A strategy for aromatic hydrocarbon bioremediation under anaerobic conditions and the impacts of ethanol: a microcosm study

Increased use of ethanol-blended gasoline (gasohol) and its potential release into the subsurface have spurred interest in studying the biodegradation of and interactions between ethanol and gasoline components such as benzene, toluene, ethylbenzene and xylene isomers (BTEX) in groundwater plumes. The preferred substrate status and the high biological oxygen demand (BOD) posed by ethanol and its biodegradation products suggests that anaerobic electron acceptors (EAs) will be required to support in situ bioremediation of BTEX. To develop a strategy for aromatic hydrocarbon bioremediation and to understand the impacts of ethanol on BTEX biodegradation under strictly anaerobic conditions, a microcosm experiment was conducted using pristine aquifer sand and groundwater obtained from Canadian Forces Base Borden, Canada. The initial electron accepter pool included nitrate, sulfate and/or ferric iron. The microcosms typically contained 400 g of sediment, 600 approximately 800 ml of groundwater, and with differing EAs added, and were run under anaerobic conditions. Ethanol was added to some at concentrations of 500 and 5000 mg/L. Trends for biodegradation of aromatic hydrocarbons for the Borden aquifer material were first developed in the absence of ethanol, The results showed that indigenous microorganisms could degrade all aromatic hydrocarbons (BTEX and trimethylbenzene isomers-TMB) under nitrate- and ferric iron-combined conditions, but not under sulfate-reducing conditions. Toluene, ethylbenzene and m/p-xylene were biodegraded under denitrifying conditions. However, the persistence of benzene indicated that enhancing denitrification alone was insufficient. Both benzene and o-xylene biodegraded significantly under iron-reducing conditions, but only after denitrification had removed other aromatics. For the trimethylbenzene isomers, 1,3,5-TMB biodegradation was found under denitrifying and then iron-reducing conditions. Biodegradation of 1,2,3-TMB or 1,2,4-TMB was slower under iron-reducing conditions. This study suggests that addition of excess ferric iron combined with limited nitrate has promise for in situ bioremediation of BTEX and TMB in the Borden aquifer and possibly for other sites contaminated by hydrocarbons. This study is the first to report 1,2,3-TMB biodegradation under strictly anaerobic condition. With the addition of 500 mg/L ethanol but without EA addition, ethanol and its main intermediate, acetate, were quickly biodegraded within 41 d with methane as a major product. Ethanol initially present at 5000 mg/L without EA addition declined slowly with the persistence of unidentified volatile fatty acids, likely propionate and butyrate, but less methane. In contrast, all ethanol disappeared with repeated additions of either nitrate or ferric iron, but acetate and unidentified intermediates persisted under iron-enhanced conditions. With the addition of 500 mg/L ethanol and nitrate, only minor toluene biodegradation was observed under denitrifying conditions and only after ethanol and acetate were utilized. The higher ethanol concentration (5000 mg/L) essentially shut down BTEX biodegradation likely due to high EA demand provided by ethanol and its intermediates. The negative findings for anaerobic BTEX biodegradation in the presence of ethanol and/or its biodegradation products are in contrast to recent research reported by Da Silva et al. [Da Silva, M.L.B., Ruiz-Aguilar, G.M.L., Alvarez, P.J.J., 2005. Enhanced anaerobic biodegradation of BTEX-ethanol mixtures in aquifer columns amended with sulfate, chelated ferric iron or nitrate. Biodegradation. 16, 105-114]. Our results suggest that the apparent conservation of high residual labile carbon as biodegradation products such as acetate makes natural attenuation of aromatics less effective, and makes subsequent addition of EAs to promote in situ BTEX biodegradation problematic.     

地下水中氯代烃污染原位生物修复精细模拟研究

原位生物修复技术是修复地下水中挥发性氯代烃（VCHs）污染的重要手段之一.精细模拟修复过程是实现高效修复的关键.然而以往多数模拟VCHs修复过程的研究并未全面考虑VCHs自然衰减性质,势必会造成预测误差.因此,本研究针对当前VCHs原位生物修复模型的局限性,提出考虑VCHs自然衰减性质的原位生物修复Monod-Natural attenuation（M-Na）模型.该模型主要将自主开发的M-Na模型的源代码以用户自定义反应模块的形式嵌入到RT3D中.通过注入乳酸提供氢气进行原位生物修复含水层中四氯乙烯（PCE）污染,验证M-Na模型的精确性,并分析了修复过程参数变化（包括乳酸的注入浓度和速率）对修复效果的影响.算例分析结果表明,VCHs的自然衰减作用可以将修复产物乙烯（ETH）浓度提高10%左右,这在VCHs污染修复模拟过程中不可忽略;同时算例中乳酸的最佳注入浓度为30 mg·L^-1,最佳注入速率为200m³·d^-1.本研究成果有望为地下水VCHs污染原位生物修复精细模拟及高效修复提供模型参考.     

中原油田石油污染土壤原位生物修复技术实验研究

通过实验室选择性富集培养,从中原油田石油污染土壤中获得了能以中原原油为碳源快速生长的石油降解菌群。结合黑麦草（Ryegrass）和苜蓿（Alfalfa）,采用该降解菌群对原油污染土壤进行了原位生物联合修复实验。接入降解菌的实验区分种植黑麦草、种植苜蓿、未种植区,另设黑麦草区和空白区。经过99 d的生物修复,石油烃累计降...     

