Bioremediation of coal tar PAH in soils using biodiesel

The addition of biodiesel together with nitrate and phosphate to soil containing coal tar, in laboratory and field experiments, resulted in degradation of coal tar polycyclic aromatic hydrocarbons (PAH) that was not apparent when the nutrients alone were added. The addition of motor diesel fuel instead of biodiesel was also tested. Over the 55 days of the field and laboratory experiments, the biodiesel resulted in an increased degradation of naphthalene in the coal tar by 52% and 85%, respectively, and motor diesel resulted in increased depletions of 85% and 96%, respectively. Other PAH containing up to four rings were depleted to lesser extents. The increases in PAH biodegradation by the diesel treatments were ascribed to tar solubilisation and dispersion thereby increasing the PAH bioavailability. The ready biodegradability and low phytotoxicity of biodiesel suggest that it may be suitable as a novel treatment for the bioremediation of coal tar contaminated soils.     

Ecorisk evaluation and treatability potential of soils contaminated with petroleum hydrocarbon-based fuels

We used a series of toxicity tests to monitor oil degradation in the Kuwaiti oil lakes. Three soils from different locations with a history of hydrocarbon contamination were treated in bench-scale microcosms with controlled nutrient amendments, moisture content, and temperature that had promoted mineralization of total hydrocarbon and oil and grease in a preliminary study. Two hundred days of bioremediation treatment lowered hydrocarbon concentration to below 2 and 5 mg g(-1) for soils A and B, respectively, while in soil C hydrocarbon concentration remained at 12 mg g(-1). Although 85% of the total petroleum hydrocarbons (TPHs) in soil A were reduced 50d after treatment, results of the seed germination and Microtox tests suggested an initial increase in toxicity, indicating that toxic intermediary metabolites may have formed during biodegradation. Also, the significant decrease of TPHs and corresponding high toxicity levels were noted in soil B 200d after bioremediation. Clearly, toxicity values, and not just hydrocarbon concentration, are a key factor in assessing the effectiveness of bioremediation techniques. Field chemistry data showed a significant reduction in hydrocarbon levels after the biological treatment. We concluded that the toxicity assessment of the contaminated soil with a battery of toxicity bioassays could provide meaningful information regarding a characterization procedure in ecological risk assessment.     

Investigation of evaporation and biodegradation of fuel spills in Antarctica. I. A chemical approach using GC-FID

Little effort has been devoted to differentiating between hydrocarbon losses through evaporation and biodegradation in treatability studies of fuel-contaminated Antarctic soils. When natural attenuation is being considered as a treatment option, it is important to be able to identify the mechanism of hydrocarbon loss and demonstrate that rates of degradation are sufficient to prevent off-site migration. Similarly, where complex thermally enhanced bioremediation schemes involve nutrient addition, water management, air stripping and active heating, it is important to appreciate the relative roles of these mechanisms for cost minimisation. Following the loss of hydrocarbons by documenting changes in total petroleum hydrocarbons offers little insight into the relative contribution of evaporation and biodegradation. We present a methodology here that allows identification and quantification of evaporative losses of diesel range organics at a range of temperatures using successively less volatile compounds as fractionation markers. We also present data that supports the general utility of so-called biodegradation indices for tracking biodegradation progress. We are also able to show that at 4 degrees C indigenous Antarctic soil bacteria degrade Special Antarctic Blend fuel components in the following order: naphthalene and methyl-napthalenes, light n-alkanes, then progressively heavier n-alkanes; whereas isoprenoids and the unresolved complex mixture are relatively recalcitrant.     

