Molecular and morphometric study of Brazilian populations of Psychodopygus davisi.

In this study, we analysed the molecular and morphometric differences of several populations of the putative sand fly vector Psychodopygus davisi (Root, 1934) (Diptera, Psychodidae, Phlebotominae) in Brazil. We amplified the 658 base pair fragments of the DNA barcoding region-cytochrome c oxidase subunit 1 (COI) gene-for 57 specimens of P. davisi and three specimens of Psychodopygus claustrei (Abonnenc, Léger & Fauran, 1979). We merged our data with public sequences of the same species available from GenBank. Then, the combined dataset-87 sequences and 20 localities-was analysed using population structure analysis and different species delimitation approaches. Geometric morphometry of wings was performed for 155 specimens of P. davisi populations from the North, Midwest and Southeast Brazilian regions, analysing the differences in centroid sizes and canonical variates. Molecular analysis indicated high intraspecific genetic distance values for P. davisi (maximum p distance = 5.52%). All algorithms identified P. davisi and P. claustrei as distinct molecular taxonomic units, despite the low interspecific distance (p distance to the nearest neighbour = 4.79%). P. davisi sequences were split into four genetic clusters by population structure analysis and at least five genetic lineages using intermediate scenarios of the species delimitation algorithms. The species validation analysis of BPP strongly supported the five-species model in our dataset. We found high genetic diversity in this taxon, which is in agreement with its wide geographic distribution in Brazil. Furthermore, the wing analysis showed that specimens from the Southeast Region of Brazil are different from those in the North and the Midwest. The evolutionary patterns of P. davisi populations in Brazil suggest the presence of candidate species, which need to be validated in future studies using a more comprehensive approach with both genomic data and morphological characters.

DNA Barcoding of genus Culicoides biting midges (Diptera: Ceratopogonidae) in the Brazilian Amazon.

Culicoides biting midges are capable to transmit Oropouche virus, Bluetongue virus and Mansonella spp. This study aimed to assess the utility of DNA barcode as an alternative method in the Culicoides species identification. The study was conducted in Jamari National Forest. Biting midges were collected using HP light traps during four months, February, April, August and October 2018. Insects were morphologically identified to the species level, and rest of the body were subjected to DNA extraction and PCR targeting a fragment of the cytochrome c oxidase subunit I (COI) gene, which were analyzed and deposited in GenBank. A phylogenetic gene tree was reconstructed using RAxML software, and the sequences were assigned at Molecular Operational Taxonomic Unit (MOTU) level by species delimitation algorithms. According to morphological approach, 18 species of 2 subgenera and 7 species groups were identified. A total of 191 new COI barcodes from 18 species were generated. Of these, fifteen species have been deposited for the first time in all datasets in the world. These sequences allowed the correct identification of 188 and 187 specimens according to the BM and BCM criteria, respectively. The intraspecific genetic distances ranged from 0 to 16.5%, while the interspecific ones ranged from 2.1 to 27.1%. The nominal species Culicoides glabellus and C. tetrathyris splitted into three and two MOTUs, respectively, except for mPTP, indicating a cryptic diversity in these species. Also, sequences of C. pseudodiabolicus formed two MOTUs using all algorithms, except for PTP and ABGD, suggesting the existence of two potential species. In contrast, some barcodes of C. quasiparaensis and C. paraensis merged into a single MOTU, which can be explained by the complex characteristics of the paraensis group, since these species have similar morphological characters. Here, we provided the first COI barcodes for biting midges in Rondônia and Brazil, and demonstrates that these are sufficient to discriminate between some species.

Updating the knowledge of sand flies (Diptera, Psychodidae) in Rondônia State, Brazil.

