Construction of highway would influence wildlife habitat being fragmented and deteriorated. It is inevitable to observe the death of animals that are killed by vehicles. Nonetheless the cause and status of animal roadkill have not been studied, and this study first attempted to analyze the roadkill in major highway in Korea. We collected 860 individuals of dead animals in Joongang Highway during 1996-2003. Among them Korean hare(Lepus sinensis) was totaled in 165 individuals(19.2%) with the highest figure; Korean racoon dog(Nyctereutes procyonoides) was 146(17.0%); Korean squirrel(Sciurus vulgaris) 56(6.5%); Korean roe deer(Capreolus capreolus) 26(3.0%). Domestic animals was totaled in 232 individuals(27.0%); domestic dog(Canis familiairis) 25, and domestic cat(Catus felis) 207. This study also categorized habitat as mountain-mountain, mountain-plain, mountain-river, plain-plain, plain-river, river-river in each side of the road. Habitat of mountain-plain was the heightest with 296(38.9%) individuals followed by mountain-mountain with 263(34.6%). This indicates that wild animals associated with mountain habitat was most influenced by the construction of roads. This study implied that we should mitigate the roadkill by designing artificial construction such as eco-bridge, fences along the sideway, and boxes under the highway, underdrain structure, etc. The monitoring and cause of habitat fragmentation with GIS approach should be followed to reduce the roadkill.