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.     

Effects of low temperature and freeze-thaw cycles on hydrocarbon biodegradation in Arctic tundra soil.

Degradation of petroleum hydrocarbons was monitored in microcosms with diesel fuel-contaminated Arctic tundra soil incubated for 48 days at low temperatures (-5, 0, and 7 degrees C). An additional treatment was incubation for alternating 24-h periods at 7 and -5 degrees C. Hydrocarbons were biodegraded at or above 0 degrees C, and freeze-thaw cycles may have actually stimulated hydrocarbon biodegradation. Total petroleum hydrocarbon (TPH) removal over 48 days in the 7, 0, and 7 and -5 degrees C treatments, respectively, was 450, 300, and 600 microg/g of soil. No TPH removal was observed at -5 degrees C. Total carbon dioxide production suggested that TPH removal was due to biological mineralization. Bacterial metabolic activity, indicated by RNA/DNA ratios, was higher in the middle of the experiment (day 21) than at the start, in agreement with measured hydrocarbon removal and carbon dioxide production activities. The total numbers of culturable heterotrophs and of hydrocarbon degraders did not change significantly over the 48 days of incubation in any of the treatments. At the end of the experiment, bacterial community structure, evaluated by ribosomal intergenic spacer length analysis, was very similar in all of the treatments but the alternating 7 and -5 degrees C treatment.     

Effects of Low Temperature and Freeze-Thaw Cycles on Hydrocarbon Biodegradation in Arctic Tundra Soil

Degradation of petroleum hydrocarbons was monitored in microcosms with diesel fuel-contaminated Arctic tundra soil incubated for 48 days at low temperatures (−5, 0, and 7°C). An additional treatment was incubation for alternating 24-h periods at 7 and −5°C. Hydrocarbons were biodegraded at or above 0°C, and freeze-thaw cycles may have actually stimulated hydrocarbon biodegradation. Total petroleum hydrocarbon (TPH) removal over 48 days in the 7, 0, and 7 and −5°C treatments, respectively, was 450, 300, and 600 μg/g of soil. No TPH removal was observed at −5°C. Total carbon dioxide production suggested that TPH removal was due to biological mineralization. Bacterial metabolic activity, indicated by RNA/DNA ratios, was higher in the middle of the experiment (day 21) than at the start, in agreement with measured hydrocarbon removal and carbon dioxide production activities. The total numbers of culturable heterotrophs and of hydrocarbon degraders did not change significantly over the 48 days of incubation in any of the treatments. At the end of the experiment, bacterial community structure, evaluated by ribosomal intergenic spacer length analysis, was very similar in all of the treatments but the alternating 7 and −5°C treatment.     

Biodegradation of aged diesel in diverse soil matrixes: impact of environmental conditions and bioavailability on microbial remediation capacity

While bioremediation of total petroleum hydrocarbons (TPH) is in general a robust technique, heterogeneity in terms of contaminant and environmental characteristics can impact the extent of biodegradation. The current study investigates the implications of different soil matrix types (anthropogenic fill layer, peat, clay, and sand) and bioavailability on bioremediation of an aged diesel contamination from a heterogeneous site. In addition to an uncontaminated sample for each soil type, samples representing two levels of contamination (high and low) were also used; initial TPH concentrations varied between 1.6 and 26.6 g TPH/kg and bioavailability between 36 and 100 %. While significant biodegradation occurred during 100 days of incubation under biostimulating conditions (64.4–100 % remediation efficiency), low bioavailability restricted full biodegradation, yielding a residual TPH concentration. Respiration levels, as well as the abundance of alkB, encoding mono-oxygenases pivotal for hydrocarbon metabolism, were positively correlated with TPH degradation, demonstrating their usefulness as a proxy for hydrocarbon biodegradation. However, absolute respiration and alkB presence were dependent on soil matrix type, indicating the sensitivity of results to initial environmental conditions. Through investigating biodegradation potential across a heterogeneous site, this research illuminates the interplay between soil matrix type, bioavailability, and bioremediation and the implications of these parameters for the effectiveness of an in situ treatment.     

