Pilot Scale Bioremediation of Creosote-Contaminated Soil—Efficacy of Enhanced Natural Attenuation and Bioaugmentation Strategies

In this study, the efficacy of bioremediation strategies (enhanced natural attenuation with nitrate and phosphate addition [ENA] and bioaugmentation) for the remediation of creosote-contaminated soil (7767 ± 1286 mg kg^?1 of the 16 EPA priority PAHs) was investigated at pilot scale. Bioaugmentation of creosote-contaminated soil with freshly grown or freeze dried Mycobacterium sp. strain 1B (a PAH degrading microorganism) was applied following bench scale studies that indicated that the indigenous soil microflora had a limited PAH metabolic activity. After 182 days, the total PAH concentration in creosote-contaminated soil was reduced from 7767 ± 1286 mg kg^?1 to 5579 ± 321 mg kg^?1, 2250 ± 71 mg kg^?1, 2050 ± 354 mg kg^?1 and 1950 ± 70 mg kg^?1 in natural attenuation (no additions) and ENA biopiles and biopiles augmented with freshly grown or freeze dried Mycobacterium sp. strain 1B respectively. In ENA and bioaugmentation biopiles, between 82% and 99% of three-ring compounds (acenaphthene, anthracene, fluorene, phenanthrene) were removed while four-ring PAH removal ranged from 33 to 81%. However, the extent of PAH degradation did not vary significantly between the ENA treatment and biopiles augmented with Mycobacterium sp. strain 1B. Four-ring PAH removal followed the order fluoranthene > pyrene > benz[a]anthracene > chrysene. The high residual concentration of some four-ring PAHs may be attributable to bioavailability issues rather than a lack of microbial catabolic activity. Comparable results between ENA and bioaugmentation at pilot scale were surprising given the limited degradative capacity of the microbial consortia enriched from the creosote-contaminated soil.     

Bioremediation of a polyaromatic hydrocarbon contaminated soil by native soil microbiota and bioaugmentation with isolated microbial consortia

Biodegradation of a mixture of PAHs was assessed in forest soil microcosms performed either without or with bioaugmentation using individual fungi and bacterial and a fungal consortia. Respiratory activity, metabolic intermediates and extent of PAH degradation were determined. In all microcosms the low molecular weight PAH’s naphthalene, phenanthrene and anthracene, showed a rapid initial rate of removal. However, bioaugmentation did not significantly affect the biodegradation efficiency for these compounds. Significantly slower degradation rates were demonstrated for the high molecular weight PAH’s pyrene, benz[a]anthracene and benz[a]pyrene. Bioaugmentation did not improve the rate or extent of PAH degradation, except in the case of Aspergillus sp. Respiratory activity was determined by CO₂ evolution and correlated roughly with the rate and timing of PAH removal. This indicated that the PAHs were being used as an energy source. The native microbiota responded rapidly to the addition of the PAHs and demonstrated the ability to degrade all of the PAHs added to the soil, indicating their ability to remediate PAH-contaminated soils.     

Molinate biodegradation in soils: natural attenuation versus bioaugmentation

The aims of the present study were to assess the potential of natural attenuation or bioaugmentation to reduce soil molinate contamination in paddy field soils and the impact of these bioremediation strategies on the composition of soil indigenous microbiota. A molinate mineralizing culture (mixed culture DC) was used as inoculum in the bioaugmentation assays. Significantly higher removal of molinate was observed in bioaugmentation than in natural attenuation microcosms (63 and 39 %, respectively) after 42 days of incubation at 22 °C. In the bioaugmentation assays, the impact of Gulosibacter molinativorax ON4^T on molinate depletion was observed since the gene encoding the enzyme responsible for the initial molinate breakdown (harboured by that actinobacterium) was only detected in inoculated microcosms. Nevertheless, the exogenous mixed culture DC did not overgrow as the heterotrophic counts of the bioaugmentation microcosms were not significantly different from those of natural attenuation and controls. Moreover, the actinobacterial clone libraries generated from the bioaugmentation microcosms did not include any 16S rRNA gene sequences with significant similarity to that of G. molinativorax ON4^T. The multivariate analysis of the 16S rRNA DGGE patterns of the soil microcosm suggested that the activity of mixed culture DC did not affect the soil bacterial community structure since the DGGE patterns of the bioaugmentation microcosms clustered with those of natural attenuation and controls. Although both bioremediation approaches removed molinate without indigenous microbiota perturbation, the results suggested that bioaugmentation with mixed culture DC was more effective to treat soils contaminated with molinate.     

