Enhanced Biodegradation of Petroleum Hydrocarbons in Contaminated Soil

Soil samples taken from a contaminated site in Northern Quebec, Canada, exhibited a low capacity for biodegradation of total petroleum hydrocarbons (TPH), despite a high capacity for the mineralization of aromatic hydrocarbons and a low toxicity of soil leachates as measured by Microtox assay. Toxicity assays directly performed on surface soil, including earthworm mortality and barley seedling emergence, indicated moderate to high levels of toxicity. Soil biostimulation did not improve the removal of petroleum hydrocarbons, while bioaugmentation of soil with a developed enrichment culture increased the efficiency of hydrocarbon removal from 20.4% to 49.2%. A considerable increase in the removal of TPH was obtained in a bioslurry process, enhancing the mass transfer of hydrocarbons from soil to the aqueous phase and increasing the efficiency of hydrocarbon removal to over 70% after 45 days of incubation. The addition of ionic or nonionic surfactants did not have a significant impact on biodegradation of hydrocarbons. The extent of hydrocarbon mineralization during the bioslurry process after 45 days of incubation ranged from 41.3% to 58.9%, indicating that 62.7% to 83.1% of the eliminated TPH were transformed into CO₂ and water.     

Enhanced Biodegradation of Petroleum Hydrocarbons in Contaminated Soil

Soil samples taken from a contaminated site in Northern Quebec, Canada, exhibited a low capacity for biodegradation of total petroleum hydrocarbons (TPH), despite a high capacity for the mineralization of aromatic hydrocarbons and a low toxicity of soil leachates as measured by Microtox assay. Toxicity assays directly performed on surface soil, including earthworm mortality and barley seedling emergence, indicated moderate to high levels of toxicity. Soil biostimulation did not improve the removal of petroleum hydrocarbons, while bioaugmentation of soil with a developed enrichment culture increased the efficiency of hydrocarbon removal from 20.4% to 49.2%. A considerable increase in the removal of TPH was obtained in a bioslurry process, enhancing the mass transfer of hydrocarbons from soil to the aqueous phase and increasing the efficiency of hydrocarbon removal to over 70% after 45 days of incubation. The addition of ionic or nonionic surfactants did not have a significant impact on biodegradation of hydrocarbons. The extent of hydrocarbon mineralization during the bioslurry process after 45 days of incubation ranged from 41.3% to 58.9%, indicating that 62.7% to 83.1% of the eliminated TPH were transformed into CO₂ and water.     

石油烃的厌氧生物降解对油藏残余油气化开采的启示

利用微生物将油藏中难以动用的原油就地转化为甲烷,以天然气的形式开采、或作为战略资源就地储备,从而大幅度提高油气资源的利用率,是当前国际上研究的前沿课题。本文综述了石油烃厌氧生物降解转化为甲烷的菌群结构、反应热力学和反应动力学等基础科学问题的最新研究进展,讨论了油藏残余油气化开采技术的可行性及开发潜力,提出了该技术进一步研究的方向。     

Aerobic and Anaerobic Biodegradation of Aged Pentachlorophenol by Indigenous Microorganisms

Pentachlorophenol (PCP) use as a general biocide, particularly for treating wood, has led to widespread environmental contamination. Biodegradation has emerged as the main mechanism for PCP degradation in soil and groundwater and a key strategy for remediation. Examining the microbial biodegrading potential for PCP at a contaminated site is crucial in determining its fate. Hundreds of studies have been published on PCP microbial degradation, but few have described the biodegradation of PCP that has been in contact with soils for many years. The bioavailability of “aged” hydrophobic organics is a significant concern. PCP- and 2,3,4,6-tetrachlorophenol (2,3,4,6-TeCP)-contaminated soil samples from several depths at a former wood treatment site were placed under varying conditions in the laboratory to determine the anaerobic and aerobic potential for biodegradation of chlorophenols at the site. PCP biodegradation occurred in both anaerobic and aerobic soil samples. Rapid aerobic degradation occurred in samples spiked with 2- and 4-chlorophenol, but not with 3-chlorophenol. Reductive dechlorination of PCP in anaerobic samples resulted in the accumulation of 3-chlorophenol. In most anaerobic replicates, 3-chlorophenol was degraded with the appearance of detectable, but not quantifiable amounts of phenol. These results indicate excellent potential for remediation at the site using the indigenous microorganisms under both aerobic and anaerobic conditions. However, a fraction of the PCP was unavailable for degradation.     

