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: 7291652
• 负责人: ROLF U HALDEN
• 金额: $ 8.63万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-28 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7291652
• 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; Condition; Count; 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; Numbers; Paper; Particle Size; Peptides; Phylogenetic Analysis; Polychlorinated Biphenyls; Polymerase Chain Reaction; Positioning Attribute; Process; Proteins; Proteomics; Public Health; Rate; Reproducibility; Research; Research Personnel; Retrieval; Risk Management; Score; Sequence Analysis; Simulate; Site; Source; Spatial Distribution; Sphingomonas; Structure; Techniques; Technology; Testing; United States Environmental Protection Agency; Work; analog; anthracene; base; cost; denaturing gradient gel electrophoresis; design; desire; 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地点包含的混合物 有机和无机化合物污染了潜在的含水层。这些的环境命运 污染物，最终是人类接触它们的人，主要受其与其互动的影响 微生物，哪些是个人或作为社区？推动原位生物修复的过程。 尽管可以在实验室中轻松确定单污染物/微生物的相互作用 存在用于预测环境中混合物命运的工具。我们的长期目标是改善 使用原位缩影在含有复杂化学混合物的位点生物修复的成功率 阵列（ISMA）技术。 ISMA是一个野外剥离，微型实验室，由大型实验室组成 并联排列的小缩影数量。部署，孵化和从 地下水井，可以分析ISMA，以揭示混合物成分对 美元降解以及微生物群落的结构和功能。我们假设 ISMA可以帮助设计生物修复策略，因为：（1）单个ISMA缩影可以是 用多种测试物质修改，以阐明混合物成分对微生物的影响； （2） 微生物对提出化合物的反应表现为生物量的变化， 社区结构和功能； （3）可以通过生化方便地检测到这些变化， 遗传和蛋白质组学策略。基于上述观察结果，该项目的具体目的是： 1。确定ISMA技术的可重复性和歧视能力。我们将探讨如何 合成地下水中不同浓度的诱导剂和共抗合体调节 通过使用大量的重复，鞘氨拟补酶的二氧酶（DDase） 结合半自动化的高通量蛋白质组学质谱法。 2。在受控的实验室条件下证明ISMA如何揭示添加剂，协同作用和 混合成分对微生物群落的拮抗作用。我们将使用定义的混合物 化学物质（例如二恶英，PCB，PAHS，TOLUALDEYDE，CD，CR，CO，CO，PB，PB，HG，HG，ZN，NI）和细菌（五个 生物修复剂）在各种排列中研究这些效果。 3。评估ISMA技术在模拟现场条件下的效用。我们将进行ISMA实验室 实验使用非地下水，沉积物和天然微生物群落的实验。 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 通过菌株RW1原位检查和DDase表达的存活，以及通过现场示范研究 阐明了这种细菌对现场土著微生物群落的影响。