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.     

Utilizing pyrene-degrading endophytic bacteria to reduce the risk of plant pyrene contamination

Aims

Endophytic bacteria are ubiquitous in plants, but little information is available on the influence of endophytic bacteria on the uptake and metabolism of PAH by plants. Thus, we seek to investigate whether the colonization of a target plant by a PAH-degrading endophytic bacterium would improve the PAH metabolism of the plant and reduce the risk of plant PAH contamination.

Methods

A pyrene-degrading endophyte was isolated from PAH-contaminated plants using enrichment culture. After root inoculation with the isolated bacterium, greenhouse container experiments were conducted. Pyrene residues in soil and plant samples were analyzed by HPLC.

Results

A pyrene-degrading endophytic bacterium, Staphylococcus sp. BJ06, was isolated from Alopecurus aequalis and could degrade 56.0 % of pyrene (50 mg?·?L^?1) within 15 days. BJ06 grew and degraded pyrene efficiently under environmental conditions. The bacterium significantly promoted ryegrass growth and pyrene removal from contaminated soil in container experiments. The pyrene concentrations in ryegrass roots and shoots in endophyte-inoculated planted soil were reduced by 31.01 % and 44.22 %, respectively, compared with endophyte-free planted soil.

Conclusions

We have provided new perspectives on the regulation and control of plant uptake of organic contaminants with endophytic bacteria. The results of this study will be valuable to risk assessments of plant PAH contamination.     

Enhancement of phenol degradation by soil bioaugmentation with Pseudomonas sp. JS150

Aims: To test whether bioaugmentation with genetically modified Pseudomonas sp. JS150 strain could be used to enhance phenol degradation in contaminated soils. Methods and Results: The efficiency of phenol removal, content of humic carbon, survival of inoculant, number of total culturable autochthonous bacteria and changes in fatty acid methyl esters (FAME) profiling obtained directly from soils were examined. Bioaugmentation significantly accelerated phenol biodegradation rate in tested soils. Phenol applied at the highest concentration (5·0 mg g^?1 soil) was completely degraded in clay soil (FC) within 65 days, whereas in sand soil (FS) within 72 days. In comparison, phenol biodegradation proceeded for 68 and 96 days in nonbioaugmented FC and FS soils, respectively. The content of humic carbon remained at the same level at the beginning and the end of incubation time in all soil treatments. The number of introduced bacteria (2·50 × 10⁹ g^?1 soil) markedly decreased during the first 4 or 8 days depending on contamination level and type of soil; however, inoculant survived over the experimental period of time. Analysis of FAME patterns indicated that changes in the percentages of cyclopropane fatty acids 17:0 cy and 19:0 cyω10c and branched fatty acids might be useful markers for monitoring the progress of phenol removal from soil. Conclusions: It was confirmed that soil bioaugmentation with Pseudomonas sp. JS150 significantly enhanced soil activity towards phenol degradation. Cyclopropane and branched fatty acids were sensitive probes for degree of phenol utilization. Significance and Impact of the Study: In future, genetically modified Pseudomonas sp. JS150 strain could be of use in the bioaugmentation of phenol‐contaminated areas.     

Isolation and characterization of pyrene and benzo[a]pyrene-degrading Klebsiella pneumonia PL1 and its potential use in bioremediation

Polycyclic aromatic hydrocarbons (PAHs), which are hard to degrade, are the main pollutants in the environment. Degradation of PAHs in the environment is becoming more necessary and urgent. In the current study, strain PL1 with degradation capability of pyrene (PYR) and benzo[a]pyrene (BaP) was isolated from soil and identified as Klebsiella pneumoniae by morphological and physiological characteristics as well as 16S rDNA sequence. With the presence of 20 mg L^?1 PYR and 10 mg L^?1 BaP in solution, the strain PL1 could degrade 63.4 % of PYR and 55.8 % of BaP in 10 days, respectively. The order of biodegradation of strain PL1 was pH 7.0?>?pH 8.0?>?pH 10.0?>?pH 6.0?>?pH 5.0. Strain PL1 degradation ability varied in different soil. The half-life of PYR in soil was respectively 16.9, 24.9, and 88.9 days in paddy soil, red soil, and fluvo-aquic soil by PL1 degradation; however, the half-lives of BaP were respectively 9.5, 9.5, and 34.0 days in paddy soil, red soil, and fluvo-aquic soil by PL1 degradation. The results demonstrate that the degradation capability on PYR and BaP by PL1 in paddy soil was relatively good, and K. pneumoniae PL1 was the new degradation bacterium of PYR and BaP. K. pneumoniae PL1 has potential application in PAH bioremediation.     

