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The introduction of nonnative salmonids into Southern Appalachia may pose a threat to resident salamander populations. In recent years, the stocking and encroachment of rainbow trout (Oncorhynchus mykiss) into headwaters where the black-bellied salamander (Desmognathus quadramaculatus) and brook trout (Salvelinus fontinalis) naturally coexist has raised concerns. In aquatic prey, predator-avoidance responses are primarily influenced through the detection of chemical cues released from predators. The objective of this study was to determine how co-occurrence with a predator influences black-bellied salamander predator recognition behavior. To evaluate this, salamander activity metrics (general activity, number of movements, and latency to move) were recorded before and after exposure to either native trout predator cue (brook), introduced trout predator cue (rainbow), or conditioned tap water (control). Larvae were collected from different streams based on their trout predator assemblage with larvae coming from brook, rainbow, rainbow/brook, and no trout stream reaches. Our results show that larvae that co-occur with trout reduced their activity when exposed to brook trout predator cue, but their response to rainbow trout predator cue depended on their previous co-occurrence. Larvae from areas with only brook trout exhibited a weak predator-avoidance when exposed to rainbow trout predator cue. A follow-up test to determine the influence of alarm cue on predator response in these larvae indicated that the alarm cue enhanced the response to the rainbow trout predator.