Partitioning of the food resources by two coexisting pufferfishes (Sphoeroides spengleri and S. testudineus) from Biscayne Bay, Florida, USA, was investigated. Gut contents from 453 bandtail and 339 checkered puffers were analyzed. The diets of both species consisted of a variety of benthic prey, but only crustaceans and molluscs were important prey groups. While differences were found in the proportions of general prey categories eaten by these fishes, both species consumed substantial quantities of brachyuran crabs, bivalves, and gastropods. Specific identification of the prey items within these three food categories revealed additional differences in prey between the two puffer species. This partitioning of the food resources by bandtail and checkered puffers was found between both species overall, between overlapping size ranges, and between both species' most abundant size group. Differences in food habits between these two fishes illustrate that congeners with virtually identical mouth structure and complete spatial overlap can significantly partition the food resources.