||Sensitive Assay, Based on Hydroxy Fatty Acids from Lipopolysaccharide Lipid A, for Gram-Negative Bacteria in Sediments.
Parker, Jeffrey H. ;
Smith, Glen A. ;
Fredrickson, Herbert L. ;
Vestal, J. Robie ;
White, David C. ;
||Florida State Univ., Tallahassee. ;Cincinnati Univ., OH. Dept. of Biological Science.;Environmental Research Lab., Gulf Breeze, FL.;National Science Foundation, Washington, DC.;National Oceanic and Atmospheric Administration, Rockville, MD. Office of Sea Grant.
||EPA-R-807321 ;NSF-OCE80-19757; EPA-600/J-82-326;
Fatty acids ;
Marine biology ;
Lipopolysaccharide lipid A
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Biochemical measures have provided insight into the biomass and community structure of sedimentary microbiota without the requirement of selection by growth or quantitative removal from the sediment grains. This study used the assay of the hydroxy fatty acids released from the lipid A of the lipopolysaccharide in sediments to provide as estimate of the gram-negative bacteria. The method was sensitive to picomolar amounts of hydroxy fatty acids. The recovery of lipopolysaccharide hydroxy fatty acids from organisms added to sediments was quantitative. The lipids were extracted from the sediments with a single-phase chloroform-methanol extraction. The lipid-extracted residue was hydrolyzed in 1 N HCL, and hydroxy fatty acids of the lipopolysaccharide were recovered in chloroform for analysis by gas-liquid chromatography. This method proved to be about fivefold more sensitive than the classical phenol-water or trichloroacetic acid methods when applied to marine sediments. By examination of the patterns of hydroxy fatty acids, it was also possible to help define the community structure of the sedimentary gram-negative bacteria.