高效耐盐碱石油烃降解菌筛选及降解特性研究

为探究长庆油田污染土壤中微生物对石油的降解特性,在该油田多个油井附近采集了10处含油污染土壤进行石油烃降解菌的筛选、分离及降解实验。通过对筛选出的四株石油烃降解菌株5-5、5-X、9-2、10-3进行革兰氏染色、菌落形态观察、生化理化试验及16S rDNA测序,鉴定出这四株菌株分别为醋酸钙不动杆菌（Acinetobacter calcoaceticus）、不动杆菌（Acinetobacter sp.）、蒙氏假单胞菌（Pseudomonas monteilii）和乳酸不动杆菌（Acinetobacter lactucae）。14 d降解实验结果显示,这四种菌株对总石油烃的降解率分别为50.92%、51.27%、78.30%和44.39%;尤其菌株Pseudomonas sp.9-2表现出优异的降解性能,且对不同组分石油烃（正构烷烃、异构烷烃及芳烃）的降解率分别达到了94.65%、69.73%和59.07%,对长链正构烷烃也体现出了较好的降解性能。另外,抗逆性试验结果表明菌株Pseudomonas sp.9-2对pH和盐度的耐受范围分别为5.0~10.0、0.5%~6.0%,表明该菌株对盐碱环境具有较好的适应性,可用于盐碱石油污染土壤的微生物修复研究。     

石油污染场地浅层地下水MNA原位修复潜能及微生物降解效益评估

监测式自然衰减（MNA）能够高效低耗地原位修复石油污染地下水的场地,微生物对污染物的降解对MNA过程起到了重要作用。在分析东北石油污染场地地下水中总石油烃（TPH）、电子受体的质量浓度分布和变化规律基础上,划分了微生物功能区。采用溶质通量计算法,对MNA原位修复的潜能及其微生物降解效果进行了评估。结果显示,场地微生物降解正在发生,利用的电子受体不同,划分为Mn、Fe和SO^2-₄还原区。污染通量模型计算显示：上游地区微生物降解强度不断增强,下游地区微生物降解强度不断降低。监测期内石油烃总量降低了394 kg,微生物降解为自然衰减过程中的主要作用,其贡献率为64%~93%,每个通量断面内微生物降解率为0.18~0.73 kg/d。由此可以证明,MNA可以有效地修复地下水中的石油污染。     

离子类型对石油污染含水层中微生物去除苯的影响

近来利用微生物原位修复受石油污染的含水层已被广泛关注,然而地下水中含有许多离子成分,这些无机离子对微生物降解有机污染物的影响机制还不清楚。本文采用批量实验研究了淄博齐鲁石化污染地下水中常见的7种无机离子(NO3-、PO4 3-、SO4 2-、Cl-、Ca2+、Mg2+、Fe3+)对微生物生长及生物降解苯的影响规律,利用高通量测序技术进一步探究了苯降解菌的种群特征。结果表明：7种离子都存在一个最适宜微生物生长的离子浓度,低于或超过该浓度苯的去除率明显降低,其中NO3-、SO42-、Fe3+最适宜浓度为0. 4mmol/L,PO4 3-、Cl-、Ca2+、Mg2+最适宜浓度分别为0. 2mmol/L、0. 1mol/L、2. 5mmol/L、2mmol/L;从微生物含量及其变化幅度来看,地下水环境中的NO3-离子对微生物的生长及苯的去除影响最显著,其他离子的影响则较小,但微生物对Cl-的耐受浓度较高。高通量测序结果显示驯化出的苯降解菌主要属于脱硫弧菌属(Desulfovibrio sp)、脱硫芽胞弯曲菌属(Desulfosporosinus sp)、不动杆菌属(Acinetobacter sp)和假单胞菌属(Pseudomonas sp)中的菌株。研究结果可为石油污染地下水的原位生物修复提供一定的科学依据。     

