DNA Barcoding Culicoides Biting Midges (Diptera: Ceratopogonidae) in Northeast Brazil.

Biting midges of the genus Culicoides are small insects associated with the transmission of several pathogens, which requires the correct identification of the species, for implementation of effective strategies against these insects. However, many species are difficult to identify only by morphological characters. Therefore, the use of molecular methods can help in the taxonomy and systematics of this group. Here, the DNA barcode approach was evaluated for nine species of Culicoides from the State of Maranhão, Brazil. We generated 39 sequences from a 476 bp (base pairs) fragment of the cytochrome c oxidase subunit I (COI) mitochondrial gene. To assess the usefulness of COI barcodes for the identification of these species, paired genetic distances from intra and interspecific comparisons and phylogenetic trees were generated in MEGA and RAxML/BEAST softwares, respectively. In addition, species delimitation was performed using the PTP, GMYC, and ABGD algorithms. The intra and interspecific genetic distances showed a clear distinction between them, demonstrating that, for the taxa studied, there can hardly be ambiguous identifications with barcodes. In the same sense, the phylogenetic reconstruction resulted in well-supported clades for all morphospecies analyzed.

Spatiotemporal Distribution of Leishmaniasis in an Endemic Area of Northeast Brazil: Implications for Intervention Actions.

This study aimed to analyze the spatiotemporal distribution of leishmaniases, and contribute to the knowledge of their epidemiological dynamics from 2007 to 2017 in the municipality of Caxias, Maranhão, Northeast Brazil. Data on American tegumentary leishmaniasis (ATL) and human visceral leishmaniasis (HVL) were obtained in the Epidemiological Surveillance Sector of Caxias, while data on canine visceral leishmaniasis (CVL) were obtained in the Zoonoses Surveillance Unit. For data analysis and spatial representation of leishmaniasis cases, the geoprocessing of the data was performed, and the geometric features of the state of Maranhão, Caxias, and the disease registration sites were obtained from the shapefile database of the Brazilian Institute of Geography and Statistics. Geostatistics was used to create maps based on the Kernel density method, starting from the points, producing a raster file for each case with several data frames, allowing the instantaneous comparison of the phenomena. During the study period, ATL, HVL, and CVL were reported in Caxias, accounting for 114,304 and 8,498 cases, respectively. The geoprocessing analysis showed that leishmaniasis is widely distributed in the urban area of Caxias. However, there are risk areas for the transmission of these diseases to humans and dogs, associated with deforestation and urban expansion, and may vary over time. Preventive measures must focus on risk areas, including conservation efforts and urban planning, in order to reduce the transmission of leishmaniases.

Postgenal ctenidum in bat flies (Diptera, Streblidae): new genus, with description of two species, and delimitation of Streblinae.

A new genus of streblid bat fly with a postgenal ctenidium is erected and two new species from Brazil are described. We tested the phylogenetic position of the new genus using morphological data, through a cladistic analysis comparing it with eight genera of Trichobiinae and four of Streblinae. Our results showed the new genus is a sister group of Streblinae. A key to genera of Streblidae with genal ctenidium and species of the new genus is provided.

Detection of Wolbachia in Mosquitoes (Diptera: Culicidae) in the State of Maranhão, Brazil.

Recently, the endobacteria Wolbachia has emerged as a biological tool for the control of arboviruses. Thus, we investigated the rate of natural infection by Wolbachia in Culicidae species from Maranhão, Brazil. For this, we amplified the Wolbachia surface protein gene (wsp) from mosquitoes collected in six localities of Maranhão, and positive samples were subjected to new analysis using group-specific primers. In total, 448 specimens comprising 6 genera and 18 species of mosquitoes were analyzed. Wolbachia DNA was PCR-detected in 7 species, three of which are new records: Aedes scapularis (Rondani, 1848), Coquillettidia juxtamansonia (Chagas, 1907) and Cq. venezuelensis (Theobald, 1912), in addition to Ae. albopictus (Skuse, 1894) and Culex quinquefasciatus Say, 1823, which are commonly described as permissive to maintain this bacterium in natural environments, and two species of the subgenera Anopheles (Nyssorhynchus) Blanchard, 1902 and Culex (Melanoconion) Theobald, 1903 which could not be identified at species level. The infection rate of all species ranged from 0 to 80%, and the average value was 16.5%. This study increases the knowledge about the prevalence of Wolbachia in the culicid fauna and may help in selecting strains for biological control purposes.

