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BACKGROUND: The sand fly Nyssomyia neivai is one of the most abundant species in Southern Brazil. It is frequently found in areas that are foci of visceral leishmaniasis in the state of Santa Catarina, caused by Leishmania infantum. In this region, the main vector of L. infantum, Lutzomyia longipalpis, has not been detected. In the absence of L. longipalpis, this study aimed to identify the sand fly fauna and diagnose any potential Leishmania spp. infection in sand flies and in dogs in a region of Southern Brazil that experienced a recent canine visceral leishmaniasis outbreak. METHODS: This report includes a survey of the sand fly fauna at the Zoonosis Control Center of the Municipality of Tubarão (Santa Catarina, Brazil). Molecular tests were conducted to investigate Leishmania spp. natural infection in sand flies using polymerase chain reaction (PCR). In positive females, in addition to morphological identification, molecular analysis through DNA barcoding was performed to determine the sand fly species. Additionally, the dogs were tested for the presence of Leishmania spp. using a non-invasive technique for the collection of biological material, to be assessed by PCR. RESULTS: A total of 3419 sand flies, belonging to five genera, were collected. Nyssomyia neivai was the most abundant species (85.8%), followed by Migonemyia migonei (13.3%), Pintomyia fischeri (0.8%), Evandromyia edwardsi (< 0.1%), and species of the genus Brumptomyia. (0.1%). Out of the 509 non-engorged females analyzed by PCR, two (0.4%) carried L. infantum DNA. The naturally infected females were identified as Ny. neivai, in both morphological and molecular analysis. In addition, two out of 47 conjunctival swabs from dogs tested positive for L. infantum, yielding an infection rate of 4.2%. CONCLUSIONS: These results confirm the presence of Ny. neivai naturally infected with L. infantum in an area where dogs were also infected by the parasite, suggesting its potential role as a vector in Southern Brazil.

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The present work assessed the experimental susceptibility of Nyssomyia antunesi and Lutzomyia longipalpis to Leishmania (Viannia) lainsoni and L. (V.) lindenbergi. A L. (Leishmania) chagasi-Lu. longipalpis combination was used as a susceptible control. Wild-caught Ny. antunesi and laboratory-bred Lu. longipalpis were membrane-fed on blood with a 5 × 106/mL log-phase promastigote culture suspension and dissected on days 2 and 8 post-blood meal (pbm) for analysis focused on the assessment of parasitoses, as well as placement and promastigote morphotyping. Survival curves were constructed. In all combinations, promastigotes were observed on day 8 pbm. For both Leishmania species, in Lu. longipalpis, the presence of parasites was observed up to the stomodeal valve, while in Ny. antunesi, the presence of parasites was observed up to the cardia. There were no significant differences in parasitosis between L. (V.) lainsoni and L. (V.) lindenbergi in either Ny. antunesi or Lu. longipalpis. Six morphological promastigote forms were distinguished in Giemsa-stained gut smears. The survival curves of all combinations decreased and were affected differently by several Lu. longipalpis-parasite combinations, as well with Lu. longipalpis-uninfected blood. These findings stress Lu. longipalpis as experimentally susceptible to Leishmania spp. and suggest the putative susceptibility of Ny. antunesi to L. (V.) lainsoni and L. (V.) lindenbergi.

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BACKGROUND: The search for attractive baits that may facilitate the capture of haematophagous insects has been epidemiologically relevant. Sand flies use chemical cues in different phases of their life cycles to find carbohydrate meals, mates, blood meals and oviposition sites. Few studies have related the behaviours of sand flies with volatile compounds that can influence their life cycles. Previous studies in our laboratory have shown that 1-hexanol released on filter paper is a good attractant for the sand fly Nyssomyia neivai, which is suspected in the transmission of the aetiologic agent of American cutaneous leishmaniasis. METHODS: In this study, we developed two release systems to modulated 1-hexanol release: system 1 contained gellan gum and pectin (4:1 ratio), 3% aluminium chloride and 1% glutaraldehyde; system 2 contained: gellan gum and pectin (4:1 ratio) and 3% aluminium chloride. After addition of 1-hexanol to each release system, trials were performed in a wind tunnel with Ny. neivai males and females (unfed, blood-fed and gravid) to evaluate activation and attraction responses. RESULTS: Males and unfed females showed the same response pattern to the systems. For both systems, the males and unfed females of the sand flies showed an activation response up to 24 h. The number of responsive gravid females was lower than unfed females for both systems. The blood-fed females showed no responses in any of the release systems. CONCLUSIONS: Our findings indicate that the state of the females (unfed, fed and gravid) can interfere with the sand fly responses to volatile compounds. Additionally, both systems evaluated with the compound showed effectiveness for sand fly attraction.