An Investigation of the Ability of a 14C-Labeled Hydrocarbon Mineralization Test to Predict Bioremediation of Soils Contaminated with Petroleum Hydrocarbons

Bioremediation is a widely accepted technology for the remediation of hydrocarbon-contaminated soil. Treatability studies are usually carried out to assess the biodegradation potential of the contaminants and to design optimal treatments. Laboratory studies measuring soil respiration are often used. One method consists of monitoring the mineralization of a ¹⁴C-labeled hydrocarbon surrogate added to the contaminated soil. This study investigates the ability of this method to properly predict the removal of the hydrocarbon contaminants initially found in soils. Mineralization of ¹⁴C-labeled hexadecane was monitored in seven soils contaminated with various hydrocarbon mixtures, both fresh and weathered, in microcosm experiments. Reduction of total petroleum hydrocarbon (TPH) concentrations was measured simultaneously in separate microcosms. Both types of microcosms were subjected to the same amendment regimes. For all soils, poor correlation was observed between the mineralization and TPH reduction data sets. Mineralization data supported contaminants removal data in only one soil. Findings indicate that the radioactive surrogate method does not reliably predict the extent of, and the effect of amendments on, the removal of the hydrocarbons initially present in soil, and may therefore predict suboptimal treatment regimes. Recommendations for soil treatability protocols are provided.     

Mass transfer and hydrocarbon biodegradation of aged soil in slurry phase

Addition of toluene into slurry phase laboratory microcosm is proposed in order to increase desorption rate of hydrocarbons and as an alternative to improve bioavailability of hydrocarbon in aged soils. Our studies showed that toluene has a positive effect on desorption of total petroleum hydrocarbons (TPH). Addition of 14,000 mg toluene/kg of soil, in highly polluted soil, increased the consumption rate of hydrocarbons three times in comparison to control without solvent. In 30 days the initial TPH concentration in soil, 292,000 mg/kg, diminished 45%. Although toluene was able to dissolve complex organic compounds such as asphaltene fraction, it probably yielded a highly toxic toluene-hydrocarbons phase. The inhibitory effect of toluene-TPH was also studied. A substrate inhibition model was used: the k(m) and k(i) constants were 57 and 490 mg TPH/L liquid phase, respectively. Experimental data were well described when the proposed model included sequential desorption and biodegradation phenomena. Damk?hler number evaluation showed that rate of mass transfer was the limiting step in overall biodegradation in nonsolvent control. When high concentration of toluene was added, then bioreaction was the limiting step, but inhibitory effect should be considered. However, toluene addition at low concentrations facilitates the biodegradation of aromatic compounds.     

Biodegradation of petroleum industry oily-sludge using Jordanian oil refinery contaminated soil

The main objective of this study was to evaluate the effect of oily sludge concentration on its biodegradability in soil. Oily sludge was collected and applied to microcosms at full-, half-, or quarter-strength concentrations equivalent to 44.2, 22.2, and 11.1 g kg^?1 soil, respectively, of total petroleum hydrocarbons (TPH) contained in oily sludge. The biodegradability of oily sludge was evaluated by measuring CO₂ evolution and by measuring removal of TPH as well as its main composing fractions; namely; alkanes, aromatics, NSO-compounds, and asphaltenes. The collected soil contained 3.63 × 10⁶ cfu g^?1 soil of hydrocarbon-degrading bacteria, which is satisfactory to drive successful biodegradation of hydrocarbons in soil. These numbers increased significantly with oily sludge addition at a rate proportional to the added TPH reaching 3.35 × 10⁷ cfu g^?1 soil in the half-strength treatment. TPH mineralization rate followed the same pattern. However, TPH-mineralization efficiency was the greatest in quarter-strength treatment at 18.3%. TPH-removal efficiency was also highest in quarter-strength treatment at 30.9%. Nutrients addition caused mineralization inhibition. Since nutrients were added as a ratio of the added carbon, inhibition was the greatest with the highest TPH treatment. While alkanes were degraded, aromatics and asphaltenes were not, and NSO-compounds were enriched. Although SDS was completely biodegradable in soil, its addition promoted mineralization and removal of TPH from soil.     

Biostimulation and Phytoremediation Treatment Strategies of Gasoline-Nickel Co-Contaminated Soil

