Amatitlania nigrofasciata is a small freshwater fish species native to Central America. Is a popular ornamental fish that has been traded and cultivated for the aquarium industry, introduced into temperate and tropical regions across the world. Two specimens of A. nigrofasciata were collected from two small creeks draining into the Guavio River, upper Meta River in Mambita, Ubalá, Cundinamarca, Colombia in the Piedmont Orinoco ecoregion. This is the first record of A. nigrofasciata in natural freshwaters of Colombia and the Orinoco River basin. Diagnostic characters of species and updated distribution at country level are provided.