Bioremediation of Oil-contaminated Soil Using Chicken Manure

In less developed countries, the prevalence of soil contaminated with used lubricating oil is high and the situation worsens with the economic advancement. The contamination has been shown to adversely affect the environment and human health. To mitigate, bioremediation could be adopted to tackle the problem of hydrocarbon-contaminated soil. Thus, this experimental research carried out the bioremediation using chicken manure in soils contaminated with 5%, 10% and 20% w/w used lubricating oil for a 42-day composting period. To compare, this research also experimented with the 5%, 10% and 20% oil-contaminated soils untreated with chicken manure. The results showed that the highest total petroleum hydrocarbons (TPHs) reduction efficiency of >60% was achieved in the 5% oil-contaminated compost remediated with chicken manure. The highest biodegradation rate of lubricating oil of 0.023–0.0025 day^?1 as measured by the first-order kinetics could also be achieved under the 5% oil contamination condition with the application of chicken manure. The findings highlight the prospect of chicken manure as a proper nutrient for enhanced remediation of hydrocarbon-contaminated soils, particularly of low contamination concentrations.     

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.     

Role of yeasts of the genus Candida in the transformation of the hydrocarbon energy in oil-contaminated gray-brown soil

Energy transformation was studied in the processes of hydrocarbon metabolism by microorganisms in oil-contaminated grey-brown soil. For such microbial populations as Candida species which use hydrocarbon substrates only as an energy source but not for growth, the criteria of biomass or incidence are presumed to have a relative significance for revealing the comparative functional role of these populations in the self-purification of oil-contaminated soils. Therefore, it is necessary to simultaneously assay both the incidence and the functional activity of microorganisms expressed in terms of respiration intensity. A diagram is proposed which shows the role of Candida yeasts in transforming the energy of aromatic and paraffin hydrocarbons in oil-contaminated soils.     

Distribution of Naphthalene Dioxygenase Genes in Crude Oil-Contaminated Soils

Polycyclic aromatic hydrocarbons (PAHs) are one of the major pollutants in soils in oil exploring areas. Biodegradation is the major process for natural elimination of PAHs from contaminated soils. Functional genes can be used as biomarkers to assess the biodegradation potential of indigenous microbial populations. However, little is known about the distribution of PAH-degrading genes in the environment. The links between environmental parameters and the distribution of PAH metabolic genes remain essentially unclear. The present study investigated the abundance and diversity of naphthalene dioxygenase genes in the oil-contaminated soils in the Shengli Oil Field (China). Spatial variations in the density and diversity of naphthalene dioxygenase genes occurred in this area. Four different sequence genotypes were observed in the contaminated soils, with the predominance of novel PAH-degrading genes. Pearson’s correlation analysis illustrated that gene abundance had positive correlations with the levels of total organic carbon and aromatic hydrocarbons, while gene diversity showed a negative correlation with the level of polar aromatics. This work could provide some new insights toward the distribution of PAH metabolic genes and PAH biodegradation potential in oil-contaminated ecosystems.     

Assessing mineralization rates of petroleum hydrocarbons in soils in relation to environmental factors and experimental scale

Mineralization rates of non-volatile petroleum hydrocarbons (HCs) in five different oil-contaminated soils with initial HC contents ranging from 0.1 to 13 g kg^-1 are estimated as a function of environmental factors. The aim of the study is threefold, (i) to study the relevance of environmental factors that may influence the mineralization rate, (ii) to compare mineralization rates estimated in two experiments at different scales, after standardizing them to environmental reference conditions, (iii) to evaluate the CO₂ production rate as a measure for the mineralization rate of HCs. Experiments were performed at laboratory scale (30–50 cm³ soil volume) in closed-jars under constant environmental conditions and in lysimeters (0.81 m³ soil volume) under dynamic climatic and hydrological conditions. A biodegradation model, coupled to transport models for soil heat, water, and gas dynamics is employed for data interpretation. The transport models are used to simulate the environmental conditions that influence the mineralization rate in the non-steady lysimeter experiments. The results show that temperature, O₂ concentration and HC content have an effect on the mineralization rates. Water content could not be identified as a direct governing environmental factor. However, an indirect effect of water content is that it influences the effective gas diffusion coefficient in soils. The CO₂ production rate seems to be a good quantity to express the mineralization rate of HCs for HC contents>1 g kg^-1. Measured CO₂ production rates standardized to reference conditions are similar for the two different experimental scales. This demonstrates that the usage of biodegradation rates obtained in the laboratory to predict the biodegradation rates under field conditions is sound, as long as the differences in environmental conditions have been taken into account.     

