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Recreational fishing with the principal motivation of leisure is associated with important economic, social and conservation values. Nonetheless, it is also a primary pathway for the introduction and spread of invasive non-native species and aquatic animal disease. Several studies have explored the overall biosecurity risk posed by recreational anglers based on self-reported behaviours and the awareness of biosecurity campaigns. Nonetheless, there has been little in-depth exploration of the characteristics of key stakeholders who are implementing biosecurity best practices and the barriers that prevent anglers from undertaking biosecurity measures in the field. This study addresses this knowledge gap using an online questionnaire to collect information on angler socio-demographic characteristics, voluntary biosecurity behaviour and barriers affecting the implementation of biosecurity best practices in Great Britain. The results indicated that cleaning behaviours vary considerably among anglers, with angling frequency and the number of pieces of equipment affecting how likely anglers were to clean and dry these items. High levels of disinfectant use were reported among anglers, potentially attributed to the concurrent advocation of disinfectant to prevent the spread of aquatic animal disease. Barriers affecting the implementation of correct cleaning behaviours included a combination of factors, such as practicality, disability, lack of available information and individual values. These findings illustrate the importance for fishery managers, water companies and policymakers to tailor prevention measures and facilities for anglers to maximise the practicality of biosecurity measures and encourage long-term implementation of best practices.