Location of study area.

<p>The location of the municipality of São João das Missões in northern Minas Gerais, Brazil. The native village of Imbaúbas located in the Xakriabá Indigenous Reserve, where the study was performed, is indicated.</p

Species identification of <i>Leishmania</i> from sand flies collected from trails in the Xakriabá Indigenous Reserve, Brazil, during the study period.

<p>Sand flies were grouped in pools of up to ten specimens of the same species, locality, and date, and ITS1 PCR-RFLP was performed with total DNA extracted from these pools. The figure represents an ethidium bromide-stained 4% agarose gel in which the amplicons were submitted to electrophoresis. Lanes: MW, molecular weight marker—100 bp; lanes 1–4, positive controls of <i>Le</i>. <i>amazonensis</i> (IFLA/BR/67/PH8), <i>Le</i>. <i>braziliensis</i> (MHOM/BR/75/M2903), <i>Le</i>. <i>infantum</i> (MHOM/BR/74/PP75), <i>Le</i>. <i>guyanensis</i> (MHOM/BR/75/M4147) respectively; 5–14, phlebotomine positive pools.</p

Detection of <i>Leishmania</i> in sand flies collected from trails in the Xakriabá Indigenous Reserve, Brazil, during the study period.

<p>Sand flies were pooled with up to ten specimens of the same locality, species and date, and ITS1 PCR was performed with total DNA extracted from these pools. The figure represents an ethidium bromide-stained 2% agarose gel in which the amplicons were submitted to electrophoresis. Lanes: MW, molecular weight marker—100 bp; lanes 1–4, positive controls of <i>Le</i>. <i>amazonensis</i> (IFLA/BR/67/PH8), <i>Le</i>. <i>braziliensis</i> (MHOM/BR/75/M2903), <i>Le</i>. <i>infantum</i> (MHOM/BR/74/PP75), <i>Le</i>. <i>guyanensis</i> (MHOM/BR/75/M4147) respectively; 5–14, phlebotomine positive pools; NC, negative control.</p

DNA Barcoding of Neotropical Sand Flies (Diptera, Psychodidae, Phlebotominae): Species Identification and Discovery within Brazil

<div><p>DNA barcoding has been an effective tool for species identification in several animal groups. Here, we used DNA barcoding to discriminate between 47 morphologically distinct species of Brazilian sand flies. DNA barcodes correctly identified approximately 90% of the sampled taxa (42 morphologically distinct species) using clustering based on neighbor-joining distance, of which four species showed comparatively higher maximum values of divergence (range 4.23–19.04%), indicating cryptic diversity. The DNA barcodes also corroborated the resurrection of two species within the shannoni complex and provided an efficient tool to differentiate between morphologically indistinguishable females of closely related species. Taken together, our results validate the effectiveness of DNA barcoding for species identification and the discovery of cryptic diversity in sand flies from Brazil.</p></div

Map showing the sampling sites across the Brazilian territory.

<p>(1) Cáceres. (2) Lagoa Santa. (3) Wenceslau Guimarães. (4) Bom Jesus do Itabapoana. (5) Afonso Cláudio. (6) Alfredo Chaves. (7) Alto Rio Novo. (8) Baixo Guandu. (9) Domingos Martins. (10) Ibitirama. (11) Itaguaçu. (12) Iúna. (13) João Neiva. (14) Manenópolis. (15) Marilândia. (16) Pancas. (17) Santa Leopoldina. (18) Santa Maria de Jetibá. (19) Santa Teresa.</p

Neighbor-joining tree of <i>COI</i> sequence divergences (K2P) obtained from 567 specimens of sand flies analyzed.

<p>Numbers next to the branches indicate Bootstrap percentages ≥80%. One black dash indicates groups partitioned by ABGD using the value of 2.15%. One white dash indicates the groups partitioned by ABGD using the cut-off of 1.29%. Two thin black dashes indicated groups that were not recognized by the ABGD partitions. The number of specimens is indicated behind each species name. PS1, PS2, and PS3 represent distinct groups that are hypothesized to represent provisional species.</p