Efficiencies of selected biotreatments for the remediation of PAH in diluted bitumen contaminated soil microcosms

Unconventional oils such as diluted bitumen from oil sands differs from most of conventional oils in terms of physiochemical properties and PAHs composition. This raises concerns regarding the effectiveness of current remediation strategies and protocols originally developed for conventional oil. Here we evaluated the efficiency of different biotreatment approaches, such as fungi inoculation (bioaugmentation), sludge addition (bioaugmentation/biostimulation), perennial grasses plantation (phytoremediation) and their combinations as well as natural attenuation (as control condition), for the remediation of soil contaminated by synthetic crude oil (a product of diluted bitumen) in laboratory microcosms. We specifically monitored the PAHs loss percentage (alkylated PAHs and unsubstituted 16 EPA Priority PAHs), the residue of PAHs and evaluated the ecotoxicity of soil after treatment. All treatments were highly efficient with more than?~?80% of ∑PAHs loss after 60 days. Distinctive loss efficiencies between light PAHs (≤?3 rings,?~?96% average loss) and heavy PAHs (4–6 rings,?~?29% average loss) were observed. The lowest average PAHs residue (0.10?±?0.02 mg·kg^?1, for an initial concentration of 0.29?±?0.12 mg·kg^?1) was achieved with the “sludge—plants (grasses)” combination. Sludge addition was the only treatment that achieved significantly lower ecotoxicity (3%?±?4% of growth inhibition of L. sativa) than the control (natural attenuation, 13%?±?4% of inhibition). Sludge addition, grasses plantation and “sludge—fungi combination” treatments could result in lower PAH exposure (than other treatments) in post-treated soil when using the Canadian Soil Quality Guidelines for the protection of environmental and human health for potentially carcinogenic and other PAHs.

     

Enhancing of Phytoremediation Efficiency Using Indole-3-Acetic Acid (IAA)

In this study, a pot experiment using Solanum nigrum L. grown in cadmium-contaminated soil was conducted in a greenhouse. Indole-3-acetic acid (IAA) was applied at three different concentrations (1 mg L^?1, 10 mg L^?1, and 100 mg L^?1) to examine the effects on phytoremediation efficiency. According to the experimental results, IAA increased the shoot biomass of S. nigrum significantly, by 124% at the highest concentration used, and increased the Cd concentration in the shoot of S. nigrum by 16%. The Cd extraction amount from a single plant was increased by up to 158%, demonstrating potential practical application for remediation practice.     

Metabolism of Atrazine in Liquid Cultures and Soil Microcosms by Nocardioides Strains Isolated from a Contaminated Nigerian Agricultural Soil

Atrazine-degrading microorganisms designated EAA-3 and EAA-4, belonging to the genus Nocardioides, were obtained from an agricultural soil in Nigeria. The degradation kinetics of the two strains revealed total disappearance of 25 mg l^?1 of atrazine in less than 72 h of incubation at the rate of 0.42 mg l^?1 h^?1 and 0.35 mg l^?1 h^?1, respectively. Screening for atrazine catabolic genes in these organisms revealed the presence of trzN, atzB, and atzC. Other genes, specifically atzA, atzD, and trzD, were not detected. Potential intermediates of atrazine catabolic route such as hydroxyatrazine, desethylatrazine, and desisopropylatrazine were utilized as sources of carbon and energy, while desisopropyl desethyl-2-hydroxyatrazine and desisopropyl-2-hydroxyatrazine were attacked but in the presence of glucose. A soil microcosm study showed that degradation was faster in microcosms contaminated with 13 mg of atrazine per g^?1 of soil compared with 480 mg g^?1 of soil. In the former, degradation was 10% higher in the inoculated soil than the non-inoculated control (natural attenuation) over the 28-day study period. Corresponding value obtained for the latter was nearly 70% higher. This study has demonstrated that the bacterial strains isolated enhanced atrazine degradation and the catabolic activities of these strains were not affected with increasing soil atrazine concentration.     

