Cutaneous leishmaniasis (CL) is a widely neglected disease in Mali, West Africa. While Phlebotomus duboscqi has been implicated as the vector, the seasonality, distribution and anthropophilic behavior of this sand fly has not been adequately studied, nor has its role in human transmission been established in his region. To understand the entomological and epidemiological aspects of CL in Mali we selected two neighboring villages, Kemena and Sougoula, in the Baraouli health district. Since 2004, light traps, sticky traps and aspiration collections have been conducted monthly, in interior and exterior locations, within and around each village. Collected flies were identified and blood meal analysis was performed using PCR to identify host meal preference. Leishmanin skin test was performed on all consenting residents of each village to determine the prevalence of infection. Similar numbers of sand flies were collected in both villages with the genus Phlebotomus representing 24% and Sergentomyia, 76% of the total collection. Of the Phlebotomus sand flies collected, 99% were P. duboscqi with P. rodhaini representing less than 1%. The seasonality of P. duboscqi appeared to be bimodal with collections peaking in April and October. P. duboscqi was primarily collected using light traps inside human dwellings. Blood fed females tested positive for human blood but no other domestic animals. Although the two villages are approximately 5 km apart, the prevalence of CL varied greatly; 40% of inhabitants in Kemena were Leishmanin skin test positive versus 20% in Sougoula. The results presented here illustrate the public health importance of CL in Mali.