Human population growth and development reduce the area and quality of natural communities and lead to a decline of associated wildlife populations. Sciurus niger avicennia (Big Cypress Fox Squirrel), a state-listed threatened subspecies endemic to south Florida, appears sensitive to habitat fragmentation and fire regime. This research assessed the distribution and habitat use of the Big Cypress Fox Squirrel from interviews with biologists, private land owners, and golf course managers and by transect sampling in natural areas on public and private lands in southwest Florida. Our findings indicate that the distribution of fox squirrel populations is influenced by land use and understory height. Conservation of this species in natural areas will require land management practices that maintain open landscapes.