Studies carried on in the last several years allow us to date the Canoa Formation as Late Pliocene. The rich paleontological record (foraminifers, mollusks, and otoliths) allowed us to outline a first articulate picture of the biogeographic relationships in the tropical eastern Pacific during the Plio-Pleistocene. The mollusk fauna shows a Panamic connotation, as the majority (88.7%) of the extant species are present between the Gulf of California and the coasts of Ecuador or northern Peru. Benthic foraminifers assemblages indicate a temperate character with a certain affinity with the Mexican Pacific microfauna. More than 7000 otoliths have been found in the Canoa Formation that are attributed to 105 taxa of 46 fish families. Only 65% of the fossil association is living today along the Ecuadorian coasts, while the association as a whole shows closer relationships with the extant Californian fish community (about 80%). In particular, a conspicuous group of fishes (15 taxa) today spread in the boreal East Pacific (from Oregonian to Californian Province or limited only to the Californian area) has been recorded in the Canoa Formation. We named these taxa “Californian guests.” After the rising of the Central American Isthmus, climatic variations may have caused changes in the pattern of surface currents and (or) activated coastal upwelling cells, supporting the diffusion of a part of the boreal biota toward more southern areas. At the present state of knowledge, both proximal causes seem to be compatible with the data presented.