Predominance of Leishmania (Leishmania) amazonensis DNA in Lutzomyia longipalpis sand flies (Diptera: Psychodidae) from an endemic area for leishmaniasis in Northeastern Brazil.

Leishmaniasis is a serious public health concern in the Northeastern region of Brazil, where the sand fly fauna is well studied, although few species have been identified as competent vectors. The detection of Leishmania spp. parasites in wild-caught sand flies could help sanitary authorities draw strategies to avoid the transmission of the parasites and, therefore, the incidence of leishmaniases. We detected Leishmania DNA in wild-caught sand flies and correlated that data with aspects of sand fly ecology in the Caxias municipality, Maranhao State, Brazil. The sand flies were sampled in the peridomicile (open areas in the vicinity of human residences) and intradomicile (inside the residences) from July/2019 to March/2020. Leishmania DNA was detected in females, targeting a fragment of the Internal Transcribed Spacer (ITS1) from ribosomal DNA. Among the fourteen species of sand flies identified, five (Lutzomyia longipalpis, Nyssomyia whitmani, Evandromyia evandroi, Micropygomyia trinidadensis, and Micropygomyia quinquefer) harbored DNA of Leishmania (Leishmania) amazonensis. The most abundant species in rural (Ny. whitmani: 35.2% and Ev. evandroi: 32.4%) and urban areas (Lu. longipalpis: 89.8%) are the permissive vectors of L. (L.) amazonensis, especially Ny. whitmani, a known vector of causative agents of cutaneous leishmaniasis. Although Lu. longipalpis is the vector of L. (L.) infantum, which was not detected in this study, its permissiveness for the transmission of L. (L.) amazonensis has been reported. We suspect that visceral leishmaniasis and cutaneous leishmaniasis are caused by L. (L.) amazonensis, and the transmission may be occurring through Lu. longipalpis, at least in the urban area.

Diversity of Culicoides (Diptera: Ceratopogonidae) in Rural Settlements and Degraded Seasonal Deciduous Forests.

Biting midges are widespread in Brazilian natural ecosystems. However, deforestation and other activities that impact the environment are reducing natural habitats where biting midges proliferate. The objective of this study was to verify whether there is variation in the composition, richness, abundance, and seasonality of biting midges between wild and rural environments, in a forest area with intense deforestation. Biting midges were captured using 6 traps installed at an average height of 1.5 m in the peridomicile, intradomicile, and deciduous seasonal forests, once a month from May 2012 to April 2013. In total, 2,182 specimens of 13 species of the genus Culicoides were captured. Species richness was similar in the intradomicile (13 species), forest (12), and peridomicile (11), but species diversity was greater in the peridomicile (H' = 0.803) compared with the intradomicile (H' = 0.717) and forest (H' = 0.687). The order of species dominance varied between the forest (Culicoides paucienfuscatus Barbosa > Culicoides leopodoi Ortiz > Culicoides foxi Ortiz > Culicoides ignacioi Forattini) and peridomicile + intradomicile habitats (C. paucienfuscatus > C. foxi > C. filariferus Hoffman > C. ignacioi). The activity of these dipterans was strongly influenced by meteorological variables, as biting midges are predominant in the rainy season (80.7% of specimens), when higher rainfall, relative humidity, and lower temperatures prevail. The abundance of biting midges was higher in the peridomicile + intradomicile (83.7% of specimens) compared with the degraded forest (16.3%), a result that reflects the loss of forest habitat due to intense and progressive deforestation.

Predominance of Leishmania (Leishmania) amazonensis DNA in Lutzomyia longipalpis sand flies (Diptera: Psychodidae) from an endemic area for leishmaniasis in Northeastern Brazil

