Abstract
Aedes aegypti is the primary vector of the dengue and chikungunya viruses. After mating, male seminal fluid molecules cause females to become unreceptive to a subsequent mating. This response is often assumed to be immediate and complete, but a growing body of evidence suggests that some females do mate more than once. It is unknown how quickly a female becomes unreceptive to a second mating. Furthermore, the degree to which she remains monandrous after laying several batches of eggs has not been rigorously tested. Therefore, we assessed the rates of polyandry in two sets of experiments using wild-type males and those with fluorescent sperm. The first experiment tested the likelihood of polyandry after postmating intervals of various durations. Most females became refractory to a second mating within 2 hours after mating, and rates of polyandry ranged from 24% immediately after mating to 3% at 20 hours after mating. The second experiment tested whether females were polyandrous after cycles of blood meals and oviposition. No re-insemination was found after one, three, or five such cycles. This study is the first to demonstrate that polyandrous behavior depends on the postmating interval. Our results will inform future applications that depend on an accurate knowledge of Ae. aegypti mating behavior, including models of gene flow, investigations of molecules that drive female mating behavior, and control strategies that deploy genetically modified mosquitoes into the field.