COMBINED NMR/OPTICAL MICROSCOPY OF BIOFILM PHYSIOLOGY STUDIES
EMSL Project ID
25694
Abstract
Bacterial biofilms are found nearly everywhere in nature. Microbial metabolism and phenotype vary rapidly within a biofilm - this is at least partly responsible for their metabolic diversity and potential for exploitation for bioenergy and environmental remediation. Detailed knowledge is lacking about the metabolic processes involved, due in part to: [i] the complexity of biofilms, which contain temporally evolving spatial concentration gradients for substrate, oxygen, organic acids, etc., and [ii] the inadequacy of current microbiological-metabolism methods which are destructive and typically yield no spatial resolution. The objectives of this two-year R21 project are to adapt, improve and apply unique combined nuclear magnetic resonance and fluorescent confocal (NMR/optical) microscopy instrumentation and techniques for time- and biofilm-depth-resolved metabolism studies of live oral biofilms. A two-step cultivation and measurement procedure will provide reproducible biofilm samples for repeatable depth-resolved metabolism measurements in a relevant (oral) growth environment. One and two-species biofilms will be studied in this fashion. NMR spectroscopic imaging and related methods will be employed to map biofilm metabolism as a function of depth with 10-micron resolution. Specific Aims are to:
1) adapt and improve combined NMR/optical microscopy to study oral biofilm physiology, and
2) employ the new capability to monitor dynamic processes relevant to caries in real time.
Oral biofilms were chosen because they constitute a logical system for developing new biofilm characterization technologies. Viable, functioning model oral biofilms containing up to 10 species have been defined - analogous environmental model communities are not yet available. As the majority of bacteria in nature exist in complex communities, knowledge gained from developing approaches to characterize oral biofilms can also be applied to DOE mission areas such as those focused on developing a fundamental understanding of microbial systems (the Genomics: GTL program) and applications of this knowledge to developing methods clean up the environment (the Natural and Accelerated Bioremediation Research (NABIR) program). Further, the non-invasive and non-contacting nature of NMR is ideal for the modeling of communities in hostile environments, such as acid mine drainage (AMD) communities.
This project will help to fulfull DOE Office of Science's investment plan to conduct comprehensive, multidisciplinary research programs focused on microbes and plants to drive scientific breakthroughs necessary for the development of cost effective biofuels and bioenergy production and to enable the design of microbes that address critical energy or waste cleanup needs
(http://www.science.doe.gov/News_Information/News_Room/2007/Budget/SC-DOE_DrOrbach_FY08BudgetTestimony_March72007-fin.pdf).
Project Details
Project type
Large-Scale EMSL Research
Start Date
2007-07-01
End Date
2010-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Chapter 22 Dynamic Metabolism Studies of Live Bacterial Films Paul D. Majors and Jeffrey S. McLean Magnetic Resonance Microscopy. Edited by Sarah Codd and Joseph D. Seymour Copyright © 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim ISBN: 978-3-527-32008-0
McLean JS, SJ Fansler, PD Majors, K Mcateer, LZ Allen, ME Shirtliff, R Lux, and W Shi. 2012. "Identifying Low pH Active and Lactate-Utilizing Taxa within Oral Microbiome Communities from Healthy Children Using Stable Isotope Probing Techniques." PLoS One 7(3):Article No. e32219. doi:10.1371/journal.pone.0032219