Potential for Microbially Mediated Natural Attenuation of Diluted Bitumen on the Coast of British Columbia (Canada).

Western Canada produces large amounts of bitumen, a heavy, highly weathered crude oil. Douglas Channel and Hecate Strait on the coast of British Columbia are two water bodies that may be impacted by a proposed pipeline and marine shipping route for diluted bitumen (dilbit). This study investigated the potential of microbial communities from these waters to mitigate the impacts of a potential dilbit spill. Microcosm experiments were set up with water samples representing different seasons, years, sampling stations, and dilbit blends. While the alkane fraction of the tested dilbit blends was almost completely degraded after 28 days, the majority of the polycyclic aromatic hydrocarbons (PAHs) remained. The addition of the dispersant Corexit 9500A most often had either no effect or an enhancing effect on dilbit degradation. Dilbit-degrading microbial communities were highly variable between seasons, years, and stations, with dilbit type having little impact on community trajectories. Potential oil-degrading genera showed a clear succession pattern and were for the most part recruited from the "rare biosphere." At the community level, dispersant appeared to stimulate an accelerated enrichment of genera typically associated with hydrocarbon degradation, even in dilbit-free controls. This suggests that dispersant-induced growth of hydrocarbon degraders (and not only increased bioavailability of oil-associated hydrocarbons) contributes to the degradation-enhancing effect previously reported for Corexit 9500A.IMPORTANCE Western Canada hosts large petroleum deposits, which ultimately enter the market in the form of dilbit. Tanker-based shipping represents the primary means to transport dilbit to international markets. With anticipated increases in production to meet global energy needs, the risk of a dilbit spill is expected to increase. This study investigated the potential of microbial communities naturally present in the waters of a potential dilbit shipping lane to mitigate the effects of a spill. Here we show that microbial degradation of dilbit was mostly limited to n-alkanes, while the overall concentration of polycyclic aromatic hydrocarbons, which represent the most toxic fraction of dilbit, decreased only slightly within the time frame of our experiments. We further investigated the effect of the oil dispersant Corexit 9500A on microbial dilbit degradation. Our results highlight the fact that dispersant-associated growth stimulation, and not only increased bioavailability of hydrocarbons and inhibition of specific genera, contributes to the overall effect of dispersant addition.

Factors that affect water column hydrocarbon concentrations have minor impacts on microbial responses following simulated diesel fuel spills.

Effects of season and mixing on hydrocarbon concentrations and the microbial community response was explored in a series of mesocosm experiments simulating surface spills of diesel into coastal waters. Mixing of any amount contributed to hydrocarbons entering the water column, but diesel fuel composition had a significant effect on hydrocarbon concentrations. Higher initial concentrations of aromatic hydrocarbons resulted in higher water column concentrations, with minimal differences among seasons due to high variability. Regardless of the concentrations of hydrocarbons, prokaryotes increased and there were higher relative abundances of hydrocarbon affiliated bacteria with indications of biodegradation within 4 d of exposure. As concentrations decreased over time, the eukaryote community shifted from the initial community to one which appeared to be composed of organisms with some resilience to hydrocarbons. This series of experiments demonstrates the wide range of conditions under which natural attenuation of diesel fuel is an effective response.

Microbial community response to simulated diluted bitumen spills in coastal seawater and implications for oil spill response.

Oil spills in coastal waters can have devastating impacts on local ecosystems, from the microscopic base through to mammals and seabirds. Increasing transport of diluted bitumen has led to concerns about how this novel product might impact coastal ecosystems. A mesocosm study determined that the type of diluent and the season can affect the concentrations of hydrocarbons entering the water column from a surface spill. Those same mesocosms were sampled to determine whether diluent type and season also affected the microbial response to a surface spill. Overall, there were no differences in impacts among the three types of diluted bitumen, but there were consistent responses to all products within each season. Although microbial abundances with diluted bitumen rarely differed from unoiled controls, community structure in these organisms shifted in response to hydrocarbons, with hydrocarbon-degrading bacteria becoming more abundant. The relative abundance of heterotrophic eukaryotes also increased with diluted bitumen, with few photosynthetic organisms responding positively to oil. Overall shifts in the microbial communities were minimal relative to spills of conventional oil products, with low concentrations of hydrocarbons in the water column. Oil spill response should focus on addressing the surface slick to prevent sinking or stranding to minimize ecosystem impacts.

Measuring the fate of different diluted bitumen products in coastal surface waters.

Diluted bitumens are produced by adding lower viscosity diluent to highly viscous bitumen to enable it to flow through pipelines and thus may behave differently than conventional oils when spilled into coastal seawater. Simulated surface spills using three different diluted bitumen products were carried out in May, July and November and water column hydrocarbons were monitored over a 14 day period. Volatile and total petroleum hydrocarbons varied in the water column depending on season and type of diluent. In summer, products diluted with synthetic crude or a mixture of condensate and crude released droplets into the water column. Diluted bitumen did not sink to the bottom of the enclosures with surface slicks showing a range of weathering after 14 d. With most of the diluted bitumen product remaining on the surface for 14 d, a rapid conventional clean up response may be effective in low energy, coastal waters.

Inorganic nutrients have a significant, but minimal, impact on a coastal microbial community's response to fresh diluted bitumen.

