The population structure of Phlebotomus ariasi, a proven vector of Leishmania infantum in the Mediterranean area, is still poorly understood. Previously, only two microsatellite loci had been developed to study the population genetics of this species. Herein we use these loci and determined fourteen novel microsatellite loci, useful for the characterization of Phlebotomus ariasi populations. These loci were tested on three populations of Phlebotomus ariasi, two from France and one from Portugal. In addition, the usefulness of these markers was also evaluated on seven other sandfly species.