This study contributes to the knowledge of central European clitellates by creating a check-list of Oligochaeta (sensu oligochaetous Clitellata; Erséus 2005) of the Czech Republic, exclusive of taxa in the family Enchytraeidae. In total, 95 aquatic oligochaete species representing 43 genera are reported for the Czech Republic. Rare species are highlighted and associated with the categories for threatened species as outlined by the International Union for the Conservation of Nature. The first records of Trichodrilus strandi Hrabě, 1936, Pristina jenkinae (Stephenson, 1931), Pristina osborni (Walton, 1906), Rhyacodrilus subterraneus Hrabě, 1963, Aulodrilus limnobius Bretscher, 1899, and Aulodrilus pigueti Kowalewski, 1914 in the Czech Republic are presented. Their ecology, morphology, and distribution are discussed.