Leishmaniases are endemic in Brazil, where Leishmania infantum has been detected in humans, dogs, cats, and phlebotomine vectors. Monitoring synanthropic vector populations is critical for leishmaniasis control-surveillance in such transmission-prone areas. Here, a suite of molecular approaches were used to assess Leishmania infection prevalence and to identify blood-meal sources in a large sample of sand flies collected in anthropic environments of a Leishmania-transmission area in Mato Grosso do Sul State (Rio Verde de Mato Grosso municipality), Central-West Brazil. We sampled sand flies monthly (January-June 2014 and 2016) in one peri-domestic site within each of six neighborhoods with recent records of human visceral and/or tegumentary leishmaniasis. kDNA-qPCR plus rDNA ITS-sequencing were used to detect and identify Leishmania in pooled female sand flies. Individual engorged females (n = 58) were used for blood-meal analyses through High-Resolution Melting (HRM) targeting the mtDNA cytb gene. Overall, 90.5% of 420 CDC trap-nights yielded vectors, for a total catch of 24,989 sand flies. We sub-sampled and identified 3088 sand flies of 12 species, including 2775 Lutzomyia longipalpis (the most abundant species at all sampling sites) and 297 Nyssomyia whitmani. Female sand flies (n = 1261) were grouped in 159 pools, of which 92 Lu. longipalpis (minimum infection rate [MIR] 8%) and 7 Ny. whitmani pools (MIR 7%) were Leishmania kDNA-positive. Most positive Lu. longipalpis were collected in the 2016 rainy season. Sequencing confirmed L. infantum in Lu. longipalpis samples. HRM analyses identified chicken DNA in 57 sand flies (98.3%), 37 of which were Leishmania DNA-positive (64.9%); human blood was found in just one (Leishmania-negative) female. Our data show ongoing risk of L. infantum transmission to humans in the study area, where Leishmania-infected sandfly vectors are common and heavily rely on chicken blood in the peri-domestic environment.

Introduction:This study describes the occurrence of trypanosomatids in phlebotomines in Brasília, Brazil. Methods: Two hundred and ten females of 13 sand fl y species were analyzed by polymerase chain reaction (PCR) using different molecular markers (D7 24Sα rRNA, kDNA, and ITS1) and sequencing. Results: PCR revealed trypanosomatid-positive samples from Nyssomyia whitmani and Evandromyia evandroi, which were negative by kDNA and ITS1 Leishmania-specifi c PCRs. DNA sequence analysis of D7 24Sα rRNA amplicons indicated the occurrence of Blastocrithidia sp. and Trypanosoma sp. in Nyssomyia whitmani and Evandromyia evandroi, respectively. Conclusions: Two trypanosomatid species other than Leishmania sp. were found to circulate in sand fl ies in Central Brazil.

BACKGROUND Knowledge on synanthropic phlebotomines and their natural infection by Leishmania is necessary for the identification of potential areas for leishmaniasis occurrence.OBJECTIVE To analyse the occurrence of Phlebotominae in gallery forests and household units (HUs) in the city of Palmas and to determine the rate of natural infection by trypanosomatids.METHODS Gallery forests and adjacent household areas were sampled on July (dry season) and November (rainy season) in 2014. The total sampling effort was 960 HP light traps and eight Shannon traps. Trypanosomatids were detected in Phlebotominae females through the amplification of the SSU rDNA region, and the positive samples were used in ITS1-PCR. Trypanosomatid species were identified using sequencing.FINDINGS A total of 1,527 sand flies representing 30 species were captured in which 949 (28 spp.) and 578 (22 spp.) were registered in July and November, respectively. In July, more specimens were captured in the gallery forests than in the HUs, and Nyssomyia whitmani was particularly frequent. In November, most of the specimens were found in the HUs, and again, Ny. whitmani was the predominant species. Lutzomyia longipalpis was commonly found in domestic areas, while Bichromomyia flaviscutellata was most frequent in gallery forests. Molecular analysis of 154 pools of females (752 specimens) identified Leishmania amazonensis, L. infantum, and Crithidia fasciculata in Ny. whitmani, as well as L. amazonensis in Lu. longipalpis, Trypanosoma sp. and L. amazonensis in Pintomyia christenseni, and L. amazonensis in both Psathyromyia hermanlenti and Evandromyia walkeri.MAIN CONCLUSIONS These results show the importance of gallery forests in maintaining Phlebotominae populations in the dry month, as well as their frequent occurrence in household units in the rainy month. This is the first study to identify Leishmania, Trypanosoma, and Crithidia species in Phlebotominae collected in Palmas, Tocantins, Brazil.

Resumo. O objetivo deste estudo foi analisar a ocorrência de flebotomíneos em matas de galeria do Distrito Federal (DF), em diferentes estações climáticas, estratos verticais e condições de preservação das matas. As capturas foram realizadas em três áreas na Fazenda Água Limpa da Universidade de Brasília. Em cada área foi analisada a ocorrência de flebotomíneos em 20 pontos visitados em dois meses, novembro de 2012 (estação chuvosa) e maio de 2013 (estação seca). Armadilhas luminosas HP foram instaladas a 1,5 m e a 10 m de altura do solo, operando entre 17:00 e 07:00 h, por três dias consecutivos.