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Use of the MIDI‐FAME technique to characterize groundwater communities
Author(s) -
Glucksman A.M.,
Skipper H.D.,
Brigmon R.L.,
Domingo J.W. Santo
Publication year - 2000
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1046/j.1365-2672.2000.01058.x
Subject(s) - groundwater , biomass (ecology) , extraction (chemistry) , pseudomonas putida , environmental science , fatty acid , filter (signal processing) , acinetobacter , environmental chemistry , chemistry , biology , chromatography , food science , microbiology and biotechnology , ecology , bacteria , biochemistry , geology , antibiotics , genetics , geotechnical engineering , computer science , computer vision
Fatty acid methyl ester (FAME) profiles were identified directly from groundwater microbial communities concentrated on and extracted with polycarbonate filters. The sensitivity of this direct extraction method was determined using pure cultures of Acinetobacter junii , Pseudomonas putida and Stenotrophomonas maltophilia . A minimum concentration of 10 7 cells filter −1 was required to identify the predominant fatty acids from each culture. However, at least 3·7 × 10 9 cells filter −1 were required to obtain fatty acid profiles that matched the signature profiles for pure cultures in a commercial database. While several saturated fatty acids (i.e. 14 : 0, 16 : 0, 18 : 0) were extracted from the polycarbonate filters, they were readily subtracted from microbial fatty acid profiles and did not interfere with the characterization of pure cultures or environmental samples. For the environmental samples, 3 l of groundwater from the Savannah River Site, Aiken, SC, (USA) contained sufficient biomass for direct extraction. A comparative analysis of FAME groundwater profiles demonstrated a qualitative difference among communities sampled from spatially discrete locations, while a groundwater well that was sampled at two time points showed strong similarities over time. Concentration of microbial biomass on polycarbonate filters coupled with the MIDI‐FAME extraction of both biomass and filter was a useful technique to characterize microbial communities from groundwater.