The objective of this study was to assess the frequency of use of marine resources in recruitment of Southern Hemisphere native riverine fish Galaxias maculatus from rivers across a latitudinal gradient. To do this, we analysed the concentrations of δ34S in vertebral column tissues from fish collected in ten Chilean river systems across latitudes 36°–47°S. The analyses of δ34S signatures in these rivers suggest that the use of marine resources by riverine populations of G. maculatus in large river systems in Chile is variable, with marine resources playing a limited role in more northern large rivers, characterised by warmer temperatures and predictable flow regimes and floodplain inundations. This is in contrast to life histories described for G. maculatus in rivers from New Zealand and Australia, where riverine populations are believed to be characterised by an obligatory recruitment phase in marine environments. Recruitment of G. maculatus in Chilean large rivers appears to depend on their freshwater productivity driven by climate as well as both longitudinal (headwaters lakes-estuary) and lateral (main channel-floodplains) hydrologic connectivities.