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In this study, we aimed to identify the current trend of novel psychoactive substance (NPS) consumption in Italy and the level of knowledge about NPS in the general population and to explore the presence of risk factors for a lifetime NPS misuse. We conducted an anonymous online survey, administered to a large sample of people of any age living in Italy, asking for their knowledge about NPS as well as their history of drug use. Descriptive statistics were reported, and regression analyses were performed to identify risk factors for lifetime use of NPS. A total of 1097 individuals gave a valid response to the survey. A lifetime use of NPS was reported by 13.5%. Multivariate regression analysis showed that male gender (OR 2.33, 95% CI 1.52–3.55), homosexual sexual orientation (OR 2.52, 95% CI 1.21–5.27), a use of emergency services due to alcohol (OR 3.38, 95% CI 2.23–5.12) and previous contact with mental health services (OR 1.58, 95% CI 1.04–2.41) were risk factors for a lifetime use of NPS (p < 0.05). An improvement of preventive strategies and simplified access to information on NPS, especially tailored to individuals at increased risk of misuse, could slow diffusion of NPS and avert the consequences of substance use.