Freshwater ecosystems and their associated biota have been negatively impacted by the human development of water resources. Fundamental to restoration activities for target species is an understanding of the factors affecting population decline or recovery. Within Australia's Murray-Darling Basin, recovery efforts to address the population decline of native freshwater fish include stock enhancement, habitat restoration, and the delivery of environmental water. Essential to guiding future management actions is information to assess the efficacy of these efforts. We undertook a study to investigate whether natural spawning and recruitment, stock enhancement, or a combination of the two is contributing to sustaining populations of golden perch (Macquaria ambigua) in the highly regulated Lachlan River, Australia. Otolith microchemistry and genetic analyses were used as complementary tools to determine the source (hatchery origin or wild-spawned) of existing populations in the catchment. We identified that natural spawning and recruitment was contributing to riverine populations in some years but that populations were heavily reliant on stocking. It was not possible to distinguish hatchery and wild-born fish using genetic tools, highlighting the value of using multiple lines of evidence to establish causal mechanisms contributing to population recovery.