Background: Sandflies are insects important for the transmission cycles of the leishmaniases. Despite being studied since the 1960s in the State of Rondônia (Brazil), several gaps exist regarding our working knowledge of these insects. This study aimed to construct an up-to-date database of sandflies using complementary information from the speciesLink database and the scientific literature, as well as to elaborate integrated abundance maps. We identified 153,155 records of sandflies captured in Rondônia between 1965-2021; after exclusion, 147,258 reports (speciesLink - 3,408, Rondônia studies - 143,850) associated with 15 genera and 140 species were mapped. The most abundant species observed were Psychodopygusdavisi (Root, 1934) (43,818 records), Nyssomyiawhitmani (Antunes & Coutinho, 1939) (12,594), Psychodopyguscarrerai (Barretto, 1946) (11,840), Psychodopygushirsutus (Mangabeira, 1942) (9,676), Nyssomyiaantunesi (Coutinho, 1939) (8,847), Trichophoromyiaubiquitalis (Mangabeira, 1942) (5,505), Psychodopygusgeniculatus (Mangabeira, 1941) (4,644), Pintomyianevesi (Damasceno & Arouck, 1956) (4,140), Trichophoromyiaauraensis (Mangabeira, 1942) (3,579), Psychodopyguscomplexus (Mangabeira, 1941) (2,659), Nyssomyiafraihai (Martins, Falcão & Silva, 1979) (2,504) and Bichromomyiaflaviscutellata (Mangabeira, 1942) (1,418). A total of 20 records of Leishmania detection corresponded to eight sand fly species. The present dataset provides updated information on the distribution of sandflies of Rondônia, including those considered potential vectors of Leishmania, which should prove useful to guide future studies. New information: The present study provides an extensive dataset built from all studies reporting phlebotomine sandflies in the Brazilian State of Rondônia. Online distribution maps can aid scientists who wish to consult the updated list of sand fly species and view the distribution of these insects, as well as those considered potential vectors of Leishmania. The results of the present study can serve as the basis for future studies on sandflies conducted in the State.

The spread of visceral leishmaniasis in Brazil: the first canine cases described in Ji-Paraná, Rondônia, Brazil.

Leishmaniases are zoonotic diseases caused by protozoa of the genus Leishmania. The disease has two clinical manifestations described in humans: visceral (VL) and cutaneous (CL) leishmaniasis. In Brazil, there has been an expansion of human VL. The participation of the dog as a reservoir of Leishmania infantum, the agent of VL, is important for the epidemiology of the disease since canine cases generally precede human cases. The present study aimed to evaluate the occurrence of Leishmania spp. infection in dogs in the municipality of Ji-Paraná by PCR assays using blood samples. Leishmania DNA was detected in two of the 105 studied dogs. The PCR products were sequenced and confirmed that the two samples (1.90%) correspond to L. infantum. The dogs had allochthonous history. Therefore, the positive results found here should serve as a warning to public health agencies. This is because Ji-Paraná is the third municipality to register cases of canine leishmaniasis (CanL) in Rondônia state. Thus, reinforcing the importance of expanding studies on the epidemiology and surveillance of VL in the region.

Occurrence of Leishmania infection in the immediate geographic region of Ji-Paraná, Rondônia State, Brazil.

INTRODUCTION: This study evaluated the epidemiology of American cutaneous leishmaniasis in the immediate region of Ji-Paraná, Rondônia State. METHODS: Samples and epidemiological data were collected from 105 patients. RESULTS: Leishmania infection was observed in 58 (55.2%) patients, and Leishmania braziliensis was present in 82.9% of the 41 sequenced samples. Infected patients were predominantly male (93.1%). Leishmania infection was twice as prevalent among rural inhabitants versus urban inhabitants. Lesions were more frequent in the upper limbs (arms/hands, 41.82%). CONCLUSIONS: The present data corroborate the zoonotic profile of cutaneous leishmaniasis; this information could help to improve surveillance and control strategies.

DNA-based blood meal analysis of Culicoides (Diptera: Ceratopogonidae) species from Jamari National Forest, Southwestern Amazon, Brazil.

