Estudo da variabilidade genética deLeishmania (Viannia) braziliensis Vianna, 1911 de diferentes regiões do Brasil

Exportado OPUSMade available in DSpace on 2019-08-09T16:22:58Z (GMT). No. of bitstreams: 1 tese_2011.pdf: 4665436 bytes, checksum: 31e608ad001e159af9b11ccf634e9cfa (MD5) Previous issue date: 21Leishmania (Viannia) braziliensis é a espécie mais prevalente nos casos humanos da leishmaniose tegumentar nas Américas. Polimorfismo genético têm sido encontrado em populações naturais de diferentes espécies de Leishmania, o que explicaria a adaptação destes parasitos às mudanças observadas nas condições ambientais. Neste estudo utilizou-se um painel significativo de isolados L. braziliensis de focos de LTA de diferentes regiões geográficas e biomas do Brasil. Os polimorfismos do DNA genômico foram analisados utilizando-se diferentes marcadores genéticos, comparou-se a variabilidade genética intra-específica das amostras e determinou-se as relações fenéticas entre elas. Na eletroforese de isoenzimas (MLEE), a maioria das amostras (87,3%) apresentou o perfil de migração idêntico ao da cepa padrão de L. braziliensis (M2903); 8 amostras (10,2%) apresentaram apenas um eletromorfo diferente para a enzima IDHNADP. Duas amostras (2,6%) apresentaram o perfil mais heterogêneo, com padrões diferentes de L. braziliensis para 3 enzimas (G6PDH, IDHNADP e MDH). Estas amostras foram designadas como variantes. Para o RAPD-PCR com o iniciador M1340F pode-se observar que o conjunto dos isolados formaram claramente dois grandes clusters, pela análise com o programa NTSYS, refletindo a diversidade genética entre as amostras de L. braziliensis testadas. A análise dos perfis gerados pela SSR-PCR evidenciou que os iniciadores CARY e K7 foram capazes de mostrar variabilidade intra-específica. Observou-se que com o iniciador CARY, as amostras agruparam-se em um clustrer, incluindo a cepa de referência de L. braziliensis. Por outro lado, com o iniciador K7 observou-se a formação de dois clusters. Uma árvore consenso foi gerada por parcimônia com os dados concatenados dos três marcadores utilizados (M1340F, CARY e K7) e observou-se a formação de dois clusters onde a maioria das amostras do bioma Amazônia /região Norte (58%) e região Nordeste (62%) estão juntas em um grupo e o bioma Cerrado /região Centro-Oeste (100%), Cerrado e Mata Atlântica (67%) e a região Sul (80%) estão em outro grupo, o que concorda com a análise feita pelo método de distância genética. Foram encontrados dois perfis para ITS1- HAEIII e após a análise dos géis foram identificados três perfis para hsp70-BstUI. Observou-se que o índice de similaridade médio entre as amostras para os testes de isoenzimas, RAPD-PCR com o iniciador M1340, SSR-PCR com o iniciador CARYe K7 foi de aproximadamente 70%, demonstrando a existência de variabilidade de L. braziliensis confirmada através das diferentes técnicas utilizadas. Combinando os resultados da análise por biomas e por região, podemos dizer que a maioria das amostras do bioma Amazonas, região Norte e bioma Caatinga estão sempre agrupadas em um mesmo clustere o bioma Cerrado, região centro-oeste em outro cluster. Este estudo refletiu a diversidade genética existente em L. braziliensis sugerindo a possibilidade de uma associação entre variabilidade e origem geográfica

Evaluation of parasitological examination, kDNA polymerase chain reaction and rK39-based immunochromatography for the diagnosis of visceral leishmaniasis in seropositive dogs from the screening-culling program in Brazil

