Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2018-09-13T13:48:58Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) amanda_oliveira_baracho.pdf: 1018229 bytes, checksum: 7da405c92d90a6056df94d878fc77113 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2018-10-02T16:26:57Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) amanda_oliveira_baracho.pdf: 1018229 bytes, checksum: 7da405c92d90a6056df94d878fc77113 (MD5) / Made available in DSpace on 2018-10-02T16:27:10Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) amanda_oliveira_baracho.pdf: 1018229 bytes, checksum: 7da405c92d90a6056df94d878fc77113 (MD5) Previous issue date: 2018 / Universidade Federal dos Vales do Jequitinhonha e Mucuri (UFVJM) / A leishmaniose visceral ? uma doen?a parasit?ria de transmiss?o vetorial causada por protozo?rios do g?nero Leishmania, com elevado grau de letalidade e que atinge milh?es de pessoas no mundo. A busca por produtos alternativos para a elimina??o do vetor tem como objetivo minimizar os impactos causados pelos inseticidas qu?micos sint?ticos utilizados pelos programas de controle, sendo cada vez mais frequente a pesquisa em produtos de origem bot?nica com atividade inseticida. Organismos vegetais s?o capazes de produzir compostos org?nicos de comprovada toxicidade frente aos insetos e, diante disto, avaliou-se neste trabalho a atividade inseticida de extratos de Caryocar brasiliense, planta nativa do Cerrado popularmente conhecida como ?pequi?, sobre flebotom?neos da esp?cie Lutzomyia longipalpis, principal transmissora da leishmaniose visceral. Extratos hidroetan?licos, etan?licos e ciclohex?nicos de folhas e cascas de C. brasiliense foram preparados e dilu?dos em solu??o de Tween 80 a 3% para a obten??o de concentra??es a 50, 100, 200 e 400 mg.mL-1, para a realiza??o dos ensaios biol?gicos. Os flebotom?neos foram coletados em uma localidade rural do munic?pio de Diamantina/MG. Um n?mero amostral de 30 flebotom?neos foi posto em contato com os extratos e a mortalidade foi avaliada ap?s 1, 2, 4, 16, 24 48 e 72 horas de exposi??o, bem como grupos controle negativos (?gua destilada e Tween) e positivo (cipermetrina). A identifica??o dos constituintes qu?micos dos extratos tamb?m foi avaliada. Ap?s 72h de experimento, foram registradas mortalidades de 93,3% pelo extrato hidroetan?lico da casca a 400 mg.mL-1, 66,3% pelo etan?lico da casca a 50 mg.mL-1 e 81,1% pelo ciclohex?nico de folhas a 200 mg.mL-1. Os extratos hidroetan?licos da casca mostraram-se estatisticamente semelhantes ? a??o da cipermetrina em 72h de experimento, indicando que este extrato foi o mais eficiente quanto ? atividade inseticida. Foram identificados triterpenos, esteroides, taninos, flavonoides, alcaloides e saponinas nos extratos. Os resultados obtidos sugerem que os extratos hidroetan?licos de C. brasiliense, sobretudo das cascas, s?o promissores na busca por compostos naturais com atividade inseticida sobre Lu. longipalpis. / Disserta??o (Mestrado) ? Programa de P?s-gradua??o em Biologia Animal, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2018. / Visceral leishmaniasis is a vector-borne disease caused by a protozoan of the genus Leishmania with high lethality rate that reaches millions of people worldwide. The impacts caused by synthetic insecticides used in control programs are well known. In this context, research on alternative products for vector control, especially products of botanical origin, has been frequent in order to minimize the impacts of synthetic ones. Plants are capable to produce organic compounds with insect toxicity. Here, the insecticidal activity of Caryocar brasiliense extracts, a native plant of Brazilian savannah, popularly known as "pequi", on Lutzomyia longipalpis, the main vector of visceral leishmaniasis, was tested. Hydroethanolic, ethanolic and cyclohexanic extracts of leaves and barks of C. brasiliense were prepared and diluted in Tween 80 (3%) to obtain concentrations of 50, 100, 200 and 400 mg.mL-1 for the biological tests. Phlebotomine sandflies were collected in a rural locality of Diamantina (Minas Gerais state, Brazil). Thirty sand flies were exposed to the extracts and mortality was evaluated after 1, 2, 4, 16, 24 48 and 72 hours of exposure, as well as negative control groups (distilled water and Tween) and positive (cypermethrin). Chemical compounds of the extracts were also evaluated. Mortalities rates of 93.3% were observed for the hydroethanolic extract of the bark at 400 mg.mL-1, 66.3% for the ethanolic bark at 50 mg.mL-1 and 81.1% for the cyclohexane of leaves at 200 mg.mL-1. The hydroethanolic extracts of the bark showed to be statistically similar to the action of cypermethrin in 72h of the experiment, indicating that this extract was the most efficient as an insecticide. Triterpenes, steroids, tannins, flavonoids, alkaloids and saponins were identified in the extracts. The results obtained suggest that the hydroethanolic extracts of C. brasiliense, especially of the bark, are promising for the use of natural compounds to control Lu. longipalpis.

