The effects of repeated synthetic fertilizer or cattle slurry applications at annual rates of 50, 100 or 200 m 3 ha À1 yr À1 over a 38 year period were investigated with respect to herbage yield, N uptake and gross soil N dynamics at a permanent grassland site. While synthetic fertilizer had a sustained and constant effect on herbage yield and N uptake, increasing cattle slurry application rates increased the herbage yield and N uptake linearly over the entire observation period. Cattle slurry applications, two and four times the recommended rate (50 m 3 ha À1 yr À1 , 170 kg N ha À1 ), increased N uptake by 46 and 78%, respectively after 38 years. To explain the long-term effect, a 15 N tracing study was carried out to identify the potential change in N dynamics under the various treatments. The analysis model evaluated process-specific rates, such as mineralization, from two organic-N pools, as well as nitrification from NH 4 þ and organic-N oxidation. Total mineralization was similar in all treatments. However, while in an unfertilized control treatment more than 90% of NH 4 þ production was related to mineralization of recalcitrant organic-N, a shift occurred toward a predominance of mineralization from labile organic-N in the cattle slurry treatments and this proportion increased with the increase in slurry application rate. Furthermore, the oxidation of recalcitrant organic-N shifted from a predominant NH 4 þ production in the control treatment, toward a predominant NO 3 À production (heterotrophic nitrification) in the cattle slurry treatments. The concomitant increase in heterotrophic nitrification and NH 4 þ oxidation with increasing cattle slurry application rate was mainly responsible for the increase in net NO 3 À production rate. Thus the increase in N uptake and herbage yield on the cattle slurry treatments could be related to NO 3 À rather than NH 4 þ production. The 15 N tracing study was successful in revealing process-specific changes in the N cycle in relationship to long-term repeated amendments.