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Abstract Nitrogen (N) is a necessary element of soil fertility and a limiting nutrient in tallgrass prairie but grazers like bison and cattle can also recycle N. Bison and cattle impact the nitrogen (N) cycle by digesting forage that is consumed, and recycled back to the soil in a more available forms stimulating soil microbial N cycling activities. Yet we do not know how both grazers comparatively affect N cycling in tallgrass prairie. Thus, we investigated if bison and cattle had similar impacts on N cycling in annually burned tallgrass prairie relative to ungrazed conditions over a 3-year period (2020–2022) at the Konza Prairie Biological Station. We examined: soil pH, soil water content, mineralized N, nitrification potential, denitrification potential and extracellular enzyme assays. Interannual variability in precipitation controlled soil water and N cycling microbial activities but grazing effects had a stronger influence on N cycling. We found significant differences and increased soil pH, nitrification and denitrification potential and less N limitation in bison vs cattle grazed soils where bison grazed soils exhibited faster N cycling. Differences between the grazers may be attributed to the different management of bison and cattle as both can impact N cycling. Overall, these data provide some evidence that bison and cattle affect N cycling differently at this study site, and improve the ecological understanding of grazer impacts on N cycling dynamics within the tallgrass prairie ecosystem.