Male-exposure (ME) feeding was conducted in a separate room away from the fly cultures. One day prior to feeding, male flies were immobilized by placing them under netting and marked on the thorax using a small dot of water-based paint. The marked males (aged thirteen to fourteen days) were then transferred to a cylindrical plexiglass cage (twenty cm twenty cm in diameter) with both openings covered by cloth mesh.
For the ME feeding procedure, zero point five ml of ME was applied onto a filter paper strip, which was subsequently placed in a petri dish (six cm in diameter) and introduced into the cage. The marked males (aged thirteen to fifteen days) were allowed to feed on the ME for one hour between zero nine thirty and ten thirty, which corresponds to the peak foraging time for ME. Individual male feeding behavior was not monitored during this exposure period.
After the feeding period, the petri dish containing the ME was removed, and the used filter paper strip was sealed in a polythene bag and discarded. The males were provided with a standard adult diet and water ad libitum.
Previous studies have shown that ME feeding enhances the mating success of B. dorsalis males one day after feeding, and this effect can persist for up to thirty-five days. Therefore, the ME-treated males were evaluated either one or two days after ME treatment, when the tested males were between fifteen and seventeen days old. Less...