Oil Sands Process-affected Water Research Articles

Potential Of Biochar And Humic Substances For Phytoremediation Of Trace Metals In Oil Sands Process Affected Water.

Oil sands process affected water (OSPW) is produced during bitumen extraction and typically contains high concentrations of trace metals. Constructed wetlands have emerged as a cost effective and green technology for the treatment of metals in wastewaters. Whether the addition of amendments to constructed wetlands can improve metal removal efficiency is unknown. We investigated the synergistic effects of carbon based amendments and wetland plant species in removal of arsenic, cadmium, cobalt, chromium, copper, nickel, and selenium from OSPW. Three native wetland species (Carex aquatilis, Juncus balticus, Scirpus validus) and two amendments (canola straw biochar, nano humus) were investigated in constructed wetland mesocosms over 60 days. Amendment effect on metal removal efficiency was not significant, while plant species effect was. Phytoremediation resulted in removal efficiencies of 78.61 to 96.31 % for arsenic, cadmium, and cobalt. Carex aquatilis had the highest removal efficiencies for all metals. Amendments alone performed well in removing some metals and were comparable to phytoremediation for cadmium, cobalt, copper, and nickel. Metals were primarily distributed in roots with negligible translocation to shoots. Our work provides insights into the role of plants and amendments during metal remediation and their complex interactions in constructed treatment wetlands.

Treatment of oil sands process-affected water using a semi pilot-scale coalescing filter: Effect of fundamental conditions and water characteristics on filtration performance

The reuse of oil sands process-affected water (OSPW) generated during oil extraction is becoming essential due to oil extraction efficiency and stringent environmental regulations. This study investigates the efficacy of a coalescing filter (CF) in removing oil components from OSPW by exploiting coalescence, which involves merging small oil droplets into larger ones using fibers. Real-time, precise oil measurement systems were employed to evaluate the oil removal efficiency of the CF across synthetic OSPW samples with varying properties. Fundamental conditions and water quality components were systematically adjusted to elucidate the influencing factors on oil removal by the CF. Results showed that despite the diesel having significantly lower viscosity than the oil contents in OSPW, CF achieved a removal efficiency of over 60% within the first 10 min of operation time, regardless of oil concentration. Total suspended solids/turbidity exhibited a decreasing trend as it passed through the filter, indicating fouling occurring within the filter. On the other hands, its increase improved CF performance by 6% compared to baseline conditions, while an increase of alkalinity decreased performance by 10%. When each water quality component exists independently, they do not significantly impact the oil removal efficiency. However, when present together, there is a more decrease in removal efficiency due to the reduced interaction between oil and filter. CF was resistant to changes in water quality. Rather, changes in fundamental factors are more sensitive to CF performance. Filter characteristics and mechanisms make residence time the primary factor affecting oil removal by the CF, with variations exceeding 30% depending on conditions. Therefore, optimizing filter number and flow rate is essential to maximize the filtration performance.

Low adsorption affinity of athabasca oil sands naphthenic acid fraction compounds to a peat-mineral mixture

Much of the toxicity in oil sands process-affected water in Athabasca oil sands tailings has been attributed to naphthenic acids (NAs) and associated naphthenic acid fraction compounds (NAFCs). Previous work has characterized the environmental behaviour and fate of these compounds, particularly in the context of constructed treatment wetlands. There is evidence that wetlands can attenuate NAFCs in natural and engineered contexts, but relative contributions of chemical, biotic, and physical adsorption with sequestration require deconvolution. In this work, the objective was to evaluate the extent to which prospective wetland substrate material may adsorb NAFCs using a peat-mineral mix (PMM) sourced from the Athabasca Oil Sands Region (AOSR). The PMM and NAFCs were first mixed and then equilibrated across a range of NAFC concentrations (5–500 mg/L) with moderate ionic strength and hardness (∼200 ppm combined Ca2+ and Mg2+) that approximate wetland water chemistry. Under these experimental conditions, low sorption of NAFCs to PMM was observed, where sorbed concentrations of NAFCs were approximately zero mg/kg at equilibrium. When NAFCs and PMM were mixed and equilibrated together at environmentally relevant concentrations, formula diversity increased more than could be explained by combining constituent spectra. The TOC present in this PMM was largely cellulose-derived, with low levels of thermally recalcitrant carbon (e.g., lignin, black carbon). The apparent enhancement of the concentration and diversity of components in PMM/NAFCs mixtures are likely related to aqueous solubility of some PMM-derived organic materials, as post-hoc combination of dissolved components from PMM and NAFCs cannot replicate enhanced complexity observed when the two components are agitated and equilibrated together.

