Few studies of post-Columbian animal economies in the Americas elaborate on the influence of traditional Indigenous knowledge on colonial economies. A vertebrate collection from Santa Elena (1566–87 CE, South Carolina, USA), the original Spanish capital of La Florida, offers the opportunity to examine that influence at the first European-sponsored capital north of Mexico. Santa Elena’s animal economy was the product of dynamic interactions among multiple actors, merging preexisting traditional Indigenous practices, particularly traditional fishing practices, with Eurasian animal husbandry to produce a new cultural form. A suite of wild vertebrates long used by Indigenous Americans living on the southeastern North Atlantic coast contributes 87% of Santa Elena’s noncommensal individuals and 63% of the noncommensal biomass. Examples of this strategy are found in vertebrate collections from subsequent Spanish and British settlements. This suggests the extent to which colonists at the Spanish-sponsored colony adopted some Indigenous animal-use practices, especially those related to fishing, and the speed with which this occurred. The new cultural form persisted into the nineteenth century and continues to characterize local cuisines.