Different lines of investigation have recently contributed to increasing the available knowledge about the invertebrates inhabiting inland waters of north Africa, but a comprehensive synopsis on Tunisian Ostracoda is missing to date. An updated checklist of Recent non-marine ostracods from Tunisia and data on their distribution is thus offered here, representing the most extensive survey on this crustacean group ever carried out in inland waters throughout the country. One-hundred-five sites covering various climate zones, from Mediterranean to desert areas, were sampled between 2002 and 2012. Most of the considered water bodies were temporary or ephemeral habitats, but a few permanent sites were sampled as well. Overall, 18 genera and 32 taxa of putative species rank were collected in the frame of this survey, among which nine species and five genera were new to Tunisian fauna. As a result of this study and based on previous investigations, nine families (Candonidae, Cyprididae, Cytherideidae, Darwinulidae, Ilyocyprididae, Leptocytheridae, Limnocytheridae, Loxoconchidae, Paradoxostomatidae), 29 genera and at least 45 species of non-marine ostracods are currently known for Tunisia, which thus prove to host the most diverse ostracod fauna among north African countries. The number of species occurring in a single sample varied from 1 to 4. The Eucypris virens complex was the most widespread taxon (58 records), followed by Heterocypris barbara (30 records), Heterocypris incongruens (22 records), and Sarscypridopsis aculeata (16 records). For some ostracod species, clear distributional gradients associated with different climatic conditions were observed. The affinities with adjacent Maghrebian ostracod faunas are discussed. This study confirms the crucial role played by marginal aquatic habitats for the conservation of biodiversity, in particular in arid and semi-arid regions.