Anopheles gambiae and Aedes aegypti are major vectors of malaria and arboviral disease (e.g. dengue and Zika), respectively. We report that the biting behavior of these mosquitoes on human hosts is abnormally altered in response to exposure to artificial light at night (ALAN). In the night active (nocturnal) and normally night biting An. gambiae, ALAN results in a decrease in biting activity that is sustained for up to 4 h following termination of the transient light exposure. Conversely, the day active (diurnal) and daytime biting Ae. aegypti respond to ALAN with an increased biting propensity. These data reveal that exposure to ALAN changes nocturnal mosquito blood-feeding behavior. These findings highlight the concern that globally increasing levels of light pollution could be impacting vector-borne disease transmission, and has implications for application of interventions for mosquito vector control.