Biodegradation of Petroleum Hydrocarbons in a Soil Polluted Sample by Oil-Based Drilling Cuttings

Microbial degradation of hydrocarbons in soils polluted by oil-based drilling mud and cuttings has been investigated by static methods such as composting or biopiling. Bioremediation of polluted soils by oil-based drilling cuttings through a slurry bioreactor has not previously been reported. The main aim of this work is to monitor hydrocarbon biodegradation in slurry of drilling cuttings and unpolluted soils and the effects of nutrients on it. Indigenous, bacterial-mixed culture isolated from a polluted soil by drilling cuttings adapted to drilling mud concentrations up to 15% (v/v) was done during a 15-month program. The total petroleum hydrocarbons’ (TPHs) removal efficiency in C/N/P 100/5/1 ratio was 90.5 and 79.85% under experimental and control conditions, respectively. The microbial count on the first day, 15 × 10⁷ CFUg^?1, reached 20 × 10⁹ CFUg^?1on the twenty-first day at experimental conditions. The TPH removal efficiency in C/N/P 100/10/2 was 92.5 and 82.25% at experiment and control, respectively. Increasing nitrogen and phosphorous amount couldn't increase microbial count in comparison with C/N/P ratio 100/5/1. The measured biomass contents and microbial counts in experiments were significantly higher than the control and confirmed hydrocarbons’ biodegradation during the time. Results showed that slurry bioreactors could accelerate the biodegradation of TPHs and reduce remediation time in soil polluted by oil-based drilling cuttings.     

A Method for Assessing Leaching Potential for Petroleum Hydrocarbons Release Sites: Multiphase and Multisubstance Equilibrium Partitioning

This article presents the rationale for the mathematical fate and transport model, which has been provided in the accompanying spreadsheet (GWProt). This spreadsheet model may be used as a simple and scientifically defensible regulatory tool for determining the risk-based soil clean up level of petroleum release sites to protect groundwater quality. The model incorporates either a three- or four-phase partitioning equilibrium mechanism, depending on the detection of Non-Aqueous Phase Liquid phase presence mathematically, as well as Raoult's Law convention and default dilution and attenuation factors. A database of contaminant-specific parameters, including solubility and organic-carbon partition-coefficient, molecular weight, and Henry's Law constant, is assembled for benzene, toluene, ethylbenzene, xylenes, and 12 other TPH equivalent carbon fractions. In addition to distributing organic chemicals among aqueous, sorbed solid, air, and NAPL phases, according to traditional partitioning equations, the algorithm incorporates equations for the conservation of mass and volume. A unique solution is obtained by solving a series of mass balance equations simultaneously using the iterative spreadsheet routine built in MICROSOFT EXCEL^TM Solver — with the restrictions that the volume is conserved and the sum of the mole fractions is equal to one. Sample calculations are presented for a range of parameter values to illustrate the use of the model and the relative leach-ability of a wide range of representative fuels. Sensitivity analysis was also performed to quantify the effects of uncertainty in the estimates of the key model parameters on model results. Model predictions were compared with the results from a water-fuel experiment. The noncar-cinogenic Hazard Index (HI) for groundwater through direct ingestion was calculated using predetermined oral reference dose (R_fd) values. Applications and limitations of the model are also discussed.     

石油污染土壤的生物修复技术

1　前　言在石油生产、贮运、炼制加工及使用过程中 ,由于事故 ,不正常操作及检修等原因 ,都会有石油烃类的溢出和排放。例如 ,油田开发过程中的井喷事故 ;输油管线和贮油罐的泄漏事故 ;油槽车和油轮的泄漏事故 ;油井清蜡和油田地面设备检修 ;炼油和石油化工生产装置检修等。石油烃类大量溢出 ,应当尽可能予以回收 ,但有的情况下回收很困难 ,即使尽力回收 ,仍会残留一部分 ,对环境 (土壤、地面和地下水 )造成污染。其进入土壤后 ,会破坏土壤结构 ,分散土粒 ,使土壤的透水性降低。其富含的反应基能与无机氮、磷结合并限制硝化作用和脱磷酸作…     

