Restoration of Petroleum-Contaminated Soil Using Phased Bioremediation

A conceptual approach is presented for the restoration of petroleum-contaminated sites by combining bioremediation with revegetation using native plants. Phased bioremediation includes active and passive treatment options for soil containing greater than 1% total petroleum hydrocarbons (TPHs). Phase I is used when initial soil TPH exceeds 1%. Phase I utilizes either active land treatment, with regular soil tillage, or passive bioremediation to attain a treatment endpoint of 1% soil TPH. Passive treatment utilizes static soil and TPH-tolerant plants. Phase II is utilized when soil contains 1% TPH or less. It combines passive bioremediation with revegetation using native plants to complete the site restoration process. The phased approach to bioremediation was developed from results of full-scale field bioremediation and laboratory treatability studies. This approach assumes that the kinetics of TPH biodegradation are initially rapid, followed by a much slower second stage. It provides active initial treatment, followed by lower-cost passive treatment. The selection of either active or passive treatment in Phase I depends on whether total cost or time of treatment is more important. Passive treatment, although less costly than active treatment, generally requires more time. Phased bioremediation may provide a flexible, cost-effective, and technically sound approach for restoration of petroleum-contaminated sites.

Vegetation used with passive bioremediation has several benefits. Plants stabilize soil, preventing erosion and thereby minimizing exposure to soil contaminants. Phytoremediation may also occur within the rhizosphere. The use of native plants has a strong ecological basis. They provide ecological diversity, are aesthetically pleasing and beneficial to wildlife, while requiring little maintenance. Phased bioremediation can provide a flexible, cost-effective, and technically sound approach for the restoration of petroleum-contaminated sites.     

Animal Waste-Enhanced Degradation of Hydrocarbon-Contaminated Soil

Land previously used for petroleum production is being converted to commercial, industrial, or residential uses and requires remediation to remove petroleum hydrocarbons (HC). Concurrently, large quantities of animal wastes are produced annually, creating waste-handling and disposal problems. A laboratory study was conducted to determine whether amending contaminated soil with animal manure and inorganic fertilizer affected the degradation rate and amount of HC in the soil. HC-impacted soil was prepared by mixing a diesel fuel: motor oil solution (1:1?v/v) with a noncontaminated loamy sand, with a target concentration of 5000?mg HC per kg soil. Treatments included ammonium sul-fate fertilizer, composted steer manure at 5, 10, and 20% on a dry weight basis, and a mixture of fertilizer and steer manure, with three replications for each treatment. Five-gram aliquot soil samples were extracted with hexane. The extracts were analyzed using a gas chromatograph with a flame ionization detector. Over a period of 41 days, control treatment HC concentrations were reduced by 32%. Hydrocarbon concentrations in (NH₄)₂SO₄ fertilizer-amended soil were decreased by nearly 54% during the same time. In contrast, HC degradation was much faster and more complete in manure-amended soils. Up to 81% of the HC in the 20% manure-treated soil were removed by day 41.     

Toxicity to Earthworms and Chemical Composition of Oil Refinery Sludge Destined for Landfarming

For landfarming of oil refinery waste to be sustainable, it is crucial to know the toxicity and chemical composition of the waste. We determined the chemical composition of waste known as American Petroleum Institute sludge and tested its toxicity in two different soils using the earthworm Eisenia andrei (Oligochaeta) in standardized bioassays. We compared the chemical composition of the sludge with available chemical profiles of the landfarming site where sludge of the same origin had been landfarmed for several years. The concentrations of Al, Mn, Pb, S and Zn and diesel range organics were higher in the landfarmed soil than in the sludge, indicating possible accumulation in the soil over time. Although no traces of volatile organic compounds were previously found in the landfarming site soil, we found high levels in the American Petroleum Institute sludge, indicating that these volatiles may not pose a long-term problem in the soil. Polycyclic aromatic hydrocarbons and gasoline range organics were present in the sludge at higher concentrations than in the site soil, indicating that their remediation through landfarming may have had some degree of success.

Earthworm biomass and reproduction were detrimentally affected at low sludge concentrations, which could have population level effects over time in a landfarming site.     

