The Great American Biotic Interchange (commonly abbreviated as GABI), also known as the Great American Interchange and the Great American Faunal Interchange, was an important late Cenozoic paleozoogeographic biotic interchange event in which land and freshwater fauna migrated from North America via Central America to South America and vice versa, as the volcanic Isthmus of Panama rose up from the sea floor and bridged the formerly separated continents. Although earlier dispersals had occurred, probably over water, the migration accelerated dramatically about 2.7 million years (Ma) ago during the Piacenzian age. It resulted in the joining of the Neotropic (roughly South American) and Nearctic (roughly North American) biogeographic realms definitively to form the Americas. The interchange is visible from observation of both biostratigraphy and nature (neontology). Its most dramatic effect is on the zoogeography of mammals, but it also gave an opportunity for reptiles, amphibians, arthropods, weak-flying or flightless birds, and even freshwater fish to migrate. Coastal and marine biota, however, was affected in the opposite manner; the formation of the Central American Isthmus caused what has been termed the Great American Schism, with significant diversification and extinction occurring as a result of the isolation of the Caribbean from the Pacific.

The occurrence of the interchange was first discussed in 1876 by the "father of biogeography", Alfred Russel Wallace. Wallace had spent five years exploring and collecting specimens in the Amazon basin. Others who made significant contributions to understanding the event in the century that followed include Florentino Ameghino, W. D. Matthew, W. B. Scott, Bryan Patterson, George Gaylord Simpson and S. David Webb. The Pliocene timing of the formation of the connection between North and South America was discussed in 1910 by Henry Fairfield Osborn.

Analogous interchanges occurred earlier in the Cenozoic, when the formerly isolated land masses of India and Africa made contact with Eurasia about 56 and 30 Ma ago, respectively.