Forest soil carbon (C) pools may act as sinks for, or sources of, atmospheric carbon dioxide, while nitrogen (N) fertilization may affect the net exchange of C in forest ecosystems. Since all major C and N processes in soil are driven by soil microorganisms, we evaluated the effects of N fertilization on biomass and bacterial and fungal activity in soils from three Norway spruce forests with different climatic and N availability conditions. N deposition and net N mineralization were higher at the sites in southern Sweden than at the site in northern Sweden. We also studied the extent to which N fertilization altered the nutrient(s) limiting bacterial growth in soil. We found that on average microbial biomass was reduced by similar to 40% and microbial activity by similar to 30% in fertilized plots. Bacterial growth rates were more negatively affected by fertilization than fungal growth rates, while fungal biomass (estimated using the phospholipid fatty acid (PLFA) 18:2 omega 6,9) decreased more than bacterial biomass as a consequence of fertilization. The microbial community structure (indicated by the PLFA pattern) was changed by fertilization, but not in the same way at the three sites. Soil bacteria were limited by a lack of carbon in all forests, with the carbon limitation becoming more evident in fertilized plots, especially in the forests that had previously been the most N-limited ones. This study thus showed that the effects of N fertilization differed depending on the conditions at the site prior to fertilization. (c) 2007 Elsevier Ltd. All rights reserved.