Impact of microbial activity on copper, lead and nickel mobilization during the bioremediation of soil PAHs

A fungal bioremediation method using P. frequentans removed up to 75% of phenanthrene with the addition of water and nutrients over a period of 30 d. During the bioremediation process, changes in metal behavior were monitored by an in situ technique (diffusive gradients in thin-films, DGT) and by soil solution chemistry. DGT provided absolute data on fluxes from the solid phase to the DGT device and relative trends of concentrations of the most labile metal species. DGT response indicated that bioremediation increases metal mobilization from the solid phase. Filtration provided data on the concentrations of solution phase (<0.45 microm) metal. In all case, metal fluxes and concentrations significantly increased after the bioremediation process began. Fluxes increased from <0.1 pg cm(-2)s(-1) before bioremediation to between 0.2 and 0.5 pg cm(-2)s(-1) after bioremediation. Metal concentrations in the soils solution (filtration at 0.45 microm) increased from 2 to 10 microg l(-1) (Cu), 1-4 microgl(-1) (Pb) and from 40 to 140 microg l(-1) (Ni) after bioremediation. Although over a short time period, these data strongly indicated that there was remobilization of metal from solid to solution (and thus to the DGT device) directly due to the bioremediation process. Although the mechanism was not unambiguously identified, it was shown not to be related to small changes in bulk pH over time and was attributed to the microbial action on the surface of the soil solid phase, releasing metal into solution. Additionally, differences in metal concentration and flux were observed in sterilized and non-sterilized soils and in the absence or presence of phenanthrene. The results indicated that the bioremediation of soil by P. frequentans increased the flux, lability and mobility of trace metal species and therefore the likely metal bioavailability to plants.     

Dynamics of carbon, nitrogen and hydrocarbons in diesel-contaminated soil amended with biosolids and maize

Contamination of soil with hydrocarbons occurs frequently when petroleum ducts are damaged. Restoration of those contaminated soils might be achieved by applying readily available organic material. An uncontaminated clayey soil sampled in the vicinity of a duct carrying diesel which ruptured recently, was contaminated in the laboratory and amended with or without maize or biosolids while production of carbon dioxide (CO(2)), dynamics of ammonia (NH(4)(+)), nitrates (NO(3)(-)), and total petroleum hydrocarbons (TPH) were monitored. The fastest mineralization of diesel, as witnessed by production of CO(2), was found when biosolids were added, but the amount mineralized after 100 days, approximately 88%, was similar in all treatments. Approximately 5 mg of the 48 mg TPH kg(-1) found in the sterilized soil at the beginning of the experiment could not be accounted for after 100 days. The concentration of TPH in the unsterilized soil decreased rapidly in all treatments, but the rate of decrease was different between the treatments. The fastest decrease was found in the soil amended with biosolids and approximately 30 mg TPH kg(-1) or 60% could not be accounted for within 7 days. The decrease in concentration of TPH at the onset of the incubation was similar in the other treatments. After 100 days, the concentration of TPH was similar in all soils and appear to stabilize at 19 mg TPH kg(-1) soil. It was concluded that biosolids accelerated the decomposition of diesel and TPH due to its large nutrient content, but after 100 days the amount of diesel mineralized and the residual concentration of TPH was not affected by the treatment applied.     

Electrokinetic enhancement of phenanthrene biodegradation in creosote-polluted clay soil

Given the difficulties caused by low-permeable soils in bioremediation, a new electrokinetic technology is proposed, based on laboratory results with phenanthrene, to afford bioremediation of polycyclic aromatic hydrocarbons (PAH) in clay soils. Microbial activity in a clay soil historically polluted with creosote was promoted using a specially designed electrokinetic cell with a permanent anode-to-cathode flow and controlled pH. The rates of phenanthrene losses during treatment were tenfold higher in soil treated with an electric field than in the control cells without current or microbial activity. Results from experiments with Tenax-assisted desorption and mineralization of 14C-labeled phenanthrene indicated that phenanthrene biodegradation was limited by mass-transfer of the chemical. We suggest that the enhancement effect of the applied electric field on phenanthrene biodegradation resulted from mobilization of the PAH and nutrients dissolved in the soil fluids.     