电场刺激技术在微生物工程中的应用

电场刺激是利用电解法将惰性电极插入电解液中,形成一种电解池系统,电解液对细胞培养物会产生不同影响,并导致生物过程发生改变。电场刺激对微生物会产生促进和杀灭两种不同作用,并在微生物工程中已有所应用,如促进微生物生长和代谢、强化废水处理、进行生物修复和用于杀菌消毒等。研究表明电场刺激在微生物工程和环境生物工程等方面有广泛的应用前景。     

Influence of biochar and compost on phytoremediation of oil-contaminated soil

The use of pyrolyzed carbon, biochar, as a soil amendment is of potential interest for improving phytoremediation of soil that has been contaminated by petroleum hydrocarbons. To examine this question, the research reported here compared the effects of biochar, plants (mesquite tree seedlings), compost and combinations of these treatments on the rate of biodegradation of oil in a contaminated soil and the population size of oil-degrading bacteria. The presence of mesquite plants significantly enhanced oil degradation in all treatments except when biochar was used as the sole amendment without compost. The greatest extent of oil degradation was achieved in soil planted with mesquite and amended with compost (44% of the light hydrocarbon fraction). Most probable number assays showed that biochar generally reduced the population size of the oil-degrading community. The results of this study suggest that biochar addition to petroleum-contaminated soils does not improve the rate of bioremediation. In contrast, the use of plants and compost additions to soil are confirmed as important bioremediation technologies.     

Bioremediation of creosote-contaminated soil: a pilot-scale landfarming evaluation

A pilot-scale landfarming investigation of the effects of biostimulation and bioaugmentation on a creosote-contaminated (258.3 g kg^–1) mispah form (FAO: lithosol) soil, with a view to developing a cost-effective bioremediation methodology for creosote-contaminated soils was conducted in nine duplicate reactors, including two controls (Treatments 1 and 2). Treatments 3–9 were watered and aerated daily and Treatment 4–9 were monthly amended with mono-ammonium phosphate. Treatment 5–9 received further amendments as follows: Treatment 5, hydrogen peroxide; Treatment 6, indigenous microbial biosupplement; Treatment 7, sewage sludge; Treatment 8, cow manure; Treatment 9, poultry manure. Residual concentrations of creosote ranged between 29 and 215 g kg^–1 after sixteen weeks. The phenolics and the 2- and 3-ringed polyaromatic hydrocarbons (PAHs) were removed below detectable levels or to very low levels. The 4- and 5-ringed PAHs were removed by between 68 and 83%. Indigenous microbial biosupplement and sewage sludge were the most effective in creosote removal. Hydrogen peroxide did not significantly enhance microbial population and creosote removal. There was no significant difference between the results obtained from the treatments amended with organic manures. However, there was a significant difference between the effects of the organic manures and the indigenous microbial biosupplement. Results from this study suggests that a combination of the two treatment techniques (biostimulation and bioaugmentation) would be a better approach to treating soil contaminated with very high concentrations of creosote.     

Biodegradation of Hopane Prevents Use as Conservative Biomarker During Bioremediation of PAHs in Petroleum Contaminated Soils

The pentacyclic triterpane C₃₀ 17α (H), 21β (H)-hopane, a biomarker commonly used in hydrocarbon bioremediation laboratory experiments and field studies, was found to be completely removed without the formation of the demethylated intermediate nor-hopane in a crude oil-contaminated soil undergoing slurry biotreatment, while PAHs such as benzo(e)pyrene were recalcitrant. The partial or complete biodegradation of hopane has also been previously reported in a few bioremediation studies and has been explored by petroleum geochemists in an effort to characterize crude oil deposits. It is currently not clear what conditions induce hopane biodegradation or biotransformation, although the use of microbial enrichment cultures appears to speed up the process. Considering that hopane is not necessarily conserved during a bioremediation study, the uncritical normalization of hydrocarbon concentrations using this biomarker can lead to incorrect estimates of biodegradation rates and extents. If hopane is found to be unstable in a particular case, other potential biomarkers such as pentahopane, oleanane, or vanadium may be used instead.     

