Understanding migratory requirements is critical for the conservation of diadromous fishes. Tupong (Pseudaphritis urvillii) are diadromous fishes found in freshwater and estuarine regions of south-eastern Australia. Previous studies have hypothesised that mature female tupong undertake downstream spawning migrations from freshwater to the estuary or sea, with a compensatory return of juveniles, and possibly spent fish, back upstream. We applied acoustic telemetry to test this hypothesis. We tagged 55 female tupong in two river systems in Victoria, Australia, and tracked movements for 4- to 6-month periods over 2 years. Thirty-one fish undertook rapid downstream migrations and then appeared to move through the estuary and out to sea between May and August in each year. Migration was associated with relatively high river discharges, and movement from the estuary to the sea tended to occur most frequently during intermediate moon phases. Low rates of movement between September and April suggested that tupong inhabited restricted home ranges outside of the spawning season. The apparently disparate migratory patterns of female (catadromous) and male (non-diadromous) tupong are rare amongst fishes globally. Differential exertion of a range of selective pressures may have resulted in the evolution of sexual differences in migratory modes in this species.