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Characterization of Reductive-Dechlorinating Microbial Communities Using a Combination of Fluorescent In Situ Hybridization and PCR-Based Molecular Tools

EPA Grant Number: U915559
Title: Characterization of Reductive-Dechlorinating Microbial Communities Using a Combination of Fluorescent In Situ Hybridization and PCR-Based Molecular Tools
Investigators: Richardson, Ruth E.
Institution: University of California - Berkeley
EPA Project Officer: Thompson, Delores
Project Period: December 1, 1999 through December 1, 2001
Project Amount: $73,516
RFA: STAR Graduate Fellowships (1999)
Research Category: Academic Fellowships , Fellowship - Engineering , Engineering and Environmental Chemistry

Description:

Objective:

The objective of this research project is to understand the microbial community structure of a mixed culture that fully dechlorinates chlorinated aliphatic hydrocarbons to ethene. Additional characterization of a partially dechlorinating culture (which produces vinyl chloride as the primary end-product) will provide evidence as to which populations are essential for the final and most important (from a public health standpoint) dechlorination step that converts vinyl chloride to ethene. By using a combination of molecular tools, it is possible to get a more complete picture of community structure than any single method would allow.

Approach:

In this study, two dechlorinating mixed cultures will be characterized and compared by a combination of 16S rDNA-based molecular methods: terminal restriction fragment length polymorphisms (T-RFLP), RFLP and sequencing of individual clones from clone libraries constructed from amplified community DNA, and fluorescent in situ hybridization (FISH). The cultures will originate from a soil sample from Alameda Naval Air Station that is contaminated with a combination of hydrocarbons and trichloroethene (TCE). One culture completely dechlorinates TCE and perchloroethene (PCE) to ethene, while the second, daughter culture dechlorinates only to vinyl chloride (VC), a known human carcinogen.

Expected Results:

This method of using a combination of molecular tools to characterize mixed cultures will make it possible to get a more complete picture of community structure than any single method would allow.

Supplemental Keywords:

reductive dechlorination, Dehalococcoides, chlorinated ethenes, TCE, FISH, T-RFLP, community structure, bioremediation. , Ecosystem Protection/Environmental Exposure & Risk, Water, Scientific Discipline, Waste, RFA, Remediation, Engineering, Chemistry, & Physics, Restoration, Chemical Engineering, Aquatic Ecosystem Restoration, Hazardous Waste, Environmental Engineering, Environmental Chemistry, Groundwater remediation, Hazardous, Bioremediation, dehalogenation, bioaugmentation, biodegradation, microbial degradation, biotechnology, groundwater, monitoring, contaminated aquifers, aquifer remediation design, reductive dehalogenation, groundwater pollution, fluorescent in situ hybridization, in-situ bioremediation, bioremediation model, microbiology, groundwater contamination, degrade trichloroethylene, TCE, in situ treatment, reductive dechlorination, TCE degradation, advanced treatment technologies, contaminated groundwater, hazardous waste treatment, dehalogenate, dechlorination, in situ remediation, aquifer remediation

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The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.

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