Based on wildlife and domestic herbivore waterhole use count data collected in the South Eastern Lowveld of Zimbabwe between 2008 and 2011, we tested whether there was any significant overlap by wild and domestic herbivores in the use of waterholes located; 1) inside a wildlife protected area, 2) at the boundary of the protected and agricultural areas and 3) in the agricultural areas. We also explored whether species diversity at these three land uses vary significantly. Our results showed overlap in place and time between wild and domestic herbivores especially in the use of waterholes located at the boundary of the protected areas and at one waterhole located in the agricultural areas. Results also indicated that overall, 95% of wild herbivores prefered using waterholes located inside the protected area.Results of this study further demonstrated that animal species diversity declines significantly along a gradient from protected areas to agricultural areas. Overall, results of this study imply that the current emerging disease interface between wildlife and livestock may not necessarily be due to direct physical interaction between disease reservoir hosts or even sharing of same waterholes at the same time but rather an indirect contact. These findings could provide an important avenue of investigating livestock and wildlife disease outbreaks at the domestic -wildlife interface which are disturbing current efforts to improve livestock production as well as biodiversity conservation in African savannas.