Effect of UV radiation transmitted through light-correcting films on the enzymatic activity of oil-polluted soil

The effect of photoluminescent (light-correcting) polymeric films on the enzymatic activity of microorganisms in oil-polluted soil was investigated under laboratory conditions. The efficiency of the use of films as a covering material containing organic and inorganic photoluminophores was studied under the UV-irradi-ation of the soil polluted with oil products in a concentration of up to 50 g/kg. The application of light-correcting films increases the activity of enzymes in oil-polluted soil: catalase by a factor of 2–4, dehydrogenase 2.5, urease 4.5, polyphenoloxidase and peroxidase by a factor of 2–3. In this situation, as chromatographic investigations showed, biodegradation of oil hydrocarbons polluting the soil proceeds 5–6 times faster in comparison with the reference versions for which a usual polymeric high-pressure polyethylene film was used.     

Characterisation of biodegradation capacities of environmental microflorae for diesel oil by comprehensive two-dimensional gas chromatography

In contaminated soils, efficiency of natural attenuation or engineered bioremediation largely depends on biodegradation capacities of the local microflorae. In the present study, the biodegradation capacities of various microflorae towards diesel oil were determined in laboratory conditions. Microflorae were collected from 9 contaminated and 10 uncontaminated soil samples and were compared to urban wastewater activated sludge. The recalcitrance of hydrocarbons in tests was characterised using both gas chromatography (GC) and comprehensive two-dimensional gas chromatography (GC×GC). The microflorae from contaminated soils were found to exhibit higher degradation capacities than those from uncontaminated soil and activated sludge. In cultures inoculated by contaminated-soil microflorae, 80% of diesel oil on an average was consumed over 4-week incubation compared to only 64% in uncontaminated soil and 60% in activated sludge cultures. As shown by GC, n-alkanes of diesel oil were totally utilised by each microflora but differentiated degradation extents were observed for cyclic and branched hydrocarbons. The enhanced degradation capacities of impacted-soil microflorae resulted probably from an adaptation to the hydrocarbon contaminants but a similar adaptation was noted in uncontaminated soils when conifer trees might have released natural hydrocarbons. GC×GC showed that a contaminated-soil microflora removed all aromatics and all branched alkanes containing less than C₁₅. The most recalcitrant compounds were the branched and cyclic alkanes with 15–23 atoms of carbon.     

Effects of oil spills on microbial heterotrophs in Antarctic soils

Oil spillage on the moist coastal soils of the Ross Sea region of Antarctica can impact on populations of microbial heterotrophs in these soils, as determined by viable plate counts and a most probable number technique. Elevated numbers of culturable hydrocarbon degraders, bacteria and fungi were detected in surface and subsurface soils from oil-contaminated sites, compared with nearby control sites. Culturable yeasts were not detected in soil from coastal control sites, yet reached >10⁵ organisms g^-1 dry weight in contaminated soils. The presence of hydrocarbons in soils resulted in a shift in the genera of culturable filamentous fungi. Chrysosporium dominated control soils, yet Phialophora was more abundant in oil-contaminated soils. Hydrocarbon degraders are most likely bacteria; however, fungi could play a role in degradation of hydrocarbons or their metabolites. Depleted levels of nitrate detected in some contaminated soils and decreased pH may be the result of growth of hydrocarbon degraders. Numbers and diversity of culturable microbes from Antarctic soil varied depending on whether a pristine site or a human-impacted (in this case, by fuel spills) site is studied.     

