Within freshwater fishes, closely related species produce alarm cues that are chemically similar, leading to conserved antipredator responses. Similar conservation trends are predicted for species with geographically isolated populations. Here, we tested this hypothesis with the guppy ( Poecilia reticulata Peters, 1859) from two populations within the Aripo River, Trinidad. Free-ranging guppies in the Lower Aripo (high-predation population) exhibited more risk-aversive inspection behaviour towards a fish predator model paired with the alarm cues of guppies collected from the same population versus a river water control. In comparison, when paired with the alarm cues of guppies from the Upper Aripo (low-predation population), the response was intermediate. In the laboratory, we tested Upper and Lower Aripo guppies to the alarm cues of the same or different Aripo River donors, Quaré River guppies (a high-predation population from a different drainage), or a water control. Both Upper and Lower Aripo River guppies exhibited the highest intensity response to donors from the same population and the lowest intensity response to Quaré River donors, with the response to different Aripo donors being intermediate. Collectively, these results demonstrate a trend of intraspecific conservation of chemical alarm cue production, leading to population-specific responses to conspecific cues.