ABSTRACT Leishmaniasis is a serious public health concern in the Northeastern region of Brazil, where the sand fly fauna is well studied, although few species have been identified as competent vectors. The detection of Leishmania spp. parasites in wild-caught sand flies could help sanitary authorities draw strategies to avoid the transmission of the parasites and, therefore, the incidence of leishmaniases. We detected Leishmania DNA in wild-caught sand flies and correlated that data with aspects of sand fly ecology in the Caxias municipality, Maranhao State, Brazil. The sand flies were sampled in the peridomicile (open areas in the vicinity of human residences) and intradomicile (inside the residences) from July/2019 to March/2020. Leishmania DNA was detected in females, targeting a fragment of the Internal Transcribed Spacer (ITS1) from ribosomal DNA. Among the fourteen species of sand flies identified, five (Lutzomyia longipalpis, Nyssomyia whitmani, Evandromyia evandroi, Micropygomyia trinidadensis, and Micropygomyia quinquefer) harbored DNA of Leishmania (Leishmania) amazonensis. The most abundant species in rural (Ny. whitmani: 35.2% and Ev. evandroi: 32.4%) and urban areas (Lu. longipalpis: 89.8%) are the permissive vectors of L. (L.) amazonensis, especially Ny. whitmani, a known vector of causative agents of cutaneous leishmaniasis. Although Lu. longipalpis is the vector of L. (L.) infantum, which was not detected in this study, its permissiveness for the transmission of L. (L.) amazonensis has been reported. We suspect that visceral leishmaniasis and cutaneous leishmaniasis are caused by L. (L.) amazonensis, and the transmission may be occurring through Lu. longipalpis, at least in the urban area.

Experimental transmission of Leishmania (Leishmania) amazonensis to immunosuppressed mice through the bite of Lutzomyia longipalpis (Diptera: Psychodidae) results in cutaneous leishmaniasis.

Lutzomyia longipalpis is the natural vector of Leishmania (Leishmania) infantum, but it is also permissive for several Leishmania species that are related to cutaneous leishmaniasis (CL). Maranhao State (Northeast of Brazil) is endemic for CL and has the highest number of cases of diffuse cutaneous leishmaniasis (DCL) in the country. It is a rare disease associated with a defective immune response mainly caused by L. (L.) amazonensis. Additionally, the number of immunosuppressed patients infected with the etiologic agents of CL has increased, including regions in which the main vectors of CL are rare. Therefore, we investigated whether Lu. longipalpis is able to transmit L. (L.) amazonensis to uninfected and immunosuppressed mice, resulting in CL. For that, 291 sand flies took an initial blood meal in mice infected with L. (L.) amazonensis. Of these, 17 underwent a second feeding on uninfected and immunosuppressed mice (of which 58.8% were also positive for Leishmania according to data on the dissection of the intestine). After 27 days of infection, these mice exhibited leishmaniotic lesions. The occurrence of parasites on the animal's skin was confirmed by limiting dilution and immunohistopathological analyses. Parasite DNA was also detected in paw lesions and inguinal lymph nodes. DNA sequencing confirmed the Leishmania species in insects and mice. The results confirmed the ability of Lu. longipalpis to become infected and experimentally transmit L. (L.) amazonensis to immunosuppressed rodents, resulting in leishmaniotic lesions. Our data open perspectives for the potential role of Lu. longipalpis in the epidemiology of urban cutaneous leishmaniasis, especially in immunosuppressed patients.

Molecular Evidence Suggests That Wolbachia pipientis (Rickettsiales: Anaplasmataceae) is Widely Associated With South American Sand Flies (Diptera: Psychodidae).

Wolbachia pipientis (Hertig) is an endosymbiotic microorganism widespread among arthropods and other invertebrate hosts, and employed in strategies to reduce the incidence of arthropod-borne diseases. Here, we used a PCR-based approach for 16S RNA and wsp genes to investigate the prevalence, geographical distribution, and strains of Wolbachia in sand flies (Diptera: Psychodidae: Phlebotominae), the main vectors of the causative agents of leishmaniasis, from three biomes in Brazil: Amazon, Cerrado, and Caatinga. We found that: 1) Wolbachia DNA is present in most (66.7%) of the sampled sand fly species, including vectors of Leishmania spp. (Ross, Trypanosomatida: Trypanosomatidae), 2) the prevalence of Wolbachia DNA varies among species and populations, 3) some strains of Wolbachia may have wider geographical and host range in South America, and 4) two phylogenetic distinct wsp sequences might represent two novel strains for Wolbachia in South America sand flies. Those findings increase the basic knowledge about Wolbachia in South American sand flies and might foster further researches on its use to reduce the transmission of sand fly-borne parasites.

Exploiting the Synergistic Effect of Kairomones and Light-Emitting Diodes on the Attraction of Phlebotomine Sand Flies to Light Traps in Brazil.

