In order to investigate the likely contact of the Mediterranean and Saharan faunas, we surveyed bats in six different habitats of Bou Hedma National Park for 1 year. We sampled bats monthly by mist-netting and echolocation call detection at two basins, Bordj and Nouh, the Bou Hedma wadi, and open and dense acacia forest and steppe. We identified 13 species, among which Pipistrellus kuhlii, Eptesicus isabellinus, and Miniopterus schreibersii were the most abundant and frequent, followed by Tadarida teniotis, Plecotus gaisleri, and two Saharan species, Rhinopoma cystops and Otonycteris hemprichii. We also rarely recorded four rhinolophids: Rhinolophus euryale, R. ferrumequinum, R. hipposideros, and R. mehelyi. Bat activity was recorded throughout the year in the six habitats, water bodies being more attractive than dry habitats for both Mediterranean and Saharan taxa. The wadi was the preferred habitat of R. cystops, T. teniotis, and O. hemprichii. The open acacia forest was attractive only during the flowering season. Species richness and bat activity peaked in late spring and reached the lowest values, but was still present, during winter.