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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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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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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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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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Sand flies are natural hosts of various microorganisms. Due to their epidemiological importance, sand fly colonies are kept in laboratories to be studied in terms of their biology and vector/host/parasite interactions. In order to investigate the presence of oviposition pheromones in Nyssomyia neivai, experiments using Solid Phase Microextraction (SPME) were performed. However, siloxanes which is an external class of contamination, present in breeding containers made by plaster used to maintain sand flies in colonies, may be hindered the experiments.

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Sandflies transmit pathogens of leishmaniasis. The natural infection of sandflies by Leishmania (Viannia) was assessed in municipalities, in the state of Paraná, in Southern Brazil. Sandflies were collected with Falcão and Shannon traps. After dissection in search of flagellates in digestive tubes and identification of the species, female sandflies were submitted to the Multiplex Polymerase Chain Reaction (multiplex PCR) for detection of the fragment of the kDNA of Leishmania (Viannia) and the fragment from the IVS6 cacophony gene region of the phlebotomine insects. The analysis was performed in pools containing seven to 12 guts from females of the same species. A total of 510 female sandflies were analyzed, including nine Migonemyia migonei, 17 Pintomyia fischeri, 216 Nyssomyia neivai, and 268 Nyssomyia whitmani. Although none of the females was found naturally infected by flagellates through dissection, the fragment of DNA from Leishmania (Viannia) was shown by multiplex PCR in one sample of Ny. neivai (0.46%) and three samples of Ny. whitmani (1.12%). It was concluded that Ny. neivai and Ny. whitmani are susceptible to Leishmania infection, and that multiplex PCR can be used in epidemiological studies to detect the natural infection of the sandfly vector, because of its sensitivity, specificity and feasibility.

Flebotomíneos transmitem os patógenos das leishmanioses. Foi avaliada a infecção natural de flebotomíneos por Leishmania (Viannia) em municípios do Estado do Paraná, sul do Brasil. Os flebotomíneos foram coletados com armadilhas de Falcão e Shannon. Após dissecação para pesquisa de flagelados no tubo digestório e identificação das espécies, as fêmeas de flebotomíneos foram submetidas a Multiplex Reação em Cadeia da Polimerase (multiplex PCR) para a detecção do fragmento do kDNA de Leishmania (Viannia) e do fragmento do gene IVS6 da cacofonia de flebotomíneos. A análise foi realizada em pools contendo sete a 12 tubos digestórios de fêmeas da mesma espécie. Um total de 510 fêmeas foram analisadas, incluindo nove Migonemyia migonei, 17 Pintomyia fischeri, 216 Nyssomyia neivai e 268 Nyssomyia whitmani. Embora nenhuma fêmea tenha sido encontrada naturalmente infectada com flagelados pela dissecação, o fragmento de DNA de Leishmania (Viannia) foi mostrado por multiplex PCR em uma amostra de Ny. neivai (0,46%) e três amostras de Ny. whitmani (1,12%). Conclui-se que Ny. neivai e Ny. whitmani são suscetíveis à infecção por Leishmania, e que multiplex PCR, devido à sua sensibilidade, especificidade e viabilidade, pode ser utilizada em estudos epidemiológicos para a detecção da infecção natural do inseto vetor.

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Introduction The aim of this study was to identify the blood feeding sources of Nyssomyia intermedia (Ny. intermedia) and Nyssomyia neivai (Ny. neivai), which are Leishmania vectors and the predominant sandfly species in the Ribeira Valley, State of São Paulo, Brazil, an endemic area for cutaneous leishmaniasis. Methods Specimens were captured monthly between February 2001 and December 2003 on a smallholding and a small farm situated in the Serra district in the Iporanga municipality. The blood meals of 988 engorged females were tested using the avidin-biotin immunoenzymatic enzyme-linked immunosorbent assay (ELISA). Seven blood meal sources were investigated: human, dog, chicken, bovine, pig, horse and rat. Results The results showed that among the females that fed on one or more blood sources, the respective percentages for Ny. intermedia and Ny. neivai, respectively, were as follows: human (23% and 36.8%), pig (47.4% and 26.4%), chicken (25.7% and 36.8%) and dog (3.9% and 0%), and the differences in the blood sources between the two species were statistically significant (p = 0.043). Conclusions Both species had predominant reactivity for one or two blood sources, and few showed reactivity indicating three or four sources. Many different combinations were observed among the females that showed reactivity for more than one source, which indicated their opportunistic habits and eclecticism regarding anthropic environmental conditions. .