Potential of vetiver (vetiveria zizanioides (L.) Nash) for phytoremediation of petroleum hydrocarbon-contaminated soils in Venezuela

Venezuela is one of the largest oil producers in the world. For the rehabilitation of oil-contaminated sites, phytoremediation represents a promising technology whereby plants are used to enhance biodegradation processes in soil. A greenhouse study was conducted to determine the tolerance of vetiver (Vetiveria zizanioides (L.) Nash) to a Venezuelan heavy crude oil in soil. Additionally, the plant's potential for stimulating the biodegradation processes of petroleum hydrocarbons was tested under the application of two fertilizer levels. In the presence of contaminants, biomass and plant height were significantly reduced. As for fertilization, the lower fertilizer level led to higher biomass production. The specific root surface area was reduced under the effects of petroleum. However, vetiver was found to tolerate crude-oil contamination in a concentration of 5% (w/w). Concerning total oil and grease content in soil, no significant decrease under the influence of vetiver was detected when compared to the unplanted control. Thus, there was no evidence of vetiver enhancing the biodegradation of crude oil in soil under the conditions of this trial. However, uses of vetiver grass in relation to petroleum-contaminated soils are promising for amelioration of slightly polluted sites, to allow other species to get established and for erosion control.     

Enhanced degradation of iprodione in soil after repeated treatments for controlling Sclerotinia minor

Poor field control of lettuce collar rot by iprodione was observed in southern France and was attributed to enhanced biodegradation of the fungicide. Enhanced biodegradation was obtained in vitro after repeated applications of iprodione to non-degrading soils. Normal soils became biodegrading after mixing with degrading soils (3 vol./1 vol.). Activity of the responsible microflora seemed dependent on soil physico-chemical characteristics.     

Design of bacterial defined mixed cultures for biodegradation of specific crude oil fractions, using population dynamics analysis by DGGE

Hydrocarbon-degrading bacteria isolated from oil-polluted soils, were used to design three defined mixed cultures (DMC) for biodegradation of Maya crude oil fractions. The first degrading culture, DMC A was made up with 10 strains. Design of DMC B (six strains) and DMC C (three strains) was based on DGGE profiles obtained throughout biodegradation assays of different petroleum fractions. Biodegradation of the aliphatic fraction (10 000 mg l⁻¹) and an aromatic–polar mixture (5000 mg l⁻¹) was evaluated for the DMC B. Biodegradation of total hydrocarbons (10 000 mg l⁻¹) and its fractions was evaluated for DMC B and DMC C. During biodegradation assays, O₂ consumption and CO₂ production were assessed by respirometry, while population dynamics of predominant strains was based on PCR-DGGE profiles of partial 16S rDNA. Aliphatic fraction was completely biodegraded by DMC B, while degradation of the aromatic–polar mixture was 12.5% and for total hydrocarbons 40.5%. DMC B was able to degrade the aromatic fraction (31%) and even the polar fraction (19.6%) present in total hydrocarbons. DMC C degraded the aromatic and polar fractions (5.6% and 2%, respectively) present in total hydrocarbons. DGGE profiles of the DMCs indicated that Pseudomonas sp., Gordonia rubripertincta and a non-identified strain were predominant and probably responsible of the hydrocarbons biodegradation. The use of DGGE-fingerprinting to track microbial populations, allowed selecting strains to design efficient oil-degrading defined mixed cultures.     

