Aedes aegypti mosquito is a major vector of arboviral disease. Here, we report that the biting behavior of normally daytime active anthropophilic Ae. aegypti mosquitoes on human hosts is abnormally increased at night following exposure to artificial light at night (ALAN). Biting was examined using a human host assay where caged mosquitoes were exposed to a human arm and blood-feeding measured. Mosquitoes were tested during the daytime, nighttime, or challenged with ALAN. As predicted from the Ae. aegypti diel/circadian biting cycle, maximal biting occurred during daytime and lowest level occurred at night. Biting in the ALAN group was increased compared with time-matched nighttime controls. These data reveal that exposure to ALAN increases nocturnal blood-feeding behavior. This finding highlights the concern that globally increasing levels of light pollution could be impacting arboviral disease transmission, such as dengue fever and Zika, and has implications for application of countermeasures for mosquito vector control.