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Numerous sand fly species have been reported in Lençóis Maranhenses National Park (LMNP) in Northeast Brazil, including important Leishmania vectors, making the park an endemic area for tegumentary and visceral leishmaniasis. We evaluated sand fly abundance monthly over 7 years, correlating it with environmental variables and monthly tourist numbers in LMNP. Sand fly species were observed throughout the year, with Lutzomyia longipalpis Lutz & Neiva and Nyssomyia whitmani Antunes and Coutinho (Diptera: Psychodidae) being the most abundant species, especially from April to June. In addition to seasonal abundance patterns, Lu. longipalpis showed increasing abundance throughout 2013 until April 2014, whereas Ny. whitmani exhibited a consistent increase throughout the study period. Redundancy analysis indicated that monthly sand fly abundances increased with humidity but decreased with mean temperature and wind speed. Ecotourists mainly visit the park from May to September when interdune lagoons are full, coinciding with high-frequency vector activity during the rainy months (May-June). Tourists also visit in January and February during school holidays, when the rains begin and sand fly abundance increases, and in July, when sand fly abundance decreases. To date, no instances of infected tourists have been recorded, likely because visits to LMNP occur during the day when sand flies are inactive. However, there is a potential risk of vector exposure if tourists engage in nighttime cultural visits to villages around the park, where leishmaniasis cases occur annually.

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Sand flies are very common in the region of Lençóis Maranhenses National Park, an important tourist attraction in Brazil. However, the role of some species and their relative importance locally in Leishmania Ross 1903 transmission is unclear. The objective of this study was to identify Leishmania infection in phlebotomine sand flies collected around the Lençóis Maranhenses National Park, an important conservation area and popular international/national tourist destination with a high incidence of leishmaniasis. Sand flies were collected in peridomiciliary areas on the tourist route from September 2012 to August 2013. The captured females were subjected to molecular analyses for the detection of Leishmania DNA. Sand flies were infected with four Leishmania species: Leishmania (Viannia) braziliensis (Vianna, 1911) was found in Lutzomyia whitmani (Antunes and Coutinho, 1939) (2.1%) and Lutzomyia longipalpis (Lutz and Neiva, 1912) (1.7%); Leishmania (Leishmania) infantum (Nicole, 1908) infected Lutzomyia wellcomei (Fraiha, Shaw, and Lainson, 1971) (20%), Lutzomyia sordellii (Shannon and Del Ponte, 1927) (4.3%), Lu. longipalpis (3.7%), and Lu. whitmani (0.8%); Leishmania (Leishmania) amazonensis (Lainson & Shaw, 1972) was found in Lu. whitmani (0.58%), while Leishmania (Viannia) lainsoni infected Lutzomyia evandroi (Costa Lima and Antunes, 1936) (3.4%), Lu. longipalpis (1.06%), and Lu. whitmani (0.29%). The occurrence of these parasites requires control measures to reduce the incidence of cutaneous leishmaniasis and to contain a possible epidemic of visceral leishmaniasis, the most severe form of the disease.

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To elucidate portions of the transmission cycles of American tegumentary leishmaniasis (TL) and visceral leishmaniasis (VL) occurring in the region surrounding the Lençóis Maranhenses National Park, an important tourist center in Brazil, the present study objectives were to determine the rate of natural infection by Leishmania spp. and the blood meal in caught sand flies species in the region. Sand flies were captured over 36 mo in 62 locations of the municipality of Barreirinhas, Maranhão with notifications of disease incidence. Species identification of parasites was performed with internal transcribed spacer 1 (ITS1) PCR-RFLP using HaeIII enzyme. Blood meal identification was performed with cytochrome b (cytb) gene PCR-RFLP using HaeIII and MboI enzyme. The species Lutzomyia longipalpis (Lutz and Neiva 1912) presented a positivity rate of 3.7% for Leishmania infantum. Species not considered vectors of this parasite such as Lu. lenti (Mangabeira 1938) and Lu. whitmani (Antunes & Coutinho 1939) showed infection rates of 0.6% and 0.9%, respectively. Among the vectors of Leishmania spp. was Lu. whitmani with detection rate of 0.3% for Le. braziliensis and Lu. flaviscutellata (Mangabeira 1942) with a detection rate of 8% for Le. amazonensis. After restriction of amplification product encoding a 359bp sequence of the cytb recognized in as follows: pigs (37.9%); dogs (27.4%); chickens (20.9%); horses (9%), rodents (3.3%), and humans (1.4%). The presence of Leishmania DNA in sand flies fed with human blood and domestic animals in villages with transmission of VL and TL suggests that transmission could be occurring in the locations of the infected patients.