Bioremediation of creosote-contaminated soil in South Africa by landfarming

AIMS: To determine the combined effects of biostimulation and bioaugmentation in the landfarming of a mispah form (lithosol; food and Agriculture Organisation (FAO)) soil contaminated with >310000 mg kg-1 creosote with a view to developing a bioremediation technology for soils heavily contaminated with creosote. METHODS AND RESULTS: The excavated soil was mixed with 2500 kg ha-1 dolomitic lime and 2000 kg ha-1 mono-ammonium phosphate (MAP) before spreading over a treatment bed of shale reinforced with clay. Sewage sludge (500 kg) was ploughed into 450 m3 of contaminated soil in the second and sixth months of treatment. A further 1000 kg ha-1 MAP was added to the soil at the end of the fifth month. Moisture was maintained at 70% field capacity. Total creosote was determined by the US Environmental Protection Agency (EPA) method 418.1 and concentrations of selected creosote components were determined by gas chromatography/flame ionisation detection (GC/FID). Total creosote was reduced by more than 90% by the 10th month of landfarming. The rate of reduction in creosote concentration was highest after the addition of sewage sludge. The three-ring PAHs were more slowly removed than naphthalene and the phenolic compounds. The four- and five-ring PAHs, although persist until the end of treatment, were reduced by 76-87% at the end of the experiment. CONCLUSIONS: A combination of biostimulation and bioaugmentation during landfarming could enhance the bioremediation of soils heavily contaminated with creosote. SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides information on the management of a combination of biostimulation and bioaugmentation during landfarming, and contributes to the knowledge and database necessary for the development of a technology for bioremediating creosote-contaminated land.     

Degradation of Polycyclic Aromatic Hydrocarbons in the Rhizosphere of Festuca arundinacea and Associated Microbial Community Changes

A phytoremediation growth chamber study was conducted to evaluate the contribution of soil microbial diversity to the contaminant degradation. Target contaminant removal from soil was assessed by monitoring concentrations of polycyclic aromatic hydrocarbons (PAHs), along with changes in the bacterial community structure over a time period of 10 months in the presence of tall fescue (Festuca arundinacea). Enhanced degradation of PAHs was observed in rhizosphere soil, with a maximum reduction in pyrene at a rate 36% higher than that noted for the unvegetated control. The dissipation of < 4-ring PAHs, 4-ring PAHs, and > 4-ring PAHs in unvegetated soil was 70%, 54%, and 49% respectively, whereas a higher dissipation rate was observed in tall fescue treated soil of 78%, 68%, and 61% at the end of the study. Microbial enumeration results showed greater total bacterial numbers and PAH-degrading bacteria in rhizosphere soil when compared to unvegetated soil. The results from the terminal restriction fragment length polymorphism (T-RFLP) analysis indicated that there was a shift in the rhizosphere bacterial community during the phytoremediation process.     

Multi-Species Ecotoxicity Assessment of Petroleum-Contaminated Soil

In 1992, a study was begun to compare the effect of landfarming vs. natural attenuation on the restoration of soil that had been contaminated with crude oil. Each of three lysimeters was filled with a sandy loam topsoil, and crude oil was applied to two of the lysimeters. One of the contaminated lysimeters was tilled, watered, and received a one-time application of fertilizer (N, P, K). No amendments were added to the second contaminated lysimeter, and the third was left uncontaminated. The lysimeters were monitored for 6 months and then left unattended. In 1995 and again in 1997 we sampled these lysimeters to evaluate the long-term effects of contamination and bioremediation. In 1995 we found marked effects on soil chemistry, bacterial, fungal, nematode, and plant populations and a higher rate of bioremediation in the fertilized-contaminated lysimeter (Lawlor et al., 1997). Data from 1997 and previously unreported data from 1995 are the subject of the current report. In 1997, low densities of hydrocarbon-degrading bacteria were found in all the lysimeters and little loss of TPH from the two contaminated lysimeters, suggesting a decreased rate of bioremediation. Nevertheless, there were increases in diversity and number of functional groups of bacteria, nematodes, and native plant species. However, molecular analyses revealed marked differences remained in the composition of dominant eubacterial species, and tests of soybeans indicated field conditions remained unsuitable for these plants.     

