Leaching of BTEX from Aged Crude Oil Contaminated Model Soils: Experimental and Modeling Results

It is generally assumed that soil properties such as organic matter content, porosity, and mineral surface area have a significant effect on the bioavailability and leachability of aged petroleum hydrocarbons. In order to test this hypothesis, nine model soils or sorbents (i.e., fine and coarse quartz sand, montmorillonite and kaolinite clay, peat, 60Å and 150Å silica gel, a loam soil, and non-porous glass beads) were spiked with a crude oil, aged for 27 months in the laboratory, and transferred to glass columns for the performance of continuous flow leaching experiments. The column effluents were periodically sampled for 43 days and analyzed for BTEX. A one-dimensional flow model for predicting the dissolution and dispersion of individual hydrocarbons from a multi-component NAPL such as crude oil was used to fit the leaching data (i.e., the BTEX concentration versus time curves) by adjusting the equilibrium oil-leachate partitioning coefficient (K ^ol ) for each respective hydrocarbon. The Peclet number, which is a measure of dispersion and a required modeling parameter, was measured in separate chloride tracer experiments for each soil column.

Results demonstrate that soil properties did not significantly affect the leaching kinetics of BTEX from the columns. Instead, BTEX leaching curves could be successfully fitted with the one-dimensional NAPL dissolution flow model for all sorbents with the exception of montmorillonite clay. The fitting parameter K ^ol for each hydrocarbon was found to be similar to the K ^ol values that were independently measured for the same crude oil by Rixey et al. (Journal of Hazardous Materials B, 65: 137–156, 1999 Rixey, W. G., Garg, S. and Nie, Y. 1999. Comparison of the fixed-bed and batch leaching characteristics of aromatic compounds in residually trapped crude oils and oily wastes. J. Hazard. Mat. B, 64: 137–156. [CSA][CROSSREF] [Google Scholar]). In addition, the fitted K ^ol values were very similar for BTEX leaching from aged compared to freshly spiked loam soil. These findings indicate that leaching of BTEX in the aged soils that are contaminated with crude oil at the high concentrations commonly found in the environment (i.e., > 20,000 mg/kg) was not affected by soil properties or aging but rather was governed by the equilibrium dissolution of these hydrocarbons from the crude oil NAPL that is coating the soil particles.     

Responses of Microbial Communities to Light-Hydrocarbon Microseepage and Novel Indicators for Microbial Prospecting of Oil/Gas in the Beihanzhuang Oilfield,Northern Jiangsu,China

Previous studies on microbial prospecting of oil/gas only focused on the anomalies of light hydrocarbon-oxidizing microbes as main exploratory indicators and their exploration applications. In this study, we investigated the responses of microbial communities to light-hydrocarbon microseepage in the Beihanzhuang Oilfield, eastern China using denaturing gradient gel electrophoresis (DGGE) analysis and by comparing the difference of two-type areas with high- and low-flux light-hydrocarbon seepages. The results showed that the high-flux light-hydrocarbon seepage favored the growth of Nocardioides, Aciditerrimonas, sulphate-reducing bacteria (SRBs) related to Desulfosporosinus and Desulfovibrio, and Chloroflexi bacteria (b-7), implying that their anomalies might be adopted as novel subsidiary indicators for microbial prospecting of oil/gas in the Beihanzhuang Oilfield. Based on the newly obtained results, we have proposed a general strategy for microbial prospecting of oil/gas, i.e., to determine the anomalies of light hydrocarbon-oxidizing microbes, to select subsidiary indicators for microbial prospecting of oil/gas based on an assessment of the responses of microbial communities to light-hydrocarbon microseepage, to quantitatively measure subsidiary indicators and delimit their anomalies, to comprehensively interpret all microbial anomalies, and to make a suggestion for oil/gas prospecting. This general strategy with novel indicators may provide a more comprehensive evaluation for light-hydrocarbon microseepage and the corresponding anomalies, thereby reducing the exploration risk of oil/gas.     

