DNA barcoding significantly improves resolution of invasive lionfish diet in the Northern Gulf of Mexico

Invasive Indo-Pacific red lionfish ( Pterois volitans ) have become well-established residents within reef communities across the western Atlantic Ocean where they pose substantial threats to native fish communities and reef ecosystems. Species-specific identification of prey is necessary to elucidate predator–prey interactions, but can be challenging with traditional visual identification methods given prey are often highly digested, thus not identifiable visually. To supplement visual diet analysis of lionfish (n = 934) sampled in the northern Gulf of Mexico, we applied DNA barcoding to identify otherwise unidentifiable fish prey (n = 696) via amplification of the cytochrome c oxidase subunit I (COI) of the mitochondrial genome. Barcoding nearly doubled the number of identifiable fish prey, thereby greatly enhancing our ability to describe lionfish diet. Thirty-three fish prey species were identified via barcoding, twenty-four of which were not previously detected by traditional methods. Some exploited reef fishes were newly reported (e.g., red snapper, Lutjanus campechanus ) or found to constitute higher proportions of lionfish diet than previously reported (e.g., vermilion snapper, Rhomboplites aurorubens ). Barcoding added a significant amount of new dietary information, and we observed the highest prey diversity reported to date for invasive lionfish. Potential cannibalism on juveniles also was identified via DNA barcoding, with the highest incidence corresponding to high lionfish densities, thus suggesting density-dependent prey demand may have driven this response. Overall, DNA barcoding greatly enhanced our ability to describe invasive lionfish diet in this study, suggesting that even studies with relatively large diet sample sizes could benefit from barcoding analysis.