Roads negatively impact wildlife through habitat fragmentation, loss of habitat connectivity, and wildlife-vehicle collisions, thus road mitigation structures, such as wildlife crossing structures (WCS), wildlife guards (WG), and fencing are commonly used to address this issue all over the world, including in the United States. In South Texas, such structures were built or modified along a State Highway in an effort to address road mortality for the endangered ocelot (Leopardus pardalis) and non-target wildlife species. The goal of this study was to examine temporal changes in wildlife interactions with WCS and WG during and after their construction and modification along a South Texas highway and to determine whether environmental factors influenced use of WCS. Using camera traps deployed to monitor the road mitigation structures, we compared crossing rates, repel rates, and species richness of all species that interacted with the structures, and we examined whether differential wildlife use of WCS and WG was affected by one or more structural dimensions, distance to nearby vegetation, and water presence. Crossings through WCS by wildlife decreased following the completion of construction of mitigation structures; however, repel interactions at WG increased. Overall, crossings decreased at WCS that had higher openness ratios and during periods of precipitation and higher daily temperatures, but distance to vegetation had minimal influence. These factors were shown to influence crossings of each of the five most frequently observed species differently. Lastly, the presence of pooled water at one WCS caused a decrease in crossings when the water level was highest but was not a barrier at lower water levels. By examining influences on wildlife interaction with road mitigation structures, we conclude that a variety of structures, including different WCS configurations, can be beneficial in facilitating movement and restricting entry into the right-of-way for a diversity of wildlife species beyond the target species.