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We describe a new species of subterranean amphipod (Amphipoda: Crangonyctidae) in the genusStygobromusfrom a hypotelminorheic seepage spring at Shepherd Parkway, part of National Capital East Parks, Washington, D.C., USA, part of the National Park System, using both morphological and genetic approaches. The Anacostia Groundwater Amphipod,S. anacostensissp. nov.is a member of theS. tenuisspecies group but differs from related congeners based on body size, serrate blade-like edge of both palms of gnathopods 1 and 2, presence of rastellate setae on the posterodistal margin of the carpus of gnathopod 2, and aspects of the second antennae, mandibular palp, pereopods 5–7, uropods 1 and 2, and telson. Moreover,S. anacostensissp. nov.is genetically distinct fromS. tenuisin the Washington D.C. metropolitan area. The description ofS. anacostensissp. nov.increases the number of describedStygobromusspecies to eight in the Washington D.C. area and highlights the need for continued biodiversity studies, even in regions that have received considerable attention.