The Arvicolinae from the latest Ruscinian and early Villanyian sites of the section of Tollo de Chiclana (Southern Spain) are re-examined in the light of new material and exhaustive comparisons with other European populations. The uppermost Ruscinian population from TCH-1B, previously assigned to Mimomys stehlini, is now ascribed to M. hassiacus, cited for the first time from southern Spain. The populations from the lower Villanyian localities of TCH-3 and 13, assigned to M. minor, are considered to belong to M. stehlini. In addition, the material from the uppermost Ruscinian karstic level of Mo1-A, assigned to “Mimomys” occitanus is ascribed to M. hassiacus. The presence of different species of Arvicolinae is key for delimiting the Ruscinian–Villanyian boundary in the continental deposits of southern Spain and correlating them with European biochronological schemes. In this paper we discuss the problems associated with specific identifications of upper Ruscinian and lower Villanyian arvicolines from Spain, pointing out the invalidity of many citations, and demonstrating the need for an extensive revision of these faunas.