This study investigated the potential effect of poultry dung (biostimulation) and stubborn grass (Sporobolus pyramidalis) (phytoremediation) on microbial biodegradation of gasoline and nickel uptake in gasoline-nickel-impacted soil. In addition, the potential stimulatory effects of nickel on hydrocarbon utilization were investigated over a small range of nickel concentrations (2.5–12.5 mg/kg). The results showed that an increase in nickel concentration increased hydrocarbon degraders in soil by a range of 8.4–17.2% and resulted in a relative increase in gasoline biodegradation (57.5–62.4%). Also, under aerobic conditions, total petroleum hydrocarbons’ (TPH) removal was 62.4% in the natural gasoline-nickel microcosm (natural attenuation), and a maximum of 78.5%, 85.7%, and 95.8% TPH removal was obtained in phytoremediation, biostimulation, and a combination of biostimulation- and phytoremediation-treated microcosms, respectively. First-order kinetics described the biodegradation of gasoline and nickel uptake very well. Half-life times obtained were 28.88, 18.24, 14.44, and 8.56 days for gasoline degradation under natural attenuation, phytoremediation, biostimulation, and combined biostimulation and phytoremediation treatment methods, respectively. The results indicate that these remediation methods have promising potential for effective remediation of soils co-contaminated with petroleum hydrocarbons and heavy metals.     

Kinetic Modelling and Half-Life Study on Bioremediation of Soil Co-Contaminated with Lubricating Motor Oil and Lead Using Different Bioremediation Strategies

The focus of this study was to investigate the effect of nutrient supplement (urea fertilizer) and microbial species augmentation (mixed culture of Aeromonas, Micrococcus, and Serratia sp.) on biodegradation of lubricating motor oil (LMO) and lead uptake by the autochthonous microorganism in LMO and lead-impacted soil were investigated. The potential inhibitory effects of lead on hydrocarbon utilization were investigated over a wide range of lead concentrations (25–200 mg/kg) owing to the complex co-contamination problem frequently encountered in most sites. Under aerobic conditions, total petroleum hydrocarbons (TPH) removal was 45.3% in the natural attenuation microcosm while a maximum of 72% and 68.2% TPH removal was obtained in biostimulation and bioaugmentation microcosms, respectively. Lead addition, as lead nitrate, to soil samples reduced the number of hydrocarbon degraders in all samples by a wide range (11–52%) depending on concentration and similarly, the metabolic activities were affected as observed in mineralization of LMO (3–60%) in soils amended with various lead concentrations. Moreover, the uptake of lead by the autochthonous microorganisms in the soil reduced with increase in the initial lead concentration. First-order kinetics described the biodegradation of LMO very well. The biodegradation rate constants were 0.015, 0.033, and 0.030 day^?1 for LMO degradation in natural attenuation, biostimulation and bioaugmentation treatment microcosms, respectively. The presence of varying initial lead concentration reduced the biodegradation rate constant of LMO degradation in the biostimulation treatment microcosm. Half-life times were 46.2, 21, and 23 days for LMO degradation in natural attenuation, biostimulation and bioaugmentation treatment microcosms, respectively. The half-life time in the biostimulation treatment microcosm was increased with a range between 10.7 and 39.2 days by the presence of different initial lead concentration. The results have promising potential for effective remediation of soils co-contaminated with hydrocarbons and heavy metals.     

Biostimulation and bioaugmentation for on-site treatment of weathered diesel fuel in Arctic soil

Bioremediation of weathered diesel fuel in Arctic soil at low temperature was studied both on-site in small-scale biopiles and in laboratory microcosms. The field study site was on Ellesmere Island (82°30'N, 62°20'W). Biostimulation was by fertilization with phosphorous and nitrogen. Bioaugmentation was with an enrichment culture originating from the field site. In biopiles, total petroleum hydrocarbons (TPH) were reduced from 2.9 to 0.5 mg/g of dry soil over a period of 65 days. In microcosms at 7 °C, TPH were reduced from 2.4 to 0.5 mg/g of dry soil over a period of 90 days. Inoculation had no effect on hydrocarbon removal in biopiles or in microcosms. Maximum TPH removal rates in the biopiles were approximately 90 μg of TPH g^–1 of soil day^–1, occurring during the first 14 days when ambient temperature ranged from 0 to 10 °C. The fate of three phylotypes present in the inoculum was monitored using most-probable-number PCR, targeting 16S rRNA genes. Populations of all three phylotypes increased more than 100-fold during incubation of both uninoculated and inoculated biopiles. The inoculum increased the initial populations of the phylotypes but did not significantly affect their final populations. Thus, biostimulation on site enriched populations that were also selected in laboratory enrichment cultures. Electronic Publication     

Stimulating in-soil rhamnolipid production in a bioslurry reactor by limiting nitrogen

