Résumé : Purpose: The situation of road crashes-related deaths remains problematic in low-income countries. The present study aims at analyzing the first-aid knowledge and practices of professional motorcyclists (PMs) in the city of Cotonou in Benin. Materials and Methods: This is a cross-sectional analytical study conducted from 25 March to 19 April 2019 in Cotonou and concerned PMs registered in a fleet who gave their consent to participate in the study. The World Health Organization’s two-stage adaptive cluster sampling technique was applied to select the eligible PMs while respecting the proportionality rate per fleet. A logistic regression analysis was done and the odds ratios were estimated with 95% confidence interval. Results: The 430 PMs surveyed were all middle-aged men with an average age of 38.38 (±8.70). Among them, 62.56% knew at least one of the emergency phone numbers for the ambulance, police or fire services and 49.53% of the PMs knew at least one of the 3 techniques evaluated. In addition, 33.23% of PMs who had witnessed at least an RC stated that they had alerted the emergency services, and 32.27% said they had helped the victims. The main reason given for the lack of initiative in RCs was lack of knowledge of the course of action to take (19.64%). The level of knowledge was associated with the level of education (AOR: 3.11; CI 95%: 1.79–5.43) and with the length of experience (AOR: 2.56; CI 95%: 1.58–4.18). Conclusion: This study reveals that the level of knowledge and practice of PMs in the field of first aid in Cotonou is low and demonstrates the relevance and the need to include this target group in the first-aid chain for road crashes in Benin.