The Oligocene (Rupelian) Byram Formation (Vicksburg Group) in Alabama, USA, is divided into three members, including (in ascending order) the Glendon Limestone, unnamed marl, and the Bucatunna Clay. The Oligocene marine units in Alabama have been historically under-investigated, but bulk samples recently obtained from Glendon Limestone Member exposures at site AWa-9 in Washington County yielded 20 unequivocal elasmobranch and teleost taxa. This surprisingly diverse paleofauna, based on isolated teeth, bones and otoliths, includes the new taxon, Gobiosoma? axsmithi sp. nov., as well as “Aetomylaeus” sp., Albula sp., Aplodinotus gemma Koken, 1888, Ariosoma nonsector Nolf and Stringer, 2003, Balistidae indet., Citharichthys sp., Myliobatoidei indet., Diretmus? sp., Hemipristis sp., Negaprion aff. N. gilmorei (Leriche, 1942), Pachyscyllium sp., Paralbula sp., Physogaleus sp., Preophidion meyeri (Koken, 1888), Sciaena pseudoradians (Dante and Frizzell in Frizzell and Dante, 1965), Sciaenops? sp., Sparus? elegantulus Koken, 1888, Sphyraena sp., and Syacium sp. Additional remains were recovered but could not be identified beyond undetermined Elasmobranchii or Teleostei. All these taxa represent first occurrences within the Glendon Limestone Member in Alabama, and the “Aetomylaeus” sp., Pachyscyllium sp., Paralbula sp., and Sciaenops? sp. specimens represent the first occurrences of each in the Oligocene of the Gulf Coastal Plain of the USA. We also report the first record of Oligocene Paralbula Blake, 1940 teeth, and the first occurrence of an Oligocene member of the Balistidae in the Western Hemisphere. This marine vertebrate assemblage indicates that the Glendon Limestone Member at site AWa-9 represented a subtropical to temperate, middle shelf paleoenvironment with a paleowater depth interpreted as 30–100 m.