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Biting midges in the genus Culicoides act as vectors of arboviruses throughout the world and as vectors of filariasis in Latin America, the Caribbean, and parts of Africa. Although Culicoides spp. are currently not considered to be vectors of Leishmania protozoa, the high abundance of biting midges in areas with active cutaneous leishmaniasis transmission points to the possibility of Culicoides infection by these pathogens. We used PCR to test captured Culicoides species for natural infection with Leishmania spp. We tested 450 Culicoides females, divided into 30 pools of 15 individuals each, as follows: nine pools of C. foxi (135 specimens), seven pools of C. filariferus (105), seven pools of C. insignis (105), five pools of C. ignacioi (75), and two pools of C. flavivenula (30). PCR confirmed the presence of Leishmania braziliensis DNA in C. ignacioi (0.14%), C. insignis (0.14%), and C. foxi (0.11); and Le. amazonensis DNA in C. filariferus (0.14%) and C. flavivenula (0.50%). We conclude that these Culicoides species can be naturally infected, but vector competence and transmission capability must be confirmed in future studies. Our results warrant further investigation into the role of these biting midge species in the leishmaniasis epidemiological cycle.

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BACKGROUND: The Lençóis Maranhenses National Park, located in Maranhão, Brazil, is a region of exceptional beauty and a popular tourist destination. The adjoining area has suffered from the impact of human activity and, consequently, has experienced outbreaks of leishmaniasis. This study aimed to evaluate the composition, abundance, species richness and seasonal distribution of sand flies in the region and to determine the constancy of the insect population. METHODS: The survey was conducted at three sites located in the municipalities of Barreirinhas and Santo Amaro between September 2012 and August 2013. Sampling was performed monthly using automatic light traps installed 1.5 m above the soil adjacent to 13 randomly selected rural dwellings. At each site, one trap was placed in the peridomicile near to animal enclosures and another (extradomicile) at 500 m from the peridomicile. RESULTS: A total of 4,474 individual sand flies were collected over the year with the highest abundance recorded during the rainy season (December to June). Nine species were collected: L. whitmani, L. longipalpis, L. lenti, L. sordellii, L. evandroi, L. flaviscutellata, L. wellcomei, L. termitophila and L. intermedia. Although peridomiciliary and extradomiciliary environments presented similar species richness, the Shannon diversity index was significantly lower in the former (H' = 2.4) compared with the latter (H' = 4.98). Lutzomyia whitmani and L. longipalpis were the most abundant species and were classified as constant (constancy index, CI = 100%) along with L. lenti (CI = 58.3), L. evandroi (CI = 58.3) and L. sordellii (CI = 66.7). The remaining four species presented CI values between 25 and 50% and were considered accessory. CONCLUSIONS: The present results confirm the present of L. whitmani and L. longipalpis in the peridomicile of houses in Lençóis National Park. The abundance of these species could explain, respectively, the endemicity of cutaneous leishmaniasis and sporadic cases of visceral leishmaniasis in the study area. However, in the case of cutaneous leishmaniasis, the presence of other sand fly vectors (in addition to L. whitmani) cannot be neglected. Finally, this study emphasizes the need for a more effective and permanent supervision to control the expansion of these vectors and leishmaniasis outbreaks.

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O objetivo deste estudo foi determinar as fontes de alimento sanguíneo de fêmeas de Lutzomyia whitmani, espécie de flebotomíneo incriminada no Maranhão como principal vetor da leishmaniose cutânea americana. Para isso, 70 fêmeas desta espécie coletadas no município de Axixá, área com um dos maiores números de casos de leishmaniose cutânea americana em humanos no Maranhão, foram analisadas utilizando a técnica da precipitina. Dos indivíduos analisados, 90 por cento apresentaram reação a algum tipo de antissoro e dentre estes, 73 por cento apresentaram reações do tipo simples com predominância para sangue de galinha (22,2 por cento), roedor (14,3) e humano (12,7 por cento). Nas reações duplas predominaram as combinações galinha/humano (6,3 por cento), galinha/gambá (4,8 por cento), boi/humano e gambá/humano (3,2 por cento). Assim, concluímos que seres humanos, animais domésticos e sinantrópicos constituem fonte alimentar sanguínea para Lutzomyia whitmani podendo desempenhar um papel importante no ciclo de transmissão da leishmaniose cutânea americana explicando os casos da doença em Axixá.

