Background Lutzomyia longipalpis (Diptera: Psychodidae) is the major vector of Leishmania (Leishmania) infantum and thus plays a crucial role in the epidemiology of American visceral leishmaniasis (AVL). This vector is the best studied species of sand fly in the Neotropical region. Many studies claim that this vector is in fact a species complex; however there is still no consensus regarding the number of species that belong into this complex or the geographical distribution of sibling species. The aim of the present study was to analyze the genetic relationships within Lu. longipalpis populations in the state of Mato Grosso do Sul (MS), Brazil.Methodology/Principal FindingsWe collected 30 Lu. longipalpis (15 females and 15 males) from five localities (Campo Grande, Três Lagoas, Aquidauana, Miranda and Bonito) and 30 Lu. Cruzi from Corumbá, totaling 180 sandflies from MS, and 30 Lu. longipalpis from Estrela de Alagoas, state of Alagoas (AL), Northeast Brazil. We show that eight previously described microsatellite loci were sufficient in distinguishing Lu. longipalpis from Lu. Cruzi, which is a closely related species, and in differentiating between Lu. longipalpis collected in MS versus Estrela de Alagoas. Analyses of the genotypes revealed introgression between sympatric Lu. longipalpis and Lu. Cruzi. Conclusions/SignificanceOur findings support the hypothesis of cryptic species within the Lu. longipalpis complex. Furthermore, our data revealed introgression between Lu. longipalpis and Lu. cruzi. This phenomenon should be further investigated to determine the level and incidence of hybridization between these two species. We also demonstrated that microsatellite markers are a powerful tool for differentiating sand fly populations and species. The present study has elucidated the population structure of Lu. longipalpis in MS and, by extension, the Neotropical Lu. longipalpis complex itself.

Owing to the existence of cryptic species that are difficult to distinguish morphologically, the search for new taxonomic characters and methods for identifying and classifying sand flies continues. Lutzomyia longipalpis (Lutz & Neiva, 1912) and Lutzomyia cruzi (Mangabeira, 1938) (Diptera: Psychodidae) are two such species that occur in sympatry in some regions of Mato Grosso do Sul State (MS). Twenty females and twenty males from each of the five populations of Lu. longipalpis and one population of Lu. cruzi from MS were examined. An outlying population of Lu. longipalpis from Estrela de Alagoas, State of Alagoas, was used to compare the degree of divergence among the groups in MS. Specimens were cleared, mounted on slides, identified, and measured using LAS-Leica. The principal component analysis of morphometric characters showed a high degree of variation among females, while males varied to a lower degree. The populations of Alagoas and Miranda demonstrated the greatest variation. The first region, Alagoas, is geographically distant from the others and occurs under distinctly different ecological conditions, which likely accounts for the variation. Further studies should be made to elucidate the factors that contribute to the differences found between the populations of MS.