Culicoides biting midges are insects involved in the transmission of filarial nematodes, protozoans, and viruses. Greater knowledge of Culicoides blood meal sources could improve our understanding of parasite transmission cycles. Our study used molecular tools to evaluate the blood meal sources of Culicoides biting midges from sylvatic environments. This study was conducted in Jamari National Forest, Rondônia, Brazil. Culicoides were captured using HP light traps positioned at ground level (1.5 m above ground) and in canopy (15 m above ground). To identify blood meal sources, females were subjected to DNA extraction and PCR targeting the cytb gene fragment, and the obtained sequences were analyzed and compared with sequences from GenBank. DNA extraction and PCR were performed on 455 Culicoides females, and blood meal sources were identified in 186 females. Thirty Culicoides specimens were collected from the Potosi trail and 156 were collected from the Santa Maria trail. A total of 22 species were captured; all 22 species were collected in canopy (100%) but only three species were collected at ground level (13.6%). The cytb fragment was amplified in 162 of 186 samples. Sample sequencing identified cytb DNA from nine blood-meal sources: Pauxi sp., Psophia viridis, Ramphastos tucanus tucanus, Choloepus didactylus, Choloepus hoffmanni, Tamandua tetradactyla, Ateles chamek, Homo sapiens and Pithecia irrorata. We observed that several different blood meal sources were utilized by a high diversity of Culicoides species. The abundance of Culicoides in the canopy may be related to the fact that the majority of blood meal hosts feed in treetops. We observed that C. (Hoffmania) sp. and C. coutinhoi tend to be more generalist, feeding on a range of mammals and piciform, gruiform and galliform birds. This data improves our knowledge of the feeding profile of biting midges from forest environments and should serve as a future basis for defining zoonotic transmission cycles.

Biting Midges in Jamari National Forest, in the Brazilian Amazon, With 12 New Records of Culicoides Species (Diptera: Ceratopogonidae) for the State of Rondônia.

Culicoides biting midges are small hematophagous flies, some species of which transmit pathogens to livestock and humans. In the Brazilian Amazon Basin, Culicoides diversity has remained largely unexamined. Jamari National Forest (JNF) is a conservation area in Rondônia State where research and environmental education are conducted in conjunction with the protection of natural resources, and the recovery of degraded areas. The present study augments our knowledge of Culicoides diversity in JNF. The survey collected 640 individuals from 41 species; 632 individuals were collected by HP light trap and 8 were collected by the BG Sentinel trap. Twelve species are first-time records for Rondônia: Culicoides acotylus Lutz, Culicoides albuquerquei Wirth & Blanton, Culicoides baniwa Felippe-Bauer, Culicoides bricenoi Ortiz, Culicoides brownei Spinelli, Culicoides carsiomelas Wirth & Blanton, Culicoides debilipalpis Lutz, Culicoides forattinii Ortiz, Culicoides ginesi Ortiz, Culicoides profundus Santarém, Felippe-Bauer & Trindade, Culicoides pseudoreticulatus Santarém, Felippe-Bauer & Castellón, and Culicoides spurius Santarém, Felippe-Bauer & Trindade. The most abundant species were Culicoides glabrior Macfie, Culicoides pseudodiabolicus Fox, Culicoides debilipalpis, Culicoides quasiparaensis Clastrier, and Culicoides brownei Spinelli. The present study updates the Culicoides species count in Rondônia to 55, and highlights the importance of JNF as a hotbed for biting midge diversity.

The spread of visceral leishmaniasis in Brazil: the first canine cases described in Ji-Paraná, Rondônia, Brazil / A disseminação da leishmaniose visceral no Brasil: os primeiros casos caninos descritos em Ji-Paraná, Rondônia, Brasil

Abstract Leishmaniases are zoonotic diseases caused by protozoa of the genus Leishmania. The disease has two clinical manifestations described in humans: visceral (VL) and cutaneous (CL) leishmaniasis. In Brazil, there has been an expansion of human VL. The participation of the dog as a reservoir of Leishmania infantum, the agent of VL, is important for the epidemiology of the disease since canine cases generally precede human cases. The present study aimed to evaluate the occurrence of Leishmania spp. infection in dogs in the municipality of Ji-Paraná by PCR assays using blood samples. Leishmania DNA was detected in two of the 105 studied dogs. The PCR products were sequenced and confirmed that the two samples (1.90%) correspond to L. infantum. The dogs had allochthonous history. Therefore, the positive results found here should serve as a warning to public health agencies. This is because Ji-Paraná is the third municipality to register cases of canine leishmaniasis (CanL) in Rondônia state. Thus, reinforcing the importance of expanding studies on the epidemiology and surveillance of VL in the region.