Les phlébotomes sont les vecteurs reconnus de plusieurs arbovirus, en particulier du genre Phlebovirus, ainsi que de parasites du genre Leishmania. Les infections par les phlébovirus sont responsables chez l’homme de maladies décrites depuis longtemps, pourtant ils demeurent méconnus, avec en particulier un manque de données épidémiologiques et d’outils de diagnostic.Dans une première partie, des études de séroprévalence nous ont permis d’aborder l’impact en santé publique, dans le sud-est de la France, de deux phlébovirus connus pour leur pouvoir pathogène chez l’homme, Toscana virus (TOSV) et Sandfly fever Sicilian virus (SFSV). Pour ce dernier, des anticorps spécifiques (IgG) ont été détectés dans moins de 1% des sérums testés, ce qui suggère que SFSV joue un rôle mineur dans les pathologies humaines de cette région ; ces résultats sont corroborés par l’absence, durant ces dernières décennies, de cas documentés d’infection aigüe due à SFSV en Europe occidentale. Nous avons donc pu concentrer notre travail sur le deuxième groupe de phlébovirus d’intérêt chez l’homme, le groupe des Sandfly fever Naples virus, qui inclut notamment TOSV. Nous avons démontré l’existence d’un lien épidémiologique entre les infections à Leishmania infantum et celles à TOSV, certainement dû au fait que ces pathogènes sont transmis par un vecteur commun (Phlebotomus perniciosus). Les analyses statistiques ont montré que les personnes exposées aux infections à TOSV ont plus de chance d’être aussi infectées par les parasites leishmanies (et vice versa). En admettant que ce lien épidémiologique entre leishmanioses et infections à TOSV est représenté par l’exposition à la piqûre d’un vecteur commun, cette étude confirme l’implication de Phlebotomus perniciosus en tant que vecteur principal de TOSV dans le sud de la France. Cette étude suggère également que certaines données épidémiologiques disponibles pour la leishmaniose pourraient être utilisées pour décrypter l’épidémiologie des infections à TOSV.La deuxième partie de cette thèse est consacrée à la détection, l’isolement et la caractérisation de virus, déjà connus ou inconnus, dans les populations de phlébotomes en France et en Afrique du Nord. Pour atteindre cet objectif, nous avons dû développer une plateforme d’analyse à au débit, adaptée pour la découverte de virus dans les phlébotomes, qui permette de traiter un grand nombre d’échantillons à faible coût. Cette plateforme a récemment été complétée par un outil de Next Generation Sequencing, afin de réaliser la caractérisation génétique complète des virus isolés et découverts. Au total, 12 576 phlébotomes ont été capturés au cours de 12 campagnes de capture menées en France, en Tunisie et en Algérie. Au sein d’une même zone géographique, la découverte de plusieurs nouveaux phlébovirus, ainsi que leur taux d’infection observé dans les populations de phlébotomes, ont démontré que la diversité de phlebovirus est bien plus importante qu’attendue.Dans la troisième partie de cette thèse, une étude de séroprévalence a été menée sur des sérums humains en utilisant des tests comparatifs de neutralisation de virus. Cette étude nous a permis d’exclure le virus Punique, récemment découvert, de la liste des principales menaces en santé publique au nord de la Tunisie, et de confirmer que TOSV est le principal phlebovirus pathogène ayant un impact en santé publique dans cette région du pays. Cette méthode de neutralisation est capable d’identifier précisément, parmi des virus génétiquement proches, le virus contre lequel les anticorps présents dans le sang ont été produits, ce qui permet de déterminer la capacité de chacun de ces virus à jouer un rôle en santé publique. / Sandflies are vectors of various arthropod-borne viruses, in particular viruses within the genus Phlebovirus, family Bunyaviridae, and of parasites in the genus Leishmania. Human diseases caused by infection with sandfly-borne phleboviruses are known for a long time, but they remain neglected due to the lack of epidemiological knowledge and of diagnostic tools.The first part consisted of seroprevalence studies in human sera to address the public health impact in south-eastern France of two recognized sandfly-borne phleboviruses, namely Toscana virus (TOSV) and Sandfly fever Sicilian virus (SFSV). Concerning the latter, specific IgG were detected in less than 1% of tested sera, suggesting that SFSV play a minor role in human disease in the region; this finding was corroborated with the lack of documented