Methane oxidising activity and community structure of 11, specifically targeted, methanotrophic species have been examined in an arable soil. Soils were sampled from three different field plots, receiving no fertilisation (C), compost (G) and mineral fertiliser (M), respectively. Incubation experiments were carried out with and without pre-incubation at elevated CH4 mixing ratios (100 ml CH4 l−1) and with and without ammonium (100 mg N kg−1) pre-incubation. Four months after fertilisation, plots C, G and M did not show significant differences in physicochemical properties and CH4 oxidising activity. The total number of methanotrophs (determined as the sum the 11 specifically targeted methanotrophs) in the fresh soils was 17.0×106, 13.7×106 and 15.5×106 cells g−1 for treatment C, G and M, respectively. This corresponded to 0.11 to 0.32% of the total bacterial number. The CH4 oxidising activity increased 105-fold (20–26 mg CH4 g−1 h−1), the total number of methanotrophs doubled (28–76×106 cells g−1) and the methanotrophic diversity markedly increased in treatments with a pre-incubation at elevated CH4 concentrations. In all soils and treatments, type II methanotrophs (62–91%) outnumbered type I methanotrophs (9–38%). Methylocystis and Methylosinus species were always most abundant. After pre-incubation with ammonium, CH4 oxidation was completely inhibited; however, no change in the methanotrophic community structure could be detected.