From 2002-2006, we used a variety of sampling techniques to survey the amphibians and water chemistry of Lower Suwannee National Wildlife Refuge in Florida’s Big Bend region. We recorded 23 amphibian species, 19 frogs and 4 salamanders. Species richness was lower than in areas of the coastal Big Bend region to the north. Amphibians occupied a wide variety of habitats and appeared tolerant of the generally acidic conditions of many of the wetlands. Although additional species may yet be found in LSNWR, this survey provides a historic baseline for assessing future status and trends of amphibian populations.