US EPA

9093914 

Meetings & Symposia 

Use of molecular methods for characterizing and tracking the dominant members of a chlorinated solvent cometabolizing mixed culture 

Dolan, ME; Giovannoni, SJ; Semprini, L 

2001 

Preprints of Papers, Presented at the National Meeting - American Chemical Society, Division of Environmental Chemistry
ISSN: 0093-3066 

AMER CHEMICAL SOC 

WASHINGTON 

41 

1260-1261 

English 

WOS:000168824802745 

A clone library was prepared from a frozen aliquot of a butane-utilizing mixed culture that will be used to bioaugment an aquifer test zone at Moffett Federal Airfield, CA, to enable co-oxidation of 1,1,1-trichloroethane (TCA), 1,1-dichloroethylene (DCE), and trichloroethylene (TCE). When sequences of the clones were compared to a database of known organisms, 64 out of 86 clones were found to be similar to Rhodococcus, Acidovorax, Hydrogenophaga, or an unknown bacterium found in a deep subsurface clay. Complete sequences of the 16S rDNA from representatives of the four groups were used to develop rRNA probes for fluorescence in situ hybridization analysis. Two restriction enzymes were chosen for terminal restriction fragment length (T-RFLP) polymorphism analysis based on computer analysis of the fully sequenced clones and sequences from similar organisms. Microcosms were prepared from groundwater and aquifer material obtained aseptically from the field site. T-RFLP analyses of the clones and the bioaugmentation culture were compared to samples obtained from indigenous and bioaugmented microcosms that have been enriched with butane and stressed with TCA, TCE, and DCE. An isolate was obtained from the mixed culture that utilized butane and degraded TCA and had the same T-RFLP signature as the Rhodococcus clone. This is an abstract of a paper presented at the 221st ACS National Meeting (San Diego, CA 4/1-5/2001).