Skip to main content
U.S. Department of Energy
Office of Scientific and Technical Information

Fluorescent antibody application in bioremediation procedures at the Savannah River Site

Conference ·
OSTI ID:469156

Direct Fluorescent Antibodies (DFA) and Most Probable Number (MPN) techniques are currently being employed at the Savannah River Site to monitor methanotrophic bacteria for the bioremediation of trichloroethylene (TCE) in field studies. Direct Fluorescent Antibodies were developed against various methanotrophic bacteria isolated from SRS as well as methanotrophic bacteria acquired from the American Type Culture Collection (ATCC). DFA`s are anticipated to be more efficient for monitoring methanotroph activity than MPN`s because of shorter processing time, lower cost, and the direct nature of the assay. The DFA method is a direct technique, in that samples are processed immediately and can be enumerated within an hour. The MPN method is indirect, since samples must be cultured for 6-8 weeks before measuring methane consumption and carbon dioxide production. Indirect methods are not highly selective and have limited application. The greatest advantage of a faster assay, is that bioremediation procedures utilizing methanotrophic bacteria could be amended. These amendments would be based on environmental monitoring with results in real time (1 hour). The elimination of the MPN technique and the use of DFA`s will save significantly on both materials and labor. The data obtained from the DFA`s and MPN`s were statistically compared to each other and to total bacterial counts (AODC). The statistical analysis used was Analysis of Variants (ANOVA). Using this analysis, groundwater samples were found to be not significantly different; whereas soil were significantly different. These methods were employed on soil samples from the Southern Sector and ground water samples from the TCE-contaminated Sanitary Landfill at SRS. Acridine Orange Direct Counts were compared to show relative differences between total bacterial and methanotroph population.

Research Organization:
Westinghouse Savannah River Co., Aiken, SC (United States)
Sponsoring Organization:
USDOE, Washington, DC (United States)
DOE Contract Number:
AC09-96SR18500
OSTI ID:
469156
Report Number(s):
WSRC-MS--96-0773; CONF-970170--1; ON: DE97004966
Country of Publication:
United States
Language:
English