The aim of this study was to determine the sources of blood meals for females of Lutzomyia whitmani, a phlebotomine species incriminated as the main vector for American cutaneous leishmaniasis in Maranhão. For this, 70 Lutzomyia whitmani females were collected in the municipality of Axixá, an area with one of the greatest numbers of cases of American cutaneous leishmaniasis in humans in Maranhão. They were analyzed using the precipitin technique. Ninety percent of the specimens showed a reaction to some type of antiserum positive immune reaction, among which 73 percent presented single reactions, with predominance for chicken blood (22 percent), rodent blood (14.3 percent) and human blood (12.7 percent). Among the double reactions, the predominant combinations were chicken/human (6.3 percent), chicken/opossum (4.8 percent), ox/human (3.2 percent) and opossum/human (3.2 percent). Thus, we conclude that humans and domestic and synanthropic animals are blood meal sources for Lutzomyia whitmani and may play an important role in the transmission cycle for American cutaneous leishmaniasis, thus explaining the cases of this disease in Axixá.

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The aim of this study was to determine the sources of blood meals for females of Lutzomyia whitmani, a phlebotomine species incriminated as the main vector for American cutaneous leishmaniasis in Maranhão. For this, 70 Lutzomyia whitmani females were collected in the municipality of Axixá, an area with one of the greatest numbers of cases of American cutaneous leishmaniasis in humans in Maranhão. They were analyzed using the precipitin technique. Ninety percent of the specimens showed a reaction to some type of antiserum positive immune reaction, among which 73% presented single reactions, with predominance for chicken blood (22%), rodent blood (14.3%) and human blood (12.7%). Among the double reactions, the predominant combinations were chicken/human (6.3%), chicken/opossum (4.8%), ox/human (3.2%) and opossum/human (3.2%). Thus, we conclude that humans and domestic and synanthropic animals are blood meal sources for Lutzomyia whitmani and may play an important role in the transmission cycle for American cutaneous leishmaniasis, thus explaining the cases of this disease in Axixá.

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Estudou-se a diversidade, a abundância relativa e a sazonalidade das espécies de flebotomíneos em duas reservas de floresta primária alterada, a reserva florestal do Sacavém (RFS) e reserva florestal do Itaqui (RFI), localizadas na área metropolitana de São Luís, capital do estado do Maranhão. Os flebotomíneos foram capturados com armadilhas luminosas tipo CDC, instaladas na borda e no centro da mata, a 1,5 metros de altura, das 18h00 às 6h00, uma vez por mês, durante um ano. No total foram capturados 1.356 indivíduos de 23 espécies dos gêneros Lutzomyia (21) e Brumptomyia (2). Na RFS foram capturados 1.061 espécimes, resultando num esforço de captura de 2,5 indivíduos/hora/armadilha e o predomínio de L. longipalpis (44,8 por cento), seguida por L. antunesi (36,4 por cento), L. sordelli (5,9 por cento), L. flaviscutellata (3,9 por cento) e L. whitmani (2,1 por cento). O maior número de espécies (11) e indivíduos (60,1 por cento) ocorreu na estação chuvosa. Na RFI foram obtidos 295 espécimes, o esforço de captura foi 0,2 indivíduos/hora/armadilha, com o predomínio de L. flaviscutellata (58,6 por cento), L. sordelli (14,6 por cento), L. longipalpis (7,1 por cento), L. evandroi (6,4 por cento), L. longipennis (3 por cento), L. trinidadensis (2,7 por cento) e L. whitmani (1,7 por cento). As espécies foram mais numerosas na estiagem (11) e os espécimes durante a estação chuvosa (54,6 por cento).

This study deals with the diversity, relative abundance and seasonality of sandflies species in two modified primary forest fragments, situated at São Luís city, capital of the state of Maranhão. The sandflies were captured with the use of light traps, CDC (Center Control Disease) type, installed at the edge and in the center of the forest, at the height of 1.5 meter, being there from 6 p.m. up to 6 a.m., once per month, during a year time. An amount of 1,356 specimens from 23 species, which 21 belonged to genera Lutzomyia and two belonged to genera Brumptomyia, were captured. In the Sacavém forest reserve, 1,061 specimens were captured, resulting in a capture effort of 2.5 individuals/hour/trap, most of them L. longipalpis (44.8 percent), followed by L. antunesi (36.4 percent), L. sordelli (5.9 percent), L. flaviscutellata (3.9 percent) and L. whitmani (2.1 percent). The highest concentrations of species (11) and individuals (60.1 percent) occurred during the rainy period. In the Itaqui forest reserve, 295 specimens were collected and the capture effort was 0.2 individuals/hour/trap, most of them were L. flaviscutellata, (58.6 percent), L. sordelli (14.6 percent), L. longipalpis (7.1 percent), L. evandroi (6.4 percent), L. longipennis (3 percent), L. trinidadensis (2.7 percent) and L. whitmani (1.7 percent). The species were more frequent during the dry season (11) and specimens during the rainy spell (54.6 percent).

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