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.

The effects of oil sands process-affected water naphthenic acid fraction components on GDF15 secretion in extravillous trophoblast cells.

Exposure to compounds present in petroleum and wastewaters from oil and gas extraction sites in the Alberta Oil Sands Region can impair reproductive health. It has been established that acid extractable organics found in oil sands process-affected water (OSPW) such as naphthenic acids (NA-fraction components; NAFC) can adversely affect reproductive outcomes. We have shown that NAFC exposure results in a significant upregulation of GDF15 in placental trophoblasts, a cellular stress marker known to be involved in human embryonic development and necessary for the maintenance of pregnancy. However, little is known regarding the mechanism(s) underlying NAFC-induced increases in GDF15 production during early placentation. The goal of this study was to examine the effects of NAFC exposure on the regulation of critical transcription factors of GDF15 in extravillous trophoblast cells. Of these transcription factors, inflammatory mediators including prostaglandins have been reported to inhibit proliferation and migration of trophoblast cells in vitro. Hence, the secondary goal of this study was to determine whether inflammation mediated through prostaglandin production is critical to GDF15 secretion. HTR-8/SVneo cells were exposed to an NAFC for 6 and 24 h to assess the expression of key transcriptional regulators, GDF15 secretion, and prostaglandin (PGE2) output. Treatment with NAFC (125 mg/L only) significantly increased GDF15 expression and secretion in association with upregulation of the transcription factors KLF4, EGR1, ATF3 and TP53. Similarly, PTGS2 (i.e. COX2) expression and PGE2 output were significantly increased at the same concentration. However, co-treatment with a COX2 selective antagonist (SC236) only partially blocked the NAFC-induced increase in PGE2 output and did not block GDF15 expression or secretion. These findings suggest that while NAFC may affect GDF15 production, it is not exclusively a result of prostaglandin-mediated inflammation. This study provides new insights into the mechanisms by which NAFC may adversely affect placental trophoblast cell function in mammals.

Treatment of naphthenic acids in oil sands process-affected waters with a surface flow treatment wetland: mass removal, half-life, and toxicity-reduction.

This study is the first to investigate the removal of naphthenic acids in a full-scale constructed wetland within the Alberta Oil Sands region. The average mass-removal efficiency for all O2-naphthenic acids measured in three separate deployments in the wetland ranged from 7.5% to 68.9% and appeared sensitive to physicochemical properties of the naphthenic acids, environmental conditions, and water quality. Treatment efficiency of individual naphthenic acids was found to increase with increasing carbon number and decreasing number of double bond equivalents in the molecule. Treatment efficiency was also found to increase with both higher initial turbidity in OSPW entering the wetland, and warmer average OSPW temperatures during wetland operation. Half-life times of naphthenic acids in the treatment wetland ranged between 8.9 and 39 days and were substantially lower than those in tailings ponds (i.e., 12.9-13.6 years) and laboratory studies focussed on bench-scale aerobic microbial biodegradation (i.e., 44-315 days). Using published dose-response data, biomimetic extraction measurements using solid phase microextraction fibers indicate that 14 days of wetland treatment resulted in a reduction in (4 d) deformity of Danio rerio from 50 to 16%, while exhibiting less than 1% toxic response for less sensitive toxic endpoints. The study concludes that wetland treatment is a feasible and productive treatment method for naphthenic acids in oil sands process-affected water due to a combination of sorption and biodegradation.

Naphthenic acid fraction components from oil sands process-affected water from the Athabasca Oil Sands Region impair murine osteoblast differentiation and function.

