Ecological niches hold critical information concerning the eco-evolutionary dynamics that govern biodiversity and abundance patterns. Cryptobenthic reef fishes account for approximately half of all reef fish species and are an abundant and important group on coral reefs worldwide. Yet, due to their small size and inconspicuous lifestyles, relatively little is known about the ecological niches of most cryptobenthic species. Here, we use gut content DNA metabarcoding to determine dietary niche overlap and prey richness in four sympatric species of cryptobenthic reef fishes in two genera (Acanthemblemaria aspera, A. spinosa, Enneanectes altivelis, and E. matador). Furthermore, we test whether dietary differentiation corresponds with differences in species distribution patterns across twelve sites on the Mesoamerican Barrier Reef in Belize. Our approach reveals dietary partitioning among the four species, which is further supported by low edge density and high modularity in the resulting trophic network. A. spinosa and E. matador consume a significantly higher richness of prey items than their congeners. This result corresponds with non-random distributions and co-occurrence patterns in both species pairs: the two high prey richness species (A. spinosa and E. matador) co-occur more frequently than predicted by chance, but they are exclusive to exposed forereef sites with high wave action. In contrast, their congeners occur across exposed forereef and sheltered backreef sites, but they do not increase in numbers at sheltered sites. Our findings suggest that A. spinosa and E. matador monopolize a wide variety of prey in exposed habitats, but they are unable to meet the energetic demands of their adaptation to high-flow habitats in sheltered areas, possibly due to lower prey availability. This, in turn, indicates strong ecological differentiation among closely related species of cryptobenthic fishes, driven by links between diet, physiology, prey availability, and wave exposure.
