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The United Nations Strategic Plan for Biodiversity 2011-2020 is a key initiative within global efforts to halt and eventually reverse the loss of biodiversity. The very first target of this plan states that "by 2020, at the latest, people are aware of the values of biodiversity and the steps they can take to conserve and use it sustainably." Zoos and aquariums worldwide, attracting more than 700 million visits every year, could potentially make a positive contribution to this target. However, a global evaluation of the educational impacts of visits to zoos and aquariums is entirely lacking in the existing literature. To address this gap, we conducted a large-scale impact evaluation study. We used a pre- and postvisit repeated-measures survey design to evaluate biodiversity literacy-understanding of biodiversity and knowledge of actions to help protect it-of zoo and aquarium visitors worldwide. Ours was the largest and most international study of zoo and aquarium visitors ever conducted. In total, 5661 visitors to 26 zoos and aquariums from 19 countries around the globe participated in the study. Aggregate biodiversity understanding and knowledge of actions to help protect biodiversity both significantly increased over the course of zoo and aquarium visits. There was an increase from previsit (69.8%) to postvisit (75.1%) in respondents demonstrating at least some positive evidence of biodiversity understanding. Similarly, there was an increase from previsit (50.5%) to postvisit (58.8%) in respondents who could identify actions to help protect biodiversity that could be achieved at an individual level. Our results are the most compelling evidence to date that zoo and aquarium visits contribute to increasing the number of people who understand biodiversity and know actions they can take to help protect biodiversity.