Catadromous fish species often use elevated flows to migrate from freshwater to downstream spawning grounds. However, in regulated river systems, artificial barriers and flow alteration can disrupt movement cues, affecting fish migration. Using a combination of acoustic telemetry, passive integrated transponder (PIT) tags and gonad maturity data, we assessed the migratory timing and flow requirements for downstream spawning migrations and upstream dispersal in two co-occurring mullet species in the Logan River, eastern Australia. Over 4 years, 141 adult sea mullet (Mugil cephalus) and 28 adult pinkeye mullet (Trachystoma petardi) were implanted with acoustic transmitters and broad-scale movements tracked using an array of 49 acoustic receivers. Juvenile upstream dispersal was monitored using implanted PIT tags and readers deployed in weir fishways. Sea mullet and pinkeye mullet undertook large-scale downstream migrations from February to March. Movements past weirs only occurred once sufficient flow had occurred and, in the case of pinkeye mullet, when gonads were mature. In contrast, juvenile dispersal into upstream habitats occurred primarily between November and April on low flows. This study advances our understanding of bidirectional movement ecology and flow requirements of two co-occurring mullet species to inform environmental flow management (e.g. to facilitate fish passage) in a regulated river system.