In this study, nitrous oxide (N2O) emissions from a full-scale denitrifying biofilter plant were continuously monitored over two periods (summer campaign in September 2014 and winter campaign in February 2015). Results of the summer campaign showed that the major part (N99%) of N2O flux was found in the liquid phase and was discharged with the effluent. N2O emissions were highly variable and represented in average 1.28 ± 1.99% and 0.22 ± 0.31% of the nitrate uptake rate during summer and winter campaigns, respectively. Denitrification was able to consume a large amount of dissolved N2O coming from the upstream nitrification stage. In the absence of methanol injection failure and with an influent BOD/NO3-N ratio higher than 3, average reduction of N2O was estimated to be of 93%. The control of exogenous carbon dosage is essential to minimize N2O production from denitrifying biofilters, in correlation to NO2-N concentrations in the filter.