Use of Genetically Engineered Microorganisms (GEMs) for the Bioremediation of Contaminants

ABSTRACT

This paper presents a critical review of the literature on the application of genetically engineered microorganisms (GEMs) in bioremediation. The important aspects of using GEMs in bioremediation, such as development of novel strains with desirable properties through pathway construction and the modification of enzyme specificity and affinity, are discussed in detail. Particular attention is given to the genetic engineering of bacteria using bacterial hemoglobin (VHb) for the treatment of aromatic organic compounds under hypoxic conditions. The application of VHb technology may advance treatment of contaminated sites, where oxygen availability limits the growth of aerobic bioremediating bacteria, as well as the functioning of oxygenases required for mineralization of many organic pollutants. Despite the many advantages of GEMs, there are still concerns that their introduction into polluted sites to enhance bioremediation may have adverse environmental effects, such as gene transfer. The extent of horizontal gene transfer from GEMs in the environment, compared to that of native organisms including benefits regarding bacterial bioremediation that may occur as a result of such transfer, is discussed. Recent advances in tracking methods and containment strategies for GEMs, including several biological systems that have been developed to detect the fate of GEMs in the environment, are also summarized in this review. Critical research questions pertaining to the development and implementation of GEMs for enhanced bioremediation have been identified and posed for possible future research.     

Submersible microbial fuel cell sensor for monitoring microbial activity and BOD in groundwater: Focusing on impact of anodic biofilm on sensor applicability

A sensor, based on a submersible microbial fuel cell (SUMFC), was developed for in situ monitoring of microbial activity and biochemical oxygen demand (BOD) in groundwater. Presence or absence of a biofilm on the anode was a decisive factor for the applicability of the sensor. Fresh anode was required for application of the sensor for microbial activity measurement, while biofilm‐colonized anode was needed for utilizing the sensor for BOD content measurement. The current density of SUMFC sensor equipped with a biofilm‐colonized anode showed linear relationship with BOD content, to up to 250 mg/L (～233 ± 1 mA/m²), with a response time of <0.67 h. This sensor could, however, not measure microbial activity, as indicated by the indifferent current produced at varying active microorganisms concentration, which was expressed as microbial adenosine‐triphosphate (ATP) concentration. On the contrary, the current density (0.6 ± 0.1 to 12.4 ± 0.1 mA/m²) of the SUMFC sensor equipped with a fresh anode showed linear relationship, with active microorganism concentrations from 0 to 6.52 nmol‐ATP/L, while no correlation between the current and BOD was observed. It was found that temperature, pH, conductivity, and inorganic solid content were significantly affecting the sensitivity of the sensor. Lastly, the sensor was tested with real contaminated groundwater, where the microbial activity and BOD content could be detected in <3.1 h. The microbial activity and BOD concentration measured by SUMFC sensor fitted well with the one measured by the standard methods, with deviations ranging from 15% to 22% and 6% to 16%, respectively. The SUMFC sensor provides a new way for in situ and quantitative monitoring contaminants content and biological activity during bioremediation process in variety of anoxic aquifers. Biotechnol. Bioeng. 2011;108: 2339–2347. © 2011 Wiley Periodicals, Inc.     

Enzymatic Evaluation During Biodegradation of Kerosene and Diesel by Locally Isolated Fungi from Petroleum-Contaminated Soils of Western India

The current study suggests that the fungal isolates P. decumbens PDX7, P. janthinellum SDX7, and A. terreus PKX4 degraded kerosene by 95%, 96%, and 75% and diesel by 79%, 75%, and 70% after 16 days based on the ability of utilizing these compounds as sole carbon sources. GC-MS chromatograms revealed that n-alkane fractions are easily degraded; however, the rate is lower for branched alkanes, n-alkyl aromatics, cyclic alkanes, and polynuclear aromatics displaying delayed and lower degradation. The ratio of aromatic/aliphatic hydrocarbons >0.8 indicates the efficiency of these fungi in removing the aromatic hydrocarbons of the petroleum products. All of the treated fungal strains exhibited higher MnP, laccase, and dehydrogenase activities on the twelfth and sixteenth days as compared to the initial fourth and eighth days. In addition, P. decumbens PDX7 and P. janthinellum SDX7 displayed higher enzymatic activities as compared to A. terreus PKX4. Fungal isolates were also tested for their growth on various xenobiotic compounds as sole carbon sources.     