A soil with aged contamination from lubricating oil (LO) and polychlorinated biphenyls (PCBs) was treated in a bioslurry reactor to investigate in-soil biosurfactant production by Pseudomonas aeruginosa, the most abundant indigenous, culturable, hydrocarbon-degrading microorganism. After 2 days of growth on LO, a depletion of nitrogen stimulated the production and accumulation of rhamnolipids to levels roughly 20 times the critical micelle concentration. Surface tensions and concentrations of monorhamnolipid and dirhamnolipid, PCBs, and total petroleum hydrocarbons (TPH) were measured in a slurry filtrate. Soil-bound PCBs and TPH were also quantified. Rhamnolipid production was observed within 1 to 2 days after nitrogen depletion in each of the 10 batches tested. By day 6, total rhamnolipid concentrations increased from below detection to average values over 1,000 mg/L, which caused over 98% of soil-bound PCBs and over 99% of TPH to be emulsified and recovered in the filtrate. After 70 days, rhamnolipid concentrations were only reduced by 15%, because of nitrogen-limited rates of rhamnolipid biodegradation. The results show that in-soil biosurfactant production can be stimulated in a controlled way with nutrient limitation and can be used to achieve soil washing.     

Enhancing the biodegradation of total petroleum hydrocarbons in oily sludge by a modified bioaugmentation strategy

The effect of successive inoculation with hydrocarbon-degrading bacteria on the dynamics of petroleum hydrocarbons degradation in soil was investigated in this study. Oily sludge was used as a source of mixed hydrocarbons pollutant. Two bacterial consortia composed of alkanes and polycyclic aromatic hydrocarbon degraders were constructed from bacteria isolated from soil and oily sludge. These consortia were applied to incubated microcosms either in one dose at the onset of the incubation or in two doses at the beginning and at day 62 of the incubation period, which lasted for 198 days. During this period, carbon mineralization was evaluated by respirometry while total petroleum hydrocarbons and its fractions were gravimetrically evaluated by extraction from soil and fractionation. Dosing the bacterial consortia resulted in more than 30% increase in the overall removal of total petroleum hydrocarbons from soil. While alkane removal was only slightly improved, aromatic and asphaltic hydrocarbon fraction removal was significantly enhanced by the addition of the second consortium. Polar compounds (resins) were enriched only as a result of aromatics and asphaltene utilization. Nonetheless, their concentration declined back to the original level by the end of the incubation period.     

Incomplete Hydrocarbon Biodegradation in Contaminated Soils: Limitations in Bioavailability or Inherent Recalcitrance?

Bioremediation has been used to treat soils contaminated with complex mixtures of organic compounds such as total petroleum hydrocarbons (TPH), oil and grease (O&G), or polycyclic aromatic hydrocarbons (PAHs). Despite the common use and cost-effectiveness of bioremediation for treating hydrocarbon-contaminated soils, it has been observed that a residual fraction remains undegraded in the soil even when optimal biodegradation conditions have been provided. This paper provides a brief review of the two major conceptual models that have been used to explain why a residual hydrocarbon fraction remains in the soil after bioremediation treatment. The contaminant sequestration model is based on the assumption that a certain fraction of hydrocarbons is “locked up” in small soil pores within soil particles or aggregates. These sorbed hydrocarbons are believed to be inaccessible to soil microorganisms. Consequently, limitations in bioavailability are thought to be the major reason for incomplete hydrocarbon biodegradation, particularly in aged or weathered soils. Alternatively, according to the inherent recalcitrance model, incomplete TPH biodegradation may be caused by the presence of certain hydrocarbons that are inherently recalcitrant to biodegradation or are only extremely slowly degradable even under optimal conditions. Each conceptual model provides different explanations regarding the potential risks of the residual hydrocarbon fraction. If the residual TPH is truly sequestered within the soil pore space, it is unlikely that these compounds will pose any significant risk to human or environmental receptors. By contrast, these risks may be considerably greater if the residual TPH fraction consists of inherently recalcitrant compounds that reside mostly on the surface of soil particles and therefore are much more available to potential receptors. Both conceptual models and their implications for the potential risk of the residual TPH fraction are discussed.     

