Roads have important effects on wild life, such as natural habitat fragmentation which leads to crossing the roads to gain access to crops, pasture, water or territories. Current rates of wildlife road mortality are neither sustainable for biodiversity nor a healthy reflection of our interactions with the environment and the animals who try to coexist with us. The present survey was conducted to estimate the road kills on the National Highways 161 which joins the cities of Akola, Washim, Hingoli, Nanded, Degloor, Sangareddy and Hyderabad in Central-Southern India with each other. A very small segment of 15 km was taken for the study. Road kill incidences were monitored and photographed on daily basis in morning and evening from March 2016 to February 2017. At every sighting of a road kill, information such as place, species name, number and status of the kill was recorded. The present study revealed a total of 60 road kills belonging to 20 species. Birds were the most affected individuals (32 %) followed by reptiles (27 %), mammals (23 %) and amphibians (18 %). Conservation and management implications are essential to prevent this threat to local biodiversity.