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Gnathia jimmybuffettisp. nov. is described from the Florida Keys. Morphological characterization of the male, female, and juvenile as well as the life cycle development of the female is examined. The male of this new species is distinguished from other species from the Tropical Northwestern Atlantic marine province by the slightly produced frontal margin; absent mediofrontal process; single, strong, conical superior frontolateral process, with 2 pairs of long simple setae; and rounded inferior frontolateral process, the weak distally curved mandible with weakly convex dentate blade. In addition, molecular characterization for three life stages, (males, females, and praniza), is provided, as well as evidence for linking host DNA to juvenile stages. This paper contributes to our understanding of the gnathiid biodiversity of the Tropical Atlantic realm, specifically the Floridian ecoregion. Furthermore, this paper validates the methods of extracting and identifying host DNA from the gnathiid blood meal.