Reaction engineering analysis of cellulase production with Trichoderma reesei RUT-C30 with intermittent substrate supply

The effects of varying initial concentrations of microcrystalline cellulose on cellulase production with Trichoderma reesei RUT-C30 as well as the effects of varying lactose and ammonium sulfate concentrations in the feed medium were studied simultaneously in parallel-operated shake flasks and, alternatively, in parallel-operated stirred-tank bioreactors on a 10-mL scale. Fifteen experiments were performed as triplicates in shake flasks as well as in stirred-tank bioreactors in parallel to identify the parameters of second-order polynomials for the estimation of the final filter paper activity of T. reesei RUT-C30 after a process time of 96 h. Even though parameter estimation was not possible based on the results of the shake flasks due to final enzyme activities at or below the detection limit (with the exception of one shake flask), the identification of the second-order polynomial was successful with the results of the parallel-operated stirred-tank bioreactors on a 10-mL scale. Reaction conditions with 53.3 g L^?1 microcrystalline cellulose in the initial medium, no lactose feeding and 3.3 g L^?1 day^?1 intermittent ammonium sulfate addition were estimated to be optimal. The final experimental validation of the optimum substrate supply on a L-scale resulted in the production of 4.88 filter paper units (FPU) mL^?1 with T. reesei RUT-C30 after 96 h. This is an improvement by a factor of 3.6 compared to the reference batch process (1.35 FPU mL^?1).     

Isolation and characterization of chromium(VI)-reducing Bacillus sp. FY1 and Arthrobacter sp. WZ2 and their bioremediation potential

Two native bacterial strains, FY1 and WZ2, that showed high chromium(VI)-reducing ability were respectively isolated from electroplating and tannery effluent–contaminated sites and identified as Bacillus and Arthrobacter. The objective of the present study was to evaluate their potential for future application in soil bioremediation. The results showed that both Bacillus sp. FY1 and Arthrobacter sp. WZ2 were tolerant to 1000 mg L^?1 Cr(VI) and capable of reducing 78–85% and 75–82% of Cr(VI) (100–200 mg L^?1) within 24 h, respectively. The Cr(VI) reduction rate decreased with increasing levels of Cr(VI) concentration (200–1000 mg L^?1). The optimum pH, temperature, and inoculum concentration for Cr(VI) reduction were found to be between pH 7.0 and 8.0; 30 and 35°C; and 1 × 10⁸ cells ml^?1, respectively. Further evidence for the bioremediation potential of Bacillus sp. FY1 and Arthrobacter sp. WZ2 was provided by the high capacity to reduce 100, 200, and 500 mg kg^?1 Cr(VI) in contaminated soil by 83–91%, 78–85%, and 71–78% within 7 days, respectively. These findings demonstrated the high potential of Bacillus sp. FY1 and Arthrobacter sp. WZ2 for application in future soil bioremediation.     

Coupling of bioaugmentation and phytoremediation to improve PCBs removal from a transformer oil-contaminated soil

This study was carried out to assess the dissipation of 17 selected polychlorinated biphenyl (PCB_i) congeners in a transformer oil-contaminated soil using bioaugmentation with 2 PCB-degrading bacterial strains, i.e., Pseudomonas spp. S5 and Alcaligenes faecalis, assisted or not by the maize (Zea mays L.) plantation. After 5 and 10 weeks of treatment, the remaining concentrations of the target PCB_i congeners in the soil were extracted and measured using GC-MS. Results showed that the bacterial augmentation treatments with Pseudomonas spp. S5 and A. faecalis led to 21.4% and 20.4% reduction in the total concentration of the target PCBs (ΣPCB_i), respectively, compared to non-bioaugmented unplanted control soil. The ΣPCB_i decreased by 35.8% in the non-bioaugmented planted soil compared with the control. The greatest degradation of the PCB congeners was observed over a 10-week period in the soil inoculated with Pseudomonas spp. S5 and cultivated with maize. Under this treatment, the ΣPCB_i decreased from 357 to 119 ng g^?1 (66.7% lower) and from 1091 to 520 ng g^?1 (52.3% lower). Overall, the results suggested that the combined application of phytoremediation and bioaugmentation was an effective technique to remove PCBs and remediate transformer oil-contaminated soils.     

PAH Degradation Capacity of Soil Microbial Communities—Does It Depend on PAH Exposure?

