Objective: We aimed to determine the species of the Culicidae family inhabiting the tank bromeliad Tillandsia elongata in an agroecosystem of the Colombian Andes, utilizing taxonomic and molecular methods. Scope: While Agriculture is recognized as one of the causes of biodiversity loss, agroecosystems have the potential to maintain the invertebrate biodiversity, including mosquito species. Methodology: We collected immature mosquitoes from 48 bromeliads in the agroecosystem “Montelindo” of Universidad de Caldas in Santágueda (Palestina, Caldas, Colombia). To collect the immature mosquitoes within each bromeliad, we filtered the water in the tank using a 150 µm-mesh sieve, and subsequently cleaned each bromeliad leaf with a brush. Taxonomic and molecular approaches were employed for mosquito species identification. Main results: This study presents the first report of immature mosquitoes from the genera Toxorhynchites and Wyeomyia inhabiting the bromeliad Tillandsia elongata in agroecosystems. Morphological identification led to establishing two species of Toxorhynchites, namely Toxorhynchites bambusicola (Lutz and Neiva) and Toxorhynchites haemorrhoidalis (Fabricius). DNA sequencing confirmed the presence of two species Wy. mitchelli (Theobald) and Wy. pertinans (Williston). Our results expand the known distribution range of Toxorhynchites species, as well as the bromeliad species that serve as breeding sites for Tx. bambusicola and Tx. haemorrhoidalis. Agroecosystems have the potential to contribute to Culicidae biodiversity through the conservation of breeding water bodies as tank bromeliads.