The phase of the moon can affect activity patterns of nocturnal animals, and may also affect visibility for motorists. However, surprisingly little is known about whether the risk of a wildlife-vehicle collision (WVC) is associated with lunar phase. We investigated the relationship between frequency of WVC at night and lunar phase for four large ungulate species that account for a high proportion of serious WVC along roads in agricultural and forested landscapes of two continents: wild boar Sus scrofa, roe deer Capreolus capreolus, and red deer Cervus elaphus in Castile and Leon, Spain, and white-tailed deer Odocoileus virginianus in New York State, USA. Three of the four species most frequently collided with vehicles at night during the full moon phase of the lunar cycle; this pattern was evident throughout the year but was stronger during some months. For roe deer, the species for which WVC was most closely associated with the lunar cycle, the frequency of WVC was 71.3% greater during the full than new moon period. Our results indicate that rates of ungulate WVC at night cycle on a period of a lunar month, which has implications for traffic safety planning and for motor vehicle collision emergency response preparation.