Project summary Sleep is critical for nearly all animals. This state is characterized by a specific set of parameters for each species however there is a lack of sleep-based studies in mosquitoes. This should be considered a major knowledge gap in mosquito biology and potentially hinders the development of new control methods and our understanding of factors influencing vectorial capacity. Our preliminary studies suggest that there will likely be explicit factors underlying mosquito sleep, which need to be fully characterized to define this biological state in mosquitoes. The focus of the proposed work is to provide the first extensive characterization of mosquito sleep. After sleep has been defind, studies on the manipulation of sleep will discern how reduced sleep (e.g., induced by human activity in urban areas) may alter behavioral and physiological aspects of mosquitoes such as host preference, blood- feeding, reproductive output, and viral transmission. These studies are supported by the following: 1) Historic and our preliminary observations of putative sleep postures of mosquitoes, 2) Initial activity monitoring results that establish that day and night active mosquitoes sleep at higher rates during the night and day, respectively, 3) targeted studies suggesting that sleep can be prevented by mechanical disturbance that impacts subsequent host landing, 4) Preliminary data showing a reduction of spontaneous neural activity after prolonged rest, 5) Our development of novel sensory deprivation equipment that allows for mosquito observation without host interference to pinpoint differences that could be related to mosquito sleep-like states, and 6) Integrative and innovative experimental design that ranges from basic behavioral analyses to neuronal recording that will provide an encompassing view of the mosquito sleep state. This study has two specific aims: Specific Aim 1. Establishing the characteristics associated with sleep-like states in mosquitoes. Specific Aim 2. Defining shifts in mosquito fitness, behavior, and viral transmission following sleep deprivation. Upon completion of these specific aims, our expected outcomes are to have defined sleep-like states in mosquitoes and, subsequently, how sleep deprivation impacts a range of epidemiologically relevant biological aspects. This will be transformative to the research field and will set the stage for multiple lines of research. Most importantly, these studies will create a novel paradigm, where aspects of mosquito biology should be measured under two independent periods: a non-resting (no sleep) and sleep-like status. Finally, our anticipated results are likely to inform on the adaptations of mosquitoes to urban areas where host activity patterns and light/dark conditions are decoupled from day/night successions and could impact sleep, mosquito-host interactions, and potentially patterns of disease transmission.

Project narrative Sleep-like states have been described in multiple insect systems, where these have been linked to immune functionality, stress tolerance, and reproductive aspects, but little is known about sleep-like states in mosquitoes. In this study, we will characterize this overlooked aspect of mosquito biology through combined behavioral and physiological studies, and follow this with sleep manipulation studies to determine the impact of sleep deprivation on reproduction, host preferences, and viral transmission. Results from this proposed research will be critical in understanding how periods of sleep-like states impact mosquitoes, drastically increasing our understanding of their biology, and lay the groundwork for assessing how sleep changes may alter pathogen transmission and the biology of mosquitoes in their natural environment.