Activity of Type I Methanotrophs Dominates under High Methane Concentration: Methanotrophic Activity in Slurry Surface Crusts as Influenced by Methane, Oxygen, and Inorganic Nitrogen.

Livestock slurry is a major source of atmospheric methane (CH), but surface crusts harboring methane-oxidizing bacteria (MOB) could mediate against CH emissions. This study examined conditions for CH oxidation by in situ measurements of oxygen (O) and nitrous oxide (NO), as a proxy for inorganic N transformations, in intact crusts using microsensors. This was combined with laboratory incubations of crust material to investigate the effects of O, CH, and inorganic N on CH oxidation, using CH to trace C incorporation into lipids of MOB. Oxygen penetration into the crust was 2 to 14 mm, confining the potential for aerobic CH oxidation to a shallow layer. Nitrous oxide accumulated within or below the zone of O depletion. With 10 ppmv CH there was no O limitation on CH oxidation at O concentrations as low as 2%, whereas CH oxidation at 10 ppmv CH was reduced at ≤5% O. As hypothesized, CH oxidation was in general inhibited by inorganic N, especially NO, and there was an interaction between N inhibition and O limitation at 10 ppmv CH, as indicated by consistently stronger inhibition of CH oxidation by NH and NO at 3% compared with 20% O. Recovery of C in phospholipid fatty acids suggested that both Type I and Type II MOB were active, with Type I dominating high-concentration CH oxidation. Given the structural heterogeneity of crusts, CH oxidation activity likely varies spatially as constrained by the combined effects of CH, O, and inorganic N availability in microsites.