生物修复石油污染土壤

本文概述了石油污染土壤的生物降解机制,分析了生物修复土壤污染技术及其影响因子,提出了强化生物修复的措施及其在我国的应用前景。     

石油污染土壤修复技术研究进展

概述了目前国内外石油污染土壤常用的修复技术及其研究进展,综述了物理修复、化学修复,特别是生物修复技术的优越性,并针对国内外石油污染土壤修复技术研发和实际应用过程中存在的问题,提出加强研发污染土壤综合修复技术、完善修复工程设计、加大新型功能材料的开发和应用力度、加强分子生态学技术在污染土壤修复中的应用4项建议。     

石油污染土壤的生态风险评价和生物修复Ⅲ.石油污染土壤的植物—微生物联合修复

在对石油污染土壤理化性质和微生物数量研究的基础上,通过接种含石油降解菌的菌剂、添加营养、定期翻动以及栽种植物等方式对其进行了修复。试验表明,添加氮磷等营养后,土壤中的烃降解菌明显增加,石油降解速率显著加快,但接种菌剂以及定期翻动对降解率没有显著影响。在修复试验进行到120 d后,对部分处理进行了植物修复。试验表明栽种狼尾草后,土壤中石油降解速率显著加快,生物毒性明显降低。另外,试验结束后对土壤中的总DNA进行了提取和基于16S rDNA V3区的PCR-DGGE研究。结果表明,经过修复处理改变了土壤中优势细菌的群落结构,使土壤细菌多样性增强。     

石油污染土壤菌剂修复技术研究

向石油污染土壤中投加环境适应能力强,降解效能高的菌种或菌群是提高石油类污染物降解效率的重要手段。本文应用菌剂修复技术对某污灌区石油烃污染土壤进行了处理,结果表明,混合施加两种菌剂,对石油的降解有相互促进作用,土壤中石油类污染物的降解率为54.23%。本研究为该地区石油污染土壤的治理提供了有力的技术保证。     

石油污染土壤的生物修复研究进展

对于石油污染土壤的修复,其生物修复具有环境友好、费用较低等特点,是最具应用前景的土壤修复技术。本文较全面地介绍了石油污染土壤生物修复的影响因素、石油污染土壤的生物修复技术,并对该领域今后的研究重点进行了展望。     

石油污染土壤的生物修复研究

通过田间试验研究了玉米和向日葵两种植物对石油污染土壤的修复作用,考察了外源菌（OX-9）对植物修复的强化和协同效应,对“外源菌一植物”修复效果进行了初步评价。结果表明,在10000mg·k-1污染浓度下,150d玉米、向13葵试验区土壤中石油降解率分别为42．5％和46．4％,较对照区提高了100．5％和118．9％。外源节细菌的施加可使生物修复速度显著加快,150d“DX-9-玉米”和“DX-9-向日葵”试验区石油烃降解率分别达到72．8％和76．4％,较同期单独植物修复的降解率提高了71.3％和64．7％。500d各试验区土壤中石油烃降解率分别为95．5％、96．1％、97．6％和98．9％,土壤中石油烃含量均低于国家标准规定限量（〈500mg·kg-1）;土壤主要理化性质、生物群落分布、呼吸强度及植物不同部位中石油烃的残留量与对照无显著差异。结果表明：玉米、向日葵与节细菌对石油污染土壤的联合生物修复效果显著;经过两年修复,污染土壤恢复健康状态。     

生物修复石油污染土壤研究现状

石油污染土壤的生物修复方法具有操作简便、费用低、对周围环境污染小、修复效率高等优点,应用前景广阔。从石油污染土壤生物修复过程中修复生物的选择、修复条件的优化、修复效果的评价等方面对近年来生物修复技术的研究进展进行了综述。为推动该方法的广泛应用,还应深入探究降解微生物之间的拮抗和协同机制、进一步降低修复成本、进一步完善修复评价标准,加强重石油污染土壤和特殊环境下石油污染土壤的治理研究,开展复合污染(如石油污染和重金属污染)土壤的修复研究。     

石油污染土壤的生物修复研究进展

生物修复技术是解决环境污染、恢复被人类活动破坏的生态系统、实现人类社会可持续发展的重要手段之一,它具有速度快、消耗低、效率高、成本低、反应条件温和以及无二次污染等显著优点.本文主要介绍了生物修复的原理和特点,石油污染土壤的各种生物修复技术:微生物修复技术、植物修复技术和菌根根际生物修复技术的研究和应用进展.     

