We monitored the herpetofauna of a large conservation area in peninsular Florida from 1990 to 2010, visiting the refuge on over 2000 different days in that period and using a variety of sampling techniques. Our goals were to evaluate the species richness and abundance of herpetofauna (amphibians and reptiles) on the site. During the course of our study, we documented 72 species (22 amphibian and 50 reptile species); thus, the site has impressive species richness relative to other conservation areas in the southeastern United States. Five of the 72 species (6.9%) we found were introduced species. Continued monitoring of the diverse herpetofauna of Lake Woodruff National Wildlife Refuge could facilitate a better understanding of ongoing changes in species abundances and community assemblages occurring in the ecosystems of peninsular Florida.