Background and Aim: Human dog-transmitted rabies remains a serious public health issue in Mali despite efforts to mitigate it. Indeed, several stakeholders have implemented multiple initiatives for years. However, there is still a lack of cooperation among stakeholders. This study was conducted to capture situational data on rabies and One Health practices among key actors involved in the fight against rabies in Mali. Materials and Methods: Data from veterinary and medical services were collected for analysis. Data collection was based on a review of technical documents (epidemiology of rabies in animals and humans and activities conducted to tackle the disease) and stakeholder interviews (awareness of rabies and the One Health approach). Results: With regard to the epidemiological situation, the findings revealed unsatisfactory vaccination coverage in dogs and low suspected sample collection for laboratory diagnosis. In addition, only half of the exposed individuals received complete post-exposure prophylaxis. A survey carried out by experts in the fields of animal health, human health, and the environment indicated that only 22.47% were well aware of rabies and had never heard of the One Health approach. Overall, there was no significant association between knowledge and participant sector of origin (p > 0.05), highlighting the need for operationalization of the One Health approach in Mali. Conclusion: Therefore, an integrated national action plan for rabies control should be developed with a particular focus on capacity building, particularly in the area of the One Health core competencies. Keywords: epidemiology, multisectoral engagement, One Health, Public Health priority zoonoses, Stakeholders.