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.

Isotope forensics of polycyclic aromatic compounds (PACs) in a contaminated shallow aquifer.

Diesel was accidently released into the shallow subsurface at an industrial site in the province of Québec, Canada, in the late 1980s. Subsequent remediation efforts removed much of the contamination; however, traces of petroleum hydrocarbons continue to impact the local aquifer. In addition to the historical diesel spill, more recent yet unconfirmed accidental releases from ongoing on-site and neighbouring industrial activities may have potentially contributed to elevated levels of polycyclic aromatic compounds (PACs) in groundwater. To identify the main source(s) of contamination, compound-specific stable carbon isotope ratios (δ13C) of PACs in groundwater monitoring wells were compared to those in asphalt produced from a nearby plant and in fuel oil #6 oil being used by local industry. The δ13C values of five individual compounds (biphenyl, C2-naphthalene, C1-fluorene, dibenzothiophene and phenanthrene) and two groups of combined C1-phenanthrenes/anthracenes in all groundwater samples were within analytical uncertainty (±0.5‰). Moreover, the δ13CPAC values in groundwater samples were distinct from those in asphalt and fuel oil #6, indicating negligible contributions from these sources. The similarity in δ13CPAC values across monitoring wells, including one situated in the former source zone containing a floating hydrocarbon phase, pointed to a common source of subsurface contamination that was attributed to the historical diesel spill. These results thus demonstrate that δ13CPAC values can be used for source apportionment in shallow aquifers decades after the original spill event.

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.

Paleo-biodegradation and hydrocarbon mixing in a major hybrid petroleum reservoir.

Some of the parental material for hydrocarbons produced from low-permeability reservoirs in Western Canada corresponds to thermal products from biodegraded oil. This has been proved by the occurrence of framboidal pyrite, which is often formed during microbial sulfate reduction (MSR). In addition, the identified pyrite framboids are associated with the presence of phosphorus (P). Phosphorus (as phosphate) is a key nutrient and energy carrier for sulfate-reducing bacteria. The pyrite-P assemblage occurs embedded in solid bitumen (thermal residue), which confirms that migrated hydrocarbons provided the environment for microbial growth. Molecular products of severe biodegradation such as 17-nortricyclic terpanes were also detected. Biodegradation effects have been masked not only by thermal degradation of biodegraded oil during maximum burial, but also due to hydrocarbon mixing with late gas-condensate charges. Suitable conditions for biodegradation (< 80 °C, basin uplift) occurred during the Early Cretaceous. The confirmation of paleo-biodegradation means that there was a significant hydrocarbon loss that we have not accounted for. Likewise, MSR and Early Cretaceous seawater sulfate might have played an important role in the generation of the hydrogen sulfide (H2S) detected today.

Isotopic and microbial evidence for biodegradation of diluted bitumen in the unsaturated zone.

The oil sands region in Western Canada is one of the world's largest proven oil reserves. To facilitate pipeline transport, highly viscous oil sands bitumen is blended with lighter hydrocarbon fractions to produce diluted bitumen (dilbit). Anticipated increases in dilbit production and transport raise the risk of inland spills. To understand the behaviour of dilbit in the unsaturated or vadose zone following a surface spill, we ran parallel dilbit and conventional heavy crude exposures, along with an untreated control, using large soil-filled columns over 104 days. Phospholipid fatty acids (PLFAs), biomarkers for the active microbial population, were extracted from column soil cores. Stable carbon isotope contents (δ13C) of individual PLFAs and radiocarbon contents (Δ14C) of bulk PLFAs were characterized over the course of the experiment. The Δ14CPLFA values in soils impacted by dilbit (-221.1 to -54.7‰) and conventional heavy crude (-259.4 to -97.9‰) indicated similar levels of microbial uptake of fossil carbon. In contrast, Δ14CPLFA values in the control column (-46.1 to +53.7‰) reflected assimilation of more recently fixed organic carbon. Sequencing of 16S ribosomal RNA genes extracted from soil cores revealed a significant increase in the relative abundance of Polaromonas, a known hydrocarbon-degrader, following exposure to both types of oil. This study demonstrates that in the first several months following a surface spill, dilbit has a similar potential for biodegradation by a native shallow subsurface microbial community as conventional heavy crude oil.

Comparing simulated shallow subsurface spills of diluted bitumen and conventional crude oil.

