Effect of salinity on the bioremediation of petroleum hydrocarbons in a saline-alkaline soil

Aims: The aim of this paper is to check the effect of salinity on the bioremediation process of petroleum hydrocarbons in the saline‐alkaline soil. Methods and Results: In this study, soil salinity was adjusted to different levels by water leaching method and the bioremediation process was conducted for 28 days. Soil pH increased after leaching and decreased during bioremediation process. At initial time, moderate salinity enhanced the biodegradation and addition of microbial consortium was not effective in enhancing degradation rate of petroleum hydrocarbons. At day of 28 days, higher degradation rate was found in treatments with more leaching times with a maximum value of 42·36%. Dehydrogenase activity increased with the progress of bioremediation and positive correlation was found between dehydrogenase activity and degradation rate of petroleum hydrocarbons. Denaturing gradient gel electrophoresis analysis result showed decreased microbial community diversity with increased salt content. Conclusions: The result suggested that salinity had great impact on bioremediation, and leaching and addition of inoculated consortium were effective in enhancing biodegradation of petroleum hydrocarbons in the saline‐alkaline soil. Significance and Impact of the Study: The result of this study is important for understanding the bioremediation process of petroleum in contaminated soil. New remediation method of petroleum contaminated soil can be developed based on this study.     

Self-immobilized bacterial cultures with potential for application as ready-to-use seeds for petroleum bioremediation

Two hydrocarbon-degrading bacterial isolates, an Arthrobacter sp. and a Gram-negative bacillus isolated from Kuwait oil lakes, exhibited considerable cell-surface hydrophobicity without production of exopolysaccharides in complex media. However, the bacteria produced copious amounts of exopolysaccharides in a low nutrient medium. When incubated with sawdust, Styrofoam or wheat bran, as carriers, under low nutrient conditions, stable exopolysaccharide-mediated immobilized cultures were formed. Such immobilized cultures when air-dried at room temperature survived storage for 6 weeks at 45 °C and still retained the ability to degrade hydrocarbons. Viability was retained by the immobilized Arthrobacter sp. and the Gram-negative bacterium at 45 °C storage for up to 6 and 12 months, respectively.     

Impact of surrogate selection on risk assessment for total petroleum hydrocarbons

Environmental contamination involving total petroleum hydrocarbons (TPH) is being investigated and remediated at underground storage tanks, tank farms, pipelines, and refineries across the country. Human health and environmental risk play a significant role in decision making at these sites. However, risk assessment for sites contaminated with petroleum products typically is complicated by inadequate information about the composition of TPH present at the site and the physical and chemical properties and toxicity of the components. To address these data gaps, risk assessors can select surrogate compounds to represent the movement of TPH in the environment at the site and toxicity of TPH present at the site. This article illustrates the potential impact of choice of surrogates on risk estimates, which in turn affect remediation costs.     

Assessment of in-situ bioremediation at a refinery waste-contaminated site and an aviation gasoline contaminated site

A combination of geochemical, microbiological and isotopic methods were used to evaluate in-situ bioremediation of petroleum hydrocarbons at one site contaminated with refinery waste and a second site contaminated with aviation gasoline at Alameda Point, California. At each site, geochemical and microbiological characteristics from four locations in the contaminated zone were compared to those from two uncontaminated background locations. At both sites, the geochemical indicators of in-situbiodegradation includeddepleted soil gas and groundwater oxygen, elevated groundwater alkalinity, and elevated soil gas carbon dioxide and methane in the contaminated zone relative to the background. Radiocarbon content of methane and carbon dioxide measured in soil gas at both sites indicated that they were derived from hydrocarbon contaminant degradation. Direct microscopy of soil core samples using cell wall stains and activity stains, revealed elevated microbial numbers and enhanced microbial activities in contaminated areas relative to background areas, corroborating geochemical findings. While microbial plate counts and microcosm studies using soil core samples provided laboratory evidence for the presence of some microbial activity and contaminant degradation abilities, they did not correlate well with either contaminant location, geochemical, isotopic, or direct microscopy data.     