The extraction of bitumen from surface mining in the Athabasca Oil Sands Region (AOSR) produces large quantities of oil sands process-affected water (OSPW) that needs to be stored in settling basins near extraction sites. Chemical constituents of OSPW are known to impair bone health in some organisms, which can lead to increased fracture risk and lower reproductive fitness. Naphthenic acid fraction components (NAFCs) are thought to be among the most toxic class of compounds in OSPW; however, the effect of NAFCs on osteoblast development is largely unknown. In this study, we demonstrate that NAFCs from OSPW inhibit osteoblast differentiation and deposition of extracellular matrix, which is required for bone formation. Extracellular matrix deposition was inhibited in osteoblasts exposed to 12.5-125 mg/L of NAFC for 21 days. We also show that components within NAFCs inhibit the expression of gene markers of osteoblast differentiation and function, namely, alkaline phosphatase (Alp), osteocalcin, and collagen type 1 alpha 1 (Col1a1). These effects were partially mediated by the induction of glucocorticoid receptor (GR) activity; NAFC induces the expression of the GR activity marker genes Sgk1 (12.5 mg/L) and p85a (125 mg/L) and inhibits GR protein (125 mg/L) and Opg RNA (12.5 mg/L) expression. This study provides evidence that NAFC concentrations of 12.5 mg/L and above can directly act on osteoblasts to inhibit bone formation and suggests that NAFCs contain components that can act as GR agonists, which may have further endocrine disrupting effects on exposed wildlife.

Molecular profiling of naphthenic acids in technical mixtures and oil sands process-affected water using polar reversed-phase liquid chromatography-mass spectrometry.

In this work, a reversed-phase ultra-HPLC (UHPLC) ultrahigh resolution MS (UHRMS) method was evaluated for the comprehensive profiling of NAs containing two O atoms in each molecule (O2NAs; general formula Cn H2n + z O2 , where n is the number of carbon atoms and z represents hydrogen deficiency). Using a polar cyanopropyl-bonded phase column and negative-ion electrospray ionization mass spectrometric detection at 120,000 FWHM (m/z 400), 187 and 226 O2NA species were found in two naphthenic acid technical mixtures, and 424 and 198 species with molecular formulas corresponding to O2NAs were found in two oil sands process-affected water samples (one from a surface mining operation and the other from a steam-assisted gravity drainage operation), respectively. To our knowledge, these are the highest numbers of molecular compositions of O2NAs that have been profiled thus far in environmental samples. Assignments were based on accurate mass measurements (≤3 ppm) combined with rational molecular formula generation, correlation of chromatographic behavior of O2NA homologues with their elemental compositions, and confirmation with carboxyl group-specific chemical derivatization using 3-nitrophenylhydrazine. Application of this UHPLC-UHRMS method to the quantitation of O2NAs in the surface mining operation-derived water sample showed excellent linearity (R2 = 0.9999) with external calibration, a linear range of 256-fold in concentration, and quantitation accuracies of 64.9 and 69.4% at two "standard substance" spiking levels.

Potential of capillary electrophoresis mass spectrometry for the characterization and monitoring of amine-derivatized naphthenic acids from oil sands process-affected water.

Capillary electrophoresis coupled to mass spectrometry (CE-MS) was used for the analysis of naphthenic acid fraction compounds (NAFCs) of oil sands process-affected water (OSPW). A standard mixture of amine-derivatized naphthenic acids is injected directly onto the CE column and analyzed by CE-MS in less than 15min. Time of flight MS analysis (TOFMS), optimized for high molecular weight ions, showed NAFCs between 250 and 800m/z. With a quadrupole mass analyzer, only low-molecular weight NAFCs (between 100 and 450m/z) are visible under our experimental conditions. Derivatization of NAFCs consisted of two-step amidation reactions mediated by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), or mediated by a mixture of EDC and N-hydroxysuccinimide, in dimethyl sulfoxide, dichloromethane or ethyl acetate. The optimum background electrolyte composition was determined to be 30% (V/V) methanol in water and 2% (V/V) formic acid. NAFCs extracted from OSPW in the Athabasca oil sands region were used to demonstrate the feasibility of CE-MS for the analysis of NAFCs in environmental samples, showing that the labeled naphthenic acids are in the mass range of 350 to 1500m/z.

Coagulation/flocculation process with polyaluminum chloride for the remediation of oil sands process-affected water: Performance and mechanism study.

