Increasing aridity usually results in decreasing bat abundance and species richness, and the Saharan desert is an example of such impoverishment. Moreover, the harsh climatic and field conditions in this area restrict the feasibility of surveys. Therefore, the bat fauna of the Tunisian Sahara was unstudied until an expedition was conducted in May 2021. A total of seven species were recorded using passive bat detectors set mainly at water bodies, which concentrate bat activity. Echolocation calls of these species did not depart from published records for the Mediterranean area. Our data failed to identify the two ecomorphotypes of Pipistrellus kuhlii but confirmed the adaptability of this species to local conditions. For the first time in Tunisia, we recorded echolocation calls of Taphozous nudiventris, a species that has been rarely reported in North Africa. The highest species richness was identified at the largest wetlands of Oued Daghsen (Oued Dkouk Nature Reserve) and at Bordj el Khadra (only six and four species, respectively). Most Mediterranean species were recorded in the northern locality, while desert species were more active in the most southern one. The importance of water bodies for bats was confirmed once more. No sign of competition among species was detected, as the Mediterranean and desert-dwelling bat species were active at the same time. Additional surveys should be planned in order to enlarge the list of seven species recorded in only one week.