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Recently, Wolbachia infection has been described in leishmaniasis vector sandflies. This endosymbiont bacterium is present in 60% of insects, and has been suggested as a mechanism of biological control of vector insects, because it causes a series of changes in the invertebrate host. In addition, recent studies have shown that this bacterium can prevent the development of parasites in vector insects. In this context, the present study aims to molecularly characterize the circulating strain of this bacterium in sandflies in the State of Bahia, Brazil, as well as the natural infection rate of Leishmania sp., and to evaluate the coinfection between Wolbachia and Leishmania. Seven hundred and forty-five (745) specimens of sandflies were collected in nine municipalities of Bahia, belonging to two species, Lutzomyia longipalpis (Lutz and Neiva, 1912) and Nyssomyia whitmani (Antunes and Coutinho, 1939). The results confirm infection by the protozoan Leishmania infantum and Wolbachia in both species collected. The identified strain of Wolbachia in sandflies was wStv MI, known to lead to a phenotype of cytoplasmic incompatibility in vector insects.

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Southern Brazil concentrates a considerable number of cases of cutaneous leishmaniasis reported since 1980, and Paraná is the state that most records CL cases in the region. The main sand fly species incriminated as vectors of Leishmania (Viannia) braziliensis (Vianna,1911) are Migonemyia (Migonemyia) migonei (França, 1920), Nyssomyia (Nyssomyia) neivai (Pinto, 1926) and Nyssomyia (Nyssomyia) whitmani (Antunes & Coutinho, 1936). In this study, we evaluated areas with climatic suitability for the distribution of these vectors and correlated these data with CL incidence in the state. The occurrence points of Mg. migonei, Ny. neivai, and Ny. whitmani were extracted from a literature review and field data. For CL analysis in the state of Paraná, data were obtained from the Informatics Department of the Unified Health System of Brazil (DATASUS), covering the period from 2001 to 2019. The layers of bioclimatic variables from the WorldClim database were used in the study. Species distribution modeling was developed using the MaxEnt Software version 3.4.4. ArcGIS software version 10.5 was used to develop suitability maps and the graphical representation of disease incidence. The AUC values were acceptable for all models (> 0,8). Bioclimatic variables BIO13 and BIO14 were the most influential in the distribution of Mg. migonei, while BIO19 and BIO6 were the variables that most influenced the distribution of Ny. neivai, and Ny. whitmani was most influenced by variables BIO5 and BIO9. During 19 years, 4992 cases of CL were reported in the state by 286 municipalities (71,6%). Northern Paraná showed the highest number of areas with very high and high climatic suitability for the occurrence of these species, coinciding with the highest number of CL cases. The modeling tools allowed analyzing the association between climatic variables and the geographical distribution of CL in the state. Moreover, they provided a better understanding of the climatic conditions related to the distribution of different species, favoring the monitoring of risk areas, the implementation of preventive measures, risk awareness, early and accurate diagnosis, and consequent timely treatment.

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Some ecological parameters and the distribution of vectors in the municipality of Eldorado, Vale do Ribeira Region, São Paulo, were studied. Entomological surveys were carried out from September 2019 to March 2021. It was observed that a few ecological parameters, including richness, abundance, diversity, and equitability, were typical of a modified environment, where artificial ecotopes maintain the presence of sandflies throughout the year. A total of 11,668 sandflies were captured. The presence of five taxa were observed in Eldorado, with low diversity and high dominance of Nyssomyia intermedia next to Ny. neivai, which are sympatric species. The results presented reinforce the importance of these species in anthropized areas in the transmission of cutaneous leishmaniasis (CL) agents and the need for entomological monitoring. Psathyromyia pascalei was encountered for the first time in the municipality, expanding the known area of distribution of this species in a modified environment.

