Direct fossil evidence shows that by the Middle Miocene the freshwater fish fauna of South America, including Amazonia, was essentially modern across a wide taxonomic and ecological range. Much diversification of modern Neotropical fishes occurred during at least the roughly 60 million year period from the Late Cretaceous to the Miocene. Miocene to Holocene Earth history events played little or no role in creating the great diversity of Neotropical fishes at or above the genus level. The record of Neotropical fishes contains almost no documented extinctions of distinct lines of fishes that are phylogenetically close to living groups. There are, however, many cases of Late and post-Miocene local extirpations of modern groups from areas now peripheral to the large, lowland cis-Andean (Eastern-slope) rivers. The fossil fish assemblages of the Pebas and Acre stages in the Miocene mega-wetland are markedly similar. Further, freshwater fishes of the Middle Miocene La Venta fauna and Late Miocene Urumaco fauna are similar to those of the Miocene mega-wetland assemblages and thus support a high degree of biotic and hydrological relatedness. The majority of Amazonian Neogene fishes were strictly freshwater inhabitants with little or no tolerance for brackish-water conditions. Virtually no Neogene fish fossils from the deep interior of Amazonia, including those of carcharinid sharks, sawfishes, stingrays, ariid catfishes and drums, are certain indicators of marine or even brackish waters.