Culex quinquefasciatus, Say, 1823

Culex quinquefasciatus View in CoL field collection and colony establishment

Immature mosquitoes were collected in all Santiago Island municipalities, Cape Verde (15°07 ′ 48.0 ʺ N, 23°31 ′ 48.0 ʺ W) between August and October 2022. Culex sp. larvae were collected using standard sampling techniques with dippers and pipettes [ 22]. A subsample of collected larvae were morphologically identified to species complex using the identification key of Ribeiro and Ramos [ 23]; the remaining larvae were reared to adults in trays filled with dechlorinated tap water and fed with fish food (TetraMin; Tetra Werke, Melle, Germany). Adult mosquitoes were maintained at 28 °C and 70–80% relative humidity under a 12:12-h light:dark cycle, with access to a sucrose 10% solution ad libitum. One-weekold females were blood-fed on human blood using an artificial blood-feeding protocol described in Siria et al. [ 24] and following the Standard Operational Procedures adopted by the Instituto Nacional de Saúde Pública of Cape Verde. An oviposition container with water was placed inside the cage 4 to 5 days after blood-feeding.

Te resulting F1 generation egg rafts were transported to the In Vivo Arthropod Security Facility (VIASEF) of the Institute of Hygiene and Tropical Medicine (IHMT), Portugal, where all experiments with WNV were conducted under biosafety level 3 (BSL3) conditions.