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Leishmaniasis is a public health problem worldwide. We aimed to predict ecological niche models (ENMs) for visceral (VL) and cutaneous (CL) leishmaniasis and the sand flies involved in the transmission of leishmaniasis in São Paulo, Brazil. Phlebotomine sand flies were collected between 1985 and 2015. ENMs were created for each sand fly species using Maximum Entropy Species Distribution Modeling software, and 20 climatic variables were determined. Nyssomyia intermedia (Lutz & Neiva, 1912) and Lutzomyia longipalpis (Lutz & Neiva, 1912), the primary vectors involved in CL and VL, displayed the highest suitability across the various regions, climates, and topographies. L. longipalpis was found in the border of Paraná an area currently free of VL. The variables with the greatest impact were temperature seasonality, precipitation, and altitude. Co-presence of multiple sand fly species was observed in the cuestas and coastal areas along the border of Paraná and in the western basalt areas along the border of Mato Grosso do Sul. Human CL and VL were found in 475 of 546 (86.7%) and 106 of 645 (16.4%) of municipalities, respectively. Niche overlap between N. intermedia and L. longipalpis was found with 9208 human cases of CL and 2952 cases of VL. ENMs demonstrated that each phlebotomine sand fly species has a unique geographic distribution pattern, and the occurrence of the primary vectors of CL and VL overlapped. These data can be used by public authorities to monitor the dispersion and expansion of CL and VL vectors in São Paulo state.

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BACKGROUND: In South America, cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL) are emerging diseases, expanding in the border area of Argentina, Brazil and Paraguay. Outbreaks of CL were reported since the 1990s, with Nyssomyia whitmani as the main vector in this region. Regarding VL, urban reports started in 2010 with Lutzomyia longipalpis as the main vector. The aim of this study was to evaluate environmental determinants related to the main vectors of leishmaniasis, to contribute to the prevention and control response to the emergence of VL and CL in the Argentina-Brazil-Paraguay border region. METHODS: The cross-sectional survey includes two cities and two close rural areas in the Argentinean Northeast Region, between November 2014 and January 2015, with a total of 95 sampling sites. REDILA-BL traps were set for three consecutive nights, and a total of 68 meso- and microscale environmental and landscape characteristics were surveyed. The association between vector abundance with different variables was evaluated using a generalized linear model with zero-inflated negative binomial distribution. We analyzed females for detection of Leishmania DNA. RESULTS: The analysis for Lu. longipalpis indicates an excess of absences when the mean NDWI around the sites were higher. The abundance of Lu. longipalpis at mesoscale level was higher when more urban services were present, and when blood sources such as chickens or dogs at the microscale level were present. For Ny. whitmani, no variable was found to be associated with the absences, while its abundance increased in association with the following variables: percentage of tree cover, presence of garbage collection service, hosted people and, at microscale, the presence of poultry. Leshmania infantum DNA was detected in 2/49 (4%) Lu. longipalpis. CONCLUSIONS: The abundance of both species is influenced by variables at different scales, their influence probably has a hierarchy and they are acting on different aspects of the biology of these vectors. The urban spatial segregation of Lu. longipalpis and the peri-urban and rural segregation of N. whitmani increase the risk of VL and CL. The selection of the better variables for each scale will allow the design of appropriate control strategies depending on species.

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Leishmaniases are a global health problem and in Argentina are considered emerging diseases. The new transmission scenarios of tegumentary leishmaniasis are especially important given that large areas of forest are being transformed into rural and urban systems. The aim of this study was to analyze the impact of the construction of a large public building and a neighborhood on the assemblage of Phlebotominae in a rural area with forest remnants and to correlate the changes observed in the species assemblage with characteristics of the environment. Entomological surveys with light traps were conducted on the construction campus in the northeastern region of Argentina at six sites representing different environmental situations. Structural environmental characteristics and meteorological conditions were recorded and analyzed. At least 16 species of Phlebotominae sand flies were collected, the most prevalent being Nyssomyia whitmani (Antunes & Coutinho), followed by the genus Brumptomyia (França & Parrot) and Migonemyia migonei (França). Our study provides evidence of how the structure of the assemblages and prevalent species respond to anthropogenic disturbances. As the construction progressed, both Ny. whitmani and the genus Brumptomyia were favored. The genus Brumptomyia was favored at sites surrounded by high proportions of forest, within patches of remnant vegetation, and relatively far from anthropogenic disturbances, while Ny. whitmani, the main vector of tegumentary leishmaniasis in the region, increases their abundant at short and intermediate distances from vegetation margins and areas close to anthropogenic disturbances, therefore increasing the risk of human exposure to vectors.

