An entomological study was conducted in a Leishmania infantum focus, including the identification of the sand fly species, the detection and the characterization of Leishmania DNA in female sand flies, and blood meal origins in engorged sand flies. A total of 643 sand flies (31% female, 69% male) was identified based on their morphological features or molecular markers. Ten different species were identified, with Phlebotomus perniciosus, the confirmed vector of L. infantum, being the most abundant (56%), P. papatasi in 25% of sand flies, the unique vector species of L. major, and P. longicuspis in 7% of cases, the suspected second vector of L. infantum. Moreover, the infection rate was 3.4% in P. perniciousus, P. papatasi, P. longicuspis, and Sergentomya minuta. Also, L. infantum was identified in five unfed P. perniciosus and two P. longicuspis. Our results suggest the vector role of P. perniciosus and P. longicuspis in the transmission cycle of L. infantum. The DNA of four mammalian species (human, rabbit, horse, and cow) was identified in the blood meals of sand flies, suggesting that these species are potential reservoirs of leishmaniasis, though it is not yet fully elucidated (especially for MON-24 and MON-80). We suggest the existence of different transmission cycles of L. infantum involving different species of sand flies and hosts.