The construction and expansion of roads cause significant impacts on the environment. The main potential impacts to biotic environment are vegetation suppression, reduction of the amount and composition of animal distribution due to forest fragmentation and increasing risks of animal (domestic and wildlife) vehicle collisions. The objective of this work was to establish a relationship between the different spatial patterns in wildlife-vehicle crash, by using spatial analysis and machine learning tools. Self-Organizing Maps (SOM), an artificial neural network (ANN), was selected to reorganize the multi-dimensional data according to the similarity between them. The results of the spatial pattern analysis were important to perceive that the point data pattern varies from an animal type to another. The events occur spatially clustered and are not uniformly distributed along the highway. SOM was able to analyze the relationship between multiple variables, linear and non-linear, such as ecological data, and established distinct spatial patterns per each animal type. In the studied area, most of the wildlife was run over very close to forest area and water bodies, and not so close to sugarcane fields, forestry and built environment. A considerable part of the wildlife-vehicle collisions occurred in areas with diverse landscape.