Arboreal microhabitats associated with phlebotomine sand flies in Caribbean Colombia: Integrative identification using COI barcoding.

Arboreal cavities, tabular roots and bark-associated structures provide protected daytime shelters for phlebotomine sand flies and may also contain conditions suitable for immature development. We characterized sand flies collected during a targeted three-day survey in Bellacohita, Moñitos, Córdoba, Colombia, in 2024 and used COI barcoding as an integrative identification tool. Active searches were conducted from 06:00-10:00 and 16:00-19:00 at 63 trees in 31 domiciles; 21 trees contained adult sand flies. Direct arboreal collections comprised 298 morphologically assigned specimens, dominated by Pintomyia evansi (212) and Lutzomyia gomezi (62). Complementary CDC light-trap collections in domiciles with positive trees yielded 105 adults and were analysed separately. Forty-nine COI consensus sequences showed affinities with Pintomyia evansi (23), Lutzomyia gomezi (13), Psychodopygus panamensis (7), Evandromyia dubitans (5) and Micropygomyia cayennensis (1). A rooted maximum-likelihood analysis of 114 sequences placed the local barcodes in five reference-supported groups. Males with unrotated or incompletely rotated genitalia were compatible with recent emergence, but this observation does not by itself confirm immature development in the sampled trees. The findings provide a baseline for targeted surveillance of tree-associated adult habitats.