New bacterial strain of the genus Ochrobactrum with glyphosate-degrading activity

Thirty bacterial strains with various abilities to utilize glyphosate as the sole phosphorus source were isolated from farm soils using the glyphosate enrichment cultivation technique. Among them, a strain showing a remarkable glyphosate-degrading activity was identified by biochemical features and 16S rRNA sequence analysis as Ochrobactrum sp. (GDOS). Herbicide (3 mM) degradation was induced by phosphate starvation, and was completed within 60 h. Aminomethylphosphonic acid was detected in the exhausted medium, suggesting glyphosate oxidoreductase as the enzyme responsible for herbicide breakdown. As it grew even in the presence of glyphosate concentrations as high as 200 mM, Ochrobactrum sp. could be used for bioremediation purposes and treatment of heavily contaminated soils.     

乙醇对含水层中燃油芳香烃内在生物修复的潜在风险

内在生物修复,是在没有工程措施促进的情况下利用土著微生物降解含水层内灾害性物质的一种修复技术,在燃油烃污染管理方面具有显著的成本效益。该技术需要确定自然衰减过程,并能够继续提供有效的风险保护。针对燃油污染含水层,北美与欧洲认为内在生物修复是值得优先考虑的应用技术。然而,随着乙醇燃油的推广使用,我国在应用这样的经验时需要考虑乙醇的潜在影响。现有的文献研究表明乙醇存在能够阻止燃油主要污染物芳香烃(BTEX)的生物降解,降低水环境的pH值,并可能增强BTEX在水中的溶解性,或存在对生物的毒性,或因为乙醇降解降低介质的渗透性能。因此,需要更好地认识乙醇的潜在风险,为发展乙醇燃油污染含水层修复策略提供科学依据。     

Three Different Topics for Bioremediation Application on Soils and Waters Contaminated with Aromatic Compounds, by Using Immobilized Bacteria

Molecular techniques and modelling are presented as powerful tools required in the performance of efficient soil and water bioremediation systems. An Escherichia coli CC118-D strain was constructed by inserting the Klebsiella pneumoniae hpa B gene, coding for the unstable 3,4-dihydroxyphenylacetate 2,3-dioxygenase, into its chromosome. When the constructed strain was immobilized, both enzyme stability and viability increased along the studied period, in absence of antibiotic. We proposed this strategy as an approach to overcoming plasmid instability and to enhance enzyme activity and stability, avoiding antibiotic utilization. A model was developed to understand and predict the behaviour of bacteria and pollutants in a bioreactor system, considering: fluid dynamics, molecular/cellular scale processes and biofilm formation.     

微生物强化处理与堆制强化处理含油污泥对比试验

为探索含油污泥的高效处理方法,对含油量为12.68%的污泥进行微生物菌剂强化处理和堆制强化处理现场试验.在第1、15、30d分别向微生物强化处理单元投加微生物菌剂和营养液;同时向堆制强化单元投加10kg畜禽粪便来提供共降解物质,强化堆制处理过程.结果表明,微生物强化处理单元含油量持续下降,56d后下降至6.42%,去除率达到47%;强化堆制单元处理效果较好,56d后含油量下降至6.98%,去除率达到31%;对照单元含油量则变化很小,56d后含油量为10.15%.对照单元的pH基本保持不变,最高为8.28,最低为7.93,微生物强化处理单元pH最低达到7.33,一般稳定在7.8左右,菌剂的添加对pH影响明显.堆制强化处理单元温度随着农家肥的添加而剧烈升高,最高达到54℃.试验进行过程中监测了样品中微生物总数的变化情况.通过气谱-质谱分析,可以明显地看出2种方法处理后,含油污泥中石油成分的差异,对碳原子数小于21的烃类有着良好的去除效果.     

城市土壤污染研究现状与趋势

城市土壤是城市生态环境系统的有机组成部分,对城市的可持续发展有着重要的意义.城市化和工业化进程的加快,造成城市土壤污染、侵蚀、酸化和硬化以及土壤生物和植被退化等一系列较为严重的城市环境问题,直接危及到城市居民的健康和安全;因此,城市土壤污染问题已经引起人们的高度重视.近10多年来,国内外有关城市土壤重金属污染,微有机物污染,生源要素污染和城市土壤污染的环境与健康风险评价等诸多研究取得了一系列的研究成果.本文主要就以上研究成果进行了较为系统地总结与阐述,并且在此基础上提出了今后有关城市土壤污染研究的发展重点与趋势,以期为改善城市生态环境质量提供重要的科学依据.     

近岸海域石油污染生物修复技术的研究进展

生物修复技术是治理大面积污染区域的一种有价值的修复方法。对国内外最新的生物刺激和生物强化技术进行调研,总结了该技术对治理近岸海域石油污染的研究进展,分析总结了施加营养盐技术对海洋环境的影响,以及生物修复技术尚存在的一些局限性。     

高效石油烃降解菌的筛选及其对原油污染土壤的修复

从陕北原油污染土壤中筛选出7株高效石油烃降解菌,其中黄杆菌属CC-2、不动细菌属SC-5、假单胞菌属SC-6表现出较强的石油烃降解能力。通过单因素试验和正交试验考察总石油烃(TPH)降解效果的影响因素,得出各因素对TPH降解率影响程度的大小次序为:溶液p H降解温度降解菌接种量摇床转速,且在降解菌接种量为7%(φ)、溶液p H为7、降解温度为30℃、摇床转速为150 r/min的最适处理条件下,菌株SC-6的TPH降解率可达61.23%。原油污染土壤生物修复实验结果表明:高效石油烃降解菌的投加有利于土壤TPH降解率和酶活性的提高;"菌株SC-6+营养剂"组修复处理42 d后的TPH降解率可达57.59%。