海洋石油污染物的微生物降解与生物修复

石油是海洋环境的主要污染物 ,已经对海洋及近岸环境造成了严重的危害。微生物降解是海洋石油污染去除的主要途径。海洋石油污染物的微生物降解受石油组分与理化性质、环境条件以及微生物群落组成等多方面因素的制约 ,N和P营养的缺乏是海洋石油污染物生物降解的主要限制因子。在生物降解研究基础上发展起来的生物修复技术在海洋石油污染治理中发展潜力巨大 ,并且取得了一系列成果。介绍了海洋中石油污染物的来源、转化过程、降解机理、影响生物降解因素及生物修复技术等方面内容 ,强调了生物修复技术在治理海洋石油污染环境中的优势和重要性 ,指出目前生物修复技术存在的问题。     

American petroleum institute in situ air sparging database

An evaluation of data detailing in situ air sparging (IAS) systems at 59 sites has been assembled into an American Petroleum Institute in situ Air Sparging Database (API‐IAS Database). The database was developed to provide site managers insights concerning the state‐of‐the‐art of IAS system design, operation, and evaluation. The IAS radius of influence (ROI) is often evaluated based on changes in a number of physical, chemical, or biological monitoring parameters. Measurements of groundwater dissolved oxygen levels was the technique used most often to determine the ROI. Other parameters such as pressure changes in the vadose and saturated zones, groundwater mounding, air bubbling in wells and tracer gases were also used to aid in the determination of the IAS ROI. A review of 37 pilot studies revealed that the IAS ROI is generally between 10 to 26 ft. IAS technology is generally being applied in sandy soils. The application of IAS technology was deemed infeasible at seven sites where soils contained high levels of silts or clay. Analysis of design and operation data at 40 IAS sites revealed that a typical IAS well is 2 in. in diameter, with a 2‐ft screen, positioned 5 to 10 ft beneath the water table. The wells typically were operated at an overpressure (i.e., pressure in excess of that required to overcome the hydrostatic head) of less than 5 psi with a flow rate of less than 5 cfm. At several sites when IAS system pressures and flows were doubled, only slight increases in ROI resulted. Significant reductions of dissolved volatile organic hydrocarbon (VOCs) were observed at 12 sites as a result of IAS. However, long‐term water quality data following an IAS system shutdown was very limited.     

多环芳烃污染土壤微生物修复研究进展

多环芳烃是我国土壤环境质量标准中要求严格管控的一类持久性有机污染物,利用微生物技术修复有机污染土壤具有绿色、经济等突出特点,应用前景广泛.目前多学科的协同发展和新技术的研究应用,为多环芳烃土壤微生物转化机制与污染生态过程等方面带来了新的认识,同时对修复技术的实际应用与调控提供了新的思考方向.本文以多环芳烃污染土壤微生物...     

Optimization of field scale biopiles for bioremediation of petroleum hydrocarbon contaminated soil at low temperature conditions by response surface methodology (RSM)

Ex-Situ Bioremediation has been increasingly viewed as an appropriate remediation technology for hydrocarbon contaminated soils under cold climates conditions in countries like Canada. A response surface methodology (RSM) based on a factorial design was performed to investigate and optimize the effects of the microbial consortia application rate and amount of mature compost amendment on the TPH removal (964 μg g⁻¹ initial concentration). 18 field-scale biopiles (16 m³ each) were constructed, maintained and subjected to different microbial consortium and mature compost application rates under cold climate conditions over a period of 94 days. TPHs removal rates in the range of 74–82% was observed in the treatments setups where mature compost and microbial consortia were used simultaneously, compared to an average 48% of TPHs removal in control setup.The interaction between these two factors were studied and modelled using a statistical regression model, which showed that the microbial consortia application rate, the mature compost amendment and their interactions had a significant effect on TPHs degradation with a coefficient of determination (R²) of 0.88. Furthermore, using a numerical optimization approach, the optimum rates predicted via RSM were estimated at 4.1 ml m⁻³ and 7% for microbial consortia and compost application rates to obtain a maximum TPH removal of 90.7%.     