Resumo As leishmanioses são doenças causadas por protozoários do gênero Leishmania. A doença apresenta duas manifestações clínicas: leishmaniose visceral (LV) e cutânea (LC). No Brasil, a LV está em expansão. A participação do cão como reservatório é importante para a epidemiologia da doença, pois os casos caninos geralmente precedem os humanos. O presente estudo avaliou a ocorrência de LV em cães (LVC) do município de Ji-Paraná por meio de ensaios de PCR, utilizando-se amostras de sangue. O DNA de Leishmania foi detectado em dois dos 105 cães estudados. Os produtos da PCR foram sequenciados e confirmaram que duas amostras (1,90%) eram Leishmania infantum. Os cães tinham histórico alóctone. Os resultados positivos encontrados servem de alerta aos órgãos públicos de saúde. Isso porque Ji-Paraná é o terceiro município a registrar casos de LVC no estado de Rondônia. Dessa forma, reforça-se a importância da ampliação dos estudos sobre a epidemiologia e vigilância da LV na região.

Occurrence of Leishmania infection in the immediate geographic region of Ji-Paraná, Rondônia State, Brazil

Abstract INTRODUCTION: This study evaluated the epidemiology of American cutaneous leishmaniasis in the immediate region of Ji-Paraná, Rondônia State. METHODS: Samples and epidemiological data were collected from 105 patients. RESULTS: Leishmania infection was observed in 58 (55.2%) patients, and Leishmania braziliensis was present in 82.9% of the 41 sequenced samples. Infected patients were predominantly male (93.1%). Leishmania infection was twice as prevalent among rural inhabitants versus urban inhabitants. Lesions were more frequent in the upper limbs (arms/hands, 41.82%). CONCLUSIONS: The present data corroborate the zoonotic profile of cutaneous leishmaniasis; this information could help to improve surveillance and control strategies.

Vertical stratification of sand fly diversity in relation to natural infections of Leishmania sp. and blood-meal sources in Jamari National Forest, Rondônia State, Brazil.

BACKGROUND: Almost 1000 cases of American cutaneous leishmaniasis have been registered yearly in Rondônia State, Brazil. Little is known about the Leishmania transmission cycle (vectors and reservoirs) in the state. This study aimed to evaluate sand fly fauna from two vertical stratification layers in order to identify potential vectors and their blood-meal sources. METHODS: The study was conducted in Jamari National Forest. Sand flies were collected in the canopy (15 m) and at ground level (1 m) using HP light traps during four months, February, April, August and October, 2018. Insects were identified to the species level, and females were subjected to DNA extraction and PCR targeting minicircle kDNA and hsp70 (for Leishmania detection and species identification), and cytb (to identify blood-meal sources). Exploratory data analysis was used to determine mean of abundance and species richness between stratifications. The hsp70 and cytb sequences were analyzed and compared with sequences from GenBank. RESULTS: Overall, 68 species were identified from 15,457 individuals. On the Potosi trail, 7531 individuals of 49 species were collected; canopy captures totaled 6463 individuals of 46 species, while ground captures totaled 1068 individuals of 38 species. On the Santa Maria trail, 7926 individuals of 61 species were collected; canopy captures totaled 6136 individuals of 51 species, while ground captures totaled 1790 individuals of 53 species. A total of 23 pools were positive for kDNA (canopy n = 21, ground n = 2). Only two samples were sequenced for hsp70 (both in canopy); one sequence exhibited similarity with Leishmania braziliensis (Lutzomyia davisi pool) and another with L. naiffi (Lu. antunesi pool). The cytb fragment was amplified in 11 of 86 samples. Sample sequencing identified cytb DNA from 5 blood-meal sources: Micrastur gilvicollis, Psophia viridis, Tamandua tetradactyla, Homo sapiens and Choloepus didactylus. CONCLUSIONS: Sand fly fauna is more diverse in the canopy than at ground level. Factors such as blood-meal sources, resting sites, and abiotic components probably contribute to high abundance in the canopy. Our results reinforce the possibility that Lu. antunesi and Lu. davisi participate in Leishmania transmission in forest environments and may play an important role in transmission from sylvatic to human hosts.