case of acute infection due to SFSV in Western Europe during the last decade. This pleaded for focusing on the other group of sandfly fever viruses known for their human interest, namely the group of Sandfly fever Naples virus that includes TOSV. We demonstrated an existing epidemiological relationship between Leishmania infantum and TOSV infections, presumably through the transmission by the common arthropod vector (Phlebotomus perniciosus). Statistical analysis showed that persons exposed to TOSV infection are at greater risk of being infected with Leishmania parasite (and vice versa). Assuming that epidemiological link between leishmaniasis and TOSV infection may be represented by the exposure to the bite of a common vector, this study confirms the involvement of Phlebotomus perniciosus as the major vector of TOSV in the South of France. This study also suggests that some of the epidemiological data available on Leishmaniasis may be used to decipher the epidemiology of TOSV infections..The second part of this thesis was dedicated to detection, isolation and characterization of existing and/or new phleboviruses in sandfly populations in France and in North Africa. To achieve this aim, we had to set up a high-throughput cost-effective platform amenable to virus discovery in sandflies; this sandfly-processing platform has been recently docked to a Next Generation Sequencing platform for full genetic characterization of newly isolated and discovered viruses. A total of 12,576 sandflies were trapped during 12 campaigns conducted in France, Tunisia and Algeria. The discovery of several new phleboviruses and their observed frequency in sandfly populations has clearly demonstrated that within a given geographic area, virus diversity is much higher than previously believed.In the third part of this thesis, a seroprevalence study based on comparative virus neutralization tests was performed on human sera and allowed to exclude the newly described Punique virus from the list of major public health threats in northern Tunisia, and to confirm that TOSV is the dominant phleboviral pathogen with an impact on public health in this part of the country. This neutralization method is suitable to identify precisely the virus against which antibodues were elicited, allowing to discriminate among closely related phleboviruses, and to determine their propensity to play a role in public health.

Introdução. A leishmaniose tegumentar americana (LTA) é uma endemia de importância em saúde pública na Amazônia Brasileira, onde possui diferentes perfis de transmissão com a participação de diversas espécies de vetores e de protozoários do gênero Leishmania. O Estado do Acre apresenta altos índices de LTA e, no ano de 2015, apresentou o mais alto coeficiente de detecção de casos (137,7/100.000 hab.) da doença no Brasil. Objetivo. Analisar aspectos epidemiológicos da LTA no município de Xapuri, Estado do Acre, Brasil, envolvendo população humana, cães domésticos, vetores e identificação de Leishmania. Materiais e Métodos. Para a avaliação epidemiológica, foram analisadas fichas de notificação dos casos humanos no período de 2008 a 2014 obtidas do sistema de vigilância epidemiológica do município. Foram analisadas as variáveis: sexo, grupo etário, escolaridade, forma clínica, diagnóstico, tratamento e evolução clínica. Os dados foram submetidos à análise descritiva e teste estatístico do qui-quadrado de Pearson utilizando o pacote estatístico STATA. Também foram obtidas amostras de material de pacientes (escarificação das lesões fixadas em lâminas) atendidos no centro médico do município, durante o período de novembro de 2014 a janeiro de 2016. As amostras foram submetidas à análise molecular para diagnóstico de Leishmania spp. O inquérito canino foi realizado em áreas urbanas e rurais; nestas, predominantemente, em seringais, onde a doença foi reportada em humanos. Após avaliação clínica dos animais para busca de lesões e sinais característicos de leishmanioses, foram coletadas amostras de sangue venoso por punção jugular ou cefálica, e quando havia lesões sugestivas da doença, foram anestesiados e submetidos à biopsia para colheita do fragmento de lesão. Amostras destes fragmentos foram submetidas a técnicas parasitológicas, inoculação em meio de cultura Neal, Novy e Nicolle, exame direto e técnicas moleculares para detecção do parasita. A identificação de Leishmania spp. foi realizada por técnicas moleculares. Os flebotomíneos foram coletados em ambiente domiciliar e