A number of previous studies have shown that N fertilization often reduces respiration in forest soils. However, the durability of this effect has not been fully explored. In this study, the response of soil respiration to a single fertilization with urea, applied 27 years earlier, was examined in a field experiment located in a stand of Pinus sylvestris in central Sweden. The doses that had been added were 120, 240 and 600 kg N ha–1. Samples were taken from the humus layer and the upper 7.5 cm of the mineral soil. Sieved samples were incubated in the laboratory. No effect of the previous fertilization on soil respiration was found, thus indicating that the reduction shown in earlier studies is not persistent. There was a tendency that the highest N dose had caused a higher N concentration and a lower C/N-ratio in the humus layer and a higher C concentration in the mineral soil.