Moonlight intensity influences the activity patterns of bats. Some bat species reduce their activity levels during brighter nights, a phenomenon known as “lunar phobia.” While lunar phobia of bats has been extensively studied in tropical regions, the same is not the case of bats in temperate regions. By using acoustic detectors, we examined differences in the activity of insectivorous bats on nights with different moonlight intensity in an agricultural landscape of central Chile. We also examined the hourly activity patterns throughout the night and how these varied between full and new moon nights. All bat species modified their activity based on the moonlight intensity; however, their effects were species-specific. The activity of Lasiurus varius, L. villosissimus, Myotis chiloensis, and Histiotus montanus was lower during bright nights, while Tadarida brasiliensis was the only species whose activity was higher during bright nights. Hourly activity throughout the night differed between full moon nights and new moon nights in most bat species. During full moon, bats concentrated their activities in the early hours of the nights; a more homogeneous activity pattern was exhibited during new moon night. Our study demonstrates that moonlight affects the activity of bats in Chile, a factor that should be considered when studying bats.