原油微生物群落构成多样性及降解菌DYL-1降解原油的研究

为分离得到高效原油降解菌,直接向原油中加入营养物刺激,培养一段时间后原油乳化降解,傅里叶红外光谱显示,2957cm-1、723cm-1处吸收峰消失;2855cm-1、1377cm-1处吸收峰减弱;880cm-1以及800cm-1吸收峰几乎消失,表明原油发生降解,效果明显。同时分离得到一株降解菌,经分子鉴定为芽孢杆菌属(Bacillus sp.),红外光谱法和紫外吸收法分析表明其具有较强的降解原油烃的能力。根据传统分子生物学的方法,构建原油菌16S rDNA克隆文库,限制性酶切片段长度多态性(Restriction Fragment Length Polymorphism,RFLP)分析了原油中的细菌多样性。结果表明此方法有效地评估了原油中的细菌群落和多样性。     

Microbial Factors Rather Than Bioavailability Limit the Rate and Extent of PAH Biodegradation in Aged Crude Oil Contaminated Model Soils

The rate and extent of polynuclear aromatic hydrocarbons (PAH) biodegradation in a set of aged model soils that had been contaminated with crude oil at the high concentrations (i.e.,>20,000?mg/kg) normally found in the environment were measured in 90-week slurry bioremediation experiments. Soil properties such as organic matter content, mineral type, particle diameter, surface area, and porosity did not significantly influence the PAH biodegradation kinetics among the 10 different model soils. A comparison of aged and freshly spiked soils indicates that aging affects the biodegradation rate and extent only for higher-molecular-weight PAHs, while the effects of aging are insignificant for 4-ring PAHs and total PAHs. In all model soils with the exception of kaolinite clay, the rate of abiotic desorption was faster than the rate of biodegradation during the initial phase of bioremediation treatment, indicating that PAH biodegradation was limited by microbial factors. Similarly, any of the higher-molecular-weight PAHs that were still present after 90 weeks of treatment were released rapidly during abiotic desorption tests, which demonstrates that bioavailability limitations were not responsible for the recalcitrance of these hydrocarbons. Indeed, an analysis of microbial counts indicates that a severe reduction in hydrocarbon degrader populations may be responsible for the observed incomplete PAH biodegradation. Therefore, it can be concluded that the recalcitrance of PAHs during bioremediation is not necessarily due to bioavailability limitations and that these residual contaminants therefore might pose a greater risk to environmental receptors than previously thought.     

复合菌系RSS-4腐解稻秆过程中的菌系动态变化

采用传统平扳分离培养方法和PCR—DGGE技术研究了水稻秸秆腐解复合菌系RSS-4在腐解稻秆过。程中菌种区系变化情况。结果表明：平板分离培养方法显示,在稻秆腐解过程中,微生物的数量呈现出先升后降的变化趋势,在整个腐解过程中细菌的数量占优势;DGGE图谱显示,至少有12种细菌和18种真菌的近缘种参与到稻秆的腐解过程。在其腐解过程中,不同腐解阶段真菌的组成呈现出多样性,数量变化差异也较大：细菌DGGE图谱中的条带1、9、10等以及真菌DGGE图谱中的条带8、9、13等为优势菌株,它们贯穿于稻秤腐解的整个过程;细菌中的条带12以及真菌中的条带4在腐解的前期起作用,而后迅速消失;细菌中的条带3、11等以及真菌中的条带3、10等在腐解的后期才出现而起作用;而细菌中的条带2以夏真菌中的条带1、5等仅出现在腐解的莱一时期。     

Enhancing the Removal of Sorbed Crude Oil from Soil Through Multiple Oxidation Steps in Stepwise Fenton Processes

The stepwise Fenton oxidation process, in which hydrogen peroxide (H₂O₂) is added in a step-by-step manner instead of at the beginning, can achieve better sorbed crude oil removal effects. The results showed that if a high ratio of sorbed total petroleum hydrocarbon (TPH) was present in soil samples S1 (100%, initial TPH: 10,009 mg/kg) and S2 (94.2%, initial TPH: 4850 mg/kg), the TPH was oxidized in each step. In addition, the total TPH removal efficiency was 49.6% compared with the 27.9% achieved in conventional Fenton oxidation in which all H₂O₂ was added at the beginning. Nevertheless, when the ratio of sorbed TPH in the soil sample S3 was low (45.3%, initial TPH: 2850 mg/kg), the TPH removal efficiency was 18.9%, which was slightly higher than 18.2% achieved in the conventional Fenton process because if the sorbed TPH concentration was low, the sorbed TPH was mainly removed in the first step. The second and the third step resulted in long-chain alkanes entering the aqueous phase rather than removing them from the soil, which posed environmental risk. Therefore, it is clear that stepwise Fenton oxidation could improve sorbed TPH removal efficiency when the sorbed TPH concentration in the soil is high.     

