Leptospirosis is a disease endemic to both rural and urban areas of tropical countries and resource-poor communities. Little information is available on the presence of Leptospira spp. in urban water sources. A study was conducted to detect pathogenic Leptospira in ornamental water fountains in Cali, Colombia. Twenty-seven water fountains were tested for pathogenic Leptospira using a multiplex PCR assay targeting the secY and the flaB genes. Pathogenic Leptospira was confirmed in 11 (41%) ornamental water fountains. Plazas, building exteriors, and sidewalks presented the highest proportion (67%) of pathogenic Leptospira-positive water fountains. Urban ornamental water fountains might be sources of pathogenic Leptospira and might pose a risk to humans who come into close contact, although relevance from a public health perspective is yet to be established.