Acenocoumarol is the only oral antivitamin K anticoagulant prescribed and marketed in Algeria for the prevention and treatment of thromboembolic diseases. Nevertheless, the difficulty of handling this drug linked to a narrow therapeutic index and to great inter- and intra- individual variabilities of the response to the treatment poses a major public health issue because of its iatrogenic risk. To improve the management of patients on acenocoumarol and optimize its dosage adjustment and reduce iatrogenic secondary to vitamin K antagonists, our main objective work is to assess the knowledge, attitude and practice of healthcare professionals regarding this anticoagulant agent. We conducted a multicenter descriptive cross-sectional study from April to May 2021. We interrogated 152 health professionals of all ages, all sexes and all grades from different health departments in Oran City (Algeria) prescribing anticoagulants, through a validated questionnaire made up of 26 questions. Statistical analysis was achieved by SPSS version 20 software. Thus, 59.9% were women with a sex ratio of 0.67, 92.8% knew that acenocoumarol belongs to the antivitamin K class, 45.0% chose INR and PT as follow-up parameters, 36.0% chose INR alone and 78.9% were unaware of acenocoumarol dose adjustment algorithms as well as 82.0% were unaware of the existence of pharmacogenetics testing. 54.0% use the recommendations of the HAS for dosage adjustment of patients on acenocoumarol. 07.9% suggested using pharmacogenetic tests to improve the dosage adjustment of acenocoumarol, 06.6% suggested patient follow-up and 05.9% suggested therapeutic patient education as well as 74.3%, wanted to do training. Therefore, this study highlighted gaps in the medical practice of physicians in terms of dosage adjustment, pharmacogenetics and its impact on dosage optimization. However, the evaluation of their knowledge was considered unsatisfactory. To overcome this lack of knowledge, it seems necessary to make physicians aware of the contribution of pharmacogenetics of acenocoumarol in Algeria.