Diversity, natural infection and blood meal sources of phlebotomine sandflies (Diptera, Psychodidae) in the western Brazilian Amazon.

BACKGROUND: The state of Rondônia (RO) is a hot spot for human cases of cutaneous leishmaniasis. Many sandfly species in RO are putative vectors of leishmaniasis. OBJECTIVES: This study examines the diversity patterns and the presence of Leishmania DNA and blood meal sources of sandflies in RO. METHODS: A sandfly survey was performed between 2016 and 2018 in 10 municipalities categorised into three different environment types: (i) Conservation Unit (CUN) - comprised of preserved ombrophilous forests; (ii) Forest Edge (FE) - small forest fragments; and (iii) Peridomicile (PE) - areas around dwellings. FINDINGS: A total of 73 species were identified from 9,535 sandflies. The most abundant species were Psychodopygus davisi (1,741 individuals), Nyssomyia antunesi (1,397), Trichophoromyia auraensis (1,295) and Trichophoromyia ubiquitalis (1,043). Diversity was the highest in CUN, followed by the FE and PE environments. One pool of Ps. davisi tested positive for Leishmania braziliensis, reinforcing the possibility that Ps. davisi acts as a vector. The cytochrome b (cytb) sequences were used to identify three blood meal sources: Bos taurus, Homo sapiens and Tamandua tetradactyla. MAIN CONCLUSIONS: Our results demonstrated that sandflies can switch between blood meal sources in differing environments. This study enhances the knowledge of the vector life cycle in RO and provides information relevant to leishmaniasis surveillance.

Diversity, natural infection and blood meal sources of phlebotomine sandflies (Diptera, Psychodidae) in the western Brazilian Amazon

BACKGROUND The state of Rondônia (RO) is a hot spot for human cases of cutaneous leishmaniasis. Many sandfly species in RO are putative vectors of leishmaniasis. OBJECTIVES This study examines the diversity patterns and the presence of Leishmania DNA and blood meal sources of sandflies in RO. METHODS A sandfly survey was performed between 2016 and 2018 in 10 municipalities categorised into three different environment types: (i) Conservation Unit (CUN) - comprised of preserved ombrophilous forests; (ii) Forest Edge (FE) - small forest fragments; and (iii) Peridomicile (PE) - areas around dwellings. FINDINGS A total of 73 species were identified from 9,535 sandflies. The most abundant species were Psychodopygus davisi (1,741 individuals), Nyssomyia antunesi (1,397), Trichophoromyia auraensis (1,295) and Trichophoromyia ubiquitalis (1,043). Diversity was the highest in CUN, followed by the FE and PE environments. One pool of Ps. davisi tested positive for Leishmania braziliensis, reinforcing the possibility that Ps. davisi acts as a vector. The cytochrome b (cytb) sequences were used to identify three blood meal sources: Bos taurus, Homo sapiens and Tamandua tetradactyla. MAIN CONCLUSIONS Our results demonstrated that sandflies can switch between blood meal sources in differing environments. This study enhances the knowledge of the vector life cycle in RO and provides information relevant to leishmaniasis surveillance.

Phlebotomine Sand Fly Composition (Diptera: Psychodidae) and Putative Vectors of American Cutaneous Leishmaniasis in Porto Velho Municipality, Western Amazon, Brazil.