浅层地下水中氯代烷烃生物降解的地下水化学 响应规律研究

以东北某油田石油污染场地多期地下水化学监测数据为基础,分析氯代烷烃自然生物降解中充当第一基质、电子受体、中间产物的水化学组分及相关地球化学参数在污染晕演化的不同时段、不同部位的变化趋势,了解氯代烷烃生物降解的地下水化学响应特征,并以此为依据对污染场地浅层地下水氯代烷烃生物降解程度进行定性评价。结果表明:2010年6月,事故发生约1年之际,在ZK1—ZK3—ZK6纵向剖面上生物降解过程中充当电子受体的水化学组分(SO42–)、第一基质(CH2Cl2、CHCl3)和代表地下水环境的主要理化指标(pH、Eh)从上游到下游均呈递增规律,而相关还原产物(Fe2+、Mn2+、HCO3–)则呈递减规律,这也表明天然微生物降解具有一定的滞后性。2010年6月氯代烷烃天然生物降解评价表明ZK1属微生物降解证据充足;ZK3观测井属微生物降解证据有限;ZK6观测井属微生物降解证据不足。而2011年5月,事故发生约2年之际,封堵污染源后随污染晕中心的向下游迁移,ZK3观测井中生物降解作用逐渐明显,在ZK3—ZK6纵向剖面上地下水化学指标呈现上述响应规律,而ZK1观测井诸多水化学指标经上游未污染地下水的补充更新,逐渐趋向污染前的背景值。     

微生物修复石油污染地下水的实验研究

为修复陕北黄土区石油污染地下水,采用优化土著微生物菌群的生物技术,进行了地下水中石油的降解与修复实验研究。在实验装置内加入了1.5%的优化菌群制剂,优化出的菌群初步鉴定主要有:假单胞菌属、微球菌属、放线菌属、真菌类的青霉属和曲霉属等。实验结果显示,在实验装置中人为添加石油含量为182.5 mg/l、862.5 mg/l、1695.0 mg/l时,经过28~37 d的微生物修复,地下水中石油的降解率为27.47%~92.46%,而无菌对照中的石油含量变化在5%以内,说明在无菌条件下地下水中石油降解缓慢。该实验过程验证了微生物修复技术在地下水石油污染修复中的有效性,探索了应用的可行性。     

某石化污染场地含水层自然降解BTEX能力评估

地下水污染问题是国家关注的重要环境问题之一,监测与评估含水层自然降解污染物能力是防治地下水污染的基本手段,也是国外地下水中修复技术研究热点课题。以华北平原某石油化工类场地为案例,通过调查场地水文地质条件、土壤及地下水污染现状,监测场地地下水中苯系物(BTEX)浓度及相关化学参数变化,运用微生物水文地球化学方法和水文地质方法,估算了该场地含水层自然降解苯系物量。这一研究成果为评估我国石油类场地地下水污染的自然修复能力提供了实证和基础数据。     

地下水有机污染及微生物修复技术应用

通过对国内外地下水有机污染现状和有机物污染地下水的途径及存在状态的分析,提出了适用于地下水有机污染治理的微生物修复技术———土著微生物法、生物活化法和生物添加法,并对微生物修复技术在地下水有机污染控制中应用的可能性及今后研究方向进行总结。     

地下水土石油类污染修复技术途径

石油工业在开采、加工、运输、储备过程中石油及其初加工产品泄漏,进入环境造成污染。本文根据国内外现有异位修复技术和原位修复技术研究成果,提出“加强场地调查,有效控制污染源,阻断污染途径,同时开展受污染地下水土修复,优化组合多种修复技术方法”路线,表明可修复地下水土石油类有机污染。     

土壤石油污染物微生物降解机理与修复技术研究

较为系统地概述了土壤石油污染物的微生物修复技术和近年来石油烃代谢机理以及微生物细胞膜转运石油烃机理研究等方面的进展。文中着重讨论烷烃代谢途径、影响烷烃降解酶合成的阻遏物和激活物、芳香烃代谢过程和基因方面的研究,以及细胞膜结构的改变和石油烃的迁移等重要特性,最后从宏观系统的修复技术和微观分子学方面探讨微生物降解石油污染土壤的发展方向。     