Submitted by Nuzia Santos ([email protected]) on 2015-03-26T18:29:25Z No. of bitstreams: 1 2014_176.pdf: 855927 bytes, checksum: 9b3144a17bad0ab4c438f6b558325146 (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-03-26T18:29:33Z (GMT) No. of bitstreams: 1 2014_176.pdf: 855927 bytes, checksum: 9b3144a17bad0ab4c438f6b558325146 (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-03-26T18:52:44Z (GMT) No. of bitstreams: 1 2014_176.pdf: 855927 bytes, checksum: 9b3144a17bad0ab4c438f6b558325146 (MD5)Made available in DSpace on 2015-03-26T18:52:44Z (GMT). No. of bitstreams: 1 2014_176.pdf: 855927 bytes, checksum: 9b3144a17bad0ab4c438f6b558325146 (MD5) Previous issue date: 2014Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Leishmanioses. Belo Horizonte, MG, BrasilFundação Ezequiel Dias. Diretoria de Pesquisa e Desenvolvimento. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Leishmanioses. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Departamento de Parasitologia. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Leishmanioses. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Leishmanioses. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Leishmanioses. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Leishmanioses. Belo Horizonte, MG, BrasilIntroduction: Dogs play a primary role in the zoonotic cycle of visceral leishmaniasis (VL). Therefore, the accurate diagnosis of infected dogs, primarily asymptomatic dogs, is crucial to the efficiency of VL control programs. Methods: We investigated the agreement of four diagnostic tests for canine visceral leishmaniasis (CVL): parasite detection, either after myeloculture or by direct microscopic examination of tissue imprints; kinetoplast-deoxyribonucleic acid-polymerase chain reaction (kDNA-PCR); and an immunochromatographic test (ICT). An enzyme-linked immunosorbent assay (ELISA) and an indirect immunofluorescence test (IFAT), both of which were adopted as part of the screening-culling program in Brazil, were used as reference tests. Our sample set consisted of 44 seropositive dogs, 25 of which were clinically asymptomatic and 19 were symptomatic for CVL according to ELISA-IFAT. Results: The highest and lowest test co-positivities were observed for ICT (77.3%) and myeloculture (58.1%), respectively. When analyzed together, the overall percentage of co-positive tests was significantly higher for the symptomatic group compared to the asymptomatic group. However, only ICT was significantly different based on the results of a separate analysis per test for each group of dogs. The majority (93.8%) of animals exhibited at least one positive test result, with an average of 2.66 positive tests per dog. Half of the symptomatic dogs tested positive for all four tests administered. Conclusions: The variability between test results reinforces the need for more efficient and reliable methods to accurately diagnose canine VL, particularly in asymptomatic animals

Survey of Sand Flies (Diptera: Psychodidae) in an Environmentally Protected Area in Brazil.

Submitted by Nuzia Santos ([email protected]) on 2016-02-29T13:50:29Z No. of bitstreams: 1 Survey of Sand Flies.pdf: 485619 bytes, checksum: f8d45630b2b8a572a9f83ac70b93519c (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2016-02-29T13:54:19Z (GMT) No. of bitstreams: 1 Survey of Sand Flies.pdf: 485619 bytes, checksum: f8d45630b2b8a572a9f83ac70b93519c (MD5)Made available in DSpace on 2016-02-29T13:54:19Z (GMT). No. of bitstreams: 1 Survey of Sand Flies.pdf: 485619 bytes, checksum: f8d45630b2b8a572a9f83ac70b93519c (MD5) Previous issue date: 2015Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Estudos em Leishmanioses. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Estudos em Leishmanioses. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Estudos em Leishmanioses. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Estudos em Leishmanioses. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Estudos em Leishmanioses. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Estudos em Leishmanioses. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Estudos em Leishmanioses. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Estudos em Leishmanioses. Belo Horizonte, MG, BrazilBrazil is one of the most important endemic areas for leishmaniasis worldwide. Protected areas that are tourist attractions likely present an important risk of transmission of cutaneous leishmaniasis (CL). Furthermore, with the geographical expansion of visceral leishmaniasis (VL), several studies have recorded the occurrence of its vector, Lutzomyia longipalpis, and cases of human and canine VL in such tourist areas. The Parque Estadual do Sumidouro is an environmentally protected area located in the Brazilian Cerrado biome and in an important area endemic for leishmaniasis in the state of Minas Gerais. The purpose of this study was to monitor the sand fly fauna in areas of tourist activity in the park. Sampling was performed every month, from September 2011 to August 2013, using CDC light traps at six sites of differing environmental characteristics. Sampled specimens were identified following Galati (2003), and females were submitted to molecular techniques for the detection and identification of Leishmania DNA. A total of 4,675 sand fly specimens of 25 species belonging to nine genera were collected. The most abundant species were Micropygomyia quinquefer, Lutzomyia renei and Pintomyia pessoai, although only Pi. pessoai is implicated in the transmission of Leishmania braziliensis. The species accumulation curve reached saturation on the 16th sampling event. Species richness, diversity and evenness differed among the sampled areas. The seasonal curve was not determined by a single unique species, and no single species was the most abundant in all environments sampled. The main vector of Leishmania (Leishmania) infantum, Lutzomyia longipalpis, accounted for only 5.35% of the specimens collected. Proven or suspected vectors of Leishmania (Viannia) braziliensis were recorded, and one female of the cortellezzii complex tested positive for Le. braziliensis DNA. Even with a low infection rate (0.62%), these data indicate the circulation of the parasite and reinforce the need for entomological and epidemiological surveillance in the park and its surroundings

