Definitive hosts of Neospora caninum are species of canids, such as domestic dogs (Canis lupus familiaris), coyotes (Canis latrans) and foxes (Vulpes vulpes), whereas ruminants, such as cattle (Bos taurus), sheep (Ovis aries), and deer (Cervidae) serve as intermediate hosts. We investigated the presence of N. caninum in feces of coyotes in Calgary, Alberta, Canada. One hundred coyote fecal samples were collected from five city parks. Following DNA extraction, the presence of N. caninum DNA was determined by using real-time PCR analysis with a primer and probe pair targeting the Nc5 gene. Ten of the 100 samples contained detectable amounts of N. caninum DNA. White-tailed deer (Odocoileus virginianus) and mule deer (Odocoileus hemionus) are common in Calgary's city parks, and although we did not examine them for the presence of N. caninum bradyzoites, the presence of N. caninum DNA in coyote fecal samples is indicative that a sylvatic host of N. caninum exists within this urban environment.