This study investigated the application of polyaluminum chloride (PACl) for the treatment of the oil sands process-affected water (OSPW). These coagulants are commonly used in water treatment with the most effective species reported to be Al13. PACl with 83.6% Al13 was synthesized using the slow base titration method and compared with a commercially available PACl in terms of aluminum species distribution, coagulation/flocculation (CF) performance, floc morphology, and contaminant removal. Both coagulants were effective in removing suspended solids, achieving over 96% turbidity removal at all applied coagulant doses (0.5-3.0 mM Al). The removal efficiencies of metals varied among different metals depending on their pKa values with metal cations having pKa values (Fe, Al, Ga, and Ti) below OSPW pH of 6.9-8.1 (dose dependent) being removed by more than 90%, while cations with higher pKa values (K, Na, Ca, Mg and Ni) had removals of less than 40%. Naphthenic acids were not removed due to their low molecular weights, negative charges, and hydrophilic characteristics at the OSPW pH. At the highest applied coagulant dose of 3.0 mM Al, the synthetic PACl reduced Vibrio fischeri inhibition effect to 43.3 ± 3.0% from 49.5 ± 0.4% in raw OSPW. In contrast, no reduction of toxicity was found for OSPW treated with the commercial PACl. Based on water quality and floc analyses, the dominant CF mechanism for particle removal during OSPW treatment was considered to be enmeshment in the precipitates (i.e., sweep flocculation). Overall, the CF using synthesized PACl can be a valuable pretreatment process for OSPW to create wastewater that is more easily treated by downstream processes.

Granular activated carbon for simultaneous adsorption and biodegradation of toxic oil sands process-affected water organic compounds.

Naphthenic acids (NAs) released into oil sands process-affected water (OSPW) during bitumen processing in Northern Alberta are problematic for oil sands industries due to their toxicity in the environment and resistance to degradation during conventional wastewater treatment processes. Granular activated carbon (GAC) has shown to be an effective media in removing biopersistent organics from wastewater using a combination of adsorption and biodegradation removal mechanisms. A simultaneous GAC (0.4 g GAC/L) adsorption and biodegradation (combined treatment) study was used for the treatment of raw and ozonated OSPW. After 28 days of batch treatment, classical and oxidized NAs removals for raw OSPW were 93.3% and 73.7%, and for ozonated OSPW were 96.2% and 77.1%, respectively. Synergetic effects of the combined treatment process were observed in removals of COD, the acid extractable fraction, and oxidized NAs, which indicated enhanced biodegradation and bioregeneration in GAC biofilms. A bacteria copy number >10(8) copies/g GAC on GAC surfaces was found using quantitative real time polymerase chain reaction after treatment for both raw and ozonated OSPW. A Microtox(®) acute toxicity test (Vibrio fischeri) showed effective toxicity removal (>95.3%) for the combined treatments. Therefore, the simultaneous GAC adsorption and biodegradation treatment process is a promising technology for the elimination of toxic OSPW NAs.

Removal of organic compounds and trace metals from oil sands process-affected water using zero valent iron enhanced by petroleum coke.

The oil production generates large volumes of oil sands process-affected water (OSPW), referring to the water that has been in contact with oil sands or released from tailings deposits. There are concerns about the environmental impacts of the release of OSPW because of its toxicity. Zero valent iron alone (ZVI) and in combination with petroleum coke (CZVI) were investigated as environmentally friendly treatment processes for the removal of naphthenic acids (NAs), acid-extractable fraction (AEF), fluorophore organic compounds, and trace metals from OSPW. While the application of 25 g/L ZVI to OSPW resulted in 58.4% removal of NAs in the presence of oxygen, the addition of 25 g petroleum coke (PC) as an electron conductor enhanced the NAs removal up to 90.9%. The increase in ZVI concentration enhanced the removals of NAs, AEF, and fluorophore compounds from OSPW. It was suggested that the electrons generated from the oxidation of ZVI were transferred to oxygen, resulting in the production of hydroxyl radicals and oxidation of NAs. When OSPW was de-oxygenated, the NAs removal decreased to 17.5% and 65.4% during treatment with ZVI and CZVI, respectively. The removal of metals in ZVI samples was similar to that obtained during CZVI treatment. Although an increase in ZVI concentration did not enhance the removal of metals, their concentrations effectively decreased at all ZVI loadings. The Microtox(®) bioassay with Vibrio fischeri showed a decrease in the toxicity of ZVI- and CZVI-treated OSPW. The results obtained in this study showed that the application of ZVI in combination with PC is a promising technology for OSPW treatment.

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 78.61-96.31 % 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.

The effect of rhizosphere pH on removal of naphthenic acid fraction compounds from oil sands process-affected water in a willow hydroponic system.