Assessing the impact of transgenic plant products on soil organisms

Little is known about the impact of transgenic plant products on soil organisms. However, previous research with synthetic organics, allelochemicals, and extracellular enzymes can be used to guide future research in this area. Projects designed to quantify the impact of transgenic plants on soil organisms must clearly establish that the gene products are responsible for any observed changes. This can only be achieved by determining the fate of transgenic plant gene products during the period of the soil bioassay. The overall impact of transgenic plants will be dictated by not only the primary gene product, but secondary products resulting from abiotic and biotic soil reactions. Primary and secondary products may exhibit both acute and chronic impacts. Such impacts are best quantified using a soil microcosm in which fungal populations and micro- and mesofauna are monitored.     

Accelerated biodegradation of petroleum hydrocarbon waste

Conventional landfarming approaches to bioremediation of refinery and other petroleum sludges are not acceptable environmentally and are banned in most North American jurisdictions. While initial bioreactor-based systems for treatment of these sludges required batch-cycle process-times of 1–3 months, an accelerated process has now been developed which can be completed in 10–12 days. In this process, up to 99% of total petroleum hydrocarbons are degraded and the sludges are converted from hazardous to non-hazardous according to the United States EPA's toxicity characteristic leachate procedure criteria. Understanding and exploiting mechanisms to improve hydrocarbon accession to the degrading microorganisms was a key development component of the process. Contrasting physiological mechanisms were observed for different component organisms of the mixed culture with respect to their associations with the hydrocarbon substrate; and the beneficial effects of using surfactants were demonstrated. The mixed culture used in the process exhibited a capacity for high-rate degradation of volatile organic carbons and the potential use of the culture as a liquid biofilter was demonstrated. The culture was also effective as an inoculant for the bioaugmentation of total petroleum hydrocarbon-contaminated soil and as a de-emulsifier of oilfield emulsions and could transform some other environmental contaminants which are not predominant components of crude oil.     

Microbial community changes during the bioremediation of creosote-contaminated soil

AIMS: To investigate the effects of aeration on the ex situ biodegradation of polycyclic aromatic hydrocarbons (PAHs) in creosote-contaminated soil and its effect on the microbial community present. METHODS AND RESULTS: Aerated and nonaerated microcosms of soil excavated from a former timber treatment yard were maintained and sampled for PAH concentration and microbial community changes by terminal restriction fragment length polymorphism (TRFLP) analysis. After an experimental period of just 13 days, degradation was observed with all the PAHs monitored. Abiotic controls showed no loss of PAH. Results unexpectedly showed greater loss of the higher molecular weight PAHs in the nonaerated control. This may have been due to the soil excavation causing initial decompaction and aeration and the resulting changes caused in the microbial community composition, indicated by TRFLP analysis showing several ribotypes greatly increasing in relative abundance. Similar changes in both microcosms were observed but with several possible key differences. The species of micro-organisms putatively identified included Bacilli, pseudomonad, aeromonad, Vibrio and Clostridia species. CONCLUSIONS: Excavation of the contaminated soil leads to decompaction, aeration and increased nutrient availability, which in turn allow microbial biodegradation of the PAHs and a change in the microbial community structure. SIGNIFICANCE AND IMPACT OF THE STUDY: Understanding the changes occurring in the microbial community during biodegradation of all PAHs is essential for the development of improved site remediation protocols. TRFLP allows useful monitoring of the total microbial community.     