Increased oil production in Canada has resulted in proposals to extend or develop new oil pipelines. Many of these proposals have been met by concerns from the public over potential environmental impacts related to construction and the potential for oil spills to negatively affect groundwater quality. Crude oil sourced from the Alberta oil sands represents a significant proportion of this increase in production. This crude oil is produced as bitumen, which is subsequently diluted with light hydrocarbons to lower viscosity to allow for pipeline transport producing diluted bitumen. In this study, we pumped water through tanks filled with sand to simulate groundwater flow. Tanks were injected with either conventional crude or diluted bitumen to simulate a crude oil spill from a pipeline rupture occurring below the water table representing a pipeline river crossing scenario. Water samples were collected from the downstream end of the tanks throughout the experiment period (∼two months). Compared to water quality guidelines, effluent waters from both conventional crude and diluted bitumen tanks contained elevated concentrations of dissolved organic compounds, particularly benzene, ethylbenzene, toluene and xylenes (BTEX). The effluent from each tank had similar concentrations of benzene, whereas discharge water from conventional crude tanks contained higher concentrations of ethylbenzene, toluene and xylenes. In both tanks, and as expected, the BTEX concentrations appeared to be proportional to those determined in their injected crude oils. The measured dissolved concentrations of benzene, ethylbenzene and toluene are lower than predicted which is attributed largely due to dilution along the flow path. In addition to organic constituents, effluent sampled from the diluted bitumen tank contained some metals (Co, Cr, Fe and V) which measured constituents of the oil.

Quantification of Spatial and Temporal Trends in Atmospheric Mercury Deposition across Canada over the Past 30 Years.

Mercury (Hg) is a pollutant of concern across Canada and transboundary anthropogenic Hg sources presently account for over 95% of national anthropogenic Hg deposition. This study applies novel statistical analyses of 82 high-resolution dated lake sediment cores collected from 19 regions across Canada, including nearby point sources and in remote regions and spanning a full west-east geographical range of ∼4900 km (south of 60°N and between 132 and 64°W) to quantify the recent (1990-2018) spatial and temporal trends in anthropogenic atmospheric Hg deposition. Temporal trend analysis shows significant synchronous decreasing trends in post-1990 anthropogenic Hg fluxes in western Canada in contrast to increasing trends in the east, with spatial patterns largely driven by longitude and proximity to known point source(s). Recent sediment-derived Hg fluxes agreed well with the available wet deposition monitoring. Sediment-derived atmospheric Hg deposition rates also compared well to the modeled values derived from the Hg model, when lake sites located nearby (<100 km) point sources were omitted due to difficulties in comparison between the sediment-derived and modeled values at deposition "hot spots". This highlights the applicability of multi-core approaches to quantify spatio-temporal changes in Hg deposition over broad geographic ranges and assess the effectiveness of regional and global Hg emission reductions to address global Hg pollution concerns.

Isotopic Analyses Fingerprint Sources of Polycyclic Aromatic Compound-Bearing Dust in Athabasca Oil Sands Region Snowpack.

Fugitive dust associated with surface mining activities is one of the principal vectors for transport of airborne contaminants in Canada's Athabasca oil sands region (AOSR). Effective environmental management requires quantitative identification of the sources of this dust. Using natural abundance radiocarbon (Δ14C) and dual (δ13C, δ2H) compound-specific isotope analysis (CSIA), this study investigated the sources of dust and particulate-bound polycyclic aromatic compounds (PACs) deposited in AOSR lake snowpack. Lower Δ14C values, higher particulate and PAC loadings, and lower δ13C values for phenanthrene and C1-alkylated phenanthrenes/anthracenes (C1-Phen) at sites closer to the mining operations indicated unprocessed oil sand and/or petroleum coke (petcoke-a byproduct of bitumen upgrading) as major sources of anthropogenic fugitive dust. However, a Bayesian isotopic mixing model that incorporated both δ13C and δ2H could discriminate petcoke from oil sand, and determined that petcoke comprised between 44 and 95% (95% credibility intervals) of a C1-Phen isomer at lakes <25 km from the heart of the mining operations, making it by far the most abundant source. This study is the first to demonstrate the potential of CSIA to provide accurate PAC source apportionment in snowpack and reveals that petcoke rather than oil sand is the main source of mining-related particulate PACs deposited directly to AOSR lakes.

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.

Using tree cores to evaluate historic atmospheric concentrations and trends of polycyclic aromatic compounds in the Oil Sands region of Alberta, Canada.