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants of the environment. But is their microbial degradation equally wide in distribution? We estimated the PAH degradation capacity of 13 soils ranging from pristine locations (total PAHs ≈ 0.1 mg kg^?1) to heavily polluted industrial sites (total PAHs ≈ 400 mg kg^?1). The size of the pyrene- and phenanthrene-degrading bacterial populations was determined by most probable number (MPN) enumeration. Densities of phenanthrene degraders reflected previous PAH exposure, whereas pyrene degraders were detected only in the most polluted soils. The potentials for phenanthrene and pyrene degradation were measured as the mineralization of ¹⁴C-labeled spikes. The time to 10% mineralization of added ¹⁴C phenanthrene and ¹⁴C pyrene was inversely correlated with the PAH content of the soils. Substantial ¹⁴C phenanthrene mineralization in all soils tested, including seven unpolluted soils, demonstrated that phenanthrene is not a suitable model compound for predicting PAH degradation in soils. ¹⁴C pyrene was mineralized by all Danish soil samples tested, regardless of whether they were from contaminated sites or not, suggesting that in industrialized areas the background level of pyrene is sufficient to maintain pyrene degradation traits in the gene pool of soil microorganisms. In contrast, two pristine forest soils from northern Norway and Ghana mineralized little ¹⁴C pyrene within the 140-day test period. Mineralization of phenanthrene and pyrene by all Danish soils suggests that soil microbial communities of inhabited areas possess a sufficiently high PAH degradation capacity to question the value of bioaugmentation with specific PAH degraders for bioremediation.     

Biodegradation of a PAH Mixture by Native Subsurface Microbiota

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

Isolation and characterization of heavy polycyclic aromatic hydrocarbon-degrading bacteria adapted to electrokinetic conditions

Polycyclic aromatic hydrocarbon (PAH)-degrading bacteria capable of growing under electrokinetic conditions were isolated using an adjusted acclimation and enrichment procedure based on soil contaminated with heavy PAHs in the presence of an electric field. Their ability to degrade heavy PAHs under an electric field was individually investigated in artificially contaminated soils. The results showed that strains PB4 (Pseudomonas fluorescens) and FB6 (Kocuria sp.) were the most efficient heavy PAH degraders under electrokinetic conditions. They were re-inoculated into a polluted soil from an industrial site with a PAH concentration of 184.95 mg kg^?1. Compared to the experiments without an electric field, the degradation capability of Pseudomonas fluorescens and Kocuria sp. was enhanced in the industrially polluted soil under electrokinetic conditions. The degradation extents of total PAHs were increased by 15.4 and 14.0 % in the electrokinetic PB4 and FB6 experiments (PB4 + EK and FB6 + EK) relative to the PB4 and FB6 experiments without electrokinetic conditions (PB4 and FB6), respectively. These results indicated that P. fluorescens and Kocuria sp. could efficiently degrade heavy PAHs under electrokinetic conditions and have the potential to be used for the electro-bioremediation of PAH-contaminated soil, especially if the soil is contaminated with heavy PAHs.     

Long-term assessment of natural attenuation: statistical approach on soils with aged PAH contamination

Natural attenuation processes valorization for PAH-contaminated soil remediation has gained increasing interest from site owners. A misunderstanding of this method and a small amount of data available does not encourage its development. However, monitored natural attenuation (MNA) offers a valuable, cheaper and environmentally friendly alternative to more classical options such as physico-chemical treatments (e.g., chemical oxidation, thermal desorption). The present work proposes the results obtained during a long-term natural attenuation assessment of historically contaminated industrial soils under real climatic conditions. This study was performed after a 10 year natural attenuation period on 60 off-ground lysimeters filled with contaminated soils from different former industrial sites (coking industry, manufactured gas plants) whose initial concentration of PAH varied between 380 and 2,077 mg kg^?1. The analysed parameters included leached water characterization, soil PAH concentrations, evaluation of vegetation cover quality and quantity. Results showed a good efficiency of the PAH dissipation and limited transfer of contaminants to the environment. It also highlighted the importance of the fine soil fractions in controlling PAH reactivity. PAH dissipation through water leaching was limited and did not present a significant risk for the environment. This PAH water concentration appeared however as a good indicator of overall dissipation rate, thereby illustrating the importance of pollutant availability in predicting its degradation potential.     