In situ bioremediation of hydrocarbon in soil

Laboratory and field experiments were carried out for bioremediation of soils contaminated by fuel oil and motor oil. Bioventing was combined with the application of selected bacteria and dissolved nutrients. In the field experiments, soil gas was evacuated by air pumps from the permeable boreholes. The process was followed by both soil and gas analysis. Biodegradation of oil contamination and the microbial activity was measured by the oil and cell concentration in the soil. In 2 months, the oil content decreased considerably, and the cell number increased by one order of magnitude or more. The evacuated gas was tested for CO₂, O₂ and volatilized hydrocarbon content. The CO₂ level proves the presence of biodegradation: a permanent high value about ten times higher than normal, could be measured for 2 months, followed by a slow decrease in the third month. Volatilized hydrocarbon content was the highest in the first 2 d. After a continuous decrease, it dropped under the threshold of measurability for the third month. Selective biodegradation of hydrocarbon mixtures (oily wastes) was investigated as well: gas Chromatographic oil analysis showed the changes in the oil composition. The appropriate microflora was working in an ideal commensalism, and as a result, all of the hydrocarbon components were degraded nearly to the same extent.     

Bacterial metabolism of polycyclic aromatic hydrocarbons: strategies for bioremediation

Polycyclic aromatic hydrocarbons (PAHs) are compounds of intense public concern due to their persistence in the environment and potentially deleterious effects on human, environmental and ecological health. The clean up of such contaminants using invasive technologies has proven to be expensive and more importantly often damaging to the natural resource properties of the soil, sediment or aquifer. Bioremediation, which exploits the metabolic potential of microbes for the clean-up of recalcitrant xenobiotic compounds, has come up as a promising alternative. Several approaches such as improvement in PAH solubilization and entry into the cell, pathway and enzyme engineering and control of enzyme expression etc. are in development but far from complete. Successful application of the microorganisms for the bioremediation of PAH-contaminated sites therefore requires a deeper understanding of the physiology, biochemistry and molecular genetics of potential catabolic pathways. In this review, we briefly summarize important strategies adopted for PAH bioremediation and discuss the potential for their improvement.     

基础生物化学实验教学对学生技能的培养

面对国内实验教学的新形势和新要求,改进和完善本科实验教学日益紧迫。介绍了本科基础生物化学实验教学的内容与实践,根据生化实验课的特点,对实验教学内容、方式等进行改进,强调了以基本技术围绕基本实验与综合性实验为中心,培养学生的实践能力、创新能力与科学思维,重点强化了实验技能与素质的训练,探索了生化实验教学中培养高素质、综合型创新人才之路。     

Fungal metabolism of polycyclic aromatic hydrocarbons: past, present and future applications in bioremediation

This article examines the importance of non-ligninolytic and ligninolytic fungi in the bioremediation of polycyclic aromatic hydrocarbon contaminated wastes. The research from the initial studies in Dave Gibson’s laboratory to the present are discussed. Received 10 August 1997/ Accepted in revised form 15 August 1997     

石油降解菌剂的研制及其在石油污染土壤修复中的应用

用液态和固态相结合的方式对包含2种细菌的石油降解菌剂进行培养,牛肉膏、蛋白胨作为液态培养基培齐初级种子,然后接种到草炭和麸皮的固态培养基中培养：分析温度、接种量、料水比、草炭与麸皮的比例、培养时间对固态培养的影响.制备的BC—E和BC-12种菌剂的活菌量分别达到2．47×10^11个／g和3．6×10^10个／g。采用研制的菌剂对石油污染土壤进行修复实验,1个月污染土壤中的石油降解率可达到45％。     

Assessment of the microbial potential for nitrate-enhanced bioremediation of a JP-4 fuel-contaminated aquifer

A site that was contaminated with JP-4 jet fuel was characterized microbiologically to assess the feasibility of nitrate-enhanced bioremediation. The results of microcosm studies indicated that the mean pseudo zero-order rate constants for alkylbenzene biodegradation and NO₃ ⁻-N removal were 1.2 and 2.4 mg L⁻¹ per day, respectively. Several alkylbenzenes were removed to a greater extent in samples contaminated with residual JP-4 than in unexposed samples and samples downgradient of the spill; benzene was recalcitrant in all samples. Numbers of total heterotrophs, JP-4-degraders, oligotrophs, total denitrifiers, denitrifiers growing in the presence of JP-4, estimates of cell number by analysis of phospholipid fatty acids, direct counts and aerobic and anaerobic protozoa were determined; however, numbers of microorganisms were not reliable predictors of alkylbenzene biodegradation activity. The presence of aerobic and anaerobic protozoa suggests that protozoa may be active under a variety of different electron acceptor conditions. The results of the characterization study indicated that the site was amenable to nitrate-enhanced bioremediation. Received 12 March 1996/ Accepted in revised form 17 September 1996     