Microbes capable of degrading hydrocarbons in oil are present in low abundances in coastal waters, but quickly respond to oil following a spill. When estimating potential biodegradation rates in the laboratory, high concentrations of inorganic nutrients are often added to prevent nutrient limitation. In this study, we tested the short term response of coastal microbes to fresh diluted bitumen under varying nutrient conditions in a cold water regime. Total hydrocarbon concentrations changed minimally over five days; however, oil composition changed over time and the abundance of microbes increased in all treatments. Addition of phosphate, with or without nitrogen, resulted in rapid changes in community composition, but after three days treatments no longer differed. Nutrients were never depleted in any treatment suggesting that, even at low inorganic nutrient concentrations, microbial communities can quickly respond to hydrocarbons following a spill.

Chemical dispersants enhance the activity of oil- and gas condensate-degrading marine bacteria.

Application of chemical dispersants to oil spills in the marine environment is a common practice to disperse oil into the water column and stimulate oil biodegradation by increasing its bioavailability to indigenous bacteria capable of naturally metabolizing hydrocarbons. In the context of a spill event, the biodegradation of crude oil and gas condensate off eastern Canada is an essential component of a response strategy. In laboratory experiments, we simulated conditions similar to an oil spill with and without the addition of chemical dispersant under both winter and summer conditions and evaluated the natural attenuation potential for hydrocarbons in near-surface sea water from the vicinity of crude oil and natural gas production facilities off eastern Canada. Chemical analyses were performed to determine hydrocarbon degradation rates, and metagenome binning combined with metatranscriptomics was used to reconstruct abundant bacterial genomes and estimate their oil degradation gene abundance and activity. Our results show important and rapid structural shifts in microbial populations in all three different oil production sites examined following exposure to oil, oil with dispersant and dispersant alone. We found that the addition of dispersant to crude oil enhanced oil degradation rates and favored the abundance and expression of oil-degrading genes from a Thalassolituus sp. (that is, metagenome bin) that harbors multiple alkane hydroxylase (alkB) gene copies. We propose that this member of the Oceanospirillales group would be an important oil degrader when oil spills are treated with dispersant.

Characterization of the microbial community structure and the physicochemical properties of produced water and seawater from the Hibernia oil production platform.

Hibernia is Canada's largest offshore oil platform. Produced water is the major waste byproduct discharged into the ocean. In order to evaluate different potential disposal methods, a comprehensive study was performed to determine the impact from the discharge. Microorganisms are typically the first organisms to respond to changes in their environment. The objectives were to characterize the microbial communities and the chemical composition in the produced water and to characterize changes in the seawater bacterial community around the platform. The results from chemical, physicochemical, and microbial analyses revealed that the discharge did not have a detectable effect on the surrounding seawater. The seawater bacterial community was relatively stable, spatially. Unique microorganisms like Thermoanaerobacter were found in the produced water. Thermoanaerobacter-specific q-PCR and nested-PCR primers were designed, and both methods demonstrated that Thermoanaerobacter was present in seawater up to 1000 m from the platform. These methods could be used to track the dispersion of produced water into the surrounding ocean.

Biostimulation for the treatment of an oil-contaminated coastal salt marsh.

A field study was conducted on a coastal salt marsh in Nova Scotia, Canada, during the summer of 2000. The objective of the study was to assess the effectiveness of biostimulation in restoring an oil-contaminated coastal marsh dominated by Spartina alterniflora under north-temperate conditions. Three remediation treatments were tested with two additional unoiled treatments, with and without added nutrients, serving as controls. This research determined the effectiveness of nitrogen and phosphorus addition for accelerating oil disappearance, the role of nutrients in enhancing restoration in the absence of wetland plants, and the rate at which the stressed salt marsh recovered. Petroleum hydrocarbons were analyzed by gas chromatography/mass spectrometry (GC/MS). Statistically significant treatment differences were observed for alkanes but not aromatics in sediment samples. No differences were evident in above-ground vegetation samples. GC/MS-resolved alkanes and aromatics degraded substantially (>90% and >80%, respectively) after 20 weeks with no loss of TPH. Biodegradation was determined to be the main oil removal mechanism rather than physical washout.

Assessment of sediment toxicity during anaerobic biodegradation of vegetable oil using Microtox and Hyalella azteca bioassays.

The potential ecological impacts of anaerobic degradation of vegetable oil on freshwater sediments were investigated. Sediment toxicity was evaluated using two regulatory biotests: the Microtox Solid Phase Test and an amphipod (Hyalella azteca) bioassay. The results of the Microtox test showed that the toxicity of the vegetable-oil-contaminated sediments (about 17-33 g oil/kg dry sediments) increased after 2 weeks incubation and then decreased to near background levels after incubation for 8 weeks under anaerobic conditions. The amphipod toxicity bioassay showed that the toxicity of fresh contaminated sediments decreased over time and returned to background levels within 8 weeks. These results suggest that the impact of vegetable oils on organisms within sediments may be limited. To account for the significance of environmental conditions, additional studies over a wide range of incubation conditions (e.g., temperature, nutrient concentration) and other test organisms at various trophic levels are recommended for both acute and chronic toxicity assessment.