A Light Touch: Solar Photocatalysis Detoxifies Oil Sands Process-Affected Waters Prior to Significant Treatment of Naphthenic Acids.

Environmental reclamation of Canada's oil sands tailings ponds is among the single largest water treatment challenges globally. The toxicity of oil sands process-affected water (OSPW) has been associated with its dissolved organics, a complex mixture of naphthenic acid fraction components (NAFCs). Here, we evaluated solar treatment with buoyant photocatalysts (BPCs) as a passive advanced oxidation process (P-AOP) for OSPW remediation. Photocatalysis fully degraded naphthenic acids (NAs) and acid extractable organics (AEO) in 3 different OSPW samples. However, classical NAs and AEO, traditionally considered among the principal toxicants in OSPW, were not correlated with OSPW toxicity herein. Instead, nontarget petroleomic analysis revealed that low-polarity organosulfur compounds, composing <10% of the total AEO, apparently accounted for the majority of waters' toxicity to fish, as described by a model of tissue partitioning. These findings have implications for OSPW release, for which a less extensive but more selective treatment may be required than previously expected.

Biodegradation in oil sands process-affected water: A comprehensive laboratory analysis of the in situ biodegradation of dissolved organic acids

Oil sands process-affected water (OSPW) is a by-product of the extraction of bitumen, and volumes of OSPW have accumulated across the Alberta oil sands region due to the governments zero-discharge policy. Some dissolved organics in OSPW, including toxic naphthenic acids (NAs), can be biodegraded in oxic conditions, thereby reducing the toxicity of OSPW. While there has been much focus on degradation of NAs, the biodegradation of other dissolved organic chemicals by endogenous organisms remains understudied. Here, using the HPLC–ultrahigh resolution Orbitrap mass spectrometry, we examined the microbial biodegradation of dissolved organic acids in OSPW. Non-targeted analysis enabled the estimation of biodegradation rates for unique heteroatomic chemical classes detected in negative ion mode. The microcosm experiments were conducted with and without nutrient supplementation, and the changes in the microbial community over time were investigated. Without added nutrients, internal standard–adjusted intensities of all organics, including NAs, were largely unchanged. The addition of nutrients increased the biodegradation rate of O2− and SO2− chemical classes. While anoxic biodegradation can occur in tailings ponds and end pit lakes, microbial community analyses confirmed that the presence of oxygen stimulated biodegradation of the OSPW samples studied. We detected several aerobic hydrocarbon-degrading microbes (e.g., Pseudomonas and Brevundimonas), and microbes capable of degrading sulfur-containing hydrocarbons (e.g., Microbacterium). Microbial community diversity decreased over time with nutrient addition. Overall, the results from this study indicate that toxic dissolved organics beyond NAs can be biodegraded by endogenous organisms in OSPW, but reaffirms that biological treatment strategies require careful consideration of how nutrients and dissolved oxygen may impact efficacy.

Pros and Cons of Separation, Fractionation and Cleanup for Enhancement of the Quantitative Analysis of Bitumen-Derived Organics in Process-Affected Waters—A Review

Oil sands process-affected water (OSPW) contains a diverse mixture of inorganic and organic compounds. Naphthenic acids (NAs) are a subset of the organic naphthenic acid fraction compounds (NAFCs) and are a major contributor of toxicity to aquatic species. Thousands of unique chemical formulae are measured in OSPW by accurate mass spectrometry and high-resolution mass spectrometry (MS) analysis of NAFCs. As no commercial reference standard is available to cover the range of compounds present in NAFCs, quantitation may best be referred to as “semi-quantitative” and is based on the responses of one or more model compounds. Negative mode electrospray ionization (ESI-) is often used for NAFC measurement but is prone to ion suppression in complex matrices. This review discusses aspects of off-line sample preparation techniques and liquid chromatography (LC) separations to help reduce ion suppression effects and improve the comparability of both inter-laboratory and intra-laboratory results. Alternative approaches to the analytical parameters discussed include extraction solvents, salt content of samples, extraction pH, off-line sample cleanup, on-line LC chromatography, calibration standards, MS ionization modes, NAFC compound classes, MS mass resolution, and the use of internal standards.