Heterotrophic Potentials and Hydrocarbon Biodegradation Potentials of Sediment Microorganisms Within the Athabasca Oil Sands Deposit

Techniques for the enumeration and the determination of the potential activity of disturbed sediment mixed populations at control sites and sites within the Athabasca oil sands formation were applied to August and December samples. These techniques included the determination of general heterotrophic potential for the assimilation and respiration of glutamate, which indicated no oil sand-related changes in the sediments but which indicated a significant seasonal change. Enumeration by epifluorescence direct counts, oil sand hydrocarbon plate counts, and most-probable-number determinations of [¹⁴C]hexadecane and [¹⁴C]-naphthalene degraders indicated that only the plate count was sensitive to increased numbers of oil sand-related hydrocarbon-oxidizing microorganisms within the oil sands deposit. Unlike the most probable number determinations of [¹⁴C]hexadecane and [¹⁴C]naphthalene degraders, however, the biodegradation potential results of these substrates indicated a significant increase in activity at oil sands sites. These biodegradation potentials also showed a marked seasonal fluctuation. Although the biodegradation potentials and the endogenous hydrocarbon plate counts indicated an oil sand-adapted mixed sediment population, the results of these techniques did not correlate well with the concentrations of bituminous hydrocarbons in the sediments. The results suggest that a general capability for hydrocarbon oxidation exists in the Athabasca River system and that this capability is enhanced within the natural bounds of the Athabasca oil sands.     

Oil degradation by basidiomycetes in soil and peat at low temperatures

A total of 17 basidiomycete strains causing white rot and growing on oil-contaminated substrates have been screened. Three strains with high (Steccherinum murashkinskyi), average (Trametes maxima), and low (Pleurotus ostreatus) capacities for the colonization of oil-contaminated substrates have been selected. The potential for degrading crude oil hydrocarbons has been assessed with the use of fungi grown on nonsterile soil and peat at low temperatures. Candida sp. and Rhodococcus sp. commercial strains have been used as reference organisms with oil-degrading ability. All microorganisms introduced in oil-contaminated soil have proved to be ineffective, whereas the inoculation of peat with basidiomycetes and oil-degrading microorganisms accelerated the destruction of oil hydrocarbons. The greatest degradation potential of oil-aliphatic hydrocarbons has been found in S. murashlinskyi. T. maxima turned out to be the most successful in degrading aromatic hydrocarbons. It has been suggested that aboriginal microflora contributes importantly to the effectiveness of oil-destructing microorganisms. T. maxima and S. murashkinskyi strains are promising for further study as oil-oxidizing agents during bioremediation of oil-contaminated peat soil under conditions of low temperatures.     

Microbe-aliphatic hydrocarbon interactions in soil: implications for biodegradation and bioremediation

Aliphatic hydrocarbons make up a substantial portion of organic contamination in the terrestrial environment. However, most studies have focussed on the fate and behaviour of aromatic contaminants in soil. Despite structural differences between aromatic and aliphatic hydrocarbons, both classes of contaminants are subject to physicochemical processes, which can affect the degree of loss, sequestration and interaction with soil microflora. Given the nature of hydrocarbon contamination of soils and the importance of bioremediation strategies, understanding the fate and behaviour of aliphatic hydrocarbons is imperative, particularly microbe-contaminant interactions. Biodegradation by microbes is the key removal process of hydrocarbons in soils, which is controlled by hydrocarbon physicochemistry, environmental conditions, bioavailability and the presence of catabolically active microbes. Therefore, the aims of this review are (i) to consider the physicochemical properties of aliphatic hydrocarbons and highlight mechanisms controlling their fate and behaviour in soil; (ii) to discuss the bioavailability and bioaccessibility of aliphatic hydrocarbons in soil, with particular attention being paid to biodegradation, and (iii) to briefly consider bioremediation techniques that may be applied to remove aliphatic hydrocarbons from soil.     

Structural and Functional Variation in Genetic Systems for Catabolism of Polycyclic Aromatic Hydrocarbons in Pseudomonas putida Strains

The genetic control of naphthalene, phenanthrene, and anthracene biodegradation was studied in three Pseudomonas putida strains isolated from coal tar- and oil-contaminated soils. These strains isolated from different geographical locations contained similar catabolic plasmids controlling the first steps of naphthalene conversion to salicylate (the nah1operon), functionally inoperative salicylate hydroxylase genes, and genes of the metha-pathway of catechol degradation (the nah2 operon). Salicylate oxidation in these strains is determined by genes located in trans-position relative to the nah1 operon: in strains BS202 and BS3701, they are located on the chromosome, and in the strain BS3790, on the second plasmid.     

