Download Assessment of Monitored Natural Attenuation for Environmental Restoration at SRS

Assessment of Monitored Natural
Attenuation for Environmental
Restoration at SRS
College of Science, Mathematics, and
Engineering Technology
Research Symposium 2006
Investigators
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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
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Read with standard CD reader
Possible to sample for hundreds of agents
Passive environmental monitoring
Homeland security monitoring