长庆油田延9低渗透油藏微生物群落研究

目的:研究长庆油田延9低渗透油藏微生物群落,为实施微生物提高原油采收率提供指导和依据。方法:长庆油田延9油藏三口不同油井(柳28-46、柳28-47和柳27-45)的油水样品建立16S rDNA克隆文库进行研究。结果:构建了柳28-46、柳28-47和柳27-45油井样品的微生物基因克隆文库,其分类操作单元(OUT)数分别为21、20和20个;序列分析比对表明,3口井的共同的优势微生物菌群为铜绿假单胞菌(Pseudomonas aeruginos),分别占各文库的32.8%、32%和42.9%,它是最常见最主要的采油功能菌之一。此外硫酸盐还原菌(SRB)和铁细菌也处于优势地位,它们是原油开采中的有害菌。结论:延9低渗透油藏微生物群落和其潜在功能的分析为开展微生物提高石油采收率应用提供了良好的基础资料。     

Extraction and Analysis of Microbial Phospholipid Fatty Acids in Soils

Phospholipid fatty acids (PLFAs) are key components of microbial cell membranes. The analysis of PLFAs extracted from soils can provide information about the overall structure of terrestrial microbial communities. PLFA profiling has been extensively used in a range of ecosystems as a biological index of overall soil quality, and as a quantitative indicator of soil response to land management and other environmental stressors.The standard method presented here outlines four key steps: 1. lipid extraction from soil samples with a single-phase chloroform mixture, 2. fractionation using solid phase extraction columns to isolate phospholipids from other extracted lipids, 3. methanolysis of phospholipids to produce fatty acid methyl esters (FAMEs), and 4. FAME analysis by capillary gas chromatography using a flame ionization detector (GC-FID). Two standards are used, including 1,2-dinonadecanoyl-sn-glycero-3-phosphocholine (PC(19:0/19:0)) to assess the overall recovery of the extraction method, and methyl decanoate (MeC10:0) as an internal standard (ISTD) for the GC analysis.     

青海油田微生物采油技术研究

从青海油田地层水中分离培养出4株微生物菌种,对其进行了耐温性、耐盐性等进行了探讨,分析了微生物处理前后原油组份及物性变化情况,并采用岩芯流动试验探讨了微生物的驱油效果。结果表明,这几种微生物菌种均能适应青海油田温度及高矿化度地层环境,作用于原油后对原油中的长链饱和烃类物质有较好的降解作用,能使其分子链变短,并产生有机酸或石油羧酸盐类表面活性剂等低分子物质,使原油的粘度、凝点及蜡含量均出现明显的下降,改善原油的流动性能。提高石油采收率。2口井的现场试验证明,微生物采油具有良好的提高石油采收率的效果和清蜡减阻效果。     

生物造粒流化床微生物落结构及其动态变化

为了研究生物造粒流化床污水处理反应器颗粒污泥中微生物群落结构及其动态变化,分别从生物造粒流化床10、60、110cm处取颗粒污泥,通过细胞裂解直接提取颗粒污泥细菌基因组DNA。以细菌和古细菌16S rRNA基因通用引物530F/1490R,对活性污泥中提取的细菌基因组DNA进行PCR扩增,长约1kb的PCR扩增产物纯化后经变性梯度凝胶电泳(DGGE)分离,获得微生物群落的DNA特征指纹图谱。结果显示,生物造粒流化床反应器颗粒污泥中的微生物群落非常丰富,在10cm处微生物的种属达到23种,60cm处为21种,110cm处为20种;生物造粒流化床不同高度都有一些各自的特有种属和共有种属,反应器不同高度的微生物群落演替不明显,微生物群落相似性为83.1%,群落结构较为稳定。     