Examining the immunotoxicity of oil sands process affected waters using a human macrophage cell line

Oil sands process affected water (OSPW) is produced during the surface mining of the oil sands bitumen deposits in Northern Alberta. OSPW contains variable quantities of organic and inorganic components causing toxic effects on living organisms. Advanced Oxidation Processes (AOPs) are widely used to degrade toxic organic components from OSPW including naphthenic acids (NAs). However, there is no established biological procedure to assess the effectiveness of the remediation processes. Our previous study showed that human macrophage cells (THP-1) can be used as a bioindicator system to evaluate the effectiveness of OSPW treatments through examining the proinflammatory gene transcription levels. In the present study, we investigated the immunotoxicological changes in THP-1 cells following exposure to untreated and AOP-treated OSPW. Specifically, using proinflammatory cytokine protein secretion assays we showed that AOP treatment significantly abrogates the ability of OSPW to induce the secretion of IL-1β, IL-6, IL-8, TNF-α, IL-1Ra and MCP-1. By measuring transcriptional activity as well as surface protein expression levels, we also showed that two select immune cell surface markers, CD40 and CD54, were significantly elevated following OSPW exposure. However, AOP treatments abolished the immunostimulatory properties of OSPW to enhance the surface expression of these immune proteins. Finally, a transcriptome-based approach was used to examine the proinflammatory effects of OSPW as well as the abrogation of immunotoxicity following AOP treatments. Overall, this research shows how a human macrophage cell-based biomonitoring system serves as an effective in vitro tool to study the immunotoxicity of OSPW samples before and after targeted remediation strategies.

Development and testing of a mechanistic model for wetland treatment of neutral and polar organic contaminants in oil sands process-affected water

A mechanistic model of the fate of neutral and polar organic contaminants in a free water surface flow wetland was developed to assess the removal efficiency of polycyclic aromatic hydrocarbons (PAHs) and naphthenic acids (O2-NAs), which are chemicals primarily responsible for oil sands process-affected water (OSPW) toxicity. The model simulates wetland mechanisms of chemical removal based on physicochemical properties, environmental conditions, and wetland design and operation. Good agreement between model estimated and observed concentrations of PAHs and O2-NAs in OSPW in the Kearl Treatment Wetland (Alberta, Canada) was observed over time. Model simulations indicate that the removal of PAHs and O2-NAs from OSPW is highly sensitive to biotransformation rates and that sustained wetland treatment is feasible with robust PAH and NA degrading microbial communities in the wetland. Wetland treatment efficiency is greatest for substances with a log KOW or log D between 3.0 and 5.5 whereas substances with a log KOW or log D >6.0 are not suitable for wetland treatment, even if they can be biodegraded. The log KOW of PAHs and log D for O2-NAs range from 0.7 to 6.7 suggesting most organic contaminants in OSPW are suitable for wetland treatment. Chemical removal via evapotranspiration is not significant pathway of chemical removal from wetlands for PAHs and O2-NAs due to their very low log KAW (less than −1.6). The treatment wetland model presented is the first model to evaluate the fate of both neutral and polar organic contaminants in a flow-through treatment wetland operation, making it a helpful tool to assess the feasibility of these treatment systems for the oil sands industry. The model is particularly useful in evaluating chemical removal efficiency of OSPW contaminants, whether specific water quality objectives can be achieved after treatment, and to assess trade-offs in wetland design and operation to balance treatment performance with design and operational features.

Draft genome sequences of six Pseudomonas spp. and one Rheinheimera sp. isolated from oil sands process-affected water from Alberta, Canada

ABSTRACT We report the draft genomes of seven bacterial strains (six Pseudomonas spp. and one Rheinheimera sp.) isolated from environmental water samples from oil sands tailings ponds that have accumulated a wide variety of organic compounds, salts and metals.

Complete genome sequence of Pseudomonas veronii strain OST1911 isolated from oil sand tailing pond water in Alberta, Canada.

Here, we report the complete genome sequence of Pseudomonas veronii strain OST1911, recovered from oil sand process-affected water accumulated in tailing ponds. This water contains numerous organic and inorganic compounds of environmental significance. The genome size is 6,435,955 bp with a G+C content of 61.21%.