Petroleum Degradation,Biosurfactant and Laccase Production by Fusarium neocosmosporiellum RH-10: A Microcosm Study

The aim of this study was to evaluate the potential of crude oil removal by fungal strains isolated from Arak refinery. The results showed that the RH10 strain is a potent strain as a surfactant producer and degrader of petrochemical hydrocarbons. The strain was identified as a Fusarium neocosmosporiellum and could degrade 58% of hydrocarbons in the minimal medium and reduce the surface tension from 45 to 26.5 mN m^?1. Moreover, residual crude oil analysis with Fourier transform infrared spectrophotometry showed that this strain was able to degrade 50% of aliphatic compounds. To investigate the mechanism of degradation, oxidase enzymes were assayed and it was found that F. neocosmosporiellum can produce 1.94 U L^?1 of laccase in 10 g L^?1 crude oil. Carbon, nitrogen, phosphorus and soil pattern optimization in a microcosm study showed that this strain removed 44% and 27% of the crude oil from contaminated soil in 1% and 5% crude oil concentrations, respectively. Under optimum condition, 9.66 g kg^?1 crude oil was removed by F. neocosmosporiellum when the initial oil concentration was 50 g kg^?1, at the end of 150 days microcosm experiment. The results demonstrated the promising potential of fungi strain for cleaning of contaminated soil.     

Effects of soil amendment with different carbon sources and other factors on the bioremediation of an aged PAH-contaminated soil

Carbon supplementation, soil moisture and soil aeration are believed to enhance in situ bioremediation of PAH-contaminated soils by stimulating the growth of indigenous microorganisms. However, the effects of added carbon and nitrogen together with soil moisture and soil aeration on the dissipation of PAHs and on associated microbial counts have yet to be fully assessed. In this study the effects on bioremediation of carbon source, carbon-to-nitrogen ratio, soil moisture and aeration on an aged PAH-contaminated agricultural soil were studied in microcosms over a 90-day period. Additions of starch, glucose and sodium succinate increased soil bacterial and fungal counts and accelerated the dissipation of phenanthrene and benzo(a)pyrene in soil. Decreases in phenanthrene and benzo(a)pyrene concentrations were effective in soil supplemented with glucose and sodium succinate (both 0.2 g C kg⁻¹ dry soil) and starch (1.0 g C kg⁻¹ dry soil). The bioremediation effect at a C/N ratio of 10:1 was significantly higher (P < 0.05) than at a C/N of either 25:1 or 40:1. Soil microbial counts and PAH dissipation were lower in the submerged soil but soil aeration increased bacterial and fungal counts, enhanced indigenous microbial metabolic activities, and accelerated the natural degradation of phenanthrene and benzo(a)pyrene. The results suggest that optimizing carbon source, C/N ratio, soil moisture and aeration conditions may be a feasible remediation strategy in certain PAH contaminated soils with large active microbial populations.     

CITRIC ACID PRODUCTION BY Candida SPECIES GROWN ON A SOY-BASED CRUDE GLYCEROL

Citric acid was produced by five species of the yeast Candida after growth on a medium containing soy biodiesel-based crude glycerol. After growth on a medium containing 10 g L^?1 or 60 g L^?1 crude glycerol for 168 hr at 30°C, Candida parapsilosis ATCC 7330 and C. guilliermondii ATCC 9058 produced the highest citric acid levels. On 10 g L^?1 or 60 g L^?1 crude glycerol for 168 hr at 30°C, the citric acid level produced by C. parapsilosis ATCC 7330 was 1.8 g L^?1 or 11.3 g L^?1, respectively, while C. guilliermondii ATCC 9058 produced citric acid concentrations of 3.0 g L^?1 or 10.4 g L^?1, respectively. Biomass production by C. guilliermondii ATCC 9058 on 10 g L^?1 or 60 g L^?1 crude glycerol for 168 hr at 30°C was highest at 1.2 g L^?1 or 6.9 g L^?1, respectively. The citric acid yields observed for C. guilliermondii ATCC 9058 after growth on 10 g L^?1 or 60 g L^?1 crude glycerol (0.35 g g^?1 or 0.21 g g^?1, respectively) were generally higher than for the other Candida species tested. When similar crude glycerol concentrations were present in the culture medium, citric acid yields observed for some of the Candida species utilized in this study were about the same or higher compared to citric acid yields by Yarrowia lipolytica strains. Based on the findings, it appeared that C. guilliermondii ATCC 9058 was the most effective species utilized, with its citric acid production being similar to what has been observed when citric acid-producing strains of Y. lipolytica were grown on crude glycerol under batch conditions that could be of significance to biobased citric acid production.