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Nyssomyia whitmani (Antunes & Coutinho) is the sandfly vector of causative agents of American cutaneous leishmaniasis in South and Central America. It is widely spread throughout Brazil, in a close association with domestic animals. Almost nothing is known about male mating aggregations in N. whitmani, let alone the influence of moonlight on this behavior. Thus, the influence of moonlight on male N. whitmani aggregations was investigated by using non-attractant suction traps deployed in chicken cages under full moon and new moon nights. In this 18-night study, 2,160 specimens and seven species were collected. The most frequent species was N. whitmani accounting for 91.48% of the total sample. The abundance of males of N. whitmani was significantly higher in the presence than in the absence of moon. The sex ratio on new moon nights was 1:1 (M:F) and on full moon nights was male-biased (1.7:1). Based on this study, an influence of moonlight on male mating aggregations of N. whitmani was verified. The extent of this influence on increasing sandfly aggregation at hosts, the moonlight influence on the early flight activity of male sandflies and the use of chickens as aggregation sites for males of N. whitmani, are all discussed.

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Phlebotomines are important vectors of bacteria, viruses and protozoan parasites. Protozoans of the genus Leishmania which cause visceral and cutaneous leishmaniases, are among the most important etiologic agents transmitted by sand flies. Different blends of human and animal volatiles have been evaluated for use in surveillance and/or control of hematophagous insects. With regard to phlebotomine sand flies there are few records of attraction under laboratory and field conditions. This study was carried out at two collecting sites located in a Brazilian town with a high prevalence of cutaneous leishmaniasis. The aim of the present study was to evaluate the effect of adding two kinds of commercial bait which mimic human odors to light traps to attract sand flies and compare the results with those of light traps without any additional bait. The commercial baits, BG-Lure® and BG-Sweetscent®, were developed to catch anthropophilic mosquitoes. Three treatments were evaluated: 1 - HP light trap with BG-Lure® added; 2 - HP light trap with BG-Sweetscent® added and 3- HP light trap without any attractant. A total of 3,682 sand flies were collected during the study. Constrained correspondence analysis was applied to perform ordination of the captured community of sand fly species that could be explained by attractant, sampling site and sex. The most abundant species: Pintomyia nevesi, Nyssomyia whitmani and Nyssomyia antunesi were further investigated by fitting generalized linear mixed models. Only Ny. whitmani showed a slight indication of an increase in catches with BG-Lure®.

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Abstract INTRODUCTION: Leishmaniasis constitutes a serious but neglected tropical disease. Recently, socio-environmental, biological and physical changes have altered the range of leishmaniasis, causing it to spread into urban areas. In Minas Gerais, the disease is endemic, exhibiting regional differences and reaching urban centers. The purpose of this study was to evaluate entomological aspects related to the ecoepidemiology of leishmaniasis in Itaúna. METHODS: Monthly catches with HP traps were carried out from June 2017 to May 2018, in three ecological areas (urban, rural, and forest). The adaptability of the species to anthropic environments was assessed using the synanthropy index (SI). RESULTS: We collected 1306 specimens of phlebotomine sand flies. Of the species of medical importance, Lutzomyia longipalpis, the vector of Leishmania infantum, represented 90.4% of the specimens identified at species level (n=1260). Nyssomyia whitmani, the vector of Leishmania braziliensis, represented 1.6% of the total. Lu. longipalpis displayed an SI of +95.8, a value that denotes a marked preference for human environments. For Ny. whitmani, the SI was -25, expressing the tendency of this species to occupy uninhabited areas. The population of the three most numerous species captured increased with rain, high temperatures, and high relative humidity. Although captured at low numbers, Ny. whitmani showed a different profile concerning the climate variables analyzed. CONCLUSIONS: Understanding the epidemiology of the disease may assist the health services in formulating effective control measures for improving community health and contributing to the establishment of a dynamic relationship and a global awareness of the health/disease process.

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A species list and a key to species of Psychodidae (Diptera, Nematocera) from Uruguay is presented, with the description of Laurenceomyia martinezae sp. nov., and first records of Brumptomyia guimaraesi (Coutinho Barreto), Nyssomyia neivai (Pinto), Paramormia furcata (Kincaid), Psychoda savaiiensis Edwards, Psychoda zetoscota Quate, and Trichomyia Haliday in Curtis. Discussion on the medically important species is provided.

