Prediction of leachate concentrations in petroleum‐contaminated soils

The efficacy of cleanup methods in reducing gasoline contamination at spill sites is typically determined by measuring benzene, toluene, xylene (BTX), and total petroleum hydrocarbon (TPH) concentrations in soil samples. Although these values may provide a direct measurement of soil contamination, they may not be indicative of what is transferred to percolating water. This study addresses this issue by measuring TPH, toluene, m‐ and p‐xylene, and naphthalene levels in gasoline‐contaminated soil columns before and after forced‐air venting and relating these values to the aqueous‐phase concentrations measured when water is percolated through the same columns.

Sandy soils with and without organic matter were packed into glass columns. The soils were brought to residual water and residual gasoline saturations by applying a vacuum to a ceramic pressure plate at the column bottom. Venting was performed by passing clean, moist air through the columns. The columns were subsequently leached under unsaturated conditions.

Soil samples were taken from the bottom of the columns upon completion of the venting or leaching phases of the experiments. Toluene, m‐ and p‐xylene, naphthalene, and TPH values were measured in soil samples extracted with either freon or methanol. Aqueous phase concentrations of these compounds were predicted using measured soil concentrations and either Raoult's law or organic matter‐water and fuel‐water partitioning theory (Boyd and Sun, 1990). The predicted results were compared with measured leachate concentrations from the same columns.

Mole fractions estimated from soil concentrations and TPH values used in Raoult's law gave good predictions of aqueous phase concentrations for compounds that had a high mole fraction in the residual nonaqueous phase liquid (NAPL). For compounds at low concentrations in the residual NAPL, an approach using a distribution coefficient that accounted for both the organic matter and residual NAPL in the soil provided better estimates than those based on Raoult's law.     

Effects of Petroleum Hydrocarbons on Plant Litter Microbiota in an Arctic Lake

The effects of petroleum hydrocarbons on the microbial community associated with decomposing Carex leaf litter colonized in Toolik Lake, Alaska, were examined. Microbial metabolic activity, measured as the rate of acetate incorporation into lipid, did not vary significantly from controls over a 12-h period after exposure of colonized Carex litter to 3.0 ml of Prudhoe Bay crude oil, diesel fuel, or toluene per liter. ATP levels of the microbiota became elevated within 2 h after the exposure of the litter to diesel fuel or toluene, but returned to control levels within 4 to 8 h. ATP levels of samples exposed to Prudhoe Bay crude oil did not vary from control levels. Mineralization of specifically labeled ¹⁴C-[lignin]-lignocellulose and ¹⁴C-[cellulose]-lignocellulose by Toolik Lake sediments, after the addition of 2% (vol/vol) Prudhoe Bay crude oil, motor oil, diesel fuel, gasoline, n-hexane, or toluene, was examined after 21 days of incubation at 10°C. Diesel fuel, motor oil, gasoline, and toluene inhibited ¹⁴C-[lignin]-lignocellulose mineralization by 58, 67, 67, and 86%, respectively. Hexane-treated samples displayed an increase in the rate of ¹⁴C-[lignin]-lignocellulose mineralization of 33%. ¹⁴C-[cellulose]-lignocellulose mineralization was inhibited by the addition of motor oil or toluene by 27 and 64%, respectively, whereas diesel fuel-treated samples showed a 17% increase in mineralization rate. Mineralization of the labeled lignin component of lignocellulose appeared to be more sensitive to hydrocarbon perturbations than was the labeled cellulose component.     

Degradation of gasoline aromatic hydrocarbons by two N2-fixing soil bacteria

Two bacterial strains, 3A and 5A, isolated from soil, were selected for their ability to degrade gasoline aromatic compounds and to fix N₂. Strains 3A and 5A have been ascribed to the genera Agrobacterium and Alcaligenes, respectively. Using gasoline as the sole carbon source these strains were as effective at degrading benzene, toluene and xylene as Pseudomonas putida ATCC12236, a reference biodegrading strain.     

