Recent global and regional exercises have been undertaken to identify 100 questions of relevance to policy makers that, if answered, would improve decision making and conservation actions. These were intentionally broad, but all included themes and questions of relevance to aquatic and fisheries professionals (e.g. exploitation, habitat alteration, effectiveness of protected areas, migratory connectivity and environmental effects of aquaculture). Here, the content of the global 100 question exercise relevant to aquatic and fisheries issues is summarized and a critical analysis is provided. Many of the questions addressed in apparently unrelated themes and topics (e.g. terrestrial, agriculture and energy policy) have potential relevance to fisheries and aquatic habitats, which underlines the connectivity between terrestrial and aquatic realms. Given the intimate link between aquatic environmental problems and human activities (including culture and economics), greater understanding of the human dimension is required to inform decision making. Stakeholder perspectives need to be included as a core component of the fisheries management triangle (i.e. managing fish, habitat and people). The benefits and risks of conducting a global 100 questions exercise with an exclusive focus on questions of relevance to fisheries and aquatic practitioners are also considered. There is no question that evidence-based approaches to conservation are essential for addressing the many threats that face aquatic ecosystems and reverse the imperilment trends among ichthyofauna. It is still unclear, however, as to the extent to which 100 questions exercises will help to achieve conservation and management targets for aquatic resources. A global 100 questions exercise that focused on fisheries and aquatic issues would certainly help to generate interest and awareness sufficient to justify such an exercise.