Novel Approaches to Studying the Situ Bioremediation of Complex Mixtures

研究复杂混合物原位生物修复的新方法

• 批准号: 7916299
• 负责人: ROLF U HALDEN
• 金额: $ 22.82万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-17 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7916299
• 关键词: Address; Affect; Anthracenes; Area; Aromatic Polycyclic Hydrocarbons; Bacteria; Baltimore; Behavior; Biochemical Genetics; Biological Assay; Biomass; Bioremediations; Carbon; Chemicals; Cloning; Communities; Complex; Complex Mixtures; Culture Techniques; Development; Devices; Dioxins; Environment; Environmental Pollution; Enzymes; Exposure to; Fingerprint; Genomics; Goals; Growth; Hazardous Waste Sites; Heating; Heavy Metals; In Situ; Indigenous; Individual; Knowledge; Laboratories; Maryland; Mass Spectrum Analysis; Metric; Monitor; Mono-S; Paper; Particle Size; Peptides; Phylogenetic Analysis; Polychlorinated Biphenyls; Positioning Attribute; Process; Proteins; Proteomics; Public Health; Reproducibility; Research; Research Personnel; Retrieval; Risk Management; Sequence Analysis; Simulate; Site; Source; Spatial Distribution; Sphingomonas; Structure; Techniques; Technology; Testing; Work; analog; base; cost; denaturing gradient gel electrophoresis; design; dibenzo(1,4)dioxin; dibenzofuran; dioxin dioxygenase; exposed human population; feeding; improved; innovation; interest; member; microbial; microbial community; microorganism; microorganism interaction; miniaturize; new technology; novel; novel diagnostics; novel strategies; phenanthrene; pollutant; programs; protein aminoacid sequence; remediation; research study; response; stressor; success; superfund site; tool

The vast majority of Superfund sites in the U.S and the Baltimore/Chesapeake Bay area contain mixtures of organic and inorganic compounds that contaminate underlying aquifers. The environmental fate of these contaminants, and ultimately human exposure to them, is governed primarily by their interactions with microorganisms, which?individually or as a community?drive the process of in situ bioremediation. Whereas single pollutant/microorganism interactions can be determined easily in the lab, no satisfactory tools exist for predicting the fate of mixtures in the environment. Our long-term goal is to improve the success rate of bioremediation at sites containing complex chemical mixtures by using in situ microcosm array (ISMA) technology. The ISMA is a field-deployable, miniaturized laboratory consisting of a large number of small microcosms arranged in parallel. Upon deployment, incubation, and retrieval from a groundwater well, the ISMA can be analyzed to reveal the impact of mixture components on the rates of Dollutant degradation and on the structure and function of microbial communities. We hypothesize that the ISMA can aid in the design of bioremediation strategies because: (1) individual ISMA microcosms can be amended with multiple test substances to elucidate the effect of mixture components on microorganisms; (2) the response of microorganisms to presented compounds manifests itself as changes in biomass, community structure and function; and (3) these changes can be detected conveniently by biochemical, genetic and proteomic strategies. Based on the above observations, the specific aims of the project are to: 1. Determine the reproducibility and discriminatory power of the ISMA technology. We will explore how varying concentrations of inducers and co-contaminants in synthetic groundwater modulate the expression of the dioxin dioxygenase (DDase) of Sphingomonas wittichii Strain RW1, by using a large number of replicates in conjunction with semi-automated, high-throughput proteomic mass spectrometry. 2. Demonstrate in controlled laboratory conditions how the ISMA can reveal additive, synergistic, and antagonistic effects of mixture components on microbial communities. We will use defined mixtures of chemicals (e.g., dioxins, PCBs, PAHs, tolualdeyhyde, Cd, Cr, Co, Pb, Hg, Zn, Ni) and bacteria (five bioremediation agents) to study these effects in various permutations. 3. Evaluate the utility of the ISMA technology in simulated field conditions. We will conduct ISMA laboratory experiments using nonsterile groundwater, sediment, and natural microbial communities. 4. Deploy the ISMA device at a Maryland Superfund site. We will assess the utility of the new technology in a field demonstration study by examining survival of and DDase expression by Strain RW1 in situ, and by elucidating the impact of this introduced bacterium on the indigenous microbial community at the site.

美国和巴尔的摩/切萨皮克湾地区的绝大多数超级基金站点都包含以下内容的混合物： 污染地下含水层的有机和无机化合物。这些物质的环境命运 污染物以及最终人类对它们的接触主要取决于它们与 微生物——单独或作为一个群体——驱动原位生物修复的过程。 尽管可以在实验室中轻松确定单一污染物/微生物的相互作用，但仍无法令人满意 存在用于预测环境中混合物的命运的工具。我们的长期目标是改善 使用原位微观世界在含有复杂化学混合物的地点进行生物修复的成功率 阵列（ISMA）技术。 ISMA 是一个可现场部署的小型实验室，由大型实验室组成 平行排列的小微观世界的数量。在部署、孵化和检索后 地下水井，可以分析 ISMA 以揭示混合物成分对速率的影响 轻微降解及其对微生物群落结构和功能的影响。我们假设 ISMA 可以帮助设计生物修复策略，因为：(1) 单个 ISMA 微观世界可以 用多种测试物质进行修正，以阐明混合物成分对微生物的影响； (2) 微生物对所呈现的化合物的反应表现为生物量的变化， 群落结构和功能； (3)这些变化可以通过生化方便地检测到， 遗传和蛋白质组学策略。根据上述观察，该项目的具体目标是： 1. 确定 ISMA 技术的再现性和辨别力。我们将探讨如何 合成地下水中不同浓度的诱导剂和共污染物可调节 维氏鞘氨醇单胞菌菌株 RW1 的二恶英双加氧酶 (DDase)，通过使用大量重复 与半自动、高通量蛋白质组质谱联用。 2. 在受控实验室条件下演示 ISMA 如何揭示相加、协同和 混合物成分对微生物群落的拮抗作用。我们将使用定义的混合物 化学品（例如二恶英、多氯联苯、多环芳烃、甲苯醛、镉、铬、钴、铅、汞、锌、镍）和细菌（五种 生物修复剂）来研究这些不同排列的影响。 3. 评估 ISMA 技术在模拟现场条件下的实用性。我们将进行ISMA实验室 使用非无菌地下水、沉积物和天然微生物群落进行实验。 4. 在马里兰州超级基金站点部署 ISMA 设备。我们将评估新技术的实用性 通过原位检查菌株 RW1 的存活率和 DDase 表达进行现场示范研究，并通过 阐明这种引入的细菌对该地点本土微生物群落的影响。