Cyclists are vulnerable road users and face disproportionately high rates of road trauma, especially in low- and middle-income countries. Behaviour of road users is a system outcome, and thus studying cyclist behaviours can identify problems in the whole road transport system and assist in generating long-lasting, cost-effective solutions to promote cyclist safety. This study aims to investigate the similarities and disparities of cycling behaviour among countries with different income levels and cycling prevalence, and the relationships among cyclist demographic characteristics, behaviours and crash involvement. It applies the Cycling Behaviour Questionnaire (CBQ) to measure the behaviour of cyclists in three countries: Australia (high-income, emerging cycling country), China (mid-income, traditional cycling country) and Colombia (low-income, emerging cycling country). A total of 1094 cyclists (Australia 347, China 368, and Colombia 379) with a mean age of 31.8 years completed the online questionnaire. Confirmatory Factor Analysis was used to examine the factorial structure of the CBQ. The results showed that the three-factor CBQ (i.e., Traffic Violations, Errors and Positive Behaviours) had a stable and uniform factorial structure across all three countries. Cyclists reported more frequent positive behaviours than risky behaviours across all three countries. The Australian cyclists reported more positive behaviours and fewer violations than cyclists from the other countries, which is likely to reflect the road environment, policy and enforcement differences. Male cyclists tended to engage in more risky riding than females and reported more crash involvement. Older cyclists reported less risky riding and less involvement in crashes than younger cyclists. Cyclists who rode more often reported more frequent risky riding. The study helps understand the prevalence of risky/positive cyclist behaviours among countries and provides insights for developing systematic countermeasures to improve cycling environment and cyclist safety.