Metabolism of Benzene, Toluene, and Xylene Hydrocarbons in Soil

Enrichment cultures obtained from soil exposed to benzene, toluene, and xylene (BTX) mineralized benzene and toluene but cometabolized only xylene isomers, forming polymeric residues. This observation prompted us to investigate the metabolism of ¹⁴C-labeled BTX hydrocarbons in soil, either individually or as mixtures. BTX-supplemented soil was incubated aerobically for up to 4 weeks in a sealed system that automatically replenished any O₂ consumed. The decrease in solvent vapors and the production of ¹⁴CO₂ were monitored. At the conclusion of each experiment, ¹⁴C distribution in solvent-extractable polymers, biomass, and humic material was determined, obtaining ¹⁴C mass balances of 85 to 98%. BTX compounds were extensively mineralized in soil, regardless of whether they were presented singly or in combinations. No evidence was obtained for the formation of solvent-extractable polymers from xylenes in soil, but ¹⁴C distribution in biomass (5 to 10%) and humus (12 to 32%) was unusual for all BTX compounds and especially for toluene and the xylenes. The results suggest that catechol intermediates of BTX degradation are preferentially polymerized into the soil humus and that the methyl substituents of the catechols derived from toluene and especially from xylenes enhance this incorporation. In contrast to inhibitory residues formed from xylene cometabolism in culture, the humus-incorporated xylene residues showed no significant toxicity in the Microtox assay.     

Substrate Interactions during the Biodegradation of Benzene, Toluene, Ethylbenzene, and Xylene (BTEX) Hydrocarbons by the Fungus Cladophialophora sp. Strain T1

The soil fungus Cladophialophora sp. strain T1 (= ATCC MYA-2335) was capable of growth on a model water-soluble fraction of gasoline that contained all six BTEX components (benzene, toluene, ethylbenzene, and the xylene isomers). Benzene was not metabolized, but the alkylated benzenes (toluene, ethylbenzene, and xylenes) were degraded by a combination of assimilation and cometabolism. Toluene and ethylbenzene were used as sources of carbon and energy, whereas the xylenes were cometabolized to different extents. o-Xylene and m-xylene were converted to phthalates as end metabolites; p-xylene was not degraded in complex BTEX mixtures but, in combination with toluene, appeared to be mineralized. The metabolic profiles and the inhibitory nature of the substrate interactions indicated that toluene, ethylbenzene, and xylene were degraded at the side chain by the same monooxygenase enzyme. Our findings suggest that soil fungi could contribute significantly to bioremediation of BTEX pollution.     

The Potential of Bacterial Isolates for Emulsification with a Range of Hydrocarbons

A study was undertaken to investigate the distribution of biosurfactant producing and crude oil degrading bacteria in the oil contaminated environment. This research revealed that hydrocarbon contaminated sites are the potent sources for oil degraders. Among 32 oil degrading bacteria isolated from ten different oil contaminated sites of gasoline and diesel fuel stations, 80% exhibited biosurfactant production. The quantity and emulsification activity of the biosurfactants varied. Pseudomonas sp. DS10‐129 produced a maximum of 7.5 ± 0.4 g/l of biosurfactant with a corresponding reduction in surface tension from 68 mN/m to 29.4 ± 0.7 mN/m at 84 h incubation. The isolates Micrococcus sp. GS2‐22, Bacillus sp. DS6‐86, Corynebacterium sp. GS5‐66, Flavobacterium sp. DS5‐73, Pseudomonas sp. DS10‐129, Pseudomonas sp. DS9‐119 and Acinetobacter sp. DS5‐74 emulsified xylene, benzene, n‐hexane, Bombay High crude oil, kerosene, gasoline, diesel fuel and olive oil. The first five of the above isolates had the highest emulsification activity and crude oil degradation ability and were selected for the preparation of a mixed bacterial consortium, which was also an efficient biosurfactant producing oil emulsifying and degrading culture. During this study, biosurfactant production and emulsification activity were detected in Moraxella sp., Flavobacterium sp. and in a mixed bacterial consortium, which have not been reported before.     

