Abstract Assessment of medium- and large-sized mammals’ species diversity, abundance, distribution, and habitat association is necessary to promote their conservation measures. The study was conducted in the former Dhidhessa wildlife sanctuary (FDWS), to assess the medium- and large-sized mammals’ composition, distribution, and diversity. Transect lines were used to collect the data. A total of 1,146 ± 91 individuals belonging to 25 species, seven orders, and 14 families were recorded. Among these, 607 ± 186.5 (52.97%) were recorded during the wet season, whereas 539 ± 144.5 (47.03%) during the dry season. The order Carnivora was the most diverse one, followed by Artiodactyla and Primates, respectively, while Rodentia, Hyracoidea, Tubulidentata, and Lagomorpha were the least diverse orders. Olive baboon (Papio anubis) was the most abundant species (19.2%), followed by Hippopotamus (Hippopotamus amphibious) (17.63%) and Vervet monkey (Chlorocebus aethiops) (13.18%). Species diversity was higher in the savanna-wooded than in the riverine forest during both the wet (H’ = 2.72, Fisher’s alpha = 6.33) and dry (H’= 2.79, Fisher’s alpha = 6.26) seasons. Species evenness was higher in the savanna-wooded habitat than in the riverine forest habitat during both the wet (J’ = 0.87 and J’ = 0.77, respectively) and dry (J’ = 0.9 and J’ = 0.80, respectively) seasons. The highest similarity in species composition was between the wet and dry seasons in the riverine forest habitat (SI = 85.58%). The lowest similarity was recorded between the dry and wet seasons of the riverine forest and the wet season of the savanna wooded habitat (SI = 45.46% and SI = 46.49%, respectively). The study suggests that mammals are at risk of anthropogenic factors in the area. So, an urgent conservation measure that involves the participation of the local community has paramount significance for the sustainable conservation of mammals in the former Dhidhessa wildlife sanctuary.