脱卤杆菌介导的厌氧微生物富集菌群对1,2,4-三氯苯的降解特性

1,2,4-三氯苯是我国工业污染场地土壤和地下水中典型的有机污染物,具有持久性、生物蓄积性和高毒性特点,对生态环境和人体健康危害巨大。1,2,4-三氯苯密度比水大,容易迁移至深部厌氧区域,因此,开展1,2,4-三氯苯厌氧微生物降解与修复研究具有重要实际应用价值。本文通过长期富集培养,获得一份可以稳定地将1,2,4-三氯苯还原脱氯至1,4-二氯苯,再进一步还原脱氯至氯苯的厌氧菌液。通过16S rRNA基因扩增子测序及引物特异性的定量PCR实验,证明厚壁菌门的脱卤杆菌属细菌(Dehalobacter species)是1,2,4-三氯苯和1,4-二氯苯厌氧还原脱卤的功能菌株,其生长率为(1.68±0.8)×10⁶ copies·μmol^-1(释放的氯离子)。通过PCR扩增,获得Dehalobacter菌株的16S rRNA基因序列,构建了系统发育树。本研究可为1,2,4-三氯苯污染场地开展原位厌氧微生物修复提供菌株资源和理论指导。     

季节性低温胁迫下陇东黄土高原油污土壤环境因子对耐冷混合菌场地生态修复的响应

为了揭示季节性低温胁迫下陇东黄土高原油污土壤环境因子对耐冷混合菌场地生态修复的响应机制,利用自主筛选构建的耐冷石油降解混合菌在甘肃省庆阳市庆城县马岭镇长庆油田陇东油泥处理站开展了为期7个月的场地修复实验,采用常规方法测定了不同季节土壤理化特性、酶活性和微生物群落特性等环境指标。结果表明：（1）在季节性低温胁迫下（9-11月）M2组（耐冷混合菌处理组）月平均降解率明显增加（P<0.01）,JZJ+M1组（金盏菊联合常温混合菌处理组）和M2组累计TPH降解率分别为15.37%±3.51%和28.64%±4.12%。（2）M2组土壤脱氢酶和多酚氧化酶在低温季节（LT）活性最高,且温度和处理存在显著交互作用（P<0.01）。在土壤营养元素方面,无论何种处理方式（JZJ+M1和M2）二者含量均为RT（常温季节）高于LT（P<0.05）,同时明显高于CK组（P<0.05）。（3）M2组土壤微生物群落Shannon-Wiener指数和均匀度指数在LT高于RT（P<0.05）。（4）NMDS和Pearson相关性分析结果显示,M2组在低温季节具有较高TPH降解率主要与土壤多酚氧化酶、脱氢酶、单月TPH降解率（μ2）和Shannon-Wiener指数有关,且均呈极显著正相关关系。通过分析环境因子的季节响应,优化场地修复在低温环境的降解条件、加速低温期石油烃降解速率,以期为陇东地区低温耐冷混合菌场地生态修复技术的推广应用提供参考和基础数据资料。     

Variations in the environmental characteristics of groundwater slightly contaminated with petroleum: effects of enhanced bioremediation in northeast China

The objective of this paper was to investigate a petroleum-contaminated groundwater site in northeast China. Based on the monitoring and analysis of environmental characteristics of groundwater slightly contaminated with petroleum, microbes were added into the petroleum-contaminated groundwater via single-well and multiple-well models to enhance bioremediation. The effect of enhanced bioremediation was monitored and analyzed, and variations in the environmental characteristics of groundwater containing total petroleum hydrocarbons (TPH) were identified to provide a scientific basis for controlling and remediating petroleum-contaminated groundwater. Findings confirmed that this site was slightly contaminated and TPH levels exhibited a slight decreasing trend. After enhanced remediation, the microbial degradation effect was large, and the TPH concentration significantly reduced. In particular, near the wells treated by bioremediation, the amounts of electron acceptors, such as NO₃ ^?, Fe³⁺, and SO₄ ^2?, markedly decreased. The product of the microbial degradation process, HCO₃ ^?, gradually increased in the treated wells. Activities of the enzyme catechol 1, 2-dioxygenase decreased, while those of catalase increased. Reductions in pH, from 7.4 to 6.1, were recorded, and the groundwater environment became further deoxidized.     

