Macrochelys suwanniensis (Suwannee Alligator Snapping Turtle) coexists with 2 other native large omnivorous turtle species (Chelydra serpentina [Snapping Turtle] and Trachemys scripta scripta [Yellow-bellied Slider]) in a 9-km section of the Santa Fe River in northern Florida. A major shift in dominant submersed aquatic vegetation prompted us to quantify trophic position and niche overlap among these 3 species. Here, we examine carbon and nitrogen isotopic values of these turtles and their potential food resources within the changing riverine system. We provide evidence of low isotopic niche overlap between M. suwanniensis and C. serpentina, whereas T. s. scripta occupied a discrete niche having lower carbon and nitrogen values.