Bacterial community dynamics and polycyclic aromatic hydrocarbon degradation during bioremediation of heavily creosote-contaminated soil

Bacterial community dynamics and biodegradation processes were examined in a highly creosote-contaminated soil undergoing a range of laboratory-based bioremediation treatments. The dynamics of the eubacterial community, the number of heterotrophs and polycyclic aromatic hydrocarbon (PAH) degraders, and the total petroleum hydrocarbon (TPH) and PAH concentrations were monitored during the bioremediation process. TPH and PAHs were significantly degraded in all treatments (72 to 79% and 83 to 87%, respectively), and the biodegradation values were higher when nutrients were not added, especially for benzo(a)anthracene and chrysene. The moisture content and aeration were determined to be the key factors associated with PAH bioremediation. Neither biosurfactant addition, bioaugmentation, nor ferric octate addition led to differences in PAH or TPH biodegradation compared to biodegradation with nutrient treatment. All treatments resulted in a high first-order degradation rate during the first 45 days, which was markedly reduced after 90 days. A sharp increase in the size of the heterotrophic and PAH-degrading microbial populations was observed, which coincided with the highest rates of TPH and PAH biodegradation. At the end of the incubation period, PAH degraders were more prevalent in samples to which nutrients had not been added. Denaturing gradient gel electrophoresis analysis and principal-component analysis confirmed that there was a remarkable shift in the composition of the bacterial community due to both the biodegradation process and the addition of nutrients. At early stages of biodegradation, the alpha-Proteobacteria group (genera Sphingomonas and Azospirillum) was the dominant group in all treatments. At later stages, the gamma-Proteobacteria group (genus Xanthomonas), the alpha-Proteobacteria group (genus Sphingomonas), and the Cytophaga-Flexibacter-Bacteroides group (Bacteroidetes) were the dominant groups in the nonnutrient treatment, while the gamma-Proteobacteria group (genus Xathomonas), the beta-Proteobacteria group (genera Alcaligenes and Achromobacter), and the alpha-Proteobacteria group (genus Sphingomonas) were the dominant groups in the nutrient treatment. This study shows that specific bacterial phylotypes are associated both with different phases of PAH degradation and with nutrient addition in a preadapted PAH-contaminated soil. Our findings also suggest that there are complex interactions between bacterial species and medium conditions that influence the biodegradation capacity of the microbial communities involved in bioremediation processes.     

Prediction of Petroleum Hydrocarbon Bioavailability in Contaminated Soils and Sediments

Recently, several laboratory methods have been developed for the prediction of contaminant bioavailability. So far, none of these methods has been extensively tested for petroleum hydrocarbons. In the present study we investigated solid-phase extraction and persulfate oxidation for the prediction of total petroleum hydrocarbon (TPH) bioavailability. One sediment and two soil samples were subjected to solid-phase extraction, persulfate oxidation, and biodegradation, after which hydrocarbon removal was compared. It was demonstrated that a short solid-phase extraction (168?h) provided a good method for the prediction of the extent of TPH degradation in an optimized slurry reactor (84?d). Solid-phase extraction slightly underestimated the degradation of readily biodegradable hydrocarbons, whereas it slightly overestimated the degradation of poorly biodegradable hydrocarbons. Persulfate oxidation appeared to be unfit for the prediction of TPH bioavailability as persulfate was unable to oxidize hydrocarbons with a high ionization potential. Hydrocarbons that were affected were likely to be transformed rather than completely oxidized. Nevertheless, persulfate oxidation provided a good method for the prediction of polycyclic aromatic hydrocarbon (PAH) bioavailability.     

Evaluation of inoculum addition to stimulate in situ bioremediation of oily-sludge-contaminated soil

A full-scale study evaluating an inoculum addition to stimulate in situ bioremediation of oily-sludge-contaminated soil was conducted at an oil refinery where the indigenous population of hydrocarbon-degrading bacteria in the soil was very low (10(3) to 10(4) CFU/g of soil). A feasibility study was conducted prior to the full-scale bioremediation study. In this feasibility study, out of six treatments, the application of a bacterial consortium and nutrients resulted in maximum biodegradation of total petroleum hydrocarbon (TPH) in 120 days. Therefore, this treatment was selected for the full-scale study. In the full-scale study, plots A and B were treated with a bacterial consortium and nutrients, which resulted in 92.0 and 89.7% removal of TPH, respectively, in 1 year, compared to 14.0% removal of TPH in the control plot C. In plot A, the alkane fraction of TPH was reduced by 94.2%, the aromatic fraction of TPH was reduced by 91.9%, and NSO (nitrogen-, sulfur-, and oxygen-containing compound) and asphaltene fractions of TPH were reduced by 85.2% in 1 year. Similarly, in plot B the degradation of alkane, aromatic, and NSO plus asphaltene fractions of TPH was 95.1, 94.8, and 63.5%, respectively, in 345 days. However, in plot C, removal of alkane (17.3%), aromatic (12.9%), and NSO plus asphaltene (5.8%) fractions was much less. The population of introduced Acinetobacter baumannii strains in plots A and B was stable even after 1 year. Physical and chemical properties of the soil at the bioremediation site improved significantly in 1 year.     