Enhanced Biodegradation of Casablanca Crude Oil by A Microbial Consortium in Presence of a Rhamnolipid Produced by Pseudomonas Aeruginosa AT10

The biodegradation of oil products in the environment is often limited by their low water solubility and dissolution rate. Rhamnolipids produced by Pseudomonas aeruginosa AT10 were investigated for their potential to enhance bioavailability and hence the biodegradation of crude oil by a microbial consortium in liquid medium. The characterization of the rhamnolipids produced by strain AT10 showed the effectiveness of emulsification of complex mixtures. The addition of rhamnolipids accelerates the biodegradation of total petroleum hydrocarbons from 32% to 61% at 10 days of incubation. Nevertheless, the enhancement of biosurfactant addition was more noticeable in the case of the group of isoprenoids from the aliphatic fraction and the alkylated polycyclic aromatic hydrocarbons (PHAS) from the aromatic fraction. The biodegradation of some targeted isoprenoids increased from 16% to 70% and for some alkylated PAHs from 9% to 44%.     

Crude Oil Treatment Leads to Shift of Bacterial Communities in Soils from the Deep Active Layer and Upper Permafrost along the China-Russia Crude Oil Pipeline Route

The buried China-Russia Crude Oil Pipeline (CRCOP) across the permafrost-associated cold ecosystem in northeastern China carries a risk of contamination to the deep active layers and upper permafrost in case of accidental rupture of the embedded pipeline or migration of oil spills. As many soil microbes are capable of degrading petroleum, knowledge about the intrinsic degraders and the microbial dynamics in the deep subsurface could extend our understanding of the application of in-situ bioremediation. In this study, an experiment was conducted to investigate the bacterial communities in response to simulated contamination to deep soil samples by using 454 pyrosequencing amplicons. The result showed that bacterial diversity was reduced after 8-weeks contamination. A shift in bacterial community composition was apparent in crude oil-amended soils with Proteobacteria (esp. α-subdivision) being the dominant phylum, together with Actinobacteria and Firmicutes. The contamination led to enrichment of indigenous bacterial taxa like Novosphingobium, Sphingobium, Caulobacter, Phenylobacterium, Alicylobacillus and Arthrobacter, which are generally capable of degrading polycyclic aromatic hydrocarbons (PAHs). The community shift highlighted the resilience of PAH degraders and their potential for in-situ degradation of crude oil under favorable conditions in the deep soils.     

秸秆降解菌剂对秸秆还田土壤中细菌种群数量的影响

要研究秸秆降解菌剂施用后1个生长季秸秆中细菌的数量和纤维素酶活力,了解其动态变化,为进一步研究分析土壤中微生物结构变化规律提供可靠的理论依据.利用分子生物学方法,对秸秆降解菌剂施用后不同生长阶段耕层中细菌基因组DNA纯化后PCR扩增其16S rDNA V3可变区,对扩增产物进行DGGE电泳,并对结果进行分析.初步分析表明,菌落数量呈现前期上升、中期最高、后期逐渐降低的趋势.     

油脂下脚料中残油微生物降解初步研究

以油脂降解菌Bacillus sp DE-8为出发菌,对油脂下脚料中的残油生物降解条件进行初步研究。结果表明:该菌对油脂下脚料降解条件为:起始pH值为8,接种量4%、摇床转速为150r.min-1、温度为32℃、发酵72h,该菌株对菜籽饼的降解率可达78.8%。     

Measurement of H2S in Crude Oil and Crude Oil Headspace Using Multidimensional Gas Chromatography,Deans Switching and Sulfur-selective Detection

A method for the analysis of dissolved hydrogen sulfide in crude oil samples is demonstrated using gas chromatography. In order to effectively eliminate interferences, a two dimensional column configuration is used, with a Deans switch employed to transfer hydrogen sulfide from the first to the second column (heart-cutting). Liquid crude samples are first separated on a dimethylpolysiloxane column, and light gases are heart-cut and further separated on a bonded porous layer open tubular (PLOT) column that is able to separate hydrogen sulfide from other light sulfur species. Hydrogen sulfide is then detected with a sulfur chemiluminescence detector, adding an additional layer of selectivity. Following separation and detection of hydrogen sulfide, the system is backflushed to remove the high-boiling hydrocarbons present in the crude samples and to preserve chromatographic integrity. Dissolved hydrogen sulfide has been quantified in liquid samples from 1.1 to 500 ppm, demonstrating wide applicability to a range of samples. The method has also been successfully applied for the analysis of gas samples from crude oil headspace and process gas bags, with measurement from 0.7 to 9,700 ppm hydrogen sulfide.     