florestal de dois seringais (Floresta e Cachoeira) e na área urbana, com armadilhas luminosas tipo CDC, instaladas mensalmente, no período de agosto de 2013 a julho de 2015. Neste período, apenas no seringal Cachoeira, também foram feitas coletas utilizando armadilhas de Shannon nas cores branca e preta, e de janeiro a maio de 2016, coletas em trocos de árvores com aspiradores manuais. Uma amostra de fêmeas coletadas pelas diferentes técnicas foi dissecada viii para investigação da presença de flagelados. Para a análise do comportamento da fauna flebotomínea foram utilizados índices ecológicos como de Shannon, Pielou e Abundância das Espécies Padronizado, média geométrica de Williams e análise dos componentes principais. Para estimar a atratividade dos flebotomíneos pelas cores branca e preta foi utilizado o teste de Mann-Whitney (p<0,05). Análises morfométricas utilizando o teste de Gabriel (teste F, p <0,05) foram feitas para distinguir alguns táxons. Para as análises moleculares dos parasitas foi empregada a técnica de Nested-PCR SSU rRNA utilizando os iniciadores S4/S12 e S17/S18 e sequenciamento. Resultados. No estudo de casos humanos, constatou-se que a doença ocorre predominantemente em populações rurais e isoladas do município, em indivíduos de ambos os sexos, com as incidências mais elevadas em crianças e adolescentes. Em 33 dos 45 pacientes com clínica positiva para LTA foram detectadas a presença de DNA de Leishmania spp. Nos cães domésticos, verificou-se alta taxa de infecção (20,0 por cento ) por Leishmania (Viannia) sp. Nos estudos de flebotomíneos, foram coletados 21.197 espécimes (14.210 fêmeas e 7.107 machos) com armadilhas CDC, e 6.309 (864 machos e 5.445 fêmeas) com armadilhas de Shannon. As frequências, abundâncias e densidades mais elevadas foram dos gêneros Nyssomyia, Psychodopygus e Trichophoromyia, coletados em ambientes silvestres, peri e intradomiciliares. Em ambiente rural foram coletados 99,9 por cento dos espécimes e no urbano apenas 0,1 por cento . Nyssomyia shawi predominou no Seringal Cachoeira, e Trichophoromyia spp. (Th. auraensis/Th. ruifreitasi) no Seringal Floresta. Espécies do gênero Psychodopygus predominaram no período chuvoso, enquanto as de Nyssomyia, no período seco. Infecções por Leishmania spp. foram detectadas em Brumptomyia sp., Nyssomyia antunesi, Ny. shawi, Lutzomyia sherlocki, Psathyromyia aragaoi, Psychodopygus carrerai carrerai, Ps. davisi, Ps. hirsutus hisutus, Ps. llanosmartinsi, Ps. lainsoni, Thrichophoromyia ubiquitalis e Trichophoromyia spp. Por meio de análises morfológicas e morfométricas das fêmeas de Trichophoromyia sugeriu-se a distinção de Th. octavioi de Trichophoromyia spp. (Th. auraensis/Th. ruifreitasi) e descreveu-se, Psathyromyia elizabethdorvalae sp. n. Conclusões. A LTA em Xapuri apresenta um perfil de transmissão silvestre e outro domiciliar. As populações humanas e caninas que frequentam ambientes florestais estão expostas a uma alta diversidade de vetores e de agentes etiológicos, o que aumenta o risco de infecção de LTA. As informações aqui apresentadas podem nortear as medidas de controle, planejamento das ações e definição de prioridades dos órgãos de vigilância epidemiológica do Estado do Acre, visando o diagnóstico precoce e tratamento adequado dos casos de leishmanioses da população humana de Xapuri / Introduction. American cutaneous leishmaniasis (ACL) is an endemic disease that deserves the attention of public health in the Brazilian Amazon, where it has various transmission profiles with the participation of several species of vectors and protozoa of the genus Leishmania. The state of Acre registers high rates of ACL, having in 2015 the highest coefficient of detection of cases (137.7 / 100,000 inhab.) in Brazil. Objective. To analyze the epidemiological aspects of LTA in the municipality of Xapuri, State of Acre, Brazil, involving the human population, domestic dogs, vectors and the identification of Leishmania ssp. Materials and Methods. For the epidemiological evaluation, records of the human cases notified between 2008 and 2014 obtained from the epidemiological surveillance system of the municipality, were analyzed. The following variables were selected for analysis: sex, age group, schooling, clinical form, diagnosis, treatment and clinical evolution. The data were submitted to descriptive analysis and the Pearson chi-squared statistical test using the statistical package STATA. Samples of patient material (scarification of lesions fixed on slides) were also