The giant sugarcane borer, Telchin licus, has been reported as an economically important sugarcane pest in Colombia; however, its taxonomic status has been scarcely investigated and previous reports offer an ambiguous characterization of both the immature and adult stages. The objective of this work is to identify Telchin species affecting sugarcane and alternative hosts in different departments of the country by integrating molecular analysis and conventional morphology. To date, T. licus has been found in the departments of Caquetá, Casanare, and Meta, while T. atymnius has been found in Antioquia, Caldas, Nariño, and Valle del Cauca. Sugarcane, Musaceae, and Heliconiaceae have been found to be hosts to both species. Additionally, the species T. cacica has also been registered in the department of Nariño, affecting heliconias and plantains. Genetic variation within the species allowed differentiation at the molecular level of subspecies of T. licus and T. atymnius, confirming that the subspecies present in Colombia are T. licus magdalena, T. atymnius humboldti, and T. atymnius atymnius. The haplotype diversity of populations is closely related to their geographical distribution, indicating low gene flow between populations and possible speciation inside the country. Analysis of genetic variance showed significant differences among and within T. atymnius populations, which may suggest a high genetic structure along the regions where it is found and the possible presence of additional subspecies to those previously reported. To understand the geographical and environmental conditions that determine the pest's distribution in Colombia, this information needs to be complemented with ecological considerations of possible geographical isolation and association of alternative hosts.