Chemical energy can be recovered from municipal wastewater as biogas through anaerobic treatment. Effluent from direct anaerobic wastewater treatment at low temperatures, however, still contains ammonium and considerable amounts of dissolved methane. After nitritation, methane can be used as electron donor for denitrification by the anaerobic bacterium ‘Candidatus Methylomirabilis oxyfera’. It was shown that in the presence of 0.7% O2, denitrifying methanotrophic activity slightly increased and returned to its original level after oxygen had been removed. At 1.1% O2, methane consumption rate increased 118%, nitrite consumption rate increased 58%. After removal of oxygen, methane consumption rate fully recovered, and nitrite consumption rate returned to 88%. Therefore, traces of oxygen that bacteria are likely to be exposed to in wastewater treatment are not expected to negatively affect the denitrifying methanotrophic process. 2.0% O2 inhibited denitrifying activity. Nitrite consumption rate decreased 60% and did not recover after removal of oxygen. No clear effect on methane consumption was observed. Further studies should evaluate if intermittent addition of oxygen results in increased growth rates of the slow-growing ‘Candidatus Methylomirabilis oxyfera’.