Microbial degradation of naphthenic acids using constructed wetland treatment systems: metabolic and genomic insights for improved bioremediation of process-affected water.

Naphthenic acids (NAs) are a complex mixture of organic compounds released during bitumen extraction from mined oil sands that are important contaminants of oil sands process-affected water (OSPW). NAs can be toxic to aquatic organisms and, therefore, are a main target compound for OSPW. The ability of microorganisms to degrade NAs can be exploited for bioremediation of OSPW using constructed wetland treatment systems (CWTS), which represent a possible low energy and low-cost option for scalable in situ NA removal. Recent advances in genomics and analytical chemistry have provided insights into a better understanding of the metabolic pathways and genes involved in NA degradation. Here, we discuss the ecology of microbial NA degradation with a focus on CWTS and summarize the current knowledge related to the metabolic pathways and genes used by microorganisms to degrade NAs. Evidence to date suggests that NAs are mostly degraded aerobically through ring cleavage via the beta-oxidation pathway, which can be combined with other steps such as aromatization, alpha-oxidation, omega-oxidation, or activation as coenzyme A (CoA) thioesters. Anaerobic NA degradation has also been reported via the production of benzoyl-CoA as an intermediate and/or through the involvement of methanogens or nitrate, sulfate, and iron reducers. Furthermore, we discuss how genomic, statistical, and modeling tools can assist in the development of improved bioremediation practices.

Draft genome sequence of Pseudomonas sp. ER28, a cyclohexane pentanoic acid degrader isolated from oil sands process-affected water from Alberta, Canada.

We report the draft genome sequence of Pseudomonas sp. ER28, capable of utilizing the model naphthenic acid, cyclohexane pentanoic acid, as its sole carbon source. It was recovered from oil sands process-affected water containing cyclic and acyclic naphthenic acids. The genome size is 5.7 Mbp, and the G + C content is 60%.

Evaluating the attenuation of naphthenic acids in constructed wetland mesocosms planted with Carex aquatilis

Surface oil sands mining and extraction in northern Alberta’s Athabasca oil sands region produce large volumes of oil sands process–affected water (OSPW). OSPW is a complex mixture containing major contaminant classes including trace metals, polycyclic aromatic hydrocarbons, and naphthenic acid fraction compounds (NAFCs). Naphthenic acids (NAs) are the primary organic toxicants in OSPW, and reducing their concentrations is a priority for oil sands companies. Previous evidence has shown that constructed wetland treatment systems (CWTSs) are capable of reducing the concentration of NAs and the toxicity of OSPW through bioremediation. In this study, we constructed greenhouse mesocosms with OSPW or lab process water (LPW) (i.e., water designed to mimic OSPW minus the NAFC content) with three treatments: (1) OSPW planted with Carex aquatilis; (2) OSPW, no plants; and (3) LPW, no plants. The OSPW–C. aquatilis treatment saw a significant reduction in NAFC concentrations in comparison to OSPW, no plant treatments, but both changed the distribution of the NAFCs in similar ways. Upon completion of the study, treatments with OSPW saw fewer high-molecular-weight NAs and an increase in the abundance of O3- and O4-containing formulae. Results from this study provide invaluable information on how constructed wetlands can be used in future remediation of OSPW in a way that previous studies were unable to achieve due to uncontrollable environmental factors in field experiments and the active, high-energy processes used in CWTSs pilot studies.

Online Membrane Sampling for the Mass Spectrometric Analysis of Oil Sands Process Affected Water-Derived Naphthenic Acids in Real-World Samples

