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To determine dietary supplementation practices and opinions, preferred means for dietary supplement (DS) education, and antidoping opinions among elite Canadian athletes varying in age and gender.A descriptive, cross-sectional survey.Elite athlete training centers in Calgary, Canada and surrounding area.A total of 582 high-performance athletes (314 male, 268 female).High-performance athletes representing 27 sports with a mean age of 19.96 ± 3.91 years completed a validated questionnaire assessing DS practices and opinions by recall. Sport categories included varsity, Canadian Sport Centre Calgary (CSCC), and National Sport School (NSS).There was extensive DS use, with 88.4% of participants taking ≥1 DS (mean of 3.08 ± 1.87 DS per user) during the previous 6 months. Overall, sport drinks (22.4%), sport bars (14.0%), multivitamins and minerals (13.5%), protein supplements (9.0%), and vitamin C (6.4%) were most frequently reported. Older athletes were significantly more likely to report greater DS usage; to be advised by teammates, health food store retailers, and magazines; to prefer supplementation education via individual interviews; to claim awareness of anti-doping rules; and to perceive anti-doping compliance. Relative to gender, significant differences were observed for the types of DS reported; supplementation advisors; justifications for DS use; and awareness of anti-doping regulations.Utilization of this validated and reliable questionnaire has the potential for broad use and provides insight into the factors that influence DS use in elite athletes.