone, in order to improve our understanding of the transitional sediment, chemistry and toxicological influence on aquatic health. To achieve this aim, we linked a rainfall simulator with an annular flume to allow for connectivity between terrestrial erosion to stream flow. Bulk sediments were collected from the minable McMurray Formation (MF) on the Ells (EL) and Steepbank (STB) Rivers and from the Clearwater Formation (CF) on the STB. All plots generated similar washoff rates (~25 L hr-1) and fine eroded sediment (d50 = 5 µm) regardless of strata. The CF generated the highest total eroded sediment (TES) followed by EL-MF and STB-MF. The STB-MF generated up to three orders of magnitude lower TES than the STB-CF. The EL generated the highest PAH concentrations, however, when normalizing by TES mass delivered to the flume, STB-MF PAH was one to two orders of magnitude higher per unit mass than EL-MF. The TES concentrations were inversely proportional to the degree of bitumen within the sediment (STB-MF > EL-MF > STB-CF). Once the TES was suspended in the flume, there was a change in the dominant PAH for most classes [e.g., Dibenzothiophenes C4 (parent material) to a C3 (TES)]. Further, no flocculation and the buoyant properties of the TES suggested that the sediment and associated PAHs may travel long distances. The EL-MF proved to be the most toxic to fathead minnow embryo survival due principally to the high PAH concentrations and sediment loads. The CF exhibited no toxicological effect. This work has shown the importance of assessing sediment/contaminant characteristics over the continuum (terrestrial erosion/washoff to river deposition) in order to support basin wide management strategies for the protection of aquatic health.

Assessing the effects of environmentally relevant concentrations of antidepressant mixtures to fathead minnows exposed over a full life cycle.

Antidepressant drugs have been detected in municipal wastewater effluents (MWWEs) at ng/L to low µg/L concentrations. We exposed fathead minnow (Pimephales promelas) over a full lifecycle to a mixture of five antidepressants at concentrations similar to a MWWE (1× AntiD Mix); venlafaxine at 2400 ng/L, citalopram at 240 ng/L, fluoxetine at 90 ng/L, sertraline at 20 ng/L, and bupropion at 90 ng/L, and 10× these concentrations (i.e. 10× AntiD Mix). Mean measured concentrations of venlafaxine, citalopram, fluoxetine, sertraline, and bupropion were 2300, 160, 110, 7 ng/L, and below detection limits, respectively, for the 1× AntiD Mix, and 33,000, 2900, 1000, 210, and 100 ng/L, respectively for the 10× AntiD Mix. During the life-cycle exposure, no significant changes were observed in survival of fathead minnows. When male fish from the exposed treatments reached maturity, their weights were increased compared to control males. There were no significant differences in condition factor, gonadosomatic index, or liver-somatic index in the exposed fish. Exposed fathead minnows produced similar numbers of eggs as control fish, and there were no changes in nest-defense behaviours of male minnows. Egg quality, % fertilization, and % hatching in F1 fry were unaffected by exposure to the antidepressants. Eggs hatched 0.5 d earlier, deformities in fry were 50% lower, and there were transient decreases in length of F1 larvae at 8 days post-hatch in offspring from the treatment with the 10× AntiD Mix. Overall, exposure to the antidepressant mixture at environmentally relevant concentrations (i.e. 1× AntiD Mix) caused no adverse effects in fathead minnows. Exposure to the 10× AntiD Mix increased the weight of adult male minnows and caused subtle effects in F1 offspring. This study is the first to assess sublethal effects in fish exposed to mixtures of antidepressants over a full lifecycle. CAPSULE: No effects were observed in fathead minnow exposed for a lifecycle to antidepressant mixtures at environmentally-relevant concentrations.