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Title: Novel tools for in situ detection of biodiversity and function of dechlorinating and uranium-reducing bacteria in contaminated environments

Author
item HAUER, K - Technical University Of Munich
item MEISINGER, D - Technical University Of Munich
item PAVLEKOVIC, M - Technical University Of Munich
item THOMAS, S - Georgia Institute Of Technology
item KNIGGENDORF, A - Technical University Of Munich
item Chee Sanford, Joanne
item SANFORD, ROBERT - University Of Illinois
item LEBRON, C - Technical University Of Munich
item LIEBL, W - Technical University Of Munich
item LOEFFLER, FRANK - University Of Tennessee
item LEE, N - Technical University Of Munich

Submitted to: Geochimica et Cosmochimica Acta
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/1/2012
Publication Date: 6/1/2009
Citation: Hauer, K., Meisinger, D.B., Pavlekovic, M., Thomas, S.H., Kniggendorf, A.K., Chee Sanford, J.C., Sanford, R.A., Lebron, C.A., Liebl, W., Loeffler, F.E., Lee, N. 2012. Novel tools for in situ detection of biodiversity and function of dechlorinating and uranium-reducing bacteria in contaminated environments. Geochimica et Cosmochimica Acta. 73:A502.

Interpretive Summary: Molecular biology-based methods for detection and identification of microorganisms responsible for reduction of toxic heavy metals and radionuclides provides a sensitive and accurate means to characterize populations in contaminated environmental systems. For an intelligent remediation design, reliable analytical tools for detection of relevant species are needed, such as PCR. However, PCR cannot visualize its targets and thus provide information about the morphology, distribution and association of the targeted species (e.g. to other cells or to surfaces like minerals). One key populations, Anaeromyxobacter spp., are known for their abilities to reduce metals (e.g. Fe cycling), halogenated aromatic compounds, and possess unique N-metabolism. Anaeromyxobacter serves as a useful model to develop molecular-based tools that can be applied to a variety of environments including complex soil matrices. Here, we report our efforts to employ whole-cell-targeted tools using fluorescence in situ hybridization (FISH) and microscopy, focusing on Fe/Mn and radionuclide transforming species, in particular, Anaeromyxobacter dehalogenans. The significance of this study are the useful probes that resulted in detecting several Fe/U reducing species for reliable identification. FISH is a better indicator of activity/viability than general viability stains, and provides a useful tool to complement existing PCR-based methods for detection of relevant bacteria in the environment.

Technical Abstract: Toxic heavy metals and radionuclides pose a growing, global threat to the environment. For an intelligent remediation design, reliable analytical tools for detection of relevant species are needed, such as PCR. However, PCR cannot visualize its targets and thus provide information about the morphology, distribution and association of the targeted species (e.g. to other cells or to surfaces like minerals). Here, we report our efforts to employ whole-cell-targeted tools using fluorescence in situ hybridization (FISH) and microscopy, focusing on Fe/Mn and radionuclide transforming species. Novel hierarchic FISH probe sets have been designed for several Fe/U reducing species for a reliable top to bottom identification. FISH is a better indicator of activity/viability than general viability stains, and may provide additional information to PCR based results.