Effects of Zeolite-Containing Material and Ammonium Nitrate on Biodegradation of n-Tridecane in Heavy Clay-Loam Leached Chernozem with High Organic Matter Content

Biostimulation based on usage of soil amendments is growing due to their efficiency in removing different petroleum hydrocarbons (PHC) from contaminated sand or loam-sand soils. However, the research on clay-rich soils with higher organic carbon content, in which PHC biodegradation may proceed differently and which are more difficult to clean up, has been less extensive. In a pot experiment, we studied and compared the effects of two soil amendments, natural zeolite-containing material (ZCM, 50 g kg^?1) as a bulking agent and ammonium nitrate (0.3 g N kg^?1) as a nitrogen fertilizer, on biodegradation of n-tridecane (1 wt.%) in a weakly acidic heavy clay loam leached chernozem with fairly high organic carbon content (3.71%). After 48 days, the nitrogen-amended contaminated soil showed enhancement of both respiratory activity (basal and substrate-induced respiration rates) and the number of n-tridecane- degraders. As a consequence, the extent of n-tridecane biodegradation (86.5%) was essentially higher in the presence of added nitrogen than that in the non-amended soil (73.7%). In contrast, due to the partial retention of n-tridecane molecules in its pores, ZCM retarded biodegradation to 56.0%, showed no significant effect on the number of n-tridecane-degraders and, moreover, enhanced the decomposition of the soil intrinsic organic matter. The obtained data indicate that more precautions should be considered when using porous sorbents such as ZCM for remedial arrangements in PHC-contaminated soils.     

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.     

Intrinsic capacities of soil microfloræ for gasoline degradation

A methodology to determine the intrinsic capacities of a microflora to degrade gasoline was developed, in particular for assessing the potential of autochtonous populations of polluted and non polluted soils for natural attenuation and engineered bioremediation. A model mixture (GM23) constituted of the 23 most representative hydrocarbons of a commercial gasoline was used. The capacities of the microfloræ (kinetics and extent of biodegradation) were assessed by chromatographic analysis of hydrocarbon consumption and of CO2 production. The degradation of the components of GM23 was assayed in separate incubations of each component and in the complete mixture. For the microflora of an unpolluted spruce forest soil, all hydrocarbons of GM23 except cyclohexane, 2,2,4- and 2,3,4-trimethylpentane isomers were degraded to below detection limit in 28 days. This microflora was reinforced with two mixed microbial communities selected from gasoline-polluted sites and shown to degrade cyclohexane and 2,2,4- trimethylpentane. With the reinforced microflora, complete degradation of GM23 was observed. The degradation patterns of individual components of GM23 were similar when the compounds were present individually or in the GM23 mixture, as long as the concentrations of 2-ethyltoluene and trimethylbenzene isomers were kept sufficiently low ( 35 mg.l-1) to remain below their inhibitory level.     

Microflora patterns in oil-contaminated soils

The method of glass slides was modified to study the dynamics of microbial cenoses in oil-contaminated soils. The resulting microflora patterns can be clearly distinguished between soil types and oil quality and utilization.     

A suitable model to describe bioremediation of a petroleum-contaminated soil

A critical environmental impact of the petroleum industry is the spillage of oil and related products that causes severe soil contamination. Although biodegradation of petroleum hydrocarbons may be successfully conducted under controlled conditions, the bioremediation of large volumes of contaminated soils still presents some technical challenges, particularly when contamination occurs in soils of high clay content. The main objective of this work is to evaluate the bioremediation of petroleum-contaminated clay-soil by fixed bed experiments. They were conducted in agreement with the full factorial experimental design 2³. The processes employed were shown to be highly effective in decontaminating the soil, achieving removal rates of around 80%. The model chosen to represent the bioremediation process provided satisfactory results. The values calculated by the model were consistent with the experimental results.