This study evaluated the species composition of sand flies and identified potential vectors of Leishmania Ross species in rural areas of Porto Velho, Rondônia State, Brazil. American cutaneous leishmaniasis (ACL) is one of the gravest threats to public health in this state. Sand flies were collected over the course of 2014 and 2015 using HP light traps. Polymerase chain reaction was performed by targeting the Leishmania mkDNA region. In total, 2,344 sand flies were collected, from which 45 species, nine subgenera, and five species group were identified. The most abundant species were Lutzomyia antunesi (Coutinho) (n = 597, 25.47%), Lutzomyia ubiquitalis (Mangabeira) (n = 496, 21.16%), and Lutzomyia octavioi (Vargas) (n = 199, 8.49%). The greatest diversity occurred in the forest environment where the most abundant species were L. antunesi (n = 588, 25.07%), L. ubiquitalis (n = 493, 21.02%), L. octavioi (n = 199, 8.49%), and Lutzomyia flaviscutellata (Mangabeira) (n = 132, 5.63%). Two pools of L. ubiquitalis were positive for Leishmania DNA, which suggests that L. ubiquitalis is a putative vector of leishmaniasis in the municipality of Porto Velho.

Culicoides hildebrandoi, a new species of the reticulatus species group from the Brazilian Amazon Region (Diptera, Ceratopogonidae).

A new species of biting midge (Diptera: Ceratopogonidae), Culicoides hildebrandoi sp. n., is described and illustrated based on female and male specimens from the states of Amazonas and Rondônia, Brazil. This new species belongs to the reticulatus species group and differs from the 24 other species of this group by the elongate slightly swollen 3(rd) palpal segment with scattered capitate sensilla but lacking a sensory pit.

Sandfly fauna (Diptera: Psychodidae) from caves in the state of Rondônia, Brazil.

This study had the aim of ascertaining the sandfly fauna and possible presence of Leishmania in these insects, collected in caves in the state of Rondônia, Brazil. Collections were conducted in eight caves located in two different areas of this state. Leishmania in the sandflies collected was detected using the polymerase chain reaction (PCR). This was the first study on sandflies from caves in Rondônia and, among the total of 1,236 individuals collected, 24 species and 10 genera were identified. The species Evandromyia georgii was collected for the first time in Rondônia and the most abundant species were Trichophoromyia ubiquitalis with 448 individuals (36.2%), followed by T. octavioi with 283 (22.9%) and E. georgii with 179 (14.5%). For the PCR, 17 pools were analyzed and five pools were positive (forT. auraensis in three pools and for Nyssomyia shawi and N. antunesi in one pool each). The kDNA region was amplified and the presence of Leishmania DNA was confirmed. The sandfly fauna in these caves can be considered diverse in comparison with similar studies in other regions. It may be that some species use caves as a temporary shelter and breeding site, while other species live exclusively in this environment. The detection of Leishmania DNA indicates that this pathogen is circulating in cave environments and that further studies are needed in order to ascertain the risks of infection by leishmaniasis in these locations with high touristic potential.

Sandfly fauna (Diptera: Psychodidae) from caves in the state of Rondônia, Brazil / Fauna de flebotomíneos (Diptera: Psychodidae) em cavernas do estado de Rondônia, Brasil

Abstract This study had the aim of ascertaining the sandfly fauna and possible presence ofLeishmania in these insects, collected in caves in the state of Rondônia, Brazil. Collections were conducted in eight caves located in two different areas of this state. Leishmania in the sandflies collected was detected using the polymerase chain reaction (PCR). This was the first study on sandflies from caves in Rondônia and, among the total of 1,236 individuals collected, 24 species and 10 genera were identified. The speciesEvandromyia georgii was collected for the first time in Rondônia and the most abundant species were Trichophoromyia ubiquitalis with 448 individuals (36.2%), followed by T. octavioi with 283 (22.9%) and E. georgii with 179 (14.5%). For the PCR, 17 pools were analyzed and five pools were positive (forT. auraensis in three pools and for Nyssomyia shawi and N. antunesi in one pool each). The kDNA region was amplified and the presence of Leishmania DNA was confirmed. The sandfly fauna in these caves can be considered diverse in comparison with similar studies in other regions. It may be that some species use caves as a temporary shelter and breeding site, while other species live exclusively in this environment. The detection of LeishmaniaDNA indicates that this pathogen is circulating in cave environments and that further studies are needed in order to ascertain the risks of infection by leishmaniasis in these locations with high touristic potential.

