[9] Absent from the Appalachians, but occurs west of them in Tennessee, Kentucky, and southern reaches Illinois, Indiana, and Missouri.

[11] Prefers habitats with loose, commonly sandy, soils for burrowing[12] like Longleaf pine forests with open understories from intermittent natural burn events.

It has shown preference for being close to deciduous shrub edges, low-growing pitch pine branches, and reindeer lichen.

These stripes, which begin on the upper eyelids, may diverge or converge, resulting in a pattern resembling a lyre or an hourglass.

[20] Although Scaphiopus holbrookii is both diurnal and nocturnal, most foraging for food sources, consisting of small invertebrates such as termites, insects, arachnids, worms, ants, beetles, and grasshoppers is completed during the day.

[22] Being burrowing species, spadefoot toads avoid a large amount of predation at the surface, but when exposed during foraging or breeding, they are preyed upon by mammals such as opossums and racoons, birds, and snakes.

[4] Eastern spadefoot toads are explosive breeders during sufficient rainfall and eggs are usually attached to submerged vegetation.

[26] Due to the explosive breeding, once eggs hatch often food becomes limited from the large populations of tadpoles.

In the event of food shortages some larvae adapt aggressive feeding habits consuming large animal prey including other S. holbrookii tadpoles.