鼠李糖脂对不同菌株降解柴油污染物的影响

通过一系列实验分析了鼠李糖脂对柴油污染物生物降解的影响。单菌株柴油降解实验结果表明,在添加生物表面活性剂鼠李糖脂后,各菌株细胞表面疏水性均发生不同程度的增加,并且对柴油的降解率均有所提高。在混合菌的柴油污染物降解实验中,发现当向土壤中添加了200 mg/L鼠李糖脂时,对柴油的降解才有较大的提高;而当添加100 mg/L的鼠李糖脂到水体中时,对柴油的降解就有较大的提高,而当鼠李糖脂浓度提高为200 mg/L时,柴油的降解率却没有进一步明显的提高。这说明鼠李糖脂对柴油降解的影响程度不仅与环境介质有关,还与添加的鼠李糖脂浓度有关。进一步分析表明,添加适当浓度的鼠李糖脂不仅可以提高对柴油的降解率,而且可加速其降解速度,缩短生物修复所需时间。     

Bioremediation of oil-contaminated soil using Candida catenulata and food waste

Even though petroleum-degrading microorganisms are widely distributed in soil and water, they may not be present in sufficient numbers to achieve contaminant remediation. In such cases, it may be useful to inoculate the polluted area with highly effective petroleum-degrading microbial strains to augment the exiting ones. In order to identify a microbial strain for bioaugmentation of oil-contaminated soil, we isolated a microbial strain with high emulsification and petroleum hydrocarbon degradation efficiency of diesel fuel in culture. The efficacy of the isolated microbial strain, identified as Candida catenulata CM1, was further evaluated during composting of a mixture containing 23% food waste and 77% diesel-contaminated soil including 2% (w/w) diesel. After 13 days of composting, 84% of the initial petroleum hydrocarbon was degraded in composting mixes containing a powdered form of CM1 (CM1-solid), compared with 48% of removal ratio in control reactor without inoculum. This finding suggests that CM1 is a viable microbial strain for bioremediation of oil-contaminated soil with food waste through composting processes.     

Enhancing bioremediation of diesel oil and gasoline in soil amended with an agroindustry sludge

This paper presents a study of the bioremediation of diesel oil and gasoline by a series of controlled laboratory tests. Sludge from an agroindustry was used to enhance bioremediation of both gasoline and diesel oil mixed with a soil mass to compare its efficiency with that of a mineral fertilizer. Effects of soil microbiology and soil mixtures were investigated by means of evolution of CO2, microorganism populations at 90 days, pH at 65 and 95 days, mineral nitrogen, and gas chromatographic analysis of the benzene, toluene, methyl tertiary butyl ether, C8, and C9+total aromatics at the end of the experiments. Treatments containing sludge showed better soil conditions after 170 days of treatment (inorganic nitrogen and microbiota activity) compared with gasoline and diesel oil without amendments. Samples had no detectable traces of the measured hydrocarbons at 170 days of treatment.     

The use of ozone in the remediation of polycyclic aromatic hydrocarbon contaminated soil

The potential of using ozone for the removal of phenanthrene from several different soils, both alone and in combination with biodegradation using a microbial inoculant (Pseudomonas alcaligenes PA-10), was examined. The greater the water content of the soil the less effective the ozone treatment, with air-dried soils showing the greatest removal of phenanthrene; while soils with higher levels of clay also reduced the effectiveness of the ozone treatments. However, at least a 50% reduction in phenanthrene levels was achieved in air-dried soil after an ozone treatment of 6 h at 20 ppm, with up to 85% removal of phenanthrene achieved in sandy soils. The biodegradation results indicate that P. alcaligenes PA-10 may be useful as an inoculant for the removal of PAHs from contaminated soils. Under the conditions used in our experiments, however, pre-ozonation did not enhance subsequent biodegradation of phenanthrene in the soils. Similar levels of phenanthrene removal occurred in both non-ozonated and ozonated Cruden Bay soil inoculated with P. alcaligenes PA-10. However, the biodegradation of phenanthrene in ozonated Boyndie soil was much slower. This may be due to the release of toxic products in this soil during ozonation.     