Resumo O objetivo deste estudo foi conhecer a fauna de flebotomíneos, e possível presença de Leishmania nestes insetos, coletados em cavernas do Estado de Rondônia. As coletas foram realizadas em oito cavernas localizadas em duas áreas diferentes do Estado. A detecção de Leishmania nos flebotomíneos foi realizada por reação em cadeia da polimerase (PCR). Este foi o primeiro trabalho com flebotomíneos em cavernas de Rondônia e um total de 1,236 indivíduos foram coletados e identificados em 24 espécies e 10 gêneros.Evandromyia georgii foi coletada pela primeira vez em Rondônia, e as espécies mais abundantes foram Trichophoromyia ubiquitalis com 448 indivíduos (36.2%) seguida por T. octavioi com 283 (22.9%) e E. georgii com 179 (14.5%). No estudo de PCR, 17 pools foram analisados, sendo cinco positivos (T. auraensis - 3, Nyssomyia shawi eN. antunesi - 1 cada). A região do kDNA foi amplificada confirmando a presença de DNA de Leishmania. A fauna de flebotomíneos nestas cavernas foi considerada diversa quando comparada com estudos semelhantes de outras regiões. É possível que algumas espécies utilizem cavernas como abrigo temporário e local de procriação e outras sejam exclusivas deste ambiente. A detecção de DNA de Leishmania indica que este patógeno está circulando no ambiente cavernícola, sendo necessários mais estudos para conhecer o risco de transmissão de leishmanioses nestes locais com alto potencial turístico.

Ecological aspects and molecular detection of Leishmania DNA Ross (Kinetoplastida: Trypanosomatidae) in phlebotomine sandflies (Diptera: Psychodidae) in terra firme and várzea environments in the Middle Solimões Region, Amazonas State, Brazil.

BACKGROUND: Phlebotomine sand flies (Diptera: Psychodidae) are insects of medical importance due to the role that some species play in the transmission of leishmaniasis. This work aimed to study some ecological aspects among sand flies fauna inhabiting two different environments: the várzea (lowland Amazonian forest) and terra firme (upland Amazonian forest), both located in Tefé Municipality, Amazonas State, Braziland to detect Leishmania infection in those phlebotomine populations. METHODS: Sand flies were collected using HP light traps. Collection took place over the course of six months: January, February, April, August, September, and October of 2013. To detect natural infection by Leishmania, DNA samples were extracted from female sand flies and submitted to Polymerase Chain Reaction (PCR) targeting the kDNA gene; Leishmania species were identified by PCR-RFLP targeting the hsp70 gene and genetic sequencing. RESULTS: In all, 5,716 individuals were collected, and 46 species were identified. Trichophoromyia ubiquitalis (3,330 - 58.26%) and Nyssomyia antunesi (661 - 11.26%) were the most abundant species. Species richness was greater in terra firme environments (42 species) than in the várzea environments (22 species), and forests ecotopes (43 species) were richer than peridomiciles (28 species). DNA of Leishmania was found in Th. ubiquitalis and Psychodopygus davisi, both of which inhabit the terra firme environment and sequencing analysis confirmed the presence of Leishmania (Viannia) lainsoni DNA in Th. ubiquitalis in Tefé Municipality. CONCLUSIONS: The high abundance of Th. ubiquitalis and Ps. davisi and detection of DNA of Leishmania sp. may indicate that both species could be putative vectors for American Cutaneous Leishmaniasis (ACL) in the terra firme environment of Tefé. The sand fly fauna found in várzea is rich and diverse, exhibiting several species, nevertheless the seasonal hydric stress during part of the year that could influence the local diversity, if compared with other studies. This is the first report in Amazonas State of Th. ubiquitalis with presence of L. (V.) lainsoni DNA.