Objectives: The overall objective of this work is to determine which microbial populations are present and metabolically active in aquatic environments. The specific objective of this project is to collect preliminary data demonstrating that bromodeoxyuridine can be coupled with fluorescence in situ hybridization targeting 16S ribosomal ribonucleic acid to simulaneously identify and measure the growth rate (based on DNA synthesis) of individual bacterial cells. Abstract: Currently, 16S ribosomal RNA-targeted methods are used to identify and enumerate bacterial cells in environmental samples. Although these methods provide quantitative information about the structure of microbial communities, they do not provide information about the biochemical function of these communities. DNA synthesis is a key biochemical function of actively growing microorganisms. Labeling of DNA synthesis with easy-to-detect molecular tags provides a means of studying growing microbial populations. We propose to use bromodeoxyuridine to label de novo DNA synthesis. We will combine fluorescence in situ hybridizations targeting 16S rRNA with BrdU labeling to simulataneously identify, enumerate, and measure the biochemical function (growth) of indivdual bacterial cells. This improved technology will provide critical information to assess microbial water quality.
Preliminary Evaluation of a New Technique for Linking Picoplankton Community Structure with Function in Aquatic Environments
Scope of Study
Scale of Phenomena
Research & Development
Surveillance and Monitoring