Solid-phase bioassays and soil microbial activities to evaluate PAH-spiked soil ecotoxicity after a long-term bioremediation process simulating landfarming

The residual ecotoxicity of long-term bioremediated soils concomitantly spiked with three PAHs at four levels (15, 75, 150, 300 mg Sigma 3 PAHs kg(-1) soil) was evaluated using physico-chemical analyses, solid-phase bioassays and soil microbial activities. The pot-scale bioremediation process consisted of weekly moderate waterings in the presence or absence of sewage sludge compost (SSC) under greenhouse conditions. After 15 months, anthracene and pyrene were almost completely degraded whereas benzo[a]pyrene was still persisting, most apparently in SSC-amended soil treatments. However, no apparent toxic effects of the residual PAHs could be detected. SSC application at 40 t ha(-1) was performed to valorize the biowaste and stimulate PAH biodegradation but caused soil salinization and pH reduction at the end of the bioremediation process. Consequently, SSC-amended soils were characterized by strong phytotoxicity to lettuce and had adverse effects on the ostracod Heterocypris incongruens. Despite the smaller number of culturable bacterial populations in SSC-amended soils, soil enzymatic activities were not affected by the organic amendment and residual PAHs; and the bioremediation efficiency was likely to be more limited by the bioavailability of PAHs rather than by the total number of PAH-degraders.     

Prediction of complete bioremediation periods for PAH soil pollutants in different physical states by mechanistic models

Mass-transfer models and biodegradation models were developed for three theoretical physical states of polycyclic aromatic hydrocarbons (PAHs) in soil. These mechanistic models were used to calculate the treatment periods necessary for complete removal of the PAH pollutants from the soil under batch conditions. Results indicate that the bioremediation of PAHs in such systems is mainly mass-transfer limited. The potential for bioremediation as a treatment technique for PAH contaminated soils is therefore mainly determined by the mass-transfer dynamics of PAHs. Under mass-transfer limited conditions simplified mathematical models, based on the assumption of a zero dissolved PAH concentrations, can be used to predict the period of time needed for complete bioremediation.     

生物修复剂在清除海滩石油污染中的应用

介绍了生物修复石油污染海滩时常用的修复剂类型及其特点.当实验室环境条件能较好控制时,生物强化剂一般是有效的;然而污染现场得出的证据不能表明其对生物降解有促进作用.实验室和现场的研究均表明营养型生物促进剂能有效促进石油的生物降解.水溶性营养易被波浪和潮汐冲刷掉;缓释型营养盐面临的主要挑战是如何控制其释放速率,以保证孔隙水中能较长时间维持理想的营养浓度;亲油型肥料中含有有机碳,有可能在微生物降解石油之前被优先降解.建议根据污染环境的特点选用适合的生物促进剂.     

The influence of a NAPL on the loss and biodegradation of 14C-phenanthrene residues in two dissimilar soils

This study was carried out to assess the influence of diesel, applied over a log concentration range, on the loss and extractability of phenanthrene (measured as putative 14C-phenanthrene residues) in two different soils. The influence of diesel on the ability of a cyclodextrin based extraction method to predict the microbial bioavailability of 14C-residues was also assessed. An increase in loss of 14C-residues with increasing diesel concentration from 0 to 2000 mg kg-1 was generally observed with time in both soils. It is suggested that this trend is attributable to competitive sorption for soil sorption sites and to a lesser extent to displacement of 14C-residues from soil sorption sites by diesel resulting in greater compound availability and therefore greater loss by degradation via the actions of indigenous microorganisms. However, in the 20000 mg kg-1 diesel treatments of both soils, results indicated a delayed loss. It is suggested that this retarded loss was due to the formation of a discrete NAPL-phase into which 14C-phenanthrene residues partitioned, thereby decreasing their availability and as a consequence their degradation. Furthermore, it is suggested that nutrient limitation may have slowed down degradation rates as diesel concentrations increased. Comparison between cyclodextrin-extractability and microbial mineralisation supported the use of cyclodextrin to assess microbial bioavailability of 14C-residues after 50 d or more ageing up to diesel concentrations of 2000 mg kg-1. However, results suggested that at high diesel concentrations (specifically 20000 mg kg-1) co-extraction of 14C-phenanthrene residues may have occurred as a result of the combined solvation powers of both the cyclodextrin and the diesel. Furthermore, mineralisation of 14C-phenanthrene residues may have been affected by extreme nutrient limitation in this treatment.     