The extraction and processing of bitumen from the oil sands in northern Alberta, Canada generates large volumes of oil sands process-affected water (OSPW). OSPW contains a complex mixture of inorganic and organic compounds, including naphthenic acid fraction compounds (NAFCs) that are of particular concern due to their toxicity to aquatic organisms. Phytoremediation is a cost-effective, scalable approach that has the potential to remove NAFCs from OSPW and reduce OSPW toxicity. Environmental pH influences the chemical form and bioavailability of NAFCs. However, little is known about the influence of pH on the uptake of NAFCs in plant systems. This study sought to elucidate the impact of rhizosphere pH on the uptake of NAFCs using a sandbar willow (Salix interior) hydroponic system. To mimic and maintain the naturally low pH conditions of the root, OSPW solutions in these systems were adjusted to a low pH level (pH 5.0) and their NAFC uptake from solution was compared to that of OSPW at native pH (pH 8.0). Our findings revealed that the lower pH hydroponic systems demonstrated enhanced NAFC removal from solution as determined by LC-MS analysis, where up to 26% of NAFCs were removed from OSPW over 72 h at pH 5.0 compared to 8% removed at pH 8.0. Similarly, analysis of spike-in 13C-labeled NAs demonstrated that the OSPW hydroponic system rapidly removed a relatively labile NA (13C-cyclohexane carboxylic acid) from solution at both pH levels, whereas near complete removal of a recalcitrant NA (13C-1-adamantane carboxylic acid) was observed in pH 5.0 solutions only. These results provide insight into the importance of rhizosphere pH on efficient NAFC uptake by plant root systems. Further research will determine whether OSPW phytoremediation efficiency can be enhanced using field treatment conditions that promote low rhizosphere pH levels.

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.

Chemical communication in wood frog (Rana sylvatica) tadpoles is influenced by early-life exposure to naphthenic acid fraction compounds.

Environmental pollutants can disrupt chemical communication between aquatic organisms by interfering with the production, transmission, and/or detection of, as well as responses to, chemical cues. Here, we test the hypothesis that early-life exposure to naphthenic acid fraction compounds (NAFCs) from oil sands tailings disrupts antipredator-associated chemical communication in larval amphibians. Wild adult wood frogs (Rana sylvatica) captured during their natural breeding period were combined (1 female:2 males) in six replicate mesocosms filled with either uncontaminated lakewater or with NAFCs isolated from an active tailings pond in Alberta, Canada, at nominal 5 mg/L concentrations. Egg clutches were incubated and tadpoles maintained in their respective mesocosms for ∼40 days post-hatch. Tadpoles (Gosner stage 25-31) were then transferred individually to trial arenas filled with uncontaminated water and exposed to one of six chemical alarm cue (AC) stimuli solutions following a 3 × 2 × 2 design (3 AC types × 2 stimulus carriers × 2 rearing exposure groups). Relative to control tadpoles, NAFC-exposed tadpoles demonstrated higher baseline activity levels (line crosses and direction changes) when introduced to uncontaminated water. Antipredator responses differed in graded fashion with AC type, with control ACs eliciting the greatest latency to resume activity, water the least, and NAFC-exposed ACs intermediate. Pre- to post-stimulus difference scores were non-significant in control tadpoles, while NAFC-exposed tadpoles demonstrated significantly greater variation. While this suggests that exposure to NAFCs from fertilization through hatching may have interfered with AC production, it is unclear whether the quality or quantity of cues was affected. There was also no clear evidence that NAFC carrier water interfered with ACs or the alarm response in unexposed control tadpoles. These results emphasize the importance of understanding how behavioral and physiological effects of early-life NAFC exposure on critical antipredator responses may persist across life history stages.

Detection of naphthenic acid uptake into root and shoot tissues indicates a direct role for plants in the remediation of oil sands process-affected water.

