Using geometric morphometrics, the skull and otolith of tiger catfish Pseudoplatystoma metaense were analysed to identify population structure in tributaries of the Apure River (i.e. the Sarare, Caparo, Guanare, Portuguesa and San Carlos Rivers) in the Orinoco basin, Venezuela. The analyses show uniformity in skull and otolith shapes of P. metaense within and among four tributaries, with only the Caparo River showing significant differences. Within the Apure basin, the stock of P. metaense was differentiated through spawning, refuge and nursery areas. This study concludes that populations of P. metaense from each major tributary in the Orinoco basin should be considered as part of a metapopulation system for management purposes. Human disturbances in the catchment have directly reduced the spawning areas available to this species, decreased the total biomass and changed the spatial distribution of spawning areas.