Habitat connectivity is vital to the persistence of migratory fishes. Native tropical island stream fish assemblages composed of diadromous species require intact corridors between ocean and riverine habitats. High dams block fish migration, but low-head artificial barriers are more widespread and are rarely assessed for impacts. Among all 46 drainages in Puerto Rico, we identified and surveyed 335 artificial barriers that hinder fish migration to 74.5% of the upstream habitat. We also surveyed occupancy of native diadromous fishes (Anguillidae, Eleotridae, Gobiidae, and Mugilidae) in 118 river reaches. Occupancy models demonstrated that barriers 2 meters (m) high restricted nongoby fish migration and extirpated those fish upstream of 4-m barriers. Gobies are adapted to climbing and are restricted by 12-m barriers and extirpated upstream of 32-m barriers. Our findings quantitatively illustrate the extensive impact of low-head structures on island stream fauna and provide guidance for natural resource management, habitat restoration, and water development strategies.