New family of biosensors for monitoring BTX in aquatic and edaphic environments

Benzene, toluene, ethylbenzene and xylenes (BTEX) contamination is a serious threat to public health and the environment, and therefore, there is an urgent need to detect its presence in nature. The use of whole‐cell reporters is an efficient, easy‐to‐use and low‐cost approach to detect and follow contaminants outside specialized laboratories; this is especially important in oil spills that are frequent in marine environments. The aim of this study is the construction of a bioreporter system and its comparison and validation for the specific detection of monocyclic aromatic hydrocarbons in different host bacteria and environmental samples. Our bioreporter system is based on the two component regulatory system TodS–TodT of P. putida DOT‐T1E, and the P_todX promoter fused to the GFP protein as the reporter protein. For the construction of different biosensors, this bioreporter was transferred into three different bacterial strains isolated from three different environments, and their performance was measured. Validation of the biosensors on water samples spiked with petrol, diesel and crude oil on contaminated waters from oil spills and on contaminated soils demonstrated that they can be used in mapping and monitoring some BTEX compounds (specifically benzene, toluene and two xylene isomers). Validation of biosensors is an important issue for the integration of these devices into pollution‐control programmes.     

Characterization of a diesel-degrading bacterium, <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> IU5, isolated from oil-contaminated soil in Korea

A diesel-degrading bacterium (strain IU5) isolated from oil-contaminated soil was characterized in this study. Fatty acid and 16s rDNA sequence analysis identified IU5 as a strain of Pseudomonas aeruginosa, and growth curve experiments identified the bacterium’s optimum conditions as pH 7 and 30 °C. P. aeruginosa IU5 degraded up to 60 of applied diesel (8500 mg/kg) over 13 days in a soil-slurry phase. In addition, this strain was able to grow on many other petroleum hydrocarbons as sole carbon sources, including crude oil, gasoline, benzene, toluene, xylene, and even PAHs such as naphthalene, phenanthrene and pyrene. Therefore, P. aeruginosa IU5 may be useful for bioremediation of soils and groundwater contaminated with a variety of hydrocarbons.     

Molecular analysis of the hydrolytic component of petroleum-contaminated soils and of soils remediated with chitin

Molecular genetic techniques (FISH and metagenomic analysis) were used to investigate prokaryotic complexes in native soils (gray forest soil and urbostratozema typical), soils contaminated by petroleum products (gasoline or diesel fuel), and soils subject to remediation by addition of a nitrogen-containing polysaccharide biopolymer chitin. The share of metabolically active prokaryotic cells in the hydrolytic complex of soil microcosms was determined, as well as their biomass and biodiversity. Compared to the control, in the pollutant-containing experimental microcosms, a decrease in the share of metabolically active prokaryotic cells was observed, as well as changes of the hydrolytic complex structure, such as an increase in the share of the phylum Actinobacteria (specifically of the genera Galiella and Nocardioides in the samples contaminated with gasoline and diesel fuel, respectively). Supplementing the hydrocarbon-contaminated system the biopolymer chitin resulted in processing of mixed-minerals with an increase in the number of layers of the smectite type and, as a result, in formation of aggregates and improved aeration. An increase in the number of metabolically active prokaryotic cells and decreased diversity of the soil prokaryotic complex were observed, which were probably associated with the development of a selective group of the hydrolytic complex of chitindegrading microorganisms.     

Hybridization Analysis of Microbial DNA from Fuel Oil–Contaminated and Noncontaminated Soil

