Abstract European rabbits (Oryctolagus cuniculus L.) occur over large areas of eastern Australia, where they disturb significant quantities of soil while constructing warrens. Although the effects of rabbits on vegetation are generally well understood, little is known about their effects on soil physical or chemical properties. We studied the effect of rabbit warrens on soil chemistry in a semi-arid woodland supporting a high density of rabbit warrens. Within one large warren, we examined nutrient concentrations within three microsites (mound, inter-mound and an intermediate disturbed area) representing a gradient of increasing rabbit disturbance, and supported this with a study of soil nutrients with depth within 1-m-deep trenches through five warrens. Landscape-level changes in surface chemistry were also examined at an additional 23 warrens. At both patch and landscape scales, pH, electrical conductivity, soluble and exchangeable Ca2+ and K+, and total Al and Ca generally increased with increasing rabbit disturbance, while total C and S (LECO), total P and S (ICP), and soluble Na+ declined, and total N remained unchanged. Although chemical changes with depth were generally ill defined, surface soils tended to be more similar in their composition than deeper soils. Overall, our results reinforced the view that rabbits have a negative effect on surface soils in semi-arid woodlands, and suggested that restoration of the original woodland vegetation may be hampered by changes in soil biogeochemistry associated with the warrens.