Prepared Bed Land Treatment of Soils Containing Diesel and Crude Oil Hydrocarbons

An ex situ, field-scale, prepared bed land treatment unit (LTU) was used to bio-remediate soils containing petroleum hydrocarbons. Two soils were treated in side-by-side units to compare performance: (1) a clayey silt containing crude oil hydrocarbons from releases 30 to 40 years ago and (2) a silty sand containing diesel fuel hydrocarbons from a leak about three years prior to the bioremediation. The effectiveness of the bioremediation in the LTU was evaluated over a period of 18 months. The results indicated that: (1) prepared bed bioremediation reduced the hydrocarbon concentration, mobility, and relative toxicity in the soil with the diesel fuel, and (2) chemical bioavailability appeared to limit bioremediation of the soil containing the crude oil hydrocarbons. Although the soils containing the crude oil hydrocarbons contained an average of 10,000?mg TPH/kg dry soil, these soils had limited hydrocarbon availability, nontoxic conditions, and low potential for chemical migration. For the soils containing the diesel fuel, active prepared bed bioremediation of about 15 weeks was adequate to reach an environmentally acceptable endpoint. At that time, there was little further TPH loss, no Microtox^TM toxicity, and limited hydrocarbon mobility.     

Enhancement of Hydrocarbon Removal in a Clay and Drilling-Waste Polluted Soil

The application of biological processes in restoring oil polluted sites is growing due to their efficiency in removing different classes of pollutants. The aim of this study was to determine the ability of microorganisms present in a drilling-waste polluted soil (36,200 mg TPH kg^?1 soil) to remove weathered hydrocarbons under stimulated and non-stimulated soil conditions. The hypothesis under study was whether petroleum hydrocarbons removal could be enhanced by manipulating C/N ratio, water content and addition of three agroindustrial wastes. A Box-Behnken design was employed to evaluate the effect of each variable. Results demonstrated that, for orange peels and banana trunk treatments, the variable with the largest effect (p < 0.01) on hydrocarbon removal was the C/N ratio, indicating that higher ratio (100/3) improved removal (79.5–82%). The largest effect (p < 0.001) on hydrocarbon removal for pineapple wastes was observed with higher water content (60%) achieving the highest removal (89%). After 90 days of experimentation, the type of agricultural waste and the agricultural waste/soil ratio were not statistically significant in any treatment. However, their addition was important relative to non-stimulated soil, which showed a hydrocarbon removal of 17%. Data reported in this study showed the application of bioremediation in clay and drilling waste-polluted soils.     

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.     

Effect of modified Fenton's reaction on microbial activity and removal of PAHs in creosote oil contaminated soil

This study describes the removal of polycyclic aromatic hydrocarbons (PAHs) from creosote oil contaminated soil by modified Fenton's reaction in laboratory-scale column experiments and subsequent aerobic biodegradation of PAHs by indigenous bacteria during incubation of the soil. The effect of hydrogen peroxide addition for 4 and 10 days and saturation of soil with H(2)O(2) on was studied. In both experiments the H(2)O(2) dosage was 0.4 g H(2)O(2)/g soil. In completely H(2)O(2)-saturated soil the removal of PAHs (44% within 4 days) by modified Fenton reaction was uniform over the entire soil column. In non-uniformly saturated soil, PAH removal was higher in completely saturated soil (52% in 10 days) compared to partially saturated soil, with only 25% in 10 days. The effect of the modified Fenton's reaction on the microbial activity in the soil was assessed based on toxicity tests towards Vibrio fischeri, enumeration of viable and dead cells, microbial extracellular enzyme activity, and oxygen consumption and carbon dioxide production during soil incubation. During the laboratory-scale column experiments, the toxicity of column leachate towards Vibrio fischeri increased as a result of the modified Fenton's reaction. The activities of the microbial extracellular enzymes acetate- and acidic phosphomono-esterase were lower in the incubated modified Fenton's treated soil compared to extracellular enzyme activities in untreated soil. Abundance of viable cells was lower in incubated modified Fenton treated soil than in untreated soil. Incubation of soil in serum bottles at 20 degrees C resulted in consumption of oxygen and formation of carbon dioxide, indicating aerobic biodegradation of organic compounds. In untreated soil 20-30% of the PAHs were biodegraded during 2 months of incubation. Incubation of chemically treated soil slightly increased PAH-removal compared to PAH-removal in untreated soil.