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We studied the occurrence of exposure to Upper Respiratory Tract Disease (URTD) in gopher tortoises (Gopherus polyphemus) at protected sites in southern Mississippi (Marion County Wildlife Management Area [MCWMA]) and east-central Florida (Kennedy Space Center [KSC]). Based on antibody assays, exposure to URTD was very common in both populations, with combined positive and suspect results ranging from 60% (KSC) to 100% (MCWMA). At KSC, males were significantly more likely to show positive or suspect results than were females. Tortoises with active signs of URTD were seen at KSC but not at MCWMA. Our data do not support the hypothesis that exposure to URTD is associated with human disturbance or habitat destruction; several sites that had good habitat conditions and little disturbance by humans had high rates of URTD,whereas some sites with high levels of disturbance had low rates of URTD. Long-term monitoring of populations with positive antibody test results is needed.