生物炭对农田土壤微生物生态的影响研究进展

生物炭作为新型土壤改良剂在国内外环境科学等领域受到广泛的关注.关于生物炭对土壤理化性质的改良研究较早,目前虽然已深入到土壤微生物生态的领域,但是大多数将土壤理化性质与土壤微生物生态分开考虑,缺乏对二者相互作用的系统评述.本文总结了施用生物炭后土壤理化性质的改变与土壤微生物群落变化之间的相互关系:生物炭不仅能够提高土壤pH值、增强土壤的持水能力、增加土壤有机质等,而且会影响土壤微生物的群落结构、改变细菌和真菌的丰度;施用生物炭后,土壤环境和土壤微生物之间互相影响互相制约,共同促进了土壤微生物生态系统的改良.本文旨在为生物炭改良农田土壤微生态的深入研究提供新的思路,从生态系统的角度促进生物炭环境效应影响的研究,使生物炭的应用更具有科学性和有效性,并对生物炭在相关领域的应用进行了展望.     

Characterization of Petroleum-Contaminated Soils by Thin-Layer Chromatography with Flame Ionization Detection

Petroleum hydrocarbons from 20 soils from refineries or other industrial sites were extracted with a mixture of chloroform and methanol (1:1, v/v), and the extracts were analyzed by thin layer chromatography with flame ionization detection (TLC/FID). The TLC/FID procedure has been used widely in biological and medical research but generally has been underutilized in environmental chemistry. The analysis method involved spotting a small volume of sample extract (typically 1 to 3?µl) on ten silica-coated quartz rods, and chromatographically separating constituents in the spots using solvent systems of increasing polarities (hexane, toluene, and dichloromethane + methanol). We achieved complete separation of saturated hydrocarbons, aromatic hydrocarbons, resins, and asphaltenes from the hydrocarbon-contaminated soils with this method. Analysis of the separated constituents by TLC/FID also allowed quantification of aromatic and aliphatic hydrocarbons without interference from soil biogenic lipids. A simplified version of the method permitted excellent separation of aliphatics +aromatics (forming a single peak) from resins and asphaltenes. The procedure is rapid (complete analysis of ten samples in about 1?h after extraction). Thus, the method seems well suited for synoptic surveys or screening and characterizing numerous samples prior to using more detailed and costly analyses.     

Degradation and Detoxification of Nicosulfuron by a Pseudomonas Strain Isolated from a Contaminated Cornfield Soil

ABSTRACT

The dissipation and detoxification of nicosulfuron (NS) by Pseudomonas aeruginosa B9 isolated from a cornfield soil was investigated. The fastest decline of NS occurred at 40 µg ml^?1 in liquid media with 0.25% glucose plus 0.05% yeast extract (DT₅₀ = 4 days) with a notable pH reduction (pH ? 5). Bioassay tests showed considerable phytotoxicity of NS for Cress (Lepidium sativum L.) with 50% shoot growth inhibition (SGI) at 40 µg ml^?1. The dissipation of NS (40 µg ml^?1) by the B9 isolate reduced the SGI significantly (SGI: up to 45 ± 3%) compared to the non-inoculated media (SGI: up to 58 ± 4%). In soils with the B9 isolate, NS dissipation, especially at 0.3 µg g^?1, was faster with a more significant SGI reduction (k = 0.08 ± 0.00 day^?1; SGI = 2 ± 1%) compared to non-inoculated samples (k = 0.03 ± 0.00 day^?1; SGI = 8 ± 1%). NS initially inhibited soil respiration, microbial biomass carbon, and dehydrogenase activity. The effect was however transient, and these parameters recovered within 10 days, especially in the presence of the isolate. Overall, this study proves Pseudomonas aeruginosa B9 as a suitable candidate for bioremediation of NS in contaminated sites.     