Abstract The enrichment of several genes (xylE, nahAcd, todC1C2BA, tmoABCDE, alkB) that encode enzymes responsible for key steps in the degradation of hydrocarbons, and one gene specific to rRNA group I of the genus Pseudomonas, was studied in DNA extracted from a fuel oil–contaminated field site, and in laboratory microcosms (with the exception of alkB). Toluene, ethylbenzene, xylene, and naphthalene concentrations were related to the extent of hybridization of the genes in the field studies. Significant differences were observed in the extent of hybridization of some of the genes between contaminated and noncontaminated samples. In the microcosm studies, gasoline at rates ranging from 0.5 mg to 125 mg gasoline/g of soil as applied to soils, and the changes in hybridization intensity of these genes monitored with time. The lower threshold of gene enrichment of these genes in response to gasoline addition was below 0.5 mg/g soil. Small increases were observed at the 0.5-mg exposure level, but hybridization intensity quickly decreased to levels below detection 6–8 days after addition of the gasoline. A dose-response effect was observed from treatments with gasoline concentrations ranging from 0.5 to 35 mg/g soil. Inhibition by toxic components in gasoline was observed at 75 and 125 mg/g soil levels. Hybridization of the Pseudomonas group 1 probe to field DNA was not significantly enriched in the contaminated field site, although these sequences were enriched in the microcosm studies. Among the genes tested, xylE was the most sensitive indicator of low levels of fuel oil contamination. Received: 23 July 1996; Accepted 9 October 1996     

Bioremediation Assessment of a BTEX‐Contaminated Soil

The elimination of BTEX (benzene, toluene, ethylbenzene, o‐xylene) compounds from soil was studied. After 18 days at 20 °C, 21% of the initial total BTEX contamination (400 mg/kg soil) was lost due to sorption onto soil. Biodegradation decreased in the order ethylbenzene > toluene > benzene > o‐xylene. NPK fertilisation stimulated biodegradation, particularly that of benzene and toluene, significantly, and oleophilic fertilisation inhibited biodegradation. After 18 days, the residual contamination in the NPK‐fertilised, unfertilised and with oleophilic nutrients amended soil was 96, 166 and 196 mg total BTEX/kg soil, respectively. The presence of BTEX initially inhibited the biological activity of the soil (fluorescein diacetate hydrolysis) considerably. This short‐term, reversible inhibition was significantly higher in the unfertilised soil than in the fertilised soil.     

Using LCA‐based Decomposition Analysis to Study the Multidimensional Contribution of Technological Innovation to Environmental Pressures

This article presents a general framework for macroenvironmental assessment, combining life cycle assessment (LCA) with the IPAT equation, and explores its combination with decomposition analysis to assess the multidimensional contribution of technological innovation to environmental pressures. This approach is illustrated with a case study in which carbon dioxide (CO₂) and nitrogen oxides (NO_x) air emissions from diesel passenger cars in Europe during the period 1990–2005 are first decomposed using index decomposition analysis into technology, consumption activity, and population growth effects. By a second decomposition, the contribution of a specific innovation (diesel engine) is calculated on the basis of the technology and consumption activity effects, through a technological comparison with a relevant alternative and the calculation of the rebound effect, respectively. The empirical analysis for diesel passenger cars highlights the discrepancies between the micro (LCA) and macro (IPAT‐LCA) analytical approaches. Thus, whereas diesel engines present a relatively less‐pollutant environmental product profile than their gasoline counterparts, total CO₂ and NO_x emissions would have increased partly as a consequence of their introduction, mainly driven by the increase in travel demand caused by the induced direct price rebound effect from fuel savings and fuel price differences. The counterintuitive result shows the need for such an analysis.     

Effect of isobutanol on toluene biodegradation in nitrate amended, sulfate amended and methanogenic enrichment microcosms

Isobutanol is an alternate fuel additive that is being considered because of economic and lower emission benefits. However, future gasoline spills could result in co-contamination of isobutanol with gasoline components such as benzene, toluene, ethyl-benzene and xylene. Hence, isobutanol could affect the degradability of gasoline components thereby having an effect on contaminant plume length and half-life. In this study, the effect of isobutanol on the biodegradation of a model gasoline component (toluene) was examined in laboratory microcosms. For this, toluene and isobutanol were added to six different toluene degrading laboratory microcosms under sulfate amended, nitrate amended or methanogenic conditions. While toluene biodegradation was not greatly affected in the presence of isobutanol in five out of the six different experimental sets, toluene degradation was completely inhibited in one set of microcosms. This inhibition occurred in sulfate amended microcosms constructed with inocula from wastewater treatment plant activated sludge. Our data suggest that toluene degrading consortia are affected differently by isobutanol addition. These results indicate that, if co-contamination occurs, in some cases the in situ half-life of toluene could be significantly extended.     