Evaluation of B. subtilis SPB1 biosurfactants' potency for diesel-contaminated soil washing: optimization of oil desorption using Taguchi design

Low solubility of certain hydrophobic soil contaminants limits remediation process. Surface-active compounds can improve the solubility and removal of hydrophobic compounds from contaminated soils and, consequently, their biodegradation. Hence, this paper aims to study desorption efficiency of oil from soil of SPB1 lipopeptide biosurfactant. The effect of different physicochemical parameters on desorption potency was assessed. Taguchi experimental design method was applied in order to enhance the desorption capacity and establish the best washing parameters. Mobilization potency was compared to those of chemical surfactants under the newly defined conditions. Better desorption capacity was obtained using 0.1 % biosurfacatnt solution and the mobilization potency shows great tolerance to acidic and alkaline pH values and salinity. Results show an optimum value of oil removal from diesel-contaminated soil of about 87 %. The optimum washing conditions for surfactant solution volume, biosurfactant concentration, agitation speed, temperature, and time were found to be 12 ml/g of soil, 0.1 % biosurfactant, 200 rpm, 30 °C, and 24 h, respectively. The obtained results were compared to those of SDS and Tween 80 at the optimal conditions described above, and the study reveals an effectiveness of SPB1 biosurfactant comparable to the reported chemical emulsifiers. (1) The obtained findings suggest (a) the competence of Bacillus subtilis biosurfactant in promoting diesel desorption from soil towards chemical surfactants and (b) the applicability of this method in decontaminating crude oil-contaminated soil and, therefore, improving bioavailability of hydrophobic compounds. (2) The obtained findings also suggest the adequacy of Taguchi design in promoting process efficiency. Our findings suggest that preoptimized desorption process using microbial-derived emulsifier can contribute significantly to enhancement of hydrophobic pollutants' bioavailability. This study can be complemented with the investigation of potential role in improving the biodegradation of the diesel adsorbed to the soil.     

Content and distribution of forms of organic N in soil and particle size fractions after long-term fertilization

Most of the N in surface soils occurs in organic forms, and when mineralized it plays a key role in soil fertility and plant nutrition. Our objective was to study the effect of long-term applications of organic manure on the content and distribution of forms of organic N in bulk soil and soil particle size fractions to characterize the inherent soil nitrogen fertility. Five treatments were as follows: (1) CK (no fertilizer and no manure added), (2) mineral fertilizer only, (3) straw + NPK, (4) green manure + NPK and (5) pig manure + NPK. Soil particle size fractions (0-2, 2-10, 10-50 and 50-100 microm) were isolated without chemical pretreatment by ultrasonic dispersion in water followed by sedimentation. The content of total N and forms of organic N in the bulk soil increased after long-term fertilization, and the effect varied with fertilizer type. The plot treated with only mineral fertilizer gave the highest NH3-N and the lowest amino sugar-N content in all treatments. The highest content of amino sugar-N and amino acid-N was found in the treatment of pig manure + NPK. The content (g kg(-1) fraction) of hydrolysable N within size fractions was in the order 0-2 > 2-10 > 50-100 > 10-50 microm, but the contribution of different size fraction to hydrolysable N decreased in the sequence 10-50 > 0-2 > 2-10 > 50-100 microm. Most of the applied mineral fertilizer N that remained in soils was distributed in the particle size fraction < 2 microm while most of the remaining N from manure applied with NPK was transferred into amino sugar-N in each size fraction, and amino acid-N in the size fractions > 2 microm during the process of humification.     

