Traditional medicine (TM) is a significant resource for primary healthcare management all over the world, and principally in Africa. Quality improvement activities that promote evidence-based practices and the integration of traditional medicine into primary healthcare systems can help improve the quality of patient care. In the Republic of Benin (West Africa), traditional medicine practitioners (TMPs) provide different treatments and ways of use, depending on the ailments and the medicinal plants used. The present study aimed at documenting the knowledge, attitudes and practices of Beninese TMPs regarding the use of medicinal plants and the challenges associated with their activities. A focus group survey was conducted using semi-structured interviews with a sample of 91 TMPs in 8 departments of the Republic of Benin. The respondents had an average age of 50 years old and belonged to various categories of TMPs. Medicinal plants are harvested depending on the season and time of the day, and are dried in the shade before being used as decoctions or infusions. Nevertheless, the majority of TMPs do not conduct the necessary tests for quality control, efficacy or toxicity of the proposed remedies, which raises several scientific interrogations, particularly for the treatment of mental and brain-related disorders. Among ~ 110 plants used in the treatment of several pathologies, 66 were revealed as threatened species. The challenges faced by TMPs are mainly material, financial and technical difficulties. The present study reports the importance of intervention to modernize TM practices in Benin. Quality improvement could enhance healthcare delivery and provide support for evidence-based interventions aimed at addressing behavioral, social, and environmental determinants of health.