The synergistic effect of light-emitting diodes (LEDs) and kairomones on the attraction of sand flies to light traps was evaluated. Octenol and lactic acid were used as chemical attractants. Green LEDs and the incandescent lamps were used as light attractants. Five CDC-type light traps with the respective combination of attractants (incandescent lamp, incandescent lamp + chemical attractant, green LED, green LED + chemical attractant, and chemical attractant alone [without light]) were set between 18:00 and 06:00 following a Latin square design. A total of 6,536 sand flies and 16 species were collected. The most frequent species collected was Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera, Psychodidae) accounting for 43.21% of all individuals. Order of success (mean, SD) of lactic acid attractant fly capture was as follows: LED + lactic acid (36.83 ± 4.74), LED alone (34.87 ± 4.61), incandescent lamp + lactic acid (22.80 ± 3.19), incandescent lamp alone (12.67 ± 2.03), and lactic acid (0.46 ± 0.13). Order of success of octenol attractant fly capture was as follows: LED + octenol (37.23 ± 5.61), LED alone (35.77 ± 5.69), incandescent lamp + octenol (18.63 ± 3.28), incandescent lamp alone (14.67 ± 2.86), and octenol alone (1.80 ± 0.65). With exception of lactic acid + incandescent light, chemical synergists played no part in significantly increasing light trap capture of phlebotomine sand flies. However, the use of LEDs, with or without such attractants, provided significantly higher capture compared to the incandescent lamp with or without such chemicals, showing that LEDs are suitable and efficient light sources for surveillance and monitoring of phlebotomine sand flies in Brazil.

Temporal Variation in Species Abundance and Richness of Culicoides (Diptera: Ceratopogonidae) in a Tropical Equatorial Area.

The aim of this study was to investigate whether temperature, rainfall, and humidity influence the annual distribution of Culicoides Latreille 1809, species abundance and richness in rural areas on São Luís Island in the state of Maranhão, Brazil. Biting midges were collected in housing for domestic animals in the peridomestic areas of 10 homes. A CDC light trap was installed in each peridomestic area from 6:00 p.m. to 6:00 a.m. once a month for 12 mo. In total, 23,633 individuals belonging to 26 Culicoides species were captured. The most abundant species were Culicoides diabolicus Hoffman, 1925 (27.43%), C. ignacioi Forattini, 1957 (25.9%), C. flavivenulus Costa Lima, 1937 (15.53%), C. insignis Lutz, 1913 (10.66%), C. filariferus Hoffman, 1939 (6.21%), C. boliviensis Spinelli & Wirth, 1984 (6.07%), C. foxi Ortíz, 1950 (2.83%), and C. leopoldoi Ortíz, 1951 (2.54%). Species richness and abundance were greater during the rainy season (24 species; 88% of the individuals) than during the dry season (18 species; 12% of individuals). However, persistent rain on the days the midges were collected or in the preceding 24 h adversely affected abundance. Mean monthly maximum temperature and rainfall on the day of the collection adversely affected Culicoides abundance and richness.

Influence of Deforestation on the Community Structure of Sand Flies (Diptera: Psychodidae) in Eastern Amazonia.

Variation in the structure of phlebotomine (sand fly) communities in forest fragments with different degrees of preservation and human occupation (peridomicile) in eastern Amazonia was studied. We identified 43 species of sand flies in our study, of which 38 occurred in both preserved forest areas and in the peridomiciles of short-term settlements, while another 28 species occurred in altered forest fragments and long-term settlements. The composition of the community at each site changed with the type of environment (forest or peridomicile), with the species Lutzomyia evandroi, L. whitmani, L. choti, L. serrana, L. triacantha, L. migonei, L. hirsuta, L. shannoni, and L. brachyphylla accounting for more than 54% of the differences among environments. The quality of the environment exerted a significant influence on the structure of phlebotomine communities, and affected their species composition, richness, and abundance.

DNA Barcoding Reveals Hidden Diversity of Sand Flies (Diptera: Psychodidae) at Fine and Broad Spatial Scales in Brazilian Endemic Regions for Leishmaniasis.