Bitumen extraction from surface-mined oil sands deposits results in the accumulation of large volumes of oil sands process-affected water (OSPW). Naphthenic acids (NAs) are primary contributors to OSPW toxicity and have been a focal point for the development of OSPW remediation strategies. Phytoremediation is an approach that utilizes plants and their associated microbes to remediate contaminants from soil and groundwater. While previous evidence has indicated a role for phytoremediation in OSPW treatment through the transformation and degradation of NAs, there are no reports that demonstrate the direct uptake of NAs into plant tissue. Using NAs labelled with 14C radioisotopes (14C-NAs) paired with whole-plant autoradiography, we show that NAs representing aliphatic (linear), single-ring, and diamondoid compounds were effectively removed from hydroponic solution and OSPW-treated soil by sandbar willow (Salix interior) and slender wheatgrass (Elymus trachycaulus) and their associated microbiomes. The NA-derived 14C label accumulated in root and shoot tissues of both plant species and was concentrated in vascular tissue and rapidly growing sink tissues, indicating that 14C-NAs or their metabolic derivatives were incorporated into physiological processes within the plants. Slender wheatgrass seedlings grown under axenic (sterile) hydroponic and soil conditions also effectively removed all 14C-NAs, including a highly stable diamondoid NA, demonstrating that plants can directly take up simple and complex NAs without the assistance of microbes. Furthermore, root and shoot tissue fractionation into major biomolecule groups suggests that NA-derived carbon is allocated toward biomolecule synthesis rapidly after NA treatment. These findings provide evidence of plant-mediated uptake of NAs and support a direct role for plants and their associated microbes in the development of future large-scale OSPW phytoremediation strategies.

Differential protein expression during growth on model and commercial mixtures of naphthenic acids in Pseudomonas fluorescens Pf-5.

Naphthenic acids (NAs) are carboxylic acids with the formula (Cn H2n+Z O2 ) and are among the most toxic, persistent constituents of oil sands process-affected waters (OSPW), produced during oil sands extraction. Currently, the proteins and mechanisms involved in NA biodegradation are unknown. Using LC-MS/MS shotgun proteomics, we identified proteins overexpressed during the growth of Pseudomonas fluorescens Pf-5 on a model NA (4'-n-butylphenyl)-4-butanoic acid (n-BPBA) and commercial NA mixture (Acros). By day 11, >95% of n-BPBA was degraded. With Acros, a 17% reduction in intensity occurred with 10-18 carbon compounds of the Z family -2 to -14 (major NA species in this mixture). A total of 554 proteins (n-BPBA) and 631 proteins (Acros) were overexpressed during growth on NAs, including several transporters (e.g., ABC transporters), suggesting a cellular protective response from NA toxicity. Several proteins associated with fatty acid, lipid, and amino acid metabolism were also overexpressed, including acyl-CoA dehydrogenase and acyl-CoA thioesterase II, which catalyze part of the fatty acid beta-oxidation pathway. Indeed, multiple enzymes involved in the fatty acid oxidation pathway were upregulated. Given the presumed structural similarity between alkyl-carboxylic acid side chains and fatty acids, we postulate that P. fluorescens Pf-5 was using existing fatty acid catabolic pathways (among others) during NA degradation.

Molecular profiles of naphthenic acid fraction compounds from mine lease wetlands in the Athabasca Oil Sands Region.

Naphthenic acid fraction compounds (NAFCs) are a toxicologically relevant component of oil sands process-affected materials (OSPM). For the first time, we report on differences in the concentrations and distribution of NAFCs from wetlands on an Athabasca oil sands mine site with varied histories of solid and liquid OSPM input. Sampling locations included natural and naturalized reference wetlands, a reclaimed tailings pond, wetlands supplemented with OSPM, opportunistic wetlands, and tailings ponds. Samples were prepared using solid-phase extraction, and analyzed by high-resolution Orbitrap mass spectrometry; NAFC concentrations and characteristics were evaluated for all locations. The NAFCs from tailings ponds were dominated by O3-NAFCs and classical naphthenic acids (NAs; i.e., O2 species) with double bond equivalences of 3 and 4. Reference wetlands had no dominant species, and relatively little NAFC content. The heteroatomic species in opportunistic wetlands were dominated by highly-oxidized NAFC species, where Σ [O3:O6] species constituted 55-75% of the assignable spectrum and 3-4% NAs; in tailings ponds NAs constituted 47-51%. A relatively young (4-year-old) wetland built on a former tailings pond had NAFC concentrations between 65 and 80 mg/L, and NAs constituted 47% of the assignable spectrum. There was thus little apparent oxidation of NAFCs at this young wetland. The composition of NAFCs from one wetland (≥15 years old) supplemented with OSPM contained a greater proportion of oxidized species than tailings, suggesting NAFC transformation therein. These data suggest that while NAFCs are persistent in some wetlands, there is preliminary evidence for oxidation in mature wetlands.