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Abstract INTRODUCTION: Nyssomyia intermedia is an important vector of American cutaneous leishmaniasis. METHODS: Sand flies were captured in a Shannon trap and after landing on human collectors and retrieved monthly. Nocturnal activity was estimated using a log-linear model. RESULTS: Peak sand fly collection in the Shannon trap occurred in summer and winter, and peak fly collection on humans occurred in spring and summer. CONCLUSIONS: Ny. intermedia was captured in both the hottest and coldest months, indicating its adaptability to different seasons and the possibility of disease transmission at any time of the year in Campus Fiocruz Mata Atlântica, Brazil.

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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.

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Migonemyia migonei is the predominant species in the dry ecoregion and Nyssomyia neivai in the humid region, although co-dominance in humid highly modified areas could be observed. Vector abundance seems to be modulated by environmental anthropization, as the abundance and predominance of Ny. neiviai and Mg. migonei increased in highly modified areas. In Humid Chaco, the risk of human-vector contact would be present throughout the year, being particularly high in spring/summer, and in Dry Chaco, the risk would be restricted to temperate and humid months.

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To contribute to the knowledge of the role of small mammals in the transmission cycle of tegumentary leishmaniasis caused by Leishmania braziliensis, we studied the small mammal community and its temporal and spatial association with phlebotominae, as well as small mammal infection by Leishmania spp. by PCR-RFLP analyses in an endemic area of northeastern Argentina. Ten small mammal samplings were conducted (2007-2009, 7506 Sherman trap nights and 422 cage trap nights). In two of these samplings, 16 capture stations each one consisting of a CDC light trap to capture phlebotominae, two to four Sherman traps and two cage traps were placed. We found co-occurrence of phlebotominae and small mammal captures in four stations, which were all the stations with small mammal captures and yielded 97% (2295 specimens, including 21 gravid females) of the total phlebotominae captures, suggesting that small mammals may provide a potential source of blood for phlebotominae females. One Didelphis albiventris and two Rattus rattus were associated with high captures of Nyssomyia whitmani, vector of L. braziliensis in the study area. The PCR-RFLP analyses confirm the presence of L. braziliensis in two sigmodontine small mammals (Akodon sp. and Euryoryzomys russatus) for the first time in Argentina, to our knowledge.

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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.

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In Puerto Iguazú City, Argentina, human and canine Visceral Leishmaniasis cases have been recorded since the year 2010, with Leishmania infantum as the etiological agent and Lutzomyia longipalpis as its main vector. In the present study, polymerase chain reaction and sequencing were used to detect L. infantum DNA in 3.9% of the female sandflies captured in Puerto Iguazú City. This is the first report of L. infantum DNA detection in Micropygomyia quinquefer, and the second one in Lu. longipalpis and Nyssomyia whitmani for Argentina. Although the detection of Leishmania DNA itself is not enough to determine a Phlebotomine species as a vector, these results are significant in setting the direction of further investigations of vectorial competence and capacity, necessary to define the roles of different sandflies species as specific or permissive vectors in the transmission VL cycle.

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In Brazil, American cutaneous leishmaniasis (ACL) ecology involves a diversity of Leishmania species transmitted by different sand fly species. Workers involved in agricultural activities are those mainly affected by ACL in some regions from Tocantins State (TO), Brazil, where the disease can be established in new settlements. The objective of this study was to examine the seasonal and hourly frequency of sand fly species, focusing on the potential vectors of ACL, in a settlement in Guaraí (TO), an ACL transmission area. Sand flies were captured in forested area close to Pedra Branca Agricultural Project settlement, from March 2006 until December 2007, using Shannon trap. Monthly captures were made from 06:00 p.m. to 10:00 p.m., and 24-h captures were done twice per semester, from 06:00 a.m. to 06:00 a.m. A total of 10,089 specimens from 30 species were identified. Psychodopygus complexus Mangabeira, Psychodopygus llanosmartinsi Fraiha & Ward, and Nyssomyia antunesi Coutinho were the most abundant species. Nyssomyia antunesi was more frequent during the dry period, whereas Ps. complexus and Ps. llanosmartinsi had high frequencies during the rainy season. Precipitation was positively correlated with Ps. complexus and Ps. llanosmartinsi abundance, and negatively correlated with Ny. antunesi During 24-h captures, the majority of specimens were captured during the night followed by a decrease at dawn. The behavior and previous finding of natural infection by Leishmania (Viannia) braziliensis of Ps. complexus led us to the conclusion that this species can be a potential vector of L. (V.) braziliensis during the rainy season in Guaraí.

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