石油污染土壤的生物降解研究

石油工业迅速的发展带来了许多环境问题。在原油生产与输送过程中[1] ,井喷、泄露及沉降排放等引起的原油进入土壤造成的土壤污染 ,很难治理。原油在环境中残留时间长 ,对土壤微生物和土壤 植物生态系统 ,甚至地下水都产生危害 ,影响土壤肥力 ,破坏土壤生产力 ,严重影响当地的粮食产量及产品质量。当前 ,治理土壤石油污染的方法主要有物理法、化学法和生物治理技术[2 ] 。污染土壤生物清洁技术就是利用微生物将土壤中有害有机污染物降解为无害无机物 (ＣＯ2 和Ｈ2 Ｏ)的过程。降解过程可以由改变土壤理化条件 (包括土壤ｐＨ ,温度、湿度、…     

Mobility and Persistence of Petroleum Hydrocarbons in Peat Soils of Southeastern Mexico

Spilled crude petroleum from oil wells contains numerous hydrocarbons, some of which are toxic and threaten life. We have studied the mobility and persistence of hydrocarbons in waterlogged soils that contain large proportions of fermented organic matter (Histosols) and large concentrations of dissolved organic carbon (DOC) in the State of Tabasco, Mexico. We sampled soil and phreatic water at sites polluted by oil spills for several decades, as well as at sites that had only recently (few weeks) been polluted, and compared their hydrocarbon contents with those of unaffected sites in the same area. Samples were analyzed for 16 non-alkylated polyaromatic hydrocarbons (PAHs) and n-alkanes from nC9 to nC34. The spilled hydrocarbons had remained predominantly in the organic surface horizons of the soil where spillage occurred; there was little evidence of movement within the soil. The fraction of low molecular weight compounds was larger at sites of recent spills than where spills happened several decades ago. Nevertheless, sites of old spills still contained large concentrations of hydrocarbons, among which those of low molecular weight represented from 30 to 49% of total PAHs and from 50 to 84% of total n-alkanes, indicating that volatilization or microbial degradation is slow in these soils. In the peat horizons the measured organic carbon partition coefficients (K _oc ) for the higher molecular weight PAHs were consistently smaller than those estimated by empirical equations by up to two orders of magnitude. The dissolved organic carbon of these peat soils seems to influence this behavior. At sites of old spills, partition coefficients for the PAHs were larger than at sites of recent spills.     

Biodegradation of a PAH Mixture by Native Subsurface Microbiota

Laboratory microcosm studies were conducted to estimate biodegradation rates for a mixture of five polycyclic aromatic hydrocarbon compounds (PAHs). Static microcosms were assembled using soil samples from two locations collected at a No. 2 fuel oil-contaminated site in the Atlantic Coastal Plain of Virginia. In microcosms from one location, five PAHs (acenaphthene, fluorene, phenanthrene, pyrene, and benzo(b)fluoranthene) biodegraded at net first-order rates of 1.08, 1.45, 1.13, 1.11, and 1.12 yr^?1, respectively. No observable lag period was noted and degradation in live microcosms ceased with the depletion of oxygen and sulfate after 125 days. In microcosms from a second location, net first-order biodegradation rates after an approximately 2-month lag period were 2.41, 3.28, and 2.98 yr^?1 for fluorene, phenanthrene, and pyrene, respectively. Acenaphthene and benzo(b)fluoranthene mass loss rates in the live microcosms were not statistically different from mass loss rates in control microcosms. Stoichiometric mass balance calculations indicate that the dominant PAH mass loss mechanism was aerobic biodegradation, while abiotic losses (attributed to micropore diffusion and oxidative coupling) ranged from 15 to 33% and biotic losses from sulfate-reduction accounted for 7 to 10% of PAH mass loss. Stoichiometric equations that include biomass yield are presented for PAH oxidation under aerobic and sulfate-reducing conditions.     