Distribution of total petroleum hydrocarbons in Dezful aquifer,Southwest of Iran

Determination of total petroleum hydrocarbon distribution (TPH) in groundwater of Dezful aquifer was the main purpose of this study. The study area, which is located between latitudes 32°00′ and 32°35′?N and longitudes 48°10′ and 49°35′?E, covers about 1,920 km² in the north of Khuzestan Province, Iran. Hydrocarbon pollutants in the area were being released into the aquifer, from a variety of sources. An oil pipe crash accident, which occurred on 19 Feb. 2009 in the vicinity of the northern part of the study area, released about 6,000 barrels of crude oil to the Karkhe River. Other possible sources of TPH in the region are asphalt factories, gas stations, and the Sabzab oil pump station. Since the main source of drinking water in the Dezful area is groundwater reservoirs, this study would be very crucial, especially when there is considerable agricultural activity in the area as well. In order to determine the presence of TPH and heavy metals in the groundwater, samples were taken from wells with different usage within two periods, i.e., in Nov. 2008 and May 2009. The second sampling operation was carried out to determine the effect of the accident in the water resources. In situ groundwater parameter measurements including pH, dissolved oxygen, temperature, and electrical conductivity were also carried out in the field. Based on the results, there are four zones in the study area which were contaminated with TPH from different origins: (1) southeast of Dezful City, which was contaminated by Shokati gas station; (2) southeast of Shush City, which was contaminated by an asphalt factory; (3) southwest of Dezful City, which was contaminated by Sabzab oil pump station; and (4) the shores of Karkhe River which were contaminated due to the pipeline crash accident. This could be a serious threat to the environment and human health because TPH concentration was higher than the EPA standard in the study area. Heavy metals were not distributed in a uniform pattern in the aquifer. The concentrations were lower than the contamination level based on the EPA drinking standard, and there was no meaningful relation between concentrations of TPH and the heavy metals. It was recommended that a monitoring network should be designed to monitor oil contaminants in the ground and surface water monthly because of importance of the water resources and presence of potential oil contaminant sources.     

Application research of enhanced in-situ micro-ecological remediation of petroleum contaminated soil

Experimental study of enhanced in-situ micro-ecological remediation of petroleum contaminated loess soil was carried out in Zhongyuan oil production areas, and the enhanced in-situ micro-ecological remediation technique includes optimistic in-situ microbial communities, physical chemistry methods, alfalfa planting and regulation of soil environmental elements. Experiments showed that the oil content in the contaminated soil with oil content about 2 898.25 mg/kg can be reduced about 98.61% after in-situ micro-ecological remediation for 99 days, which demonstrated the effectiveness of in-situ micro-ecological remediation methods for petroleum contaminated soil in central plains of China, and explored the practical and feasible application of these methods.     

某石油类污染场地地下水石油烃生物降解的地球化学证据

生物降解地下水石油烃会改变地下水环境的水化学组成,因此可以通过分析污染晕中电子受体、生物降解代谢产物以及重要的地球化学参数量值变化获得生物降解的地球化学证据。本次在对某石油污染场地地质、水文地质条件、污染源污染方式调查基础上,根据地下水样测试结果,详细分析了地下水石油烃污染分布特征、污染晕中指示生物降解作用的电子受体、代谢产物以及重要地球化学参数的空间变化规律,研究结果表明:污染场地内存在氧还原、硝酸盐还原、硫酸盐还原等生物降解作用,其中硫酸盐还原是污染场地地下水石油烃生物降解的优势反应; 在沿地下水流向上,TPH浓度、HCO₃^-浓度和碱度逐渐降低,Eh、电子受体(DO、NO₃^-、SO₄^2-)浓度逐渐升高; 在垂直于地下水流向上,从中心向两侧各组分也呈相似的变化规律。     