有机污染土壤和地下水生物修复研究热点和趋势——基于Web of Science数据库的文献计量学分析

生物修复作为经济有效、绿色可持续的修复技术,在有机污染土壤和地下水修复上具有广阔的应用前景。基于WebofScience核心数据库,通过文献计量可视化应用软件VOSviewer和CiteSpace,分析了1990–2020年有机污染土壤和地下水生物修复领域的研究热点及趋势。结果表明,有机污染土壤和地下水生物修复领域的论文发表数量呈增长趋势,发文总量最多的国家是美国和中国,但是2012年后中国年发文量快速增加,并位居第一。该领域的相关研究主要发表在Chemosphere、Environmental ScienceTechnology、Science of the Total Environment等top期刊上。全球研究机构中中国科学院发文量最多,但是来自美国加州大学的总被引频次和h-index最高。发文量最多的是来自英国兰卡斯特大学的学者Semple教授,我国发文量最多的是来自中国科学院南京土壤研究所的骆永明研究员。下一步研究重点和热点:针对复合污染土壤和地下水,研发新型耦合强化生物修复技术,采用先进的分子生物学方法探索功能微生物及其功能基因,阐明生物降解机理,明确原位污染土壤和地下水的靶向性调控机制。     

Some microbial contaminants and control agents in a diet and larvae of Heliothis spp

Sixteen tests were conducted with four antimicrobial agents that are routinely incorporated into lepidopterous larval diets for bacterial and fungal inhibition. The agents tested in all possible combinations were benomyl, chlortetracycline, sorbic acid, and methyl P-hydroxybenzoate. Those combinations that contained benomyl and sorbic acid in the formula were effective in complete microbial suppression during the larval portion of the 35-day testing period. Insect development in plastic cups of test diets with benomyl and sorbic acid averaged 97 and 93% pupation and adult emergence, respectively. The 12 tests that did not contain benomyl and sorbic acid were contaminated with some of the following fungi: Aspergillus niger, A. flavus, Rhizopus nigricans, Cladosporium, Fusarium, yeasts, or the bacterium, Bacillus subtilis. Microorganisms in Heliothis spp. larvae from mass-rearing diets were tested, examined, and identified. Bacteria recovered from laboratory-reared Heliothis larvae included Pseudomonas aeruginosa, P. maltophilia, Micrococcus luteus, α-hemolytic Streptococcus, Serratia marcescens, and S. rubidaea. Tests were conducted with sensi-discs for the selection of antimicrobials to arrest contamination in the larval digestive systems.     

Characterization of a Polycyclic Aromatic Hydrocarbon-Degrading Microbial Consortium from a Petrochemical Sludge Landfarming Site

Anthracene, phenanthrene, and pyrene are polycyclic aromatic hydrocarbon (PAHs) that display both mutagenic and carcinogenic properties. They are recalcitrant to microbial degradation in soil and water due to their complex molecular structure and low solubility in water. This study presents the characterization of an efficient PAH (anthracene, phenanthrene, and pyrene)-degrading microbial consortium, isolated from a petrochemical sludge landfarming site. Soil samples collected at the landfarming area were used as inoculum in Warburg flasks containing soil spiked with 250 mg kg^-1 of anthracene. The soil sample with the highest production of CO₂-C in 176 days was used in liquid mineral medium for further enrichment of anthracene degraders. The microbial consortium degraded 48%, 67%, and 22% of the anthracene, phenanthrene, and pyrene in the mineral medium, respectively, after 30 days of incubation. Six bacteria, identified by 16S rRNA sequencing as Mycobacterium fortuitum, Bacillus cereus, Microbacterium sp., Gordonia polyisoprenivorans, two Microbacteriaceae bacteria, and a fungus identified as Fusarium oxysporum were isolated from the enrichment culture. The consortium and its monoculture isolates utilized a variety of hydrocarbons including PAHs (pyrene, anthracene, phenanthrene, and naftalene), monoaromatics hydrocarbons (benzene, ethylbenzene, toluene, and xylene), aliphatic hydrocarbons (1-decene, 1-octene, and hexane), hydrocarbon mixtures (gasoline and diesel oil), intermediary metabolites of PAHs degradation (catechol, gentisic acid, salicylic acid, and dihydroxybenzoic acid) and ethanol for growth. Biosurfactant production by the isolates was assessed by an emulsification index and reduction of the surface tension in the mineral medium. Significant emulsification was observed with the isolates, indicating production of high-molecular-weigh surfactants. The high PAH degradation rates, the wide spectrum of hydrocarbons utilization, and emulsification capacities of the microbial consortium and its member microbes indicate that they can be used for biotreatment and bioaugumentation of soils contaminated with PAHs.