High cattle densities, expanding human settlements and the conversion of miombo woodland into farms and teak plantations are threatening wildlife populations in the Kilombero Valley, Tanzania, and conservation research on this internationally important wetland is required as part of an integrated approach to its future management. The effect of land-use change on antelopes (family Bovidae) was investigated by surveying tracks and dung during three seasons over 1999–2000 in an area of mixed land-use. Use of miombo woodland, grassland and farmland habitats by antelopes was highest during the wet season (April–May), probably representing the movements of animals away from the floodplain. Duiker, puku Kobus vardoni and reedbuck Redunca spp. predominantly used the farmland during the wet season, at which time buffalo Syncerus caffer were more common in the miombo woodland. The findings of this study have three main implications for the conservation of the valley. Firstly, the inadvertent provision of suitable wet season habitats for puku and other small-medium antelopes by rice farmers could lead to higher levels of illegal hunting, and may increase the potential for conflict between agriculture and wildlife. Secondly, the loss of miombo vegetation will most strongly affect the larger species of antelope (sable Hippotragus niger and waterbuck Kobus ellipsiprymnus), which favour open-woodland habitats; future work should therefore determine levels of habitat use by antelopes in and around maturing teak plantations. Thirdly, any management prescriptions to conserve the Kilombero Valley should include the land on the edge of the floodplain.