Substrate interactions during the biodegradation of benzene,toluene, ethylbenzene,and xylene (BTEX) hydrocarbons by the fungus Cladophialophora sp. strain T1

The soil fungus Cladophialophora sp. strain T1 (= ATCC MYA-2335) was capable of growth on a model water-soluble fraction of gasoline that contained all six BTEX components (benzene, toluene, ethylbenzene, and the xylene isomers). Benzene was not metabolized, but the alkylated benzenes (toluene, ethylbenzene, and xylenes) were degraded by a combination of assimilation and cometabolism. Toluene and ethylbenzene were used as sources of carbon and energy, whereas the xylenes were cometabolized to different extents. o-Xylene and m-xylene were converted to phthalates as end metabolites; p-xylene was not degraded in complex BTEX mixtures but, in combination with toluene, appeared to be mineralized. The metabolic profiles and the inhibitory nature of the substrate interactions indicated that toluene, ethylbenzene, and xylene were degraded at the side chain by the same monooxygenase enzyme. Our findings suggest that soil fungi could contribute significantly to bioremediation of BTEX pollution.     

Empirical Models Estimating Carbon Dioxide Accumulation in Two Petroleum Hydrocarbon–Contaminated Soils

Bioremediation is a popular method in degrading diesel fuel contaminants from soil. Bioremediation can be enhanced by estimating the effect of important environmental parameters on microbial activity. Respirometry was used to develop empirical models describing the effects of temperature, moisture, nitrogen, and phosphorus concentration on microbial activity in a diesel-contaminated soil from Wyoming. Carbon dioxide (CO₂) data were analyzed using a base equation where its coefficient values were functions of each parameter. Two physiologically different groups of microorganisms were identified from the results under different operating temperatures. The empirical correlations were combined into one model and this model was tested against a hydrocarbon-contaminated soil collected from a site in Egypt with similar history of contamination. The predicted CO₂ evolution agreed well with the actual data obtained from the Egyptian soil samples, showing a sound predicting power of the empirical model for petroleum hydrocarbon biodegradation. Overall, the empirical correlations developed from the respirometric data provide a method to describe microbial activity in diesel-contaminated soils.     

Lab-Scale Biodegradation Study of BTEX under the Unsaturated Condition and Its Effect on Soil Matric Potential

Benzene, toluene, ethylbenzene, and xylene are collectively known as BTEX which contributes to volatile environmental contaminants. This present study investigates the microbial degradation of BTEX in batch and continuous soil column experiments and its effects on soil matric potential. Batch degradation experiments were performed with different initial concentrations of BTEX using the BTEX tolerant culture isolated from petroleum-contaminated soil. In batch study, the degradation pattern for single substrate showed that xylene was degraded much faster than other compounds followed by ethylbenzene, toluene, and benzene with the highest μ_max = 0.140 h^?1 during initial substrate concentration of 100 mg L^?1. Continuous degradation experiments were performed in a soil column with an inlet concentration of BTEX of about 2000 mg L^?1 under unsaturated flow in anaerobic condition. BTEX degradation pattern was studied with time and the matric potential of the soil at different parts along the length of the column were determined at the end of the experiment. In continuous degradation study, BTEX compounds were degraded with different degradation pattern and an increase in soil matric potential was observed with an increase in depth from top to bottom in the column with applied suction head. It was found that column biodegradation contributed to 69.5% of BTEX reduction and the bacterial growth increased the soil matric potential of about 34% on an average along the column height. Therefore, this study proves that it is significant to consider soil matric potential in modeling fate and transport of BTEX in unsaturated soils.     

