The Timber Rattlesnake (Crotalus horridus) has a large distribution spanning much of the eastern United States. Because temperature, habitat type, prey composition and abundance, and a variety of other factors may dictate reptile behavior, populations of conspecific species may exhibit behavioral differences across latitudinal and elevational gradients. Using radio telemetry, we tracked 10 adult Timber Rattlesnakes (7 males, 3 females) from May 2016 to June 2017 in southeastern Louisiana to examine the spatial ecology of male and non-gravid female snakes. Mean annual and seasonal home ranges of non-gravid female Timber Rattlesnakes were not statistically different from that of males. Mean seasonal home range sizes and average distances travelled of both sexes was smallest in winter, and had a general increasing trend beginning in spring with a peak in fall. These increases seemed to coincide with the breeding season, taking place from early July until the end of November. Comparison of this study with other studies throughout its distribution could have implications towards future management of conservation for other southern populations of Timber Rattlesnakes.