基于多相流数值模拟的某石油污染场地地下水中VOCs自然衰减过程识别及能力评估

监测自然衰减(monitoring natural attenuation,MNA)技术是目前普遍认可的去除地下水中挥发性有机污染物(volatile organic compounds,VOCs)的技术。但受其修复周期长、监测费用昂贵等因素的影响,实地开展MNA技术修复污染场地具有一定的局限性。基于此,本研究运用多相流数值模拟手段识别了某石油污染场地内典型VOCs污染物(苯、甲苯、萘)在地下水中的自然衰减过程并评估了其自然衰减能力。结果表明:采用TMVOC所建立的多相流数值模拟模型能较好地预测和识别VOCs在地下水中的衰减规律;在研究区中,苯、甲苯和萘由于理化性质差异,在地下水中的污染羽分布特征不同,其自然衰减过程受挥发、吸附和生物降解作用的影响程度也不同;挥发和生物降解作用对VOCs自然衰减的影响程度均为苯>甲苯>萘,而吸附作用对VOCs自然衰减的影响程度为萘>甲苯>苯;在污染源被阻断的前提下,苯、甲苯和萘分别在泄漏发生7.0、6.5和6.0年后通过自然衰减达到理想去除效果。本文研究成果可以为水文地质条件类似的VOCs污染场地MNA修复方案的制定和修复效果评估提供理论支撑。     

Reducing leachable petroleum hydrocarbon concentration in weathered fuel oil contaminated soil by chemical oxidation with hydrogen peroxide

Number 6 fuel oil is one of the most used energy sources for electricity generation. However, leaks can contaminate soil and also groundwater due to leaching. At old sites, the oil may have low toxicity but still contaminate groundwater with foul-tasting compounds even at low concentrations. The purpose of this study was to evaluate the feasibility of applying H₂O₂ to reduce the leaching potential of a fuel oil contaminated soil. A silt-loam soil was collected from a contaminated thermal-electric plant with a hydrocarbon concentration of 3.2% in soil producing 4.3 mg/l in leachate. Hydrogen peroxide was applied (0.1, 0.2, 0.3, 0.6, 1.2% dry weight basis), and petroleum hydrocarbons were measured in soil and leachate pre- and post-treatment (72 h). At first, the soil and leachate concentrations diminished linearly (24.4 and 27.3% in soil and leachate, respectively). This was followed by a phase in which the concentration in leachate diminished greatly (75.8%) although the concentration in soil was reduced only moderately (15.1%). Overall, hydrocarbons in leachates were reduced 82.4% even though concentrations in soil were only reduced 35.8%. Correlation analysis showed that at only 1.0% w/w H₂O₂ a concentration of petroleum hydrocarbons in leachate safe for human consumption (≤ 1 mg/l) could be obtained even with a final hydrocarbon concentration in soil > 2%. Thus, this study presents an alternative strategy for remediation of fuel oil contaminated soils in urban environments that protects water sources by focusing on contamination in leachates, without spending extra financial resources to reduce the hydrocarbon concentration in low-toxicity soil.     

Use of a forensic geochemical technique and a hydrogeological assessment to determine the source of petroleum hydrocarbon contamination at a foreshore site in Korea

Investigations undertaken in 2013 at a foreshore site that was historically used for ship repairs in Korea indicated that soil and groundwater were contaminated with petroleum hydrocarbons over an area of 10,000 m². Groundwater investigations and modelling were undertaken to determine the characteristics of aquifers beneath the site, including the direction and rate of groundwater flow and the extent of dispersion of total petroleum hydrocarbon contamination. To identify petroleum products residing in contaminated soils, the hump of an unresolved complex mixture on the baseline found in the data of gas chromatography was used as a new method. Based on objective facts obtained from basic investigation, the current state of contamination, and discrimination of oil species, the polluter as specified in the Korean Soil Environment Conservation Act was identified.