菌剂-菌根联合修复石油污染土壤的实验研究

植物根际是一个能降解土壤中污染物的生物活跃区。本文应用菌根修复技术对某污灌区石油烃污染土壤进行了处理。在污染土壤中种植玉米和黄豆,通过施加不同的菌剂,采取菌剂和菌根强化修复措施,在运行一个生长季节后,土壤中石油类污染物降解率可达53%~78%。本研究为该地区石油污染土壤的治理提供了有力的技术保证。     

石油污染盐碱土壤的淋洗施肥修复

采用淋洗施肥修复方法处理石油污染盐碱土壤,评价该修复方法对石油污染盐碱土壤的修复效果,并且采用最大或然数法和Biolog方法对土壤微生物数量和微生物群落水平生理特性进行研究。结果表明,经过182d的培养,淋洗施肥处理中油和脂的降解率分别比对照处理和施肥处理高（19.7±4.3）%和（13.8±3.4）%,土壤盐分去除率分别比对照处理和施肥处理高（66.5±2.9）%和（41.3±6.2）%,说明该处理是一种修复石油污染盐碱土壤的有效方式。淋洗施肥处理明显提高异养细菌、石油烃降解菌、烷烃降解菌和多环芳烃降解菌数量和土壤微生物活性,促进了微生物对土壤中油和脂的降解。此外,淋洗施肥处理提高了土壤微生物Shannon多样性指数和Simpson指数,促进了微生物种群的稳定,这暗示着土壤微生物种群正在逐渐恢复。     

Engineered in situ bioremediation of soil and groundwater polluted with weathered hydrocarbons

In this work, we present our experience in the engineered in situ biostimulation of a hydrocarbon-polluted subsoil of an old metal-working plant. The site had a long history of fuel, lubricant oil, and diesel spills and leakages that were initially treated by means of physico-chemical techniques. After one year of treatment, weathered hydrocarbons were firmly sorbed in the unsaturated zone, limiting the effectiveness of the physico-chemical methods being applied. Also, low nutrient and dissolved oxygen levels limited natural attenuation yields. Therefore, a bioremediation approach based on the injection of hydrogen peroxide, an oleophilic fertilizer, and a surfactant was applied. Total petroleum hydrocarbons and gas chromatography-mass spectrometry determinations were performed as a chemical means of monitoring the process. Microbial populations, including the presence of hydrocarbon-degrading bacteria, were simultaneously analyzed during the process, using enrichment techniques and confocal laser scanning microscopy observations with fluorescent indicators. Hydrocarbon-degrading bacteria were already present in the polluted subsoil prior to the implementation of this bioremediation technique and increased noticeably during the first 2 months of treatment. Hence, the suitability of the nutrient and oxygen amending approach was confirmed as further demonstrated by chemical determinations.     

二(2-乙基己基)邻苯二甲酸酯降解菌的分离和鉴定及其在污染土壤生物修复中的作用

Di-（2-ethylhexyl） phthalate（DEHP） is a high-molecular-weight phthalate ester（PAE） that has been widely used in the manufacture of polyvinylchloride and contributes to environmental pollution.The objectives of the present study were to isolate a DEHP degrader that can utilize DEHP as a carbon source and to investigate its capacity to biodegrade DEHP in both liquid culture and soil.A bacterial strain WJ4 was isolated from an intensively managed vegetable soil,which was contaminated with PAEs.The strain WJ4 was affiliated to the genus Rhodococcus and was able to remove DEHP from soil effectively.A period of only 7 d was required to degrade about 96.4%of DEHP(200 mg L^-1) in the liquid culture,and more than 55%of DEHP(1.0 g kg^-1) in the artificially contaminated soil was removed within 21 d.Furthermore,Rhodococcus sp.strain WJ4 had a strong ability to degrade DEHP without additional nutrients in liquid minimal medium culture and DEHP-contaminated soil and to degrade the homologue of DEHP in both liquid culture and soil.Strain WJ4 represents a novel tool for removing PAEs from contaminated soils and it may have great potential for application in the remediation of environmental pollution by PAEs.     