Importance of Environmental Black Carbon to Dissolved Petroleum Hydrocarbons Sorption on Soil

Importance of environmental black carbon (BC) to sorption of dissolved petroleum hydrocarbons (DPH) on two soils with high BC:TOC ratios (33% and 11%, respectively) was evaluated at a relatively high concentrations (mg/L ～ μ g/L range). Sorption isotherms of DPH were determined for the two original soils and soils combusted at 375°C (only BC). The sorption isotherms of the original soils were linear, whereas the isotherms of the combusted soils were highly nonlinear (n _F = 0.45, 0.60). It is indicated that intrinsic BC-water sorption coefficient is not possible to be used to estimate total sorption to the original soil, even in our relatively high concentrations. From the sorption isotherms, Freundlich coefficient of environmental BC sorption, K _F,BC ^env of 10 ^{2.55 ± 0.21} was calculated and could be used as a generic starting point for environmental modeling purposes. From the data, it could be deduced that BC was responsible for 50% of the total sorption at concentrations of 45 and 4 μ g/L (μ g/L range), which were significantly higher than literature concentrations (ng/L range). These results demonstrate that in soil with high BC:TOC ratio BC is the most important geosorbent constituent with respect to sorption of DPH at relatively high concentrations ranged in μ g/L.     

Biotransformation and Dissolution of Petroleum Hydrocarbons in Natural Flowing Seawater at Low Temperature

The objective of this study was to establish methods for controlled studies of hydrocarbon depletion from thin oil films in cold natural seawater, and to determine biotransformation in relation to other essential depletion processes. Mineral oil was immobilized on the surface of hydrophobic Fluortex fabrics and used for studies of microbial biodegradation in an experimental seawater flow-through system at low temperatures (5.9-7.4 degrees C) during a test period of 42 days. The seawater was collected from a depth of 90 m, and microbial characterization by epifluorescence microscopy, fluorescence in situ hybridization, and most-probable number analysis showed relatively larger fractions of archaea and oil-degrading microbes than in the corresponding surface water. Chemical analysis of hydrocarbons attached to the fabrics during the test period showed that n-alkanes (C10-C36) were decreased by 98% after 21 days, while naphthalenes were depleted by 99-100% during the same period. At the end of the period 4-5 ring polyaromatic hydrocarbon (PAH) compounds were removed by 82% from the fabrics. Analysis of the recalcitrant pentacyclic triterpane C30 17alpha(H),21beta(H)-hopane showed that the oil remained adsorbed to the fabrics during the test period. Comparison of depletion analysis with calculation of hydrocarbon dissolution in a flow-through system indicated that naphthalenes and smaller PAH compounds (alkylated 2-ring and 3-ring compounds) were removed from the fabrics by dissolution. The data further implied that depletion of n-alkanes and 4-5 ring PAH hydrocarbons were the result of biotransformation processes. PCR amplification of bacterial 16S rRNA genes from microbes adhering on the immobilized oil surfaces showed the dominance of a few bands when analysed in denaturing gradient gel electrophoresis (DGGE). Sequence analysis of DGGE bands revealed phylogenetic affiliation to the alpha- and gamma-subdivisions of proteobacteria and to the Chloroflexus-Flavobacterium-Bacteroides group.     

Biotreatment of Total Petroleum Hydrocarbons from an oily Sludge Using Co-Composting Approach

Discharging of non-treated oily sludge from oil refineries has undesirable impacts on the environment. In this research, the biotreatment of total petroleum hydrocarbons (TPHs) from Abadan Petroleum Refinery, Iran was done using co-composting method. A 5 kg mixture of oily sludge and compost (oily sludge: compost ratios 1:0, 1:0.1, 1:0.3, 1:0.5, and 1:0.7 w/w) and a bulking agent of wheat straw were used as treatments. All treatments were placed in a wooden box inside a laboratory and aerated every 2–3 days by mixing during two months of the experiments. The variation of TPHs concentration, bacterial density, C/N ratio, pH, and temperature were assessed during the 63 days- experiments. Results showed that the majority of TPHs of samples was removed at first 30 days. The maximum TPHs removal (65%) was obtained in the sludge: compost = 1:0.5 at the operation time of 63 day. The significant different between removal efficiency of oily sludge: compost ratios and the control sample (p value < 0.05) indicated the appropriate density of TPHs degrader-bacteria in treated samples. The highest variation in C/N ratio was observed 15± 0.58 in the oily sludge: compost = 1:0.1. At the beginning of experiments, the pH values of all treatments was alkaline, but became neutral at the end of the reaction time. The TPHs degradation kinetic for all oily sludge: compost ratios followed pseudo second-order model. In general, co-composting has high efficiency in oily sludge remediation and can be potentially applied as a simple and cost-effective approach for remediation of oily sludge.     

