Abstract
Lutzomyia longipalpis is the main vector of Leishmania infantum, the causative agent of visceral leishmaniasis in the Neotropical realm. Its taxonomic status has been widely discussed once it encompasses a complex of species. The knowledge about the genetic structure of insect vector populations helps the elucidation of components and interactions of the disease ecoepidemiology. Thus, the objective of this study was to genotypically analyze populations of the Lu. longipalpis complex from a macrogeographic perspective using Next Generation Sequencing. Polymorphism analysis of three molecular markers was used to access the levels of population genetic structure among nine different populations of sand flies. Illumina Amplicon Sequencing Protocol® was used to identify possible polymorphic sites. The library was sequenced on paired-end Illumina MiSeq platform. Significant macrogeographical population differentiation was observed among Lu. longipalpis populations via PCA and DAPC analyses. Our results revealed that populations of Lu. longipalpis from the nine municipalities were grouped into three clusters. In addition, it was observed that the levels of Lu. longipalpis population structure could be associated with distance isolation. This new sequencing method allowed us to study different molecular markers after a single sequencing run, and to evaluate population and inter-species differences on a macrogeographic scale.
Highlights
The history of visceral leishmaniasis (VL) in the Americas is closely related to the Lutzomyia longipalpis species, which was described by Lutz and Neiva [1] and identified as a Leishmania infantum vector based on consistent evidence from various studies on sand fly vector competence [2, 3, 4, 5, 6]
Forty-five DNA samples extracted from sand flies specimens representing all Brazilian regions were PCR amplified for three gene fragments
After processing the sequencing data, we observed that in 3 specimens (1 from São Borja, 1 from Fortaleza and 1 from Palmas) we obtained very few sequences for at least one of the amplified fragments, for this reason we decided to exclude them for further analysis
Summary
The history of visceral leishmaniasis (VL) in the Americas is closely related to the Lutzomyia longipalpis species, which was described by Lutz and Neiva [1] and identified as a Leishmania infantum vector based on consistent evidence from various studies on sand fly vector competence [2, 3, 4, 5, 6]. Macrogeographic genetic structure of Lutzomyia longipalpis complex populations and analysis, decision to publish, or preparation of the manuscript
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