obtained from the medical center of the city during the period from November 2014 to January 2016. These samples were submitted to molecular analysis for the diagnosis of Leishmania spp. The canine survey was carried out in both urban and rural areas, in the latter rubber plantations where the disease had been reported in humans predominated. After a clinical evaluation to search for lesions and characteristic signs of leishmaniasis, samples of venous blood were collected by jugular or cephalic puncture, and when the animals presented lesions suggestive of cutaneous leishmaniasis, they were anesthetized and submitted to biopsy to harvest the lesion fragment. Samples of these fragments were submitted to parasitological techniques, inoculation in Neal, Novy and Nicolle culture medium, direct examination and molecular techniques to detect Leishmania spp. The samples were submitted to molecular analysis for the diagnosis of Leishmania spp. The phlebotomine survey was carried out in forest and the domiciliary environment of the city and in two rubber plantation areas (Seringal Cachoeira and Seringal Floresta), monthly using CDC light traps, from August 2013 to July 2015. In this period, only in the Seringal Cachoeira were collections also made using black and white Shannon traps. Collections in tree trunks and among tree roots with manual aspirators were also undertaken from January to May 2016. xi Samples of females collected by different techniques were dissected to investigate the presence of flagellates. For the analysis of the behavior of the phlebotomine fauna, ecological indexes such as Shannon, Pielou, and Standardized Species Abundance, Williams geometric mean and main component analysis were used. The Mann-Whitney test (p <0.05) was used to estimate the attractiveness of the black and white colors to sandflies. Morphometric analyses using the Gabriel test (test F, p <0.05) were made to distinguish between some taxa. For the molecular analyses of the parasites the Nested-PCR SSU rRNA technique was used using primers S4 / S12 and S17 / S18 and sequencing. Results. In the study of human cases, it was found that the disease occurs in rural and isolated populations and in individuals of both sexes, especially in children and adolescents. In 33 of the 45 patients with positive ACL, the presence of Leishmania spp DNA was detected. The domestic dogs have shown a high infection rate (20.0 per cent ) attributed to Leishmania (Viannia) sp. In the sandfly studies, 21,197 specimens (14,210 females and 7,107 males) were collected in CDC traps, and 6,309 (864 males and 5,445 females) were collected in Shannon traps. The highest frequencies, abundances and densities were of the genera Nyssomyia, Psychodopygus and Trichophoromyia collected in wild, peri and intradomiciliary environments. Nyssomyia shawi predominated in the Seringal Cachoeira, and the Trichophoromyia spp. (Th. auraensis/Th. ruifreitasi) in the Seringal Floresta. Species of the genus Psychodopygus predominated in the rainy season while those of Nyssomyia in the dry period. Natural infections by Leishmania spp were detected in Brumptomyia sp., Nyssomyia antunesi, Ny. shawi, Lutzomyia sherlocki, Psathyromyia aragaoi, Psychodopygus carrerai carrerai, Ps. davisi, Ps. hirsutus hirsutus, Ps. llanosmartinsi, Ps. lainsoni, Trichophoromyia ubiquitalis and Trichophoromyia sp. Morphological and morphometric analyses of Trichophoromyia females were suggested to distinguish Th. octavioi from Trichophoromyia spp (Th. auraensis/Th. ruifreitasi), Pa. elizabethdorvalae being described for the first time. Conclusions. ACL in Xapuri presents one profile of wild transmission and another of domiciliar transmission. Both populations, human and canine, because they live in forest environments are exposed to a high diversity of vectors and etiological agents, which increases the risk of ACL infection. The information presented here may guide the measures of control, planning of actions and definition of priorities taken by the organs of surveillance and epidemiology of the State of Acre, aiming at the early diagnosis and appropriate treatment of the human cases in Xapuri

