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There is a perception that sustained frequent fires cause nitrogen limitation over the long term (50–100 y) by volatilizing the nitrogen in soil, plant biomass and litter. Here we test this perception in a South African savanna located in the Kruger National Park. At our study site we compare the effects of 50 y of fire exclusion, season (August and February) and frequency (triennial and annual August and triennial February) of burn on nitrogen cycling and availability. We do this using three different methods to determine nitrogen mineralization; in situ incubations, laboratory incubations and ion-exchange resin bags. On each treatment we established two parallel transects 100 m apart with 10 sampling points per treatment along these transects. Daily mineralization rates for in situ incubations were determined monthly from August 2004 to June 2005 at each of the sampling points. Ion-exchange resin bags were buried (5 cm) at the same points and left in the field from August 2004 to August 2005. In February 2005 five randomly located soil samples from each of the four treatments were collected for laboratory incubations using a 7-cm-diameter soil auger. Regardless of method used our results show that there are no significant differences in daily nitrogen mineralization rates after 50 y of different burning treatments from annual burning to fire exclusion. In fact, both in situ and laboratory incubations show that nitrogen availability is higher on the annual burn than the fire exclusion (0.16 ?g g?1 soil d?1 vs. 0.11 ?g g?1 soil d?1 and 0.46 ?g g?1 soil d?1 vs. 0.30 ?g g?1 soil d?1 respectively). Perceived negative effects of fire on ecosystem functioning has curbed the use of fire as a management tool with fire often actively suppressed in savanna. The results of our study show that fire can be used more vigorously in mesic African savanna to manipulate tree:grass ratios without negatively affecting the nitrogen cycle.