The effects of a technical mixture of naphthenic acids on placental trophoblast cell function.

There is considerable concern that naphthenic acids (NA) related to oil extraction can negatively impact reproduction in mammals, yet the mechanisms are unknown. Since placental dysfunction is central to many adverse pregnancy outcomes, the goal of this study was to determine the effects of NA exposure on placental trophoblast cell function. HTR-8/SVneo cells were exposed to a commercial technical NA mixture for 24 hours to assess transcriptional regulation of placentation-related pathways and functional assessment of migration, invasion, and angiogenesis. Pathway analysis suggests that NA treatment resulted in increased epithelial-to-mesenchymal transition. However, there was reduced migration and invasive potential. NA treatment increased angiogenesis-related pathways with a concomitant increase in tube formation. Since decreased trophoblast invasion/migration and aberrant angiogenesis have been associated with placental dysfunction, these findings suggest that it is biologically plausible that exposure to NA may result in altered placental development and/or function.

Profiling low molecular weight organic compounds from naphthenic acids, acid extractable organic mixtures, and oil sands process-affected water by SPME-GC-EIMS.

Naphthenic acids (NAs) are complex mixtures of carboxylic acids from petroleum that have industrial applications and that may be released to the environment after oil spills. There is significant research on the chemical composition and toxicity of water-soluble NAs derived from oil sands mining in Alberta, Canada. Yet, little is known about low molecular weight organic compounds (LMWOC) from these sources. Headspace solid-phase microextraction coupled to gas chromatography-electron impact mass spectrometry was used for LMWOC profiling of commercial NA blends, and an acid-extractable organics (AEOs) mixture from a tailings pond. From Sigma 1, Sigma 2, Merichem NAs and the AEO extract, 54, 56, 40 and 4 compounds were identified, respectively. These include aliphatic and cyclic hydrocarbons, carboxylic acids, alkylbenzenes, phenols, naphthalene and alkyl-naphthalene, and decalin compounds. A sample of oil sands process-affected water (OSPW) and aqueous solutions of the NA blends were evaluated for matrix effects on LMWOC profiles. Principal component and clustering analyses revealed that LMWOC profiles of commercial extracts were closely related but distinct from the AEO and OSPW samples. Some of the identified LMWOC are reported to be genotoxic or carcinogenic, and therefore the NA mixtures and AEOs should be considered hazardous materials and further evaluated.

Engineering Microbes for Remediation of Oil Sands Tailings.

Synthetic biology and adaptive laboratory evolution are key tools for developing biotechnology platforms for the remediation of oil sands tailings. However, field deployment and subsequent regulation of engineered and/or evolved strains is rife with uncertainties and risks. Here, we detail an innovation strategy to derisk and deploy engineered bioremediation platforms.

Persistent and transgenerational effects of raw and ozonated oil sands process-affected water exposure on a model vertebrate, the zebrafish.

Exposure to oil sands process-affected water (OSPW), a by-product of Canadian oil sands mining operations, can cause both acute and chronic adverse effects in aquatic life. Ozonation effectively degrades naphthenic acids in OSPW, mitigating some of the toxicological effects of exposure. In this study we examined the effect of developmental exposure to raw and ozonated OSPW had on the breeding success, prey capture, and alarm cue response in fish months/years after exposure and the transgenerational effect exposure had on gene expression, global DNA methylation, and larval basal activity. Exposure to raw and ozonated OSPW had no effect on breeding success, and global DNA methylation. Exposure altered the expression of vtg and nkx2.5 in the unexposed F1 generation. Exposure to both raw and ozonated OSPW had a transgenerational impact on larval activity levels, anxiety behaviors, and maximum swim speed compared to the control population. Prey capture success was unaffected, however, the variability in the behavioral responses to the introduction of prey was decreased. Fish developmentally exposed to either treatment were less active before exposure and did not have an anxiety response to the alarm cue hypoxanthine-3-n-oxide. Though ozonation was able to mitigate some of the effects of OSPW exposure, further studies are needed to understand the transgenerational effects and the implications of exposure on complex fish behaviors.