Anastrepha manizaliensis Norrbom & Korytkowski, new species, is described from Colombia. It breeds in fruit of Juglans neotropica Diels (Juglandaceae), commonly known in Colombia as “cedro negro”. The new species was previously confused with Mexican fruit fly, Anastrepha ludens (Loew), a major pest of citrus and mango, leading to quarantine problems, but true A. ludens does not occur in Colombia.