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Nyssomyia intermedia and Nyssomyia neivai constitute a species complex associated with Leishmania transmission. The aim of this study was to analyse the ecological profiles of the Ny. intermedia and Ny. neivai populations in a sympatric area in the Brazilian savannah along the banks of the Velhas River. Captures were performed from July 2003-June 2005 in two distinct environments: a gallery forest with various degrees of anthropogenic modification and animal shelters. A total of 20,508 Ny. neivai (86%) and Ny. intermedia (14%) sandflies were collected. The difference between the proportions of the sandflies that were collected (Ny. neivai/Ny. intermedia) per bank was significant. The right bank presented a greater number of sandflies (65%) and more preserved vegetation. The abundance of Ny. neivai was higher than that of Ny. intermedia on both banks. The results demonstrate that anthropic activities can affect the sandfly populations in this area, thereby leading to a reduction in species abundance. Nevertheless, the environments with higher levels of antropogenic modification displayed sandfly population numbers that favour the Leishmania transmission cycle.

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O objetivo deste trabalho foi descrever a composição da fauna e a frequência de flebotomíneos em seis localidades rurais do município de Prudentópolis, estado do Paraná. Os flebotomíneos foram capturados com o emprego de armadilhas de Falcão, entre 18 h e 1 h, em diferentes meses de 2008 e2009. Capturou-se um total de 1.429 flebotomíneos de sete espécies, com predomínio de Nyssomyia neivai em todas as localidades. As médias horárias (MH) de flebotomíneos foram mais elevadas em residências (10,7), galinheiros (10,3), matas ciliares (6,5) e pocilgas (5,8). As localidades ondeos flebotomíneos foram capturados apresentavam características ambientais apropriadas para atransmissão da Leishmaniose Tegumentar Americana. O conhecimento da fauna, da frequência de flebotomíneos e da forma de ocupação do ambiente pode auxiliar na escolha do método mais apropriado para reduzir a densidade de flebotomíneos no peridomicílio e domicílio.

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The kairomone octenol is known as attractive to hematophagous Diptera such as mosquitoes, tsetse flies, and midges. There is little evidence that traps baited with octenol are also effective in attracting phlebotomine sand flies. The present report evaluated octenol in modified Centers for Disease Control and Prevention (CDC) traps in two experiments: 1) modified CDC trap without light and 2) modified CDC trap with light. The traps were baited with octenol at concentrations of 0.5, 27, and 43 mg/h in Rincão locality, São Paulo, Brazil. Traps without octenol were used as controls. The sand fly Nyssomyia neivai (Pinto) (= Lutzomyia neivai) (Diptera: Psychodidae: Phlebotominae) was the prevalent species (99.9%) in both experiments. The results of the experiments showed that traps baited with octenol at 27 and 43 mg/h caught significantly more N. neivai than control and octenol at 0.5 mg/h with and without light. This is the first report that shows that octenol itself is attractive to N. neivai and associated with light traps significantly increases the catches...

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São apresentados no presente estudo os resultados de coletas de flebotomíneos com armadilhas de Falcão e Shannon, em 53 localidades rurais de 19 municípios do Estado do Paraná. Foram coletados 27.441 flebotomíneos de 13 espécies, com predomínio de Nyssomyia whitmani. Constatou-se que há grande concentração de flebotomíneos nos abrigos de animais domésticos e nas matas.

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The survival, absolute population size, gonotrophic cycle duration, and temporal and spatial abundance of Nyssomyia neivai (Pinto) were studied in a rural area endemic for American cutaneous leishmaniasis (ACL) in Conchal, Sõo Paulo State, southeastern Brazil, using mark-release-recapture techniques and by monitoring population fluctuation. The monthly abundance exhibited a unimodal pattern, with forest and domicile habitats having the highest relative abundances. A total of 1,873 males and 3,557 females were marked and released during the six experiments, of which 4.1–13.0% of males and 4.1–11.8% of females were recaptured. Daily survivorship estimated from the decline in recaptures per day was 0.681 for males and 0.667 for females. Gonotrophic cycle duration was estimated to be 4.0 d. Absolute population size was calculated using the Lincoln Index and ranged from 861 to 4,612 males and from 2,187 to 19,739 females. The low proportion of females that reach the age when they are potentially infective suggests that N. neivai has a low biological capacity to serve as a vector and that factors such as high biting rates and opportunistic feeding behavior would be needed to enable Leishmania (Viannia) braziliensis Vianna transmission. This agreed with the epidemiological pattern of ACL in southeastern Brazil that is characterized by low incidence, with isolated cases acquired principally within domiciliary habitats...

