The river supports abundant wildlife, including otters, alligators, many species of turtles and fish, and every variety of water bird—waders, divers and dabblers. Osprey, hawks and swallowtail kites soar along the river corridor while smaller birds and animals hide in the lush vegetation.

The Rainbow River in Florida is a vibrant ecosystem home to a diverse array of freshwater and semi-aquatic wildlife. In 2026, the most iconic residents are the North American river otters, which are frequently seen playing near the headsprings. The river is also a sanctuary for West Indian Manatees, particularly during the cooler winter months when they seek the constant 72°F (22°C) spring water. Beneath the crystal-clear surface, you will find various fish species including largemouth bass, bluegill, and gar, alongside numerous turtles like the Florida red-bellied cooter. Reptiles are well-represented by alligators, though they tend to stay in the quieter, marshy edges of the river away from the main tubing paths. Birdwatchers in 2026 can spot ospreys, anhingas (often seen drying their wings on branches), great blue herons, and the rare wood stork. The surrounding cypress trees and lush vegetation also provide habitat for white-tailed deer and various species of water snakes, making it one of Florida's most biodiverse springs.