Human-bat interactions are common in rural areas across the tropics. Over 40 bat species occur in Madagascar, most of which are endemic. Forest loss is changing the distribution of bats throughout the island, with potential increases in both the abundance of synanthropic species and human-bat interactions. We set out to study knowledge of, interactions with, and attitudes towards bats in rural Madagascar, including reports of food and ethnomedicinal uses of bats, their cultural representations in folklore, and the existence of culturally enforced taboos in relation to them. We administered 108 surveys with open- and closed-ended questions with adults from the Tanala and Betsileo ethnic groups living around Ranomafana National Park. Most interviewees mentioned at least two types of bats. Over 10% of the interviewees had consumed bats and ∼20% used bat guano as a fertilizer. Around one-fifth recognized cultural taboos inhibiting bat hunting and consumption and most considered bats not to be dangerous. However, some informants mentioned that bats could carry diseases and complained about the bad smell and noise associated with bat roosts in houses and public buildings. Nearly 25% of the respondents could identify cultural representations of bats in local folklore. Malagasy rural communities interact closely with bats, but severely underestimate the diversity of bat species around them. Taken together, our results greatly increase the understanding of social-ecological complexities of human-bat relationships in rural Madagascar, offer possible pathways for biocultural approaches to conservation, and yield insights applicable to other communities coexisting with bats across the humid tropics.