Sand fly (Diptera: Psychodidae) taxonomy is complex and time-consuming, which hampers epidemiological efforts directed toward controlling leishmaniasis in endemic regions such as northeastern Brazil. Here, we used a fragment of the mitochondrial cytochrome c oxidase I (COI) gene to identify sand fly species in Maranhão State (northeastern Brazil) and to assess cryptic diversity occurring at different spatial scales. For this, we obtained 148 COI sequences of 15 sand fly species (10 genera) from Maranhão (fine spatial scale), and joined them to COI sequences from other Brazilian localities (distant about 2,000 km from Maranhão, broad spatial scale) available in GenBank. We revealed cases of cryptic diversity in sand flies both at fine (Lutzomyia longipalpis (Lutz and Neiva) and Evandromyia termitophila (Martins, Falcão and Silva)) and broad spatial scales (Migonemyia migonei (França), Pressatia choti (Floch and Abonnenc), Psychodopygus davisi (Root), Sciopemyia sordellii (Shannon and Del Ponte), and Bichromomyia flaviscutellata (Mangabeira)). We argue that in the case of Bi. flaviscutellata, the cryptic diversity is associated with a putative new species. Cases in which DNA taxonomy was not as effective as morphological identification possibly involved recent speciation and/or introgressive hybridization, highlighting the need for integrative approaches to identify some sand fly species. Finally, we provide the first barcode sequences for four species (Brumptomyia avellari (Costa Lima), Evandromyia infraspinosa (Mangabeira), Evandromyia evandroi (Costa Lima and Antunes), and Psychodopygus complexus (Mangabeira)), which will be useful for further molecular identification of neotropical species.

Molecular Identification of Leishmania spp. in Sand Flies (Diptera: Psychodidae: Phlebotominae) in the Lençóis Maranhenses National Park, Brazil.

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.

Detection of Leishmania DNA and Blood Meal Identification in Sand Flies (Diptera: Psychodidae) From Lençois Maranhenses National Park Region, Brazil.

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.

Leishmania infection and blood food sources of phlebotomines in an area of Brazil endemic for visceral and tegumentary leishmaniasis.

The aims of the study were to determine the blood feeding preferences of sandflies and to identify species of Leishmania that infected phlebotomines in Caxias, Maranhão, Brazil, an area that is highly endemic for leishmaniasis. Sandflies were captured in light traps located in the peridomiciliary environments of randomly selected houses in urban and rural settings between 1800 and 0600 hours on new moon days between March 2013 and February 2015. DNA extracts from 982 engorged female sandflies were submitted to fragment length polymorphism analysis to identify infecting species of Leishmania, and blood sources were identified for 778 of these specimens. Infection by Leishmania infantum was detected in Lutzomyia longipalpis, Lu. whitmani and Lu. termitophila; L. infantum/L. braziliensis in Lu. longipalpis, Lu. whitmani and Lu. trinidadensis; L. shawi in Lu. longipalpis; L. mexicana in Lu. longipalpis; L. braziliensis in Lu. longipalpis and Lu. whitmani; L. guyanensis in Lu. longipalpis and Lu. termitophila; L. amazonensis in Lu. longipalpis and L. lainsoni or L. naiffi in Lu. longipalpis, while Lu. longipalpis and Lu. trinidadensis were infected with unidentified Leishmania sp. Blood sources were identified in 573 individual phlebotomines and the preferred hosts were, in decreasing order, chicken, dog, rodent and human with lower preferences for pig, horse, opossum and cattle. Lu. longipalpis and Lu. whitmani performed mixed feeding on man, dog and rodent, while Lu. longipalpis was the most opportunistic species, feeding on the blood of all hosts surveyed, but preferably on dog/chicken, dog/rodent and rodent/chicken. Our findings reveal the concomitant circulation of Leishmania species that cause visceral leishmaniasis and tegumentary leishmaniasis in the study area, and explain the occurrence of autochthonous human cases of both clinical forms of leishmaniasis in Caxias, Maranhão. The results support our hypothesis that, in the municipality of Caxias, transmission of Leishmania occurs in close proximity to humans.

Detection of Leishmania amazonensis and Leishmania braziliensis in Culicoides (Diptera, Ceratopogonidae) in an endemic area of cutaneous leishmaniasis in the Brazilian Amazonia.

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.

An ecological study of sand flies (Diptera: Psychodidae) in the vicinity of Lençóis Maranhenses National Park, Maranhão, Brazil.

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.