Diadromous fishes are known biotransport vectors that can move nutrients, energy and contaminants in an upstream direction in lotic ecosystems. This function has been demonstrated repeatedly in anadromous salmonids, but the role of other diadromous species, especially tropical taxa, as biotransport vectors is less studied. Amphidromous fish species exhibit potential to act as upstream vectors of nutrients and contaminants in their postlarval and juvenile stages, but this role is largely unknown because of limited understanding of larval growth habitats. Moreover, because some species are harvested in artisanal fisheries as postlarvae, and postlarvae are consumed by riverine and estuarine predators, heavy contaminant loads may present a human or wildlife health concern. This research incorporates stable isotope and contaminant analyses to infer larval habitats and contaminant accumulation of amphidromous fishes on the Caribbean island of Puerto Rico. The isotopic signatures of postlarval amphidromous fishes indicated marine basal sources and food web components, rather than those from riverine habitats. Additionally, postlarvae did not contain concentrations of anthropogenic pollutants that would be of ecological or human health concern. These findings are the first and strongest evidence that amphidromous fish postlarvae function as biotransport vectors of marine nutrients into and up river ecosystems without posing a health threat to the receiving food web or human consumers.