Degradation of straight-chain aliphatic and high-molecular-weight polycyclic aromatic hydrocarbons by a strain of Mycobacterium austroafricanum

AIMS: Our goal was to characterize a newly isolated strain of Mycobacterium austroafricanum, obtained from manufactured gas plant (MGP) site soil and designated GTI-23, with respect to its ability to degrade polycyclic aromatic hydrocarbons (PAHs). METHODS AND RESULTS: GTI-23 is capable of growth on phenanthrene, fluoranthene, or pyrene as a sole source of carbon and energy; it also extensively mineralizes the latter two in liquid culture and is capable of extensive degradation of fluorene and benzo[a]pyrene, although this does not lead in either of these cases to mineralization. Supplementation of benzo[a]pyrene-containing cultures with phenanthrene had no significant effect on benzo[a]pyrene degradation; however, this process was substantially inhibited by the addition of pyrene. Extensive and rapid mineralization of pyrene by GTI-23 was also observed in pyrene-amended soil. CONCLUSIONS: Strain GTI-23 shows considerable ability to mineralize a range of polycyclic aromatic hydrocarbons, both in liquid and soil environments. In this regard, GTI-23 differs markedly from the type strain of Myco. austroafricanum (ATCC 33464); the latter isolate displayed no (or very limited) mineralization of any tested PAH (phenanthrene, fluoranthene or pyrene). When grown in liquid culture, GTI-23 was also found to be capable of growing on and mineralizing two aliphatic hydrocarbons (dodecane and hexadecane). SIGNIFICANCE AND IMPACT OF THE STUDY: These findings indicate that this isolate of Myco. austroafricanum may be useful for bioremediation of soils contaminated with complex mixtures of aromatic and aliphatic hydrocarbons.     

Enhanced anaerobic bioremediation of groundwater contaminated by fuel hydrocarbons at Seal Beach, California

Enhanced anaerobic biodegradation of groundwater contaminated by fuel hydrocarbons has been evaluated at a field experiment conducted at the Naval Weapons Station, Seal Beach, California. This experiment included the establishment of three different remediation zones in situ: one zone was augmented with sulfate, one was augmented with sulfate and nitrate, and the third was unaugmented. This enables a comparison of hydrocarbon biodegradation under sulfate-reducing, sequential denitrifying/sulfate-reducing, and methanogenic conditions, respectively. In general, the results from the field experiment are: (1) Certain fuel hydrocarbons were removed preferentially over others, but the order of preference is dependent upon the geochemical conditions; and (2) In the zones that were augmented with sulfate and/or nitrate, the added electron acceptors were consumed quickly, indicating that enhancement via electron acceptor injection accelerates the biodegradation process. More specifically, in the sulfate-reducing zone, sulfate was utilized with an apparent first-order rate coefficient of approximately 0.1 day^-1. In the combined denitrifying/sulfate-reducing zone, nitrate was utilized preferentially over sulfate, with an apparent first-order rate coefficient of 0.1–0.6 day^-1. However, the data suggest that slow sulfate utilization does occur in the presence of nitrate, i.e., the two processes are not strictly sequential. With regard to the aromatic BTEX hydrocarbons, toluene was preferentially removed under intrinsic conditions; biodegradation of benzene was slow if it occurred at all; augmentation with sulfate preferentially stimulated biodegradation of o-xylene; and ethylbenzene appeared recalcitrant under sulfate-reducing conditions but readily degradable under denitrifying conditions.     

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.