Effects of nutrient and temperature on degradation of petroleum hydrocarbons in contaminated sub-Antarctic soil

Mesocosm studies using sub-Antarctic soil artificially contaminated with diesel or crude oil were conducted in Kerguelen Archipelago (49 degrees 21' S, 70 degrees 13' E) in an attempt to evaluate the potential of a bioremediation approach in high latitude environments. All mesocosms were sampled on a regular basis over six months period. Soils responded positively to temperature increase from 4 degrees C to 20 degrees C, and to the addition of a commercial oleophilic fertilizer containing N and P. Both factors increased the hydrocarbon-degrading microbial abundance and total petroleum hydrocarbons (TPH) degradation. In general, alkanes were faster degraded than polyaromatic hydrocarbons (PAHs). After 180 days, total alkane losses of both oils reached 77-95% whereas total PAHs never exceeded 80% with optimal conditions at 10 degrees C and fertilizer added. Detailed analysis of naphthalenes, dibenzothiophenes, phenanthrenes, and pyrenes showed a clear decrease of their degradation rate as a function of the size of the PAH molecules. During the experiment there was only a slight decrease in the toxicity, whereas the concentration of TPH decreased significantly during the same time. The most significant reduction in toxicity occurred at 4 degrees C. Therefore, bioremediation of hydrocarbon-contaminated sub-Antarctic soil appears to be feasible, and various engineering strategies, such as heating or amending the soil can accelerate hydrocarbon degradation. However, the residual toxicity of contaminated soil remained drastically high before the desired cleanup is complete and it can represent a limiting factor in the bioremediation of sub-Antarctic soil.     

多环芳烃污染土壤生物修复的强化方法

生物降解是去除环境中多环芳烃(PAHs)的重要途径,通过采取一些强化措施,如使用表面活性剂,添加营养物质和提供共代谢底物等,可显著提高PAHs降解速度和程度,为生物修复技术的成功应用提供前提。在分析中,对近年来国内外在PAHs污染土壤生物修复强化方面的研究进展进行了综述。     

In-situ surfactant/surfactant-nutrient mix-enhanced bioremediation of NAPL (fuel)-contaminated sandy soil aquifers

SCOPE AND BACKGROUND: Contamination of soils, aquifers and groundwater by nonaqueous phase liquid (NAPL) pollutants constitutes a major environmental issue of concern, worldwide. The residual (biodegradation-resistant) hydrophobic fuel hydrocarbons entrapped in the soil porous matrix, possess a particular bioremediation challenge due to their becoming virtually immobile, nor desorbable, or water dispersible. Consequently, they are not available as substrates to the micro-organism-based biodegradation. MATERIALS AND METHODS: Our research involves the development of economically feasible, surfactant/surfactant-nutrient mix (SSNM)-enhanced bioremediation methodologies for sustainable, in situ bioremediation of fuel-contaminated aquifers. This requires, methodologically, (a) the optimization, via in vitro 'flow' (columns) lab experiments and screening processes, of an effective mixture for the intended SSNM-enhanced bioremediation; and (b) the study of the combined effect of the optimized SSNM on the solubilization/mobilization and biodegradation of NAPL (fuel) in in vitro site/aquifer-simulated bioremediation. RESULTS AND DISCUSSION: The essence of our findings: (1) kerosene's maximum enhanced mobilization - f = 3.6, compared with that of deionized water, was achieved with an SSNM having the composition of linear alkylbenzene sulfonate (LABS): coco-amphodiacetate (containing N): surfactant-nutrient X (containing both N and P) = 0.15: 0.15: 0.05 g/L, respectively; (2) 62-64% of the initial amount of kerosene in the initially saturated soil matrix, 'packed' in a column, has been eluted from it during approximately 30 days, compared with 68% of kerosene biodegradation in 'vessel' settings, in 21 days. CONCLUSIONS: (1) The indigenous microorganisms present in th vadose zones of fuel-contaminated sandy soil aquifers are potentially capable of unassisted removal of approximately 80% of the initially contained fuel (kerosene), during a period of about 42 days; (2) the major effects of the SSNM addition are (a) enhanced mobilization of the bulky NAPL; and (b) enhanced desorbtion/ solubilization/dispersion of the entrapped NAPL which, in turn, facilitate their enhanced biodegradation. RECOMMENDATIONS AND PERSPECTIVE: Our findings suggest that pre-optimized, biodegradable SSNM is essential for surfactants-based bioremediation of NAPL-contaminated aquifers, in order to make this in-situ methodology both technologically and economically feasible.