Tree cores and bark were sampled from jack pine trees at 18 sites in the Athabasca Oil Sands Region (AOSR) of Alberta, Canada, to investigate spatial and temporal trends of polycyclic aromatic compounds (PACs). Spatial trends were investigated in the bark samples, where ΣPAC concentrations ranged from 75 to 3615 ng/g. Highest concentrations were observed from trees within 40 km of the nearest mining or upgrading facility perimeter fence, in line with previous deposition studies in the AOSR. The sampled tree cores were separated into segments representing 5 years of growth/atmospheric collection by counting tree rings. A significant increase in PAC concentrations over the lifetime of the tree was observed at sites with the highest PAC concentrations, and the average % increase in concentration from 1970 to 2015 was in line with average % growth in bitumen extraction in the AOSR. Finally, the concentrations in the tree core segments representing collection from 2010 to 2015 were converted into an atmospheric PAC concentration using previously published wood-air partition coefficients. The calculated atmospheric concentrations were within the same range as concentrations reported from the passive atmospheric sampling network in this region. The importance of site location is highlighted, with forest edge sites providing an improved comparison for atmospheric exposure and deposition. This is the first study to use tree cores to calculate an atmospheric concentration of PACs, demonstrating the applicability of this methodology for providing historic atmospheric data.

Advances in Distinguishing Groundwater Influenced by Oil Sands Process-Affected Water (OSPW) from Natural Bitumen-Influenced Groundwaters.

The objective of this study was to advance analytical methods for detecting oil sands process-affected water (OSPW) seepage from mining containments and discriminating any such seepage from the natural bitumen background in groundwaters influenced by the Alberta McMurray formation. Improved sampling methods and quantitative analyses of two groups of monoaromatic acids were employed to analyze OSPW and bitumen-affected natural background groundwaters for source discrimination. Both groups of monoaromatic acids showed significant enrichment in OSPW, while ratios of O2/O4 containing heteroatomic ion classes of acid extractable organics (AEOs) did not exhibit diagnostic differences. Evaluating the monoaromatic acids to track a known plume of OSPW-affected groundwater confirmed their diagnostic abilities. A secondary objective was to assess anthropogenically derived artificial sweeteners and per- and polyfluoroalkyl substances (PFAS) as potential tracers for OSPW. Despite the discovery of acesulfame and PFAS in most OSPW samples, trace levels in groundwaters influenced by general anthropogenic activities preclude them as individual robust tracers. However, their inclusion with the other metrics employed in this study served to augment the tiered, weight of evidence methodology developed. This methodology was then used to confirm earlier findings of OSPW migrations into groundwater reaching the Athabasca River system adjacent to the reclaimed pond at Tar Island Dyke.

Distinguishing Natural from Anthropogenic Sources of Acid Extractable Organics in Groundwater near Oil Sands Tailings Ponds.

Distinguishing between naphthenic acids (NAs) associated with oil sands process-affected water (OSPW) and those found naturally in groundwaters in contact with the bituminous McMurray Formation poses a considerable analytical challenge to environmental research in Canada's oil sands region. Previous work addressing this problem combined high-resolution Orbitrap mass spectrometry with carbon isotope values generated by online pyrolysis (δ13Cpyr) to characterize and quantify the acid extractable organics (AEOs) fraction containing NAs in the subsurface near an oil sands tailings pond. Here, we build upon this work through further development and application of these techniques at two different study sites near two different tailings ponds, in conjunction with the use of an additional isotopic tool-sulfur isotope analysis (δ34S) of AEOs. The combined use of both δ13Cpyr and δ34S allowed for discrimination of AEOs into the three end-members relevant to ascertaining the NA environmental footprint within the region: (1) OSPW; (2) McMurray Formation groundwater (i.e., naturally occurring bitumen), and; (3) naturally occurring non-bitumen. A Bayesian isotopic mixing model was used to determine the relative proportions of these three sources in groundwater at both study sites. Although background levels of OSPW-derived AEOs were generally low, one sample containing 49-99% (95% credibility interval) OSPW-derived AEOs was detected within an inferred preferential flow-path, highlighting the potential for this technique to track tailings pond seepage.

Evaluating in situ biodegradation of 13C-labelled naphthenic acids in groundwater near oil sands tailings ponds.

