Big Cypress National Preserve constitutes approximately one-third of the range of the endangered Florida panther (Puma concolor coryi). Because recreational hunting is allowed in Big Cypress National Preserve, we examined 8 response variables (activity rates, movement rates, predation success, home-range size, home-range shifts, proximity to off-road vehicle trails, use of areas with concentrated human activity, and habitat selection) to evaluate how Florida panthers respond to human activity associated with deer and hog hunting. Data consisted of panther radiolocations collected since 1981 by the Florida Fish and Wildlife Conservation Commission and the National Park Service, which we augmented with radiolocations and activity monitoring from 1994 to 1998. A split-plot (treatment and control) study design with repeated measures of the variables for each panther taken before, during, and after the hunting season was used. We did not detect responses to hunting for variables most directly related to panther energy intake or expenditure (i.e., activity rates, movement rates, predation success of females; P>0.10). However, panthers reduced their use of Bear Island (P=0.021), an area of concentrated human activity, and were found farther from off-road vehicle trails (P≤0.001) during the hunting season, which was indicative of a reaction to human disturbance. Whereas the reaction to human activity on off-road vehicle trails probably has minor biological implications and may be linked to prey behavior, the decreased use of Bear Island is most likely a direct reaction to human activity and resulted in increased use of adjacent private lands. Future habitat loss on those private lands could exacerbate the negative consequences of this response by panthers.