Introduction Emerging evidence suggests that psychedelic compounds, including the Amazonian botanical decoction ayahuasca, may provide clinical benefit in the treatment of alcohol or other drug use disorders. This study investigates associations between ayahuasca consumption in naturalistic settings and current alcohol and other drug use. Methods Online cross-sectional study of people who have consumed ayahuasca in religious, traditional and non-traditional settings in over 40 countries. A total of 8629 participants (53% male, average age 40 years) were included in the analysis. Logistic regressions were used to explore associations between ayahuasca drinking variables and the current use of alcohol and other drugs, as well as the influence of confounding factors, such as church or community membership. Results The number of times ayahuasca had been consumed was strongly associated with increased odds of never or rarely drinking alcohol, never or rarely engaging in ‘risky drinking’ and having not consumed a range of drugs in the past month, with these effects greater for those with a prior substance use disorder compared to those without. The strength of ayahuasca drinkers subjective spiritual experience, number of personal self-insights obtained and drinking ayahuasca with an ayahuasca church were also associated with lower substance use in some models. Discussion and Conclusions Consumption of ayahuasca in naturalistic settings is associated with lower self-reported current consumption of alcohol and other drugs for those with and without prior substance use disorders, with such effects present after adjusting for religious or social group effects.