Phlebotomine Sand Fly Fauna and Leishmania Infection in the Vicinity of the Serra do Cipó National Park, a Natural Brazilian Heritage Site.

Submitted by Nuzia Santos ([email protected]) on 2016-01-25T18:20:42Z No. of bitstreams: 1 Phlebotomine Sand Fly Fauna and Leishmania Infection in the Vicinity of the Serra do Cipó National Park.pdf: 776437 bytes, checksum: b210d45a854694e346bdaf87e0560e12 (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2016-01-25T18:25:24Z (GMT) No. of bitstreams: 1 Phlebotomine Sand Fly Fauna and Leishmania Infection in the Vicinity of the Serra do Cipó National Park.pdf: 776437 bytes, checksum: b210d45a854694e346bdaf87e0560e12 (MD5)Made available in DSpace on 2016-01-25T18:25:24Z (GMT). No. of bitstreams: 1 Phlebotomine Sand Fly Fauna and Leishmania Infection in the Vicinity of the Serra do Cipó National Park.pdf: 776437 bytes, checksum: b210d45a854694e346bdaf87e0560e12 (MD5) Previous issue date: 2015Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Leishmanioses. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Leishmanioses. Belo Horizonte, MG, BrasilFundação Ezequiel Dias. Diretoria de Pesquisa e Desenvolvimento. Belo Horizonte, MG, BrasilUniversidade Federal Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Parasitologia. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Leishmanioses. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Leishmanioses. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Leishmanioses. Belo Horizonte, MG, BrasilSecretaria Municipal de Saúde. Jaboticatubas, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Leishmanioses. Belo Horizonte, MG, BrasilIn the New World, the leishmaniases are primarily transmitted to humans through the bites of Leishmania-infected Lutzomyia (Diptera: Psychodidae) phlebotomine sand flies. Any or both of two basic clinical forms of these diseases are endemic to several cities in Brazil—the American cutaneous leishmaniasis (ACL) and the American visceral leishmaniasis (AVL). The present study was conducted in the urban area of a small-sized Brazilian municipality (Jaboticatubas), in which three cases of AVL and nine of ACL have been reported in the last five years. Jaboticatubas is an important tourism hub, as it includes a major part of the Serra do Cipó National Park. Currently, no local data is available on the entomological fauna or circulating Leishmania. During the one-year period of this study, we captured 3,104 phlebotomine sand flies belonging to sixteen Lutzomyia species. In addition to identifying incriminated or suspected vectors of ACL with DNA of the etiological agent of AVL and vice versa, we also detected Leishmania DNA in unexpected Lutzomyia species. The expressive presence of vectors and natural Leishmania infection indicates favorable conditions for the spreading of leishmaniases in the vicinity of the Serra do Cipó National Par

Environmental characteristics of the sand fly collection sites in PES from September 2011 to August 2013.

<p>Environmental characteristics of the sand fly collection sites in PES from September 2011 to August 2013.</p

Map showing the location of PES and the location of sampling sites in the park.

<p>(1)—Park surroundings—peridomestic areas; (2)—Cerrado 1; (3)—Cerrado 2; (4)—Rupicolous vegetation; (5)—Cave entrance