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Assessment of Monitored Natural Attenuation for Environmental Restoration at SRS College of Science, Mathematics, and Engineering Technology Research Symposium 2006 Investigators • • • • • Dr. Jim Payne Dr. Waltena Simpson Dr. John Williams 1 Graduate Student 7 Undergraduate Studnets Contract Research Project • Funding by DoE Savannah River Operations Office • Period of Performance – 1 year with 3 month extension completed 12/31/06 • Funding Level - $178,798 Project Deliverables • Task 1: Phytoremediation Contributions to Monitored Natural Attentuation of TCE in C- Area Plume Seeps • Task 2: Assessment of Technologies for Monitoring Organic and Inorganic Contaminants and Environmental Bacteria Dr. John Williams’ Co-PI efforts included training two SCSU Interns and overseeing their field work, collaboratively at SREL with Dr. Gary Mills. The hyporheic zone is the groundwater zone beneath a stream bed. Their focus was to measure contaminant degradation products and to collect microbial samples to analyze what bacteria were most important to natural attenuation in this important hyporheic zone. • Degradation of perchloroethylene (PCE) generates trichloroethylene (TCE) and dichloroethylene (DCE) •The following graph documents that MNA degradation is occurring in the deeper hyporheic zone. • VOC concentrations are much higher on the plume side of the stream compared to the opposing ‘clean’ side. •The presence of some PCE, TCE, and DCE in the hyporheic zone on the ‘clean’ side is probably due to differences in subsurface flow rates. The slope on the plume side is much steeper. Dr. Simpson’s duties included training an SCSU student in her laboratory in the sandwich ELISA technique. The goal of the student’s project was to determine if FITC-labeled antibodies could by used to detect the bacterium Methylosinus trichosporium. M. trichosporium is a methanotroph that is capable of degrading the environmental contaminant trichloroethylene (TCE). Due to its potential role in bioremediation, a rapid, cost-effective means of detecting it in soils is needed. A microplate fluorescence reader was utilized to detect bound FITC-labeled antibodies. Detection of M. trichosporium using FITC-labeled antibodies 3500 3241 3099.1 3000 2810.1 3294.85 3189.8 2898 2788.9 Relative Light Units (RLU) 2500 2000 Averages (RLU) 1500 1000 500 0.001 11.769 Blank Blank 0 (1:500) 1:250 Fitc (1:500) (1:500) No (1:250) 1:100 Fitc Bacteria A- 1:250 Fitc D 1:250 Fitc E-H 1:100 Fitc (1:250) 1:100 (1:250) No bacteria AB 1:250 Fitc G-H 1:100 Fitc • We were unable to detect Methylosinus trichosporium using the sandwich ELISA technique. • Background Relative Light Units for samples without M. trichosporium were comparable to those of samples containing M. trichosporium. • This may be due to possible strain differences between the M. trichosphorium used in our studies and that used in studies by SRNL. • Alternatively, the antibodies used may not be specific enough to detect M. trichosporium. Thin Film Bio-detector A Solid Substrate Immunological Assay: Detection of the Antigen-Antibody Reaction Ms. Adella Francis and Mr. James Ross undergraduate students at SCSU worked with Dr. Jim Payne on development and characterization of a passive detector suitable for monitoring air or liquid streams. THE DEVICE • Based on an existing concept patent with SRNL • Thin metallic film of indium on a plastic microscope slide • Antibody adheres to the metallic film forming the detector • Interaction of the antibody with the appropriate antigen effects the optical properties of the detector RESULTS • Optimum indium thickness – 100 nm • No sample degradation over long periods of time – samples from 1998 still work • Detection in the ppm range on these samples using FTIR • Presence of antigen effects reflectance thus making “ go-no go device” quite easy APPLICATIONS • • • • Read with standard CD reader Possible to sample for hundreds of agents Passive environmental monitoring Homeland security monitoring