Establishing Correlations and Scale-Up Factor for Estimating the Petroleum Biodegradation Rate in Soil

The proper design of a bioremediation strategy for petroleum-contaminated sites requires a reasonable estimate of the biodegradation rate constant, which is not easy due to spatial heterogeneity. Accordingly, predictive models were developed by completing a bioventing study at the meso-scale. Reactors holding 4 kg of disturbed soil were tested using five different types of soils. Using statistical analysis, a two-stage process was observed, with a fast rate of hydrocarbon degradation in the first 8 days and a slower rate in the remaining 22 days. Review of the correlations showed that the initial population of petroleum-degrading bacteria and increasing silt content had a positive effect on the degradation. A negative impact on the degradation rate was seen by increasing the fraction of organic matter and clay content. Comparison of previously completed micro-scale and meso-scale degradation rates gave a scale-up factor (SF) of 1.8 ± 0.5. Soils with an increased sand fraction had slightly higher SF values, whereas soils high in organic matter content had lower SF values. The measured SF values and developed correlations will help practitioners with site closure decisions, indicating the need for additional SF work to allow better transfer of meso-scale data to the field.     

Degradation of Polycyclic Aromatic Hydrocarbons in the Rhizosphere of Festuca arundinacea and Associated Microbial Community Changes

A phytoremediation growth chamber study was conducted to evaluate the contribution of soil microbial diversity to the contaminant degradation. Target contaminant removal from soil was assessed by monitoring concentrations of polycyclic aromatic hydrocarbons (PAHs), along with changes in the bacterial community structure over a time period of 10 months in the presence of tall fescue (Festuca arundinacea). Enhanced degradation of PAHs was observed in rhizosphere soil, with a maximum reduction in pyrene at a rate 36% higher than that noted for the unvegetated control. The dissipation of < 4-ring PAHs, 4-ring PAHs, and > 4-ring PAHs in unvegetated soil was 70%, 54%, and 49% respectively, whereas a higher dissipation rate was observed in tall fescue treated soil of 78%, 68%, and 61% at the end of the study. Microbial enumeration results showed greater total bacterial numbers and PAH-degrading bacteria in rhizosphere soil when compared to unvegetated soil. The results from the terminal restriction fragment length polymorphism (T-RFLP) analysis indicated that there was a shift in the rhizosphere bacterial community during the phytoremediation process.     

Long-Term Evolution of Biodegradation and Volatilization Rates in a Crude Oil-Contaminated Aquifer

Volatilization and subsequent biodegradation near the water Table make up a coupled natural attenuation pathway that results in significant mass loss of hydrocarbons. Rates of biodegradation and volatilization were documented twice 12 years apart at a crude-oil spill site near Bemidji, Minnesota. Biodegradation rates were determined by calibrating a gas transport model to O₂, CO₂, and CH₄ gas-concentration data in the unsaturated zone. Reaction stoichiometry was assumed in converting O₂ and CO₂ gas-flux estimates to rates of aerobic biodegradation and CH₄ gas-flux estimates to rates of methanogenesis. Model results indicate that the coupled pathway has resulted in significant hydrocarbon mass loss at the site, and it was estimated that approximately 10.52 kg/day were lost in 1985 and 1.99 kg/day in 1997. In 1985 3% of total volatile hydrocarbons diffusing from the floating oil were biodegraded in the lower 1 m of the unsaturated zone and increased to 52% by 1997. Rates of hydrocarbon biodegradation above the center of the floating oil were relatively stable from 1985 to 1997, as the primary metabolic pathway shifted from aerobic to methanogenic biodegradation. Model results indicate that in 1997 biodegradation under methanogenenic conditions represented approximately one-half of total hydrocarbon biodegradation in the lower 1 m of the unsaturated zone. Further downgradient, where substrate concentrations have greatly increased, total biodegradation rates increased by greater than an order of magnitude from 0.04 to 0.43 g/m²-day. It appears that volatilization is the primary mechanism for attenuation in early stages of plume evolution, while biodegradation dominates in later stages.     