百菌清降解菌株H4的分离与表征及其修复污染土壤的潜能研究

A chlorothalonil(CTN)-degrading bacterial strain H4 was isolated in this study from a contaminated soil by continuous enrichment culture to identify its characteristics and to investigate its potential for remediation of CTN in contaminated soil. Based on the morphological, physiological and biochemical tests and 16 S r DNA sequence analysis, the strain was identified as Stenotrophomonas sp. After liquid culture for 7 d, 82.2% of CTN was removed by strain H4. The isolate could degrade CTN over a broad range of temperatures and p H values, and the optimum conditions for H4 degradation were p H 7.0 and 30℃. Reintroduction of the bacteria into artificially contaminated soil resulted in substantial removal of CTN( 50%) after incubation for 14 d. Soil samples treated by H4 showed significant increases(P 0.05) in soil dehydrogenase activity, soil polyphenol oxidase activity, average well-color development obtained by the Biolog Eco plate TM assay and Shannon-Weaver index, compared with the control. Strain H4 might be a promising candidate for application in the bioremediation of CTN-contaminated soils.     

2种菌联合修复农田土壤镉污染的研究

采用盆栽试验,运用BCR四步法、火焰原子吸收法和稀释涂布平板法等方法,研究了沼泽红假单胞菌(Rhodopseudanonas palustris)和枯草芽孢杆菌(Bacillus subtilis)联合作用对修复农田土壤重金属镉污染盆栽种植小白菜(Brassica campestris L. ssp.chinensis Makino)的效果及安全性,旨在探讨降低土壤镉生物有效性的有效途径,为新型生物菌肥研究提供理论依据。结果表明,沼泽红假单胞菌、枯草芽孢杆菌单独作用以及联合作用均能使农田土壤中镉的固定态增加,降低农田土壤中镉的生物有效性,联合菌处理组镉的生物有效性降低最显著,为32.70%,均对农田土壤中微生物各生理类群数量有显著性影响(P0.05),其中,细菌的数量达到10~5 CFU/g;均显著降低了小白菜根、茎中镉含量,联合菌处理组根、茎中镉的去除率分别达到58.06%～71.37%和73.66%～76.70%;均增加了小白菜株高、茎鲜重、叶绿素a、b的含量和叶绿素a/b的比值,联合菌处理组小白菜的生物量增加18.13%～69.55%。研究结果说明,沼泽红假单胞菌和枯草芽孢杆菌联合作用,对农田土壤镉污染的修复效果显著,且促进了小白菜的生长,具有修复农田土壤重金属污染的潜力。     

一株烟草秸秆降解菌的分离、鉴定及酶学性质研究

从皖南地区烟稻轮作田土壤中经CMC-Na初筛获得5株纤维素降解菌,经DNS法测定纤维素酶活性复筛得到一株降解活性较高的降解菌YC-2。根据该菌16S r DNA序列比对结果,结合形态和生理生化特征,确定YC-2为枯草芽孢杆菌(Bacillus subtilis)。对YC-2酶学性质进行相关研究,并分析YC-2对烟碱的耐受情况及对烟草秸秆的降解情况,结果表明:在7天内,YC-2对烟草秸秆降解率为10.14%;酶学特性表现为最适反应温度为60℃且在15~60℃之间具有稳定性;最适反应pH为7.0,在pH 4.0~7.5范围内稳定性较好;YC-2在浓度为1~2 g/L烟碱中能快速生长,而在高浓度的烟碱中生长受到抑制。因此,菌株YC-2产纤维素酶活性较高、相对耐热耐碱且对烟杆有一定分解作用,通过进一步诱变选育和发酵条件优化有较好的田间应用潜力。     

Mechanisms of production of soil protease by proteolytic Bacillus subtilis in wetland rice soil

In sterile soil inoculated with proteolytic Bacillus subtilis, the correlation between variations in protease activity and cell numbers was investigated during incubation, and the results were compared with those obtained in different growth media. In sterile soil inoculated with B. subtilis, the extracted soil z-FLase activity (hydrolytic activity towards benzyloxycarbonyl-L-phenylalanyl-L-leucine extracted with 0.1 M phosphate buffer) was correlated with the extracted soil protein content (r=0.95; P<0.01) and the number of vegetative cells (r=0.88, P<0.05), while the extracted soil caseinase activity was well correlated with the number of spores (r=0.82, P<0.05) and not with the number of vegetative cells. Extracellular z-FLase activity in different growth media, inoculated with the proteolytic B. subtilis, was correlated with the extracellular protein concentration (r=0.73, P<0.01) and the number of cells (r=0.73, P<0.01), but extracellular caseinase activity was not correlated with either of these measurements. From these results, we concluded that soil z-FLase production might relate to vegetative growth of the proteolytic B. subtilis and soil caseinase might relate to B. subtilis sporulation.     