Study of Heavy Metal Distribution in Soils Impacted with Crude Oil in Southern Nigeria

The study presents the levels and enrichment factors of heavy metals in soils of southern Nigeria that have received significant impact of crude oil spillage. The results revealed that the concentrations of heavy metals in the examined soils fitted into levels found in agricultural soils except for cadmium. Heavily impacted sites showed elevated levels of heavy metals compared to less impacted sites and background levels. The mean enrichment factors for Cd, Cu, Cr, Pb, Mn, Ni, and Zn were 37.3, 2.8, 14.4, 14.0, 0.77, 5.4, and 1.27 for topsoil and 37.5, 1.30, 7.81, 1.59, 4.12, and 1.28 for subsoil, respectively. This clearly indicates that there is gradual build-up of heavy metals in these soils as a result of the oil spillage and related anthropogenic activities in this area.     

中国喀斯特地区微生物群落结构和功能的研究进展

频繁的人类生产活动使植被遭到破坏,造成基岩裸露的石漠化现象,严重制约了喀斯特地区自然和社会经济的发展,随着生态修复工程的大力开展,微生物群落的结构和功能在生态修复中逐渐受到重视,因为微生物作为喀斯特生态系统的重要组成部分,不仅在物质循环过程中起着重要作用,也在喀斯特生态系统修复中占有十分重要的地位。所以微生物群落结构和功能的研究可以作为衡量生态系统稳定性的重要指标。就中国喀斯特地区的典型植被恢复的不同阶段、成土过程、不同的土地利用方式、矿山修复过程以及不同水域中的微生物群落结构及功能等方面的研究状况进行系统梳理,结合实例综合论述喀斯特地区生态修复过程中微生物作用的研究进展,以期为喀斯特地区生态修复提供参考。     

贵州山区土壤中微生物担是能源物质碳流动的源与汇

在传统的农业生态系统的研究中 ,主要精力放在营养物 (如Ｎ)上 ,认为它们是限制生产力的因素 ;而往往忽略了土壤中碳的重要性 ,认为收获不受Ｃ限制的影响。然而 ,碳循环中的有机碳的分解作用部分控制着出现在地表下和显露在地表上的农业过程[4]。土壤中所储存的有机质 ,其数量既反映土壤从植物残留物的输入所获得的有机质与微生物群落的能量和营养需求之间的平衡 ,又反映植物对营养物的需求与有机质分解作用之间的平衡。因此 ,土壤中碳的平衡能反映出有机质中能量物质的储存[5]。大部分由光合作用形成的碳 ,是通过地表下的生态系统来流动的[…     

The Role of Heterotrophic Microbial Communities in Estuarine C Budgets and the Biogeochemical C Cycle with Implications for Global Warming: Research Opportunities and Challenges

Estuaries are among the most productive and economically important marine ecosystems at the land–ocean interface and contribute significantly to exchange of CO₂ with the atmosphere. Estuarine microbial communities are major links in the biogeochemical C cycle and flow of C in food webs from primary producers to higher consumers. Considerable attention has been given to bacteria and autotrophic eukaryotes in estuarine ecosystems, but less research has been devoted to the role of heterotrophic eukaryotic microbes. Current research is reviewed here on the role of heterotrophic eukaryotic microbes in C biogeochemistry and ecology of estuaries, with particular attention to C budgets, trophodynamics, and the metabolic fate of C in microbial communities. Some attention is given to the importance of these processes in climate change and global warming, especially in relation to sources and sinks of atmospheric CO₂, while also documenting the current paucity of research on the role of eukaryotic microbes that contribute to this larger question of C biogeochemistry and the environment. Some recommendations are made for future directions of research and opportunities of applying newer technologies and analytical approaches to a more refined analysis of the role of C in estuarine microbial community processes and the biogeochemical C cycle.