Enrichment of anaerobic benzene-degrading microorganisms by in situ microcosms

Microcosms filled with different solids (sand, lava, Amberlite XAD-7) were exposed for 67 days in the sulfidic part of a groundwater monitoring well downstream of the source zone of a benzene-contaminated aquifer and subsequently incubated in the laboratory. Benzene was repeatedly degraded in several microcosms accompanied by production of sulfide, leading to stable benzene-degrading enrichment cultures. In control microcosms without filling material, benzene was initially degraded, but the benzene-degrading capacity could not be sustained. The results indicate that long-term physiologically active benzene-degrading microorganisms were attached to surfaces of the solids. The biodiversity and attachment behavior of microorganisms in the in situ microcosms was assessed by confocal laser scanning microscopy and single-strand conformation polymorphism (SSCP) analysis, followed by sequencing of dominant SSCP bands. The microbial community was composed of several different Bacteria, representing members of Clostridia, Bacteroidales, all subgroups of the Proteobacteria, Verrucomicrobia, Nitrospira, Chloroflexi and Chlorobi. Only a few archaeal sequences could be retrieved from the communities. The majority of phylotypes were affiliated to bacterial groups with a possible functional relationship to the bacterial sulfur cycle, thus indicating that the microbial community in the investigated aquifer zone depends mainly on inorganic sulfur compounds as electron donors or acceptors, a finding that corresponds to the geochemical data.     

Anaerobic Biodegradation and Hydrogeochemical Controls on Natural Attenuation of Trichloroethene in an Inland Forested Wetland

A field and laboratory investigation of natural attenuation, focusing on anaerobic biodegradation, was conducted in a forested wetland where a plume of trichloroethene discharges from a sand aquifer through organic-rich wetland and stream-bottom sediments. The rapid response of the wetland hydrology to precipitation events altered groundwater flow and geochemistry during wet conditions in the spring compared to the drier conditions in the summer and fall. During dry conditions, partial reductive dechlorination of trichloroethene to cis-1,2-dichloroethene occurred in methanogenic wetland porewater. Influx of oxygenated recharge during wet conditions resulted in a change from methanogenic to iron-reducing conditions and a lack of 1,2-dichloroethene production in the wet spring conditions. During these wet conditions, dilution was the primary attenuation mechanism evident for trichloroethene in the wetland porewater. Trichloroethene degradation was insignificant in anaerobic microcosms constructed with the shallow wetland sediment, and microbiological analyses showed a low microbial biomass and absence of known dehalorespiring microorganisms. Despite the typically organic-rich characteristic of wetland sediments, natural attenuation by anaerobic degradation may not be an effective groundwater remediation for chlorinated solvents at all sites.     

Treatment of Jet Fuel-Contaminated Runoff Water by Subsurface Infiltration

Subsurface infiltration of jet fuel-contaminated surface runoff from airport activities has been shown to be a feasible treatment method. Complete removal of monoaromatic hydrocarbons and naphthalene was observed in 1-m-long (0.2-m-diameter) soil columns. Mineral soil mixed with 5% peat (MinP) showed the best hydrocarbon removal at variable hydraulic loads (50 to 375 mm/d). Natural mineral soil from the C-horizon (Min) showed good hydrocarbon removal after an initial phase, during which hydrocarbons were found in the leachate. This is believed to be a result of adaptation of the soil microflora to the applied hydrocarbons. When hydrocarbon application was stopped and resumed, a new adaptation phase was observed. Cleaned soil (335 mg/kg total petroleum hydrocarbons [TPH]) from a former diesel-polluted site (MinO) showed good hydrocarbon removal but poor hydraulic conductivity. Removal rates of 4.7 and 3.0 mg/kg/d total hydrocarbons were found for Min and MinP, respectively, over the total column length. The highest removal rates (20 and 18 mg/kg/d total hydrocarbons) are observed in the top 20 cm for Min and MinP. Respectively, Potassium acetate (KAc), used as a deicing chemical on airport runways, caused breakthrough of hydrocarbons in the soil columns. The observed breakthrough was caused by oxygen depletion in MinP and MinO. No oxygen depletion was observed in Min, but substrate competition might have limited the hydrocarbon degradation. Based on the results from this study, a full-scale experimental infiltration plant is being constructed and will be tested during 1997. The full-scale tests will focus on the observed relation between oxygen availability and hydrocarbon removal.