Introdução. A leishmaniose tegumentar americana (LTA) é uma endemia de importância em saúde pública na Amazônia Brasileira, onde possui diferentes perfis de transmissão com a participação de diversas espécies de vetores e de protozoários do gênero Leishmania. O Estado do Acre apresenta altos índices de LTA e, no ano de 2015, apresentou o mais alto coeficiente de detecção de casos (137,7/100.000 hab.) da doença no Brasil. Objetivo. Analisar aspectos epidemiológicos da LTA no município de Xapuri, Estado do Acre, Brasil, envolvendo população humana, cães domésticos, vetores e identificação de Leishmania. Materiais e Métodos. Para a avaliação epidemiológica, foram analisadas fichas de notificação dos casos humanos no período de 2008 a 2014 obtidas do sistema de vigilância epidemiológica do município. Foram analisadas as variáveis: sexo, grupo etário, escolaridade, forma clínica, diagnóstico, tratamento e evolução clínica. Os dados foram submetidos à análise descritiva e teste estatístico do qui-quadrado de Pearson utilizando o pacote estatístico STATA. Também foram obtidas amostras de material de pacientes (escarificação das lesões fixadas em lâminas) atendidos no centro médico do município, durante o período de novembro de 2014 a janeiro de 2016. As amostras foram submetidas à análise molecular para diagnóstico de Leishmania spp. O inquérito canino foi realizado em áreas urbanas e rurais; nestas, predominantemente, em seringais, onde a doença foi reportada em humanos. Após avaliação clínica dos animais para busca de lesões e sinais característicos de leishmanioses, foram coletadas amostras de sangue venoso por punção jugular ou cefálica, e quando havia lesões sugestivas da doença, foram anestesiados e submetidos à biopsia para colheita do fragmento de lesão. Amostras destes fragmentos foram submetidas a técnicas parasitológicas, inoculação em meio de cultura Neal, Novy e Nicolle, exame direto e técnicas moleculares para detecção do parasita. A identificação de Leishmania spp. foi realizada por técnicas moleculares. Os flebotomíneos foram coletados em ambiente domiciliar e florestal de dois seringais (Floresta e Cachoeira) e na área urbana, com armadilhas luminosas tipo CDC, instaladas mensalmente, no período de agosto de 2013 a julho de 2015. Neste período, apenas no seringal Cachoeira, também foram feitas coletas utilizando armadilhas de Shannon nas cores branca e preta, e de janeiro a maio de 2016, coletas em trocos de árvores com aspiradores manuais. Uma amostra de fêmeas coletadas pelas diferentes técnicas foi dissecada viii para investigação da presença de flagelados. Para a análise do comportamento da fauna flebotomínea foram utilizados índices ecológicos como de Shannon, Pielou e Abundância das Espécies Padronizado, média geométrica de Williams e análise dos componentes principais. Para estimar a atratividade dos flebotomíneos pelas cores branca e preta foi utilizado o teste de Mann-Whitney (p<0,05). Análises morfométricas utilizando o teste de Gabriel (teste F, p <0,05) foram feitas para distinguir alguns táxons. Para as análises moleculares dos parasitas foi empregada a técnica de Nested-PCR SSU rRNA utilizando os iniciadores S4/S12 e S17/S18 e sequenciamento. Resultados. No estudo de casos humanos, constatou-se que a doença ocorre predominantemente em populações rurais e isoladas do município, em indivíduos de ambos os sexos, com as incidências mais elevadas em crianças e adolescentes. Em 33 dos 45 pacientes com clínica positiva para LTA foram detectadas a presença de DNA de Leishmania spp. Nos cães domésticos, verificou-se alta taxa de infecção (20,0 por cento ) por Leishmania (Viannia) sp. Nos estudos de flebotomíneos, foram coletados 21.197 espécimes (14.210 fêmeas e 7.107 machos) com armadilhas CDC, e 6.309 (864 machos e 5.445 fêmeas) com armadilhas de Shannon. As frequências, abundâncias e densidades mais elevadas foram dos gêneros Nyssomyia, Psychodopygus e Trichophoromyia, coletados em ambientes silvestres, peri e intradomiciliares. Em ambiente rural foram coletados 99,9 por cento dos espécimes e no urbano apenas 0,1 por cento . Nyssomyia shawi predominou no Seringal Cachoeira, e Trichophoromyia spp. (Th. auraensis/Th. ruifreitasi) no Seringal Floresta. Espécies do gênero Psychodopygus predominaram no período chuvoso, enquanto as de Nyssomyia, no período seco. Infecções por Leishmania spp. foram detectadas em Brumptomyia sp., Nyssomyia antunesi, Ny. shawi, Lutzomyia sherlocki, Psathyromyia aragaoi, Psychodopygus carrerai carrerai, Ps. davisi, Ps. hirsutus hisutus, Ps. llanosmartinsi, Ps. lainsoni, Thrichophoromyia ubiquitalis e Trichophoromyia spp. Por meio de análises morfológicas e morfométricas das fêmeas de Trichophoromyia sugeriu-se a distinção de Th. octavioi de Trichophoromyia spp. (Th. auraensis/Th. ruifreitasi) e descreveu-se, Psathyromyia elizabethdorvalae