Large volumes of oil sands process-affected waters (OSPW) result from heavy oil extraction in Alberta, Canada. Currently, a toxic legacy of ca. 500 Mm3 is stored in tailings ponds under a zero-discharge policy. OSPW is a complex mixture of suspended and dissolved materials including a wide range of inorganic and organic contaminants. Classically defined naphthenic acids (NAs; CnH2n+ZO2) are one of the primary toxic fractions in OSPW and have therefore been the subject of considerable research interest. Most studies employ considerable sample cleanup followed by liquid chromatography and/or high-resolution mass spectrometry (HRMS) for the characterization of these complex mixtures. However, these strategies can be time- and cost-intensive, limiting the scope of research and adoption for regulatory purposes. Condensed phase membrane introduction mass spectrometry (CP-MIMS) is emerging as a “fit-for-purpose” approach for the analysis of NAs. This technique directly interfaces the mass spectrometer with an aqueous sample using a hydrophobic semi-permeable membrane, requiring only pH adjustment to convert NAs to a membrane-permeable form. Here, we examine the perm-selectivity of classical NAs (O2) relative to their more oxidized counterparts (O3–O7) and heteroatomic (N, S) species collectively termed naphthenic acid fraction compounds (NAFCs). The investigation of 14 model compounds revealed that classically defined NAs are greater than 50-fold more membrane permeable than their oxidized/heteroatomic analogs. HRMS analysis of real OSPW extracts with and without membrane clean-up further supported selectivity towards the toxic O2 class of NAs, with &gt;85% of the overall signal intensity attributable to O2 NAs in the membrane permeate despite as little as 34.7 ± 0.6% O2 NAs observed in the directly infused mixture. The information collected with HRMS is leveraged to refine our method for analysis of NAs at unit mass resolution. This new method is applied to 28 archived real-world samples containing NAs/NAFCs from constructed wetlands, OSPW, and environmental monitoring campaigns. Concentrations ranged from 0–25 mg/L O2 NAs and the results measured by CP-MIMS (unit mass) and SPE-HRMS (Orbitrap) showed good agreement (slope = 0.80; R2 = 0.76).

Persistent Cytotoxicity and Endocrine Activity in the First Oil Sands End-Pit Lake

Oil sands process-affected water (OSPW) is a byproduct of bitumen extraction that has persistent toxicity owing to its complex mixture of organics. A prominent remediation strategy that involves aging OSPW in end-pit lakes and Base Mine Lake (BML) is the first full-scale test. Its effectiveness over the first 5 years was investigated here using real-time cell analysis, yeast estrogenic and androgenic screens (YES/YAS), and ultra-high-resolution mass spectrometry. HepG2 cytotoxicity per volume of BML organics extracted decreased with age; however, the toxic potency (i.e., toxicity per mass of extract) was not significantly different between years. This was consistent with mass spectral evidence showing no difference in chemical profiles, yet lower total abundance of organics in field-aged samples, suggestive that dilution explains the declining cytotoxicity in BML. The IC50’s of BML extracts for YES/YAS antagonism were at environmental concentrations and were similar despite differences in field-age. Persistent YES/YAS antagonism and cytotoxicity were detected in experimental pond OSPW field-aged >20 years, and while organic acids were depleted here, non-acid chemical classes were enriched compared to BML, suggesting these contribute to persistent toxicity of aged OSPW. To avoid a legacy of contaminated sites, active water treatment may be required to accelerate detoxification of end-pit lakes.

Adsorption of single‐ring model naphthenic acid from oil sands tailings pond water using physically activated petroleum coke

AbstractPetroleum coke‐derived activated cokes were prepared and used for the adsorptive removal of a single‐ring naphthenic acid (NA) from synthetic oil sands process affected water (OSPW). CO2 activation produced carbon with a larger mesopore volume fraction (0.67) than steam activation (0.25). Interestingly, prolonging the activation time of CO2 from 6 to 9 h led to a simultaneous increase in specific surface area (276–405 m2/g) and mesopore volume (0.51–0.67). Furthermore, a positive relationship was found between the pseudo‐second‐order kinetic rate constant and the mesoporous volume of the activated coke. This suggests both the importance of pore size on kinetics and the fact that physical activation with a reagent such as CO2 may be better suited than chemical activation due to its ability to create mesopores. Although enlarging the pores and accelerating the adsorption rate, post‐oxidation had detrimental effects on adsorption capacity, resulting in a decrease in equilibrium adsorbed amount from 115 to 34 mg/g, a 70% decrease, when post‐oxidized with O2, due to the negative charge of oxygen‐containing functional groups. On the other hand, the measured adsorption capacity increased by over 60% when activated coke was treated with ammonia, a result of the positively charged nitrogen‐containing surface groups. Finally, in real OSPW, the activated coke had a much lower capacity for total acid‐extractable organics than for the model NA. Therefore, activated petroleum coke may not be the best choice for treating raw tailings pond water and may be better suited for polishing.

Water quality monitoring with in vitro bioassays to compare untreated oil sands process-affected water with unimpacted rivers

The selected battery of in vitro bioassays may be used to monitor exceedances of effect-based trigger (EBT) values in environments potentially receiving treated oil sands process-affected water.

