The American crocodile Crocodylus acutus occurs across the Americas, with its northernmost distribution being in South Florida, USA. This species has undergone severe declines across its range and is categorized globally as Vulnerable on the IUCN Red List and as Threatened on the U.S. Federal Endangered Species List. Long-term monitoring studies in the USA have documented a shift in American crocodile nesting activity and an expansion of its range throughout the southern and eastern coasts of South Florida. However, no successful American crocodile nests have been recorded until now on the west coast of South Florida. Here we document the American crocodile nest monitoring conducted during 1997–2021 at Rookery Bay National Estuarine Research Reserve and the first successful nest from the west coast of South Florida for C. acutus. Marco Airport and McIlvane Marsh are the two main American crocodile nesting areas identified at the Reserve, with 92 nests and 3,586 eggs recorded during 1997–2021. We found most nests at Marco Airport (95.7%) and only four nests (4.3%) at McIlvane Marsh. To date, none of the nests found at Marco Airport have produced successful hatchlings. In contrast, hatchlings have been produced at McIlvane Marsh since nests were first documented there in 2020. We discuss the implications of our findings in terms of the future conservation of the species.