sp. n. Conclusões. A LTA em Xapuri apresenta um perfil de transmissão silvestre e outro domiciliar. As populações humanas e caninas que frequentam ambientes florestais estão expostas a uma alta diversidade de vetores e de agentes etiológicos, o que aumenta o risco de infecção de LTA. As informações aqui apresentadas podem nortear as medidas de controle, planejamento das ações e definição de prioridades dos órgãos de vigilância epidemiológica do Estado do Acre, visando o diagnóstico precoce e tratamento adequado dos casos de leishmanioses da população humana de Xapuri / Introduction. American cutaneous leishmaniasis (ACL) is an endemic disease that deserves the attention of public health in the Brazilian Amazon, where it has various transmission profiles with the participation of several species of vectors and protozoa of the genus Leishmania. The state of Acre registers high rates of ACL, having in 2015 the highest coefficient of detection of cases (137.7 / 100,000 inhab.) in Brazil. Objective. To analyze the epidemiological aspects of LTA in the municipality of Xapuri, State of Acre, Brazil, involving the human population, domestic dogs, vectors and the identification of Leishmania ssp. Materials and Methods. For the epidemiological evaluation, records of the human cases notified between 2008 and 2014 obtained from the epidemiological surveillance system of the municipality, were analyzed. The following variables were selected for analysis: sex, age group, schooling, clinical form, diagnosis, treatment and clinical evolution. The data were submitted to descriptive analysis and the Pearson chi-squared statistical test using the statistical package STATA. Samples of patient material (scarification of lesions fixed on slides) were also obtained from the medical center of the city during the period from November 2014 to January 2016. These samples were submitted to molecular analysis for the diagnosis of Leishmania spp. The canine survey was carried out in both urban and rural areas, in the latter rubber plantations where the disease had been reported in humans predominated. After a clinical evaluation to search for lesions and characteristic signs of leishmaniasis, samples of venous blood were collected by jugular or cephalic puncture, and when the animals presented lesions suggestive of cutaneous leishmaniasis, they were anesthetized and submitted to biopsy to harvest the lesion fragment. Samples of these fragments were submitted to parasitological techniques, inoculation in Neal, Novy and Nicolle culture medium, direct examination and molecular techniques to detect Leishmania spp. The samples were submitted to molecular analysis for the diagnosis of Leishmania spp. The phlebotomine survey was carried out in forest and the domiciliary environment of the city and in two rubber plantation areas (Seringal Cachoeira and Seringal Floresta), monthly using CDC light traps, from August 2013 to July 2015. In this period, only in the Seringal Cachoeira were collections also made using black and white Shannon traps. Collections in tree trunks and among tree roots with manual aspirators were also undertaken from January to May 2016. xi Samples of females collected by different techniques were dissected to investigate the presence of flagellates. For the analysis of the behavior of the phlebotomine fauna, ecological indexes such as Shannon, Pielou, and Standardized Species Abundance, Williams geometric mean and main component analysis were used. The Mann-Whitney test (p <0.05) was used to estimate the attractiveness of the black and white colors to sandflies. Morphometric analyses using the Gabriel test (test F, p <0.05) were made to distinguish between some taxa. For the molecular analyses of the parasites the Nested-PCR SSU rRNA technique was used using primers S4 / S12 and S17 / S18 and sequencing. Results. In the study of human cases, it was found that the disease occurs in rural and isolated populations and in individuals of both sexes, especially in children and adolescents. In 33 of the 45 patients with positive ACL, the presence of Leishmania spp DNA was detected. The domestic dogs have shown a high infection rate (20.0 per cent ) attributed to Leishmania (Viannia) sp. In the sandfly studies, 21,197 specimens (14,210 females and 7,107 males) were collected in CDC traps, and 6,309 (864 males and 5,445 females) were collected in Shannon traps. The highest frequencies, abundances and densities were of the genera Nyssomyia, Psychodopygus and