Purpose Ayahuasca ceremonies are currently practiced all over the world. This study aims to investigate ayahuasca ceremonies in Colombia (where ayahuasca use is culturally entrenched) and Italy (where ayahuasca use has only recently spread). Design/methodology/approach In total, 32 Italian (53% males; 47% females) and 28 Colombian (79% males; 21% females) ayahuasca ceremony participants completed an online survey. Ceremonial setting and set (motivations for ayahuasca use, ayahuasca perceived effects and features of participants, i.e. drug use and problematic use, interpersonal dependency, spiritual orientation and quality of life) were investigated. Mann–Whitney U test, Pearson’s χ 2 test and Fisher’s exact test were used to compare the Italian and Colombian samples. Findings No differences emerged in the ceremonial setting. Slight differences were observed in motivations for ayahuasca intake and religious beliefs, but not in the preparation for ceremonies, quality of life, interpersonal dependence and spiritual orientation. Italians showed greater use of other psychoactive substances and more drug use problems. Practical implications Prevention regarding safer practices of consumption should be promoted to avoid improper uses. Originality/value Few studies have explored ayahuasca ceremonies (set and setting characteristics) in countries with different traditions on ayahuasca consumption. The results highlight that the use and experience of ayahuasca can take on different meanings depending on the cultural context.