苏云金芽孢杆菌菌剂对棉花根际土壤细菌数量及多样性的影响

采用稀释平板培养法与PCR-DGGE技术, 以阿维菌素为阳性对照, 水为阴性对照, 研究了大田喷施推荐剂量(0.1 kg·hm^-2)和高剂量(10 kg·hm^-2)苏云金芽孢杆菌(Bt)菌剂对棉花根际土壤细菌种群数量及结构的影响。结果表明, 喷雾处理后1~3 d, 不同处理组土壤细菌数量间无显著性差异, 在3 d时细菌数量均值达到最大, 之后开始下降, 12 d后清水对照、推荐剂量Bt菌剂、高剂量Bt菌剂处理组土壤细菌数量均值维持在4.0×107 CFU·g^-1左右; 推荐剂量Bt菌剂处理样品土壤细菌数量在6 d时显著高于清水对照, 其余时间与清水对照间无显著性差异; 高剂量Bt菌剂处理与清水对照在整个试验期间均无显著性差异; 阿维菌素处理组土壤细菌数量在0~6 d内与清水对照无显著性差异, 而在12~45 d内显著低于其他3个处理组。DGGE图谱显示, Bt菌剂处理对棉花根际土壤17种细菌均无显著抑制作用。聚类分析结果表明, Bt菌剂对土壤细菌群落结构的扰动在12 d后得到恢复。与阴性对照组相比, Bt菌剂对土壤细菌多样性指数无显著影响, 而阳性对照阿维菌素对土壤细菌种群消长和多样性指数有较强的影响。对DGGE图谱中17条电泳条带的序列分析, 证明棉花根际土壤中存在起固氮作用的慢生根瘤菌属细菌和具有污染修复与净化活性的鞘脂菌属、鞘氨醇单胞菌属和红球菌属细菌。Bt菌剂与阿维菌素处理均对这些土壤有益菌群无明显不利影响。总体结果表明, Bt菌剂无论是在正常推荐剂量下还是在较高剂量(推荐剂量的100倍)下使用, 对棉花根际土壤微生态环境产生的冲击都较小, 是一种生态安全性较高的生物农药。     

有机溶剂对有机氯农药污染场地土壤淋洗修复效用研究

Problems associated with organochlorine pesticide (OCP)-contaminated sites in China have received wide attention. To solve such problems, innovative ex-situ methods of site remediation are urgently needed. We investigated the feasibility of the extraction method with different organic solvents, ethanol, 1-propanol, and three fractions of petroleum ether, using a soil collected from Wujiang (WJ), China, a region with long-term contamination of dichlorodiphenyltrichloroethanes (DDTs). We evaluated different influential factors, including organic solvent concentration, washing time, mixing speed, solution-to-soil ratio, and washing temperature, on the removal of DDTs from the WJ soil. A set of relatively better parameters were selected for extraction with 100 mL L 1 petroleum ether (60-90℃): washing time of 180 min, mixing speed of 100 r min 1 , solution-to-soil ratio of 10:1, and washing temperature of 50℃. These selected parameters were also applied on three other seriously OCP-polluted soils. Results demonstrated their broad-spectrum effectiveness and excellent OCP extraction performance on the contaminated soils with different characteristics.     

生态毒理学工具在评价石油化工厂区土地耕作适宜性中的运用

The demand for simple and rapid bioassays in ecotoxicological evaluations is of paramount importance in order to speed up environmental monitoring programs. In this study we performed bioassays with lettuce seeds and two species of terrestrial isopods （Armadillidium vulgare and Porcellio dilatatus） for the ecotoxicological assessment of a landfarming soil from a petrochemical complex area. The solubilized content of test soil demonstrated a concentration-response type toxic effect on seed germination rate, and a delay on germination, but showed toxic effect on seedlings wet weight only at the highest concentration. Toxic effects were also observed in mortality rate and avoidance behavior of the two woodlice species. These results demonstrated the sensitiveness of the organisms studied, and highlighted the possibility to use these bioassays in environmental monitoring programs in areas contaminated with fossil fuels.     

应用新引物DGGE分析土壤中硝酸盐还原细菌群落

The narG gene is frequently used as a molecular marker for bacterial nitrate-reducing community analysis. In this study, a new set of primers targeting the narG gene was designed and applied to semi-nested polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) assay. The potential of the new primers was verified on DNA directly extracted from soils from five different experimental sites distributed in Central and Southern Italy. Specificity of the primers was determined by excision, amplification, and sequencing of bands resolved by DGGE. A phylogenetic analysis showed the correlation between the sequences retrieved from the soils studied and the narG sequences from β and γ-Proteobacteria. These primers expanded the existing molecular tools for ecological study on the size and diversity of nitrate-reducing bacterial community in soil.