A Vegetation Assessment of the Kearl Treatment Wetland following Exposure to Oil Sands Process-Affected Water

Treatment wetlands have emerged as a potential option for the treatment of oil sands process-affected water (OSPW). The long-term viability of these treatment systems relies, in part, on the health and productivity of wetland vegetation. The aim of this study is to investigate the physiological and community-level effects on wetland vegetation at the Kearl Treatment Wetland (KTW) following exposure to different OSPW sources. Annual vegetation assessments were performed during 2016–2021 to monitor species composition, total percent cover, species richness, species morphology (plant stem density, leaf length, and leaf width), and plant vigor in the KTW. Cattail was found to dominate the deep-water zones whereas water sedge was found to dominate the shallow-water zones of the wetland. Species richness in the KTW decreased with time which is typical of constructed wetlands receiving industrial effluents. No changes in plant stem density of cattails or water sedge were observed; however, leaf length decreased from 2019 to 2021, and leaf width decreased from 2020 to 2021. Plant vigor ratings increased in the KTW each year suggesting that the vegetation does not show any major signs of phytotoxicity from OSPW exposure. These results demonstrate the complex dynamics and resiliency of the vegetative community in treatment wetlands, but continued efforts to monitor plant uptake and accumulation are needed to fully assess the phytotoxic effects of OSPW exposure.

Augmentation of field fluorescence measures for improved in situ contaminant detection.

This research proposes a new method that fuses data from the field and lab-based optical measures coupled with machine learning algorithms to quantify the concentrations of toxic contaminants found in fuels and oil sands process-affected water. Selected pairs of excitation/emission intensities at key wavelengths are inputs to an augmentation neural network (NN), trained using lab-based measurements, that generates synthetic high-resolution spectra. Then, an image processing NN is used to estimate the contaminant concentrations from the spectra generated from a few key wavelengths. The presented approach is tested using naphthenic acids, phenol, fluoranthene and pyrene spiked into natural waters. The spills or loss of containment of these contaminants represent a significant risk to the environment and public health, requiring accurate and rapid detection methods to protect the surrounding aquatic environment. Results were compared with models based on only the corresponding peak intensities of each contaminant and with an image processing NN using the original spectra. Naphthenic acids, fluoranthene and pyrene were easy to detect by all methods; however, performance for more challenging signals to identify, such as phenol, was optimized by the proposed method (peak picking with mean absolute error (MAE) of 30.48µg/L, generated excitation-emission matrix with MAE of 8.30µg/L). Results suggested that data fusion and machine learning techniques can improve the detection of contaminants in the aquatic environment at environmentally relevant concentrations.

Naphthenic acid fraction compounds reduce the reproductive success of wood frogs (Rana sylvatica) by affecting offspring viability

Understanding the toxicity of organic compounds in oil sands process-affected water (OSPW) is necessary to inform the development of environmental guidelines related to wastewater management in Canada's oil sands region. In the present study, we investigated the effects of naphthenic acid fraction compounds (NAFCs), one of the most toxic components of OSPW, on mating behaviour, fertility, and offspring viability in the wood frog (Rana sylvatica). Wild adult wood frogs were exposed separately from the opposite sex to 0, 5, or 10 mg/L of OSPW-derived NAFCs for 24 h and then combined in outdoor lake water mesocosms containing the same NAFC concentrations (n = 2 males and 1 female per mesocosm, n = 3 mesocosms per treatment). Mating events were recorded for 48 h and egg masses were measured to determine adult fertility. NAFC exposure had no significant effect on mating behaviour (probability of amplexus and oviposition, amplexus and oviposition latency, total duration of amplexus and number of amplectic events) or fertility (fertilization success and clutch size). Tadpoles (50 individuals per mesocosm at hatching, and 15 individuals per mesocosm from 42 d post-hatch) were reared in the same mesocosms under chronic NAFC exposure until metamorphic climax (61–85 d after hatching). Offspring exposed to 10 mg/L NAFCs during development were less likely to survive and complete metamorphosis, grew at a reduced rate, and displayed more frequent morphological abnormalities. These abnormalities included limb anomalies at metamorphosis, described for the first time after NAFC exposure. The results of this study suggest that NAFCs reduce wood frog reproductive success through declines in offspring viability and therefore raise the concern that exposure to NAFCs during reproduction and development may affect the recruitment of native amphibian populations in the oil sands region.