Rationale: Ayahuasca is a South American psychoactive beverage that contains the naturally occurring psychedelic agent N,N-dimethyltryptamine (DMT). This “tea” has been used for centuries in religious and medicinal contexts in the rain forest areas of South America and is presently gaining the attention of psychedelic users in North America and Europe. Objectives: In the present study, the psychological effects and tolerability of ayahuasca were assessed. Methods: Three increasing doses of encapsulated freeze-dried ayahuasca (0.5, 0.75, and 1.0 mg DMT/kg body weight) were administered to six healthy male volunteers with prior experience in the use of this tea, in a single-blind crossover placebo-controlled clinical trial. Results: Ayahuasca produced significant dose-dependent increases in five of the six subscales of the Hallucinogen Rating Scale, in the LSD, MBG, and A scales of the Addiction Research Center Inventory, and in the “liking”, “good effects” and “high” visual analogue scales. Psychological effects were first noted after 30–60 min, peaked between 60–120 min, and were resolved by 240 min. The tea was well tolerated from a cardiovascular point of view, with a trend toward increase for systolic blood pressure. Modified physical sensations and nausea were the most frequently reported somatic-dysphoric effects. The overall experience was regarded as pleasant and satisfactory by five of the six volunteers, while one volunteer experienced an intensely dysphoric reaction with transient dis- orientation and anxiety at the medium dose and voluntarily withdrew from the study. Conclusions: Ayahuasca can be described as inducing changes in the perceptual, affective, cognitive, and somatic spheres, with a combination of stimulatory and visual psychoactive effects of longer duration and milder intensity than those previously reported for intravenously administered DMT.