In Western cultures, some combinations of musical notes are consonant (pleasant) and others are dissonant (unpleasant). Although the aesthetic contrast between consonance and dissonance is commonly thought to have biological roots, and thus to be universally present in humans, a definitive test has remained elusive. Here we report experiments with the Tsimane’—a native Amazonian society with minimal exposure to Western culture—and comparison populations elsewhere in Bolivia and the USA that varied in exposure to Western music. Participants rated the pleasantness of sounds. Despite exhibiting Western-like preferences for several other aesthetic contrasts, and Western-like discrimination abilities, the Tsimane’ rated consonant and dissonant chords and vocal harmonies as equally pleasant. In contrast, Bolivian city- and town-dwellers exhibited significant preferences for consonance, albeit to a lesser degree than U.S. residents. The results indicate that consonance preferences dramatically vary across cultures, and can be fully absent in some cases. The observed variation in preferences is plausibly determined by exposure to Western music, suggesting that culture plays a dominant role in shaping aesthetic responses to consonance.