5. WHO cone assays
The bio-efficacy of several bed nets was evaluated using the WHO cone assay protocol (WHO, 2013). Cohorts of 50 mosquitoes were exposed to 25cm x 25cm pieces of the netting of PermaNet 3.0, PermaNet 2.0, Olyset, and Olyset Plus. PermaNet 3.0 had 2 pieces from the roof and the side, as the top is impregnated with insecticide and PBO while the side has only the insecticide. Insecticide-free nets were used as controls to which mosquitoes were also exposed. Mosquitoes were exposed to the nets for 3 minutes in 5 replicates (each of 10 mosquitoes) and then transferred into paper cups. The knockdown effect was recorded at 1-hour post-exposure, and the mortality rate was scored 24 hours post-exposure.