Stable isotope pulse-chasing and compound specific stable carbon isotope analysis of phospholipid fatty acids to assess methane oxidizing bacterial populations in landfill cover soils

The oxidation of methane by bacteria residing in soils constitutes an important terrestrial methane sink. These bacteria are particularly abundant in the covering soils of landfill caps due to the supply of high concentrations of methane from the landfill below. Only about 0.1% of soil bacteria are amenable to available methods of culturing, resulting in the need for a method of in situ analysis. A combination of phospholipid fatty acid (PLFA) analysis and stable isotopic labeling has been employed in this investigation as a means of cultivation-independent bacterial analysis. Soil samples taken from the profiles of two landfill caps, one of clay and one of sand, were incubated with C-13-labeled methane. PLFAs were analyzed by gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) in order to determine their C-13 content, from which the PLFA distribution of the methane-oxidizing bacteria was calculated. Neither landfill cap supported communities of bacteria capable of oxidizing ambient levels of methane but only those elevated levels that are usually attributable to landfills. The clay-capped landfill profile exhibited a change in the methane-oxidizing bacterial community with depth, whereas the sand-capped landfill site displayed a mixture of both type I and II methanotrophs throughout the profile. Two additional samples, taken from sites where methane production was evident, were particularly dominated by type II methanotrophic bacteria.