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Realizou-se coletas de flebotomíneos de maio de 2005 a abril de 2006. Os resultados foram comparados com os das coletas realizadas entre abril de 2001 e setembro de 2002, para avaliar as medidas empregadas para diminuir a densidade destes insetos, no Recanto Marista, município de Doutor Camargo, Estado do Paraná. As coletas foram feitas com armadilhas de Falcão, em domicílios e galinheiros, das 22 às 2 horas, semanalmente, quatro vezes ao mês. Em 2005 e 2006, coletaram-se 213.195 flebotomíneos, 1.113,8 em média por hora (MH) e em 2001 e 2002, 199.821 (MH=1.653,5). Nyssomyia neivai prevaleceu (75,4 por cento) em todos os ecótopos que, juntamente com Nyssomyia whitmani (23,4 por cento), Migonemyia migonei (0,8 por cento) e Pintomyia fischeri (0,4 por cento), representaram 99,7 por cento do total coletado. Nos ecótopos representados por galinheiros foram coletados 88,7 por cento dos flebotomíneos. Constatou-se que houve queda na densidade de flebotomíneos no período de coletas 2005 e 2006 em relação à de 2001 e 2002, especialmente no domicílio.

Collections of sandflies were made between May 2005 and April 2006. The results were compared with those from collections undertaken between April 2001 and September 2002, in order to evaluate the measures used to decrease the density of these insects in Recanto Marista, municipality of Doutor Camargo, State of Paraná. The collections were carried out by Falcão traps inside domiciles and hen sheds, from 10 p.m. to 2 a.m. once a week, four times a month. In 2005 and 2006, 213,195 sandflies were collected (average of 1,113.8 per hour), compared with 199,821 (average of 1,653.5 per hour) in 2001 and 2002. Nyssomyia neivai predominated (75.4 percent) in all the ecotopes. Nyssomyia neivai, Nyssomyia whitmani, Migonemyia migonei and Pintomyia fischeri accounted for 99.7 percent of all the sandflies collected. The ecotope of hencoops accounted for 88.7 percent of the sandflies collected. It was observed that the sandfly density had decreased between the 2001-2002 and 2005-2006 collections, especially in homes.

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A leishmaniose tegumentar americana tem sido notificada em todos os estados do Brasil e no Paraná essa doença é endêmica. O objetivo deste trabalho foi detectar a infecção natural de flebotomíneos para verificar a competência vetorial destes insetos e a identificação da espécie parasitária. Os flebotomíneos foram coletados com armadilhas de Falcão e Shannon, nos municípios de Doutor Camargo, Fênix e Mandaguari, de novembro de 2005 a agosto de 2006. Coletaram-se 12.930 flebotomíneos, dos quais 2.487 fêmeas foram dissecadas e destes 1.230 fêmeas foram submetidas à reação em cadeia da polimerase. Pelo método da dissecação, foi detectada uma fêmea de Nyssomyia whitmani com infecção natural por flagelados e pela reação em cadeia da polimerase não se detectou a presença de DNA de Leishmania em nenhuma das fêmeas. Apesar de não ter sido detectada a infecção natural de Nyssomyia neivai nas localidades em apreço e ainda que os requisitos de incriminação vetorial não tenham sido atendidos, não se deve negligenciar o potencial vetorial desta espécie.

American cutaneous leishmaniasis has been reported in all Brazilian states and in the Paraná this disease is endemic. The objective of this work was to detect natural infections in phlebotomines to verify the vector competence of these insects and the identification of the parasite species. Phlebotomines were collected using Falcão and Shannon traps, in the municipalities of Doutor Camargo, Fênix and Mandaguari, between November 2005 and August 2006. from 12,930 phlebotomines were collected, 2,487 females were dissected and 1,230 dissected females had been submitted to polymerase chain reaction. Flagellates were detected in a female Nyssomyia whitmani that had been dissected and for polymerase chain reaction failed to detect Leishmania DNA in any females. Even though flagellates were not detected in Nyssomyia neivai it should still be considered as a potencial vector.

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Nyssomyia intermedia (Lutz & Neiva 1912) and N. neivai (Pinto 1926) are possible vectors of tegumentary leishmaniasis in some regions of Brazil. Further, the latter was until recently, considered a junior synonym of the former. This study has the purpose of updating our knowledge of the geographical distribution of these species, based on specimens deposited at the collection of the Centro de Pesquisas René Rachou-Fiocruz, Faculdade de Saúde Pública-Universidade de São Paulo, and on data presented by literature as also to associate this distribution with the cutaneous leishmaniasis cases reported. It has been reported that N. intermedia occurs in the states of the Northeastern Region, in Rio de Janeiro, Espírito Santo, on the northern coast of São Paulo, in eastern Minas Gerais, Mato Grosso do Sul, and Goiás, close to the border with Minas Gerais and Bahia. N. neivai occurs in the Southern Region, southern coast and in western São Paulo, southern and western Minas Gerais, southern Goiás, and southern Pará, beyond Argentina, Bolivia, and Paraguay. It is important to highlight that N. intermedia and N. neivai occur in sympatry in Minas Gerais and São Paulo. N. intermedia or N. neivai are predominant or are captured abundantly in several cutaneous leishmaniasis foci in the Southeastern and Southern regions of Brazil.

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