The aim of this study was to assess the seasonal dynamics of mycobacterial genes from aquatic ecosystems. To achieve this goal, two field trips were conducted in four towns endemic to Buruli ulcer in Côte d’Ivoire: Abidjan, Daloa, Tiassalé, and Yamoussoukro; one during the dry season and the other during the rainy season. Water was analyzed for the presence of the insertion sequence IS2404 using real-time PCR. Conventional PCR was then used to highlight MIRU/VNTR sequences. Following the analyses, six samples presented the MIRU 1 sequence during the dry season, with one copy found in three samples and two copies in another three. In contrast, during the rainy season, seven samples contained the MIRU 1 sequence, with two copies in one sample and three copies in two others. It is important to note that all samples were negative for the Locus 6 gene. This study highlights the significant impact of seasonal climatic conditions on the genetic composition of water samples and suggests a close correlation between genetic markers and mycobacterial contamination. These findings could have important public health and water resource management implications, as they provide additional information to understanding the risks associated with water contamination based on seasons. This seasonal approach to studying genetic markers in water samples opens new avenues for further research aimed at deepening our understanding of the dynamics of microbiological contaminants caused by mycobacteria in aquatic ecosystems.