Abstract The conservation of fish assemblages depends on the longitudinal and lateral connectivity between riverine habitats, in particular during the breeding season and the initial development. This study investigated the composition and spatio-temporal structure in the ichthyoplankton of the hydrographic basin of the Paraguay River in western Brazil to identify the local spawning grounds and the importance of the longitudinal connectivity of economically valuable migratory species. Data were collected at 10 sites between two breeding seasons (2017/2018 and 2018/2019). Were collected 8,635 larvae, representing 55 taxa in 25 families, including the migratory species Brycon hilarii, Hemisorubim platyrhynchos, Piaractus mesopotamicus, Prochilodus lineatus, Pseudoplatystoma spp., Salminus brasiliensis, Sorubim lima, and Zungaro jahu, which are important fishery resources, with the highest larval densities being recorded between November and January. The Sepotuba, Paraguay, Jauru, and Cabaçal rivers were the areas of greatest connectance, and should thus be considered critical for the conservation of the longitudinal connectivity of this fluvial system, indicating that the migratory species spawn upriver. More efficient fisheries management mechanisms are needed, respecting the spawning period of migratory species, maintaining quality and longitudinal connectivity between habitats, and characteristics necessary for successful larval recruitment.
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