Identifying and Quantifying Active Denitrifiers in Complex Environments Using Functional Gene Expression Analysis
使用功能基因表达分析识别和量化复杂环境中的活性反硝化菌
基本信息
- 批准号:0853864
- 负责人:
- 金额:$ 30万
- 依托单位:
- 依托单位国家:美国
- 项目类别:Standard Grant
- 财政年份:2009
- 资助国家:美国
- 起止时间:2009-05-15 至 2012-12-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
0853864de los Reyes IIIDetermining the identity of microorganisms responsible for specific biotransformation processes remains one of the major challenges in environmental microbiology and environmental engineering. The classic approach of using 16S rRNA gene sequences to detect and analyze bacterial communities in environmental samples without isolation and cultivation is not possible when studying denitrifying bacteria, because they are phylogenetically diverse. This is a major issue, given the importance of denitrification in water and wastewater pollution, greenhouse gas emission, and ozone layer destruction. This proposal is to develop and to optimize a novel molecular technique to determine the phylogenetic identity of functionally active nitrate-, nitrite-, nitric oxide-, and nitrous oxide-reducers in activated sludge and soil. Specific objectives are: (1) to optimize the Sequential mRNA FISH Flow Cytometry (SmRFF) method (developed previously in our lab) and investigate its potential for other microbial processes; (2) to determine the effect of floc structure, DO and carbon and nitrogen levels on the active denitrifying community; (3) to identify ammonia oxidizing bacteria (AOB) that denitrify; and (4) to examine the denitrifying ecology of a bioreactor that uses denitrifying AOB for treating high ammonia, low COD wastewater. The Sequential mRNA FISH-Flow Cytometry (SmRFF) method holds tremendous promise and may lead to significant results, as well as open new areas of research. This proposed project will be among the first studies to perform the following: (1) separate cells that are actively expressing the nitrate-, nitrite-, nitric oxide-, and nitrous oxide ?reducing genes; (2) capture nearly full-length 16S rRNA genes of active denitrifiers; and (3) apply combinatorial flow cytometry to identify AOB that are expressing the nirS and nirK genes. Determining the minimum level of mRNA copies needed for mRNA FISH will allow the more general application of SmRFF to other microbial ecology areas, such as bioremediation. The proposed research if successful would be a potentially significant methodological advance that would impact general microbial ecology as well as biological processes in environmental engineering. This project will contribute to the multidisciplinary education in the areas of microbiology, molecular biology and engineering. The PI has mentored 24 REU students in the last 7 years, and will continue to expose undergraduates to research and the excitement of discovery. This project will also potentially benefit municipal and industrial wastewater treatment plants that perform nitrogen removal. In addition, results from this project will be incorporated in summer workshops for high school students, and microbiology workshops for wastewater treatment professionals that are regularly organized by the PI.
洛斯雷耶斯III确定负责特定生物转化过程的微生物的身份仍然是环境微生物学和环境工程的主要挑战之一。使用16 S rRNA基因序列来检测和分析环境样品中的细菌群落而无需分离和培养的经典方法在研究细菌时是不可能的,因为它们具有遗传多样性。这是一个主要问题,因为脱氮在水和废水污染、温室气体排放和臭氧层破坏中的重要性。该建议是开发和优化一种新的分子技术,以确定活性污泥和土壤中功能活性硝酸盐,亚硝酸盐,一氧化氮和一氧化二氮还原剂的系统发育身份。具体目标是:(1)优化SmRFF方法(2)确定絮体结构、溶解氧和碳氮水平对活性硝化菌群的影响;(3)鉴定硝化菌群中的氨氧化细菌(AOB);(4)考察了厌氧氨氧化生物反应器处理高氨氮、低COD废水的厌氧生态学。顺序mRNA FISH流式细胞术(SmRFF)方法具有巨大的前景,可能会导致显着的结果,以及打开新的研究领域。这个拟议的项目将是第一批研究执行以下:(1)分离细胞,积极表达硝酸盐,亚硝酸盐,一氧化氮和一氧化二氮?减少基因;(2)捕获活性细菌的几乎全长16 S rRNA基因;和(3)应用组合流式细胞术鉴定表达nirS和nirK基因的AOB。确定mRNA FISH所需的mRNA拷贝的最低水平将允许SmRFF更普遍地应用于其他微生物生态学领域,如生物修复。这项研究如果成功,将是一个潜在的重大方法学进步,将影响一般的微生物生态学以及环境工程中的生物过程。该项目将有助于微生物学、分子生物学和工程学领域的多学科教育。PI在过去的7年里指导了24名REU学生,并将继续让本科生接触研究和发现的兴奋。该项目还将使进行脱氮的市政和工业废水处理厂受益。此外,该项目的成果将纳入高中生暑期讲习班和由PI定期组织的废水处理专业人员微生物学讲习班。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Francis de los Reyes其他文献
Francis de los Reyes的其他文献
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{{ truncateString('Francis de los Reyes', 18)}}的其他基金
NSF IRES Track I: Filling the gaps in WaSH research through field-based student research experiences in Malawi
NSF IRES 第一轨:通过马拉维学生的实地研究经验填补 WaSH 研究的空白
- 批准号:
2246372 - 财政年份:2023
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$ 30万 - 项目类别:
Standard Grant
RAPID: Monitoring for SARS-CoV-2 in municipal wastewater and sewage to elucidate infection dynamics across major metropolitan areas of the United States
RAPID:监测城市废水和污水中的 SARS-CoV-2,以阐明美国主要大都市区的感染动态
- 批准号:
2029025 - 财政年份:2020
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$ 30万 - 项目类别:
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Planning Grant: Engineering Research Center on Sanitation and Water Infrastructure of the Future for Marginalized Communities (SWIFt-MC)
规划拨款:边缘化社区未来卫生和水基础设施工程研究中心 (SWIFt-MC)
- 批准号:
1937085 - 财政年份:2019
- 资助金额:
$ 30万 - 项目类别:
Standard Grant
Using Microbial Ecology Theory to Understand Microbial Community Dynamics and Improve Function of Anaerobic Bioreactors
利用微生物生态学理论了解微生物群落动态并改善厌氧生物反应器的功能
- 批准号:
1805666 - 财政年份:2018
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$ 30万 - 项目类别:
Standard Grant
Inducing Aerobic Granulation in Continuous-Flow Reactors using Shear Variability
利用剪切变化在连续流反应器中诱导好氧造粒
- 批准号:
1336544 - 财政年份:2013
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$ 30万 - 项目类别:
Standard Grant
Ecophysiology of Nitrifying and Denitrifying Microbial Communities and their Interactions in Microbial Flocs
硝化和反硝化微生物群落的生态生理学及其在微生物絮体中的相互作用
- 批准号:
0348392 - 财政年份:2004
- 资助金额:
$ 30万 - 项目类别:
Continuing Grant
CAREER: Molecular and Engineering Approaches for Analyzing Microbial Selection in Activated Sludge: Competition between Filaments and Floc-formers
职业:分析活性污泥中微生物选择的分子和工程方法:细丝和絮凝剂之间的竞争
- 批准号:
0092851 - 财政年份:2001
- 资助金额:
$ 30万 - 项目类别:
Continuing Grant
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