Kinetics of biodegradation of gasoline and its hydrocarbon constituents

Aerobic biodegradation of gasoline and its constituents, benzene, toluene and ethylbenzene were studied by an enrichment from soil indigenous microbial population. The enrichment culture completely degraded 16.1–660 mg/l gasoline in 2.5–16 days respectively, without accumulation of any by-products. The kinetics of gasoline as well as benzene, toluene and ethylbenzene biodegradation was investigated with initial gasoline concentrations of 16.1–62.6 mg/l. The maximum specific rates of biodegradation of benzene, toluene and ethylbenzene were 0.12, 0.38 and 0.19 mg mg biomass⁻¹ day⁻¹ respectively. When benzene and toluene were used as sole substrate, the maximum specific rates of their biodegradation were 62.9 and 16.4 times greater than the corresponding values for a mixture (gasoline). The microbial culture was able to mineralize up to 200 mg/l pure toluene and benzene. Maximum mineralization efficiencies of benzene and toluene were 76.7 ± 5.1% and 76.8 ± 1.3% respectively. Self-inhibition and competitive inhibition patterns were observed during the biodegradation of benzene and toluene alone and in the mixture respectively. The observed kinetics was modeled according to Andrews' inhibition model. Received: 6 August 1997 / Received revision: 18 November 1997 / Accepted: 29 November 1997     

Distribution of Petroleum and Aromatic Hydrocarbons at a Former Crude Oil and Natural Gas Production Facility

Site characterization and remediation activities were performed at a former crude oil and natural gas production facility prior to redevelopment of the site. Field activities included delineation, excavation and segregation of approximately 1,250,000 m³ of soil impacted by total petroleum hydrocarbons (TPH) and the aromatic volatile organic compounds (VOCs) benzene, toluene, ethylbenzene, and xylenes (hereafter, collectively referred to as BTEX). Petroleum hydrocarbon chain length information was used to determine whether remediation was required in impacted areas, because the site-specific cleanup values for TPH compounds, established by the California State Regional Water Quality Control Board (RWQCB), were based on hydrocarbon chain length. Site-specific cleanup levels were also established by the RWQCB for BTEX. Subsurface investigation activities performed at the site indicated that the mean percentage of condensate and TPH compounds in the gasoline range was significantly greater at depths ranging from 4.6 to 18 m than in shallower samples. There was no significant difference in the mean concentration of BTEX compounds and mean percentage of diesel range and heavier hydrocarbons with depth. The occurrence of BTEX, diesel range, and heavier hydrocarbons at depth may result from preferential pathways for downward migration of contaminants, including blown out wells, abandoned wellbores, and the presence of faults. Vapor phase diffusion may also be a major transport mechanism controlling movement of BTEX compounds beneath the site.     

Assessment of C:N Ratios and Water Potential for Nitrogen Optimization in Diesel Bioremediation

Sandy clay loam soil contaminated with 5000, 10,000 or 20,000 mg/kg of diesel fuel no. 2 was amended with 0 (ambient nitrogen only), 250, 500, or 1000 mg/kg nitrogen (NH₄Cl) to evaluate the role of C:N ratios and soil water potential on diesel biodegradation efficacy. The soil was incubated at 25°C for 41 days and microbial O₂ consumption measured respirometrically. Highest microbial respiration was observed in the 250 mg N/kg soil treatments regardless of diesel concentration. Higher levels of nitrogen fertilization decreased soil water potential and resulted in an extended lag phase and reduced respiration. Application of 1000 mg/kg nitrogen reduced maximum respiration by 20% to 52% depending on contaminant levels. Optimal C:N ratios among those tested were 17:1, 34:1, and 68:1 for the three diesel concentrations, respectively, and were dependent on contaminant concentration. Nitrogen fertilization on the basis of soil pore water nitrogen (mg N/kg soil H₂O) is independent of hydrocarbon concentration but takes into account soil moisture content. This method accounts for both the nutritional and osmotic aspects of nitrogen fertilization. In the soil studied the best nitrogen augmentation corresponded to a soil pore water nitrogen level of 1950 mg N/kg H₂O at all diesel concentrations.