Trichophoromyia collected in wild, peri and intradomiciliary environments. Nyssomyia shawi predominated in the Seringal Cachoeira, and the Trichophoromyia spp. (Th. auraensis/Th. ruifreitasi) in the Seringal Floresta. Species of the genus Psychodopygus predominated in the rainy season while those of Nyssomyia in the dry period. Natural infections by Leishmania spp were detected in Brumptomyia sp., Nyssomyia antunesi, Ny. shawi, Lutzomyia sherlocki, Psathyromyia aragaoi, Psychodopygus carrerai carrerai, Ps. davisi, Ps. hirsutus hirsutus, Ps. llanosmartinsi, Ps. lainsoni, Trichophoromyia ubiquitalis and Trichophoromyia sp. Morphological and morphometric analyses of Trichophoromyia females were suggested to distinguish Th. octavioi from Trichophoromyia spp (Th. auraensis/Th. ruifreitasi), Pa. elizabethdorvalae being described for the first time. Conclusions. ACL in Xapuri presents one profile of wild transmission and another of domiciliar transmission. Both populations, human and canine, because they live in forest environments are exposed to a high diversity of vectors and etiological agents, which increases the risk of ACL infection. The information presented here may guide the measures of control, planning of actions and definition of priorities taken by the organs of surveillance and epidemiology of the State of Acre, aiming at the early diagnosis and appropriate treatment of the human cases in Xapuri

Circulation des Phlébovirus en Turquie, l'Iran et l'Algérie a été étudiée. L'isolement, la caractérisation génomique, les relations phylogénétiques de six virus ont été présentées: le virus Adana (ADAV), deux souches de virus Toros (TORV), le virus Zerdali (ZERV) de la Turquie; le virus Dashli (DASHV) de l'Iran; le virus Toscana (TOSV) d'Algérie. Cette étude a commencé avec la collection de 38,131 phlébotomes de la nature. La méthode de séquençage de nouvelle génération (NGS) à haut débit nous à été utilisée pour l’analyse des génomes complet des virus isoles. En conclusion, cette étude a d'importantes contributions sur phlébovirus négligées. Voici quelques-unes des contributions significatives; (i) ZERV et TORV qui sont étroitement apparentés au virus Tehran (THEV) et le virus Corfou (CFUV), respectivement, ont été isolés depuis 56 et 30 ans des premiers isolements de THEV et CFUV, respectivement, (ii) Détection du virus ADAV un animal domestique et sur quelques sérums humain par test de neutralisation. Ce virus ADAV constitue avec le virus le virus (SALV), le virus Arbia (ARBV), et le virus (ADRV) le groupe Salehabad. Seul le virus ADRV a été détectée dans le liquide cérébro-spinal auparavant landais que avec les autres, aucune preuve pathogène n’a été détectée, (iii) Nous avons découvert la plus récente circulation phlébovirus en Iran après 56 années, (iv) TOSV a été isolé en Algérie pour la première fois et la circulation a été confirmée par séropositivités dans le sérum humain. / Sandfly-borne phlebovirus circulation in Turkey, Iran, and Algeria was investigated. The isolation, genomic characterization, phylogenetic relationships of 6 viruses was presented: Adana virus (ADAV), two strains of Toros virus (TORV), Zerdali virus (ZERV) from Turkey; Dashli virus (DASHV) from Iran; Toscana virus (TOSV) from Algeria. This study has begun with the collection of 38,131 sandflies from nature. The well established, high-throughput methodology was applied for the discovery of viruses including PCR tools and cell culture methods. Next generation sequencing (NGS) technology facilitated to perform complete genome analysis of the isolated viruses. In conclusion, this study has contributions to the neglected sandfly-borne phlebovirus group and filled some gaps about the circulation of these agents in Turkey, Iran, and Algeria. Following are some significant contributions; (i) ZERV and TORV which are closely related to Tehran virus (THEV) and Corfou virus (CFUV), respectively were isolated after 56 and 30 years of the first isolations of THEV and CFUV, respectively, (ii) There was no evidence of the pathogenicity of Salehabad virus (SALV) and Arbia virus (ARBV) except the detection of Adria virus (ADRV) in CSF until ADAV which belongs to the Salehabad serocomplex was detected in domestic animal and very few human sera by neutralization assay, (iii) We have discovered the most recent sandfly-borne phlebovirus circulation in Iran after 56 years, (iv) TOSV was isolated in Algeria for the first time and circulation was confirmed by seropositivities in human sera.