Potential seepage of naphthenic acids (NAs) from tailings ponds into surface water and groundwater is one of the main environmental concerns associated with the Canadian Athabasca oil sands mining operations. Here we report the application of 13C-labelled NA surrogate compounds to evaluate intrinsic biodegradation along groundwater flow-paths originating from oil sands tailings ponds at two different sites: a glacio-fluvial aquifer (Site 1) and a low-lying wetland (Site 2). Microcosms containing the carboxyl group labelled (99%) NA surrogates (cyclohexanecarboxylic acid, CHCA; 1,2-cyclohexanedicarboxylic acid, CHDCA; 1-adamantanecarboxylic acid, ACA) were lowered into monitoring wells for several months to allow sufficient time for substrate degradation and formation of a biofilm in conditions characteristic of the local aquifer. Phospholipid fatty acids (PLFAs), biomarkers for the active microbial population, were extracted from the biofilms for stable carbon isotope (δ13C) analysis. At Site 1, highly 13C-enriched δ13C values (up to ~+7100‰) confirmed the in situ microbial breakdown of CHCA and CHDCA. At Site 2, δ13C-PLFA values from -60.6 to -24.5‰ indicated uptake of a 13C-depleted substrate such as biogenic methane and not 13C-labelled ACA. Determination of the microbial community using 16s RNA sequencing confirmed the presence of methane-oxidizing bacteria in the subsurface at Site 2. The in situ biodegradation of NAs at Site 1 demonstrates that the indigenous microbial population in the shallow subsurface near tailings ponds can readily break down some of these compounds prior to surface water discharge. The lack of evidence for microbial uptake of 13C-labelled ACA at Site 2 demonstrates that other NAs, in particular tricyclic diamondoid acids, may persist in the environment following seepage from tailings ponds or natural sources.

Naphthenic acids in groundwater overlying undeveloped shale gas and tight oil reservoirs.

The acid extractable organics (AEOs) containing naphthenic acids (NAs) in groundwater overlying undeveloped shale gas (Saint-Édouard region) and tight oil (Haldimand sector, Gaspé) reservoirs in Québec, Canada, were analysed using high resolution Orbitrap mass spectrometry and thermal conversion/elemental analysis - isotope ratio mass spectrometry. As classically defined by CnH2n+ZO2, the most abundant NAs detected in the majority of groundwater samples were straight-chain (Z = 0) or monounsaturated (Z = -2) C16 and C18 fatty acids. Several groundwater samples from both study areas, however, contained significant proportions of presumably alicyclic bicyclic NAs (i.e., Z = -4) in the C10-C18 range. These compounds may have originated from migrated waters containing a different distribution of NAs, or are the product of in situ microbial alteration of shale organic matter and petroleum. In most groundwater samples, intramolecular carbon isotope values generated by pyrolysis (δ13Cpyr) of AEOs were on average around 2-3‰ heavier than those generated by bulk combustion (δ13C) of AEOs, providing further support for microbial reworking of subsurface organic carbon. Although concentrations of AEOs were very low (<2.0 mg/L), the detection of potentially toxic bicyclic acids in groundwater overlying unconventional hydrocarbon reservoirs points to a natural background source of organic contaminants prior to any large-scale commercial hydrocarbon development.

Lake-sediment record of PAH, mercury, and fly-ash particle deposition near coal-fired power plants in Central Alberta, Canada.

We report a historical record of atmospheric deposition in dated sediment cores from Hasse Lake, ideally located near both currently and previously operational coal-fired power plants in Central Alberta, Canada. Accumulation rates of spheroidal carbonaceous particles (SCPs), an unambiguous marker of high-temperature fossil-fuel combustion, in the early part of the sediment record (pre-1955) compared well with historical emissions from one of North America's earliest coal-fired power plants (Rossdale) located ∼43 km to the east in the city of Edmonton. Accumulation rates in the latter part of the record (post-1955) suggested inputs from the Wabamun region's plants situated ∼17-25 km to the west. Increasing accumulation rates of SCPs, polycyclic aromatic hydrocarbons (PAHs) and Hg coincided with the previously documented period of peak pollution in the Wabamun region during the late 1960s to early 1970s, although Hg deposition trends were also similar to those found in western North American lakes not directly affected by point sources. A noticeable reduction in contaminant inputs during the 1970s is attributed in part to technological improvements and stricter emission controls. The over one hundred-year historical record of coal-fired power plant emissions documented in Hasse Lake sediments has provided insight into the impact that both environmental regulations and changes in electricity output have had over time. This information is crucial to assessing the current and future role of coal in the world's energy supply.

Isotopic Evidence for Oil Sands Petroleum Coke in the Peace-Athabasca Delta.

The continued growth of mining and upgrading activities in Canada's Athabasca oil sands (AOS) region has led to concerns about emissions of contaminants such as polycyclic aromatic hydrocarbons (PAHs). Whereas a recent increase in PAH emissions has been demonstrated within around 50 km of the main center of surface mining and upgrading operations, the exact nature of the predominant source(s) and the geographical extent of the deposition are still under debate. Here, we report a century-long source apportionment of PAHs using dual (δ(2)H, δ(13)C) compound-specific isotope analysis on phenanthrene deposited in a lake from the Athabasca sector of the Peace-Athabasca Delta situated ∼150 km downstream (north) of the main center of mining operations. The isotopic signatures in the core were compared to those of the main potential sources in this region (i.e., unprocessed AOS bitumen, upgrader residual coke, forest fires, coal, gasoline and diesel soot). A significant concurrent increase (∼55.0‰) in δ(2)H and decrease (∼1.5‰) in δ(13)C of phenanthrene over the last three decades pointed to an increasingly greater component of petcoke-derived PAHs. This study is the first to quantify long-range (i.e., >100 km) transport of a previously under-considered anthropogenic PAH source in the AOS region.

Source Apportionment of Background PAHs in the Peace-Athabasca Delta (Alberta, Canada) Using Molecular Level Radiocarbon Analysis.

The downstream accumulation of polycyclic aromatic hydrocarbons (PAHs) in the Peace-Athabasca Delta (PAD), an ecologically important landscape, is a key issue of concern given the rapid development of the oil sands industry in Northern Alberta, Canada. In addition to PAHs derived from industrial activity (i.e., oil sands mining) within the Athabasca watershed, however, forest fires and erosion of fossil fuel deposits within both the Athabasca and Peace watersheds are two potentially important natural sources of PAHs delivered to the PAD. Consequently, evaluating the environmental impact of mining activities requires a quantitative understanding of natural, background PAHs. Here, we utilize molecular-level natural-abundance radiocarbon measurements on an amalgamated sediment record from a Peace River flood-susceptible oxbow lake in the northern Peace sector of the PAD to quantitatively discriminate sources of naturally occurring alkylated PAHs (fossil and modern biomass). A radiocarbon mass balance quantified a predominantly natural petrogenic source (93% petrogenic, 7% forest fire) for alkylated PAHs during the past ∼50 years. Additionally, a significant petrogenic component determined for retene, a compound usually considered a biomarker for softwood combustion, suggests that its use as a unique forest fire indicator may not be suitable in PAD sediments receiving Peace watershed-derived fluvial inputs.

Direct evaluation of in situ biodegradation in Athabasca oil sands tailings ponds using natural abundance radiocarbon.

Compound-specific stable (δ(13)C) and radiocarbon (Δ(14)C) isotopes of phospholipid fatty acids (PLFAs) were used to evaluate carbon sources utilized by the active microbial populations in surface sediments from Athabasca oil sands tailings ponds. Algal-specific PLFAs were absent at three of the four sites investigated, and δ(13)CPLFA values were generally within ~3‰ of that reported for oil sands bitumen (~-30‰), suggesting that the microbial communities growing on petroleum constituents were dominated by aerobic heterotrophs. Δ(14)CPLFA values ranged from -906 to -586‰ and pointed to significant uptake of fossil carbon, particularly in PLFAs (e.g., cy17:0 and cy19:0) often associated with petroleum hydrocarbon degrading bacteria. The comparatively heavier Δ(14)C values found in other, less specific PLFAs (e.g., 16:0) indicated the preferential uptake of younger organic matter by the general microbial population. Since the main carbon pools in tailings sediment were essentially "radiocarbon dead" (i.e., Δ(14)C ~ -1000‰), the principal source for this relatively modern carbon is considered to be the Athabasca River, which provides the bulk of the water used in the bitumen extraction process. The preferential utilization of the minor amount of younger and presumably more labile material present in systems otherwise dominated by petroleum carbon has important implications for remediation strategies, since it implies that organic contaminants may persist long after reclamation has begun. Alternatively, this young organic matter could play a vital and necessary role in supporting the microbial utilization of fossil carbon via cometabolism or priming processes.