Ecological and biogeographical studies of Neotropical non-marine ostracods are rare, although such information is needed to develop reliable paleoecological and paleoclimatic reconstructions for the region. An extensive, yet little explored South American area of paleoclimatic interest, is the arid-semiarid ecotone (Arid Diagonal) that separates arid Patagonia from subtropical/tropical northern South America, and lies at the intersection of the Pacific and Atlantic atmospheric circulation systems. This study focused on the Laguna Llancanelo basin, Argentina, a Ramsar site located within the Arid Diagonal, and was designed to build a modern dataset using ostracods (diversity, spatial distribution, seasonality, habitat preferences) and water chemistry. Cluster and multivariate analysis of the data indicated that salinity is the most significant variable segregating two ostracod groups. Limnocythere aff. staplini is the only species that develops abundant populations in the saline ephemeral Laguna Llancanelo during almost all seasons, and is accompanied by scarce Cypridopsis vidua in summer. The latter species is abundant in freshwater lotic sites, where Ilyocypris ramirezi, Herpetocypris helenae, and Cyprididae indet. are also found in large numbers. Darwinula stevensoni, Penthesilenula incae, Heterocypris incongruens, Chlamydotheca arcuata, Chlamydotheca sp., Herpetocypris helenae, and Potamocypris smaragdina prefer freshwater lentic conditions (springs), with C. arcuata and Chlamydotheca sp. found only in the Carapacho warm spring, which has a year-round constant temperature of ~20 °C. Seasonal sampling was necessary because some taxa display a highly seasonal distribution. Species that were recorded have either subtropical or Patagonian affinities, although a few taxa are endemic or common to both regions. These data can serve as modern analogues for reconstructing the late Quaternary history of the area, and to investigate the extent and position of the arid/semiarid ecotone (Arid Diagonal) during past glacial/interglacial cycles.Instituto de Limnología "Dr. Raúl A. Ringuelet