SOP for testing the susceptibility of adult sand flies to insecticides in WHO bottle bioassays

This SOP describes the process for evaluating the susceptibility of adult sand fly vectors (Phlebotomus spp. and Lutzomyia longipalpis) to insecticides in the WHO bottle bioassay. The WHO bottle bioassay is a direct response-to-exposure test, in which sand fly mortality is measured after 1 h of exposure to a known standard concentration (e.g., the discriminating concentration, DC) of an insecticide. The mortality of test sand flies is recorded at 24 h (or more for slow-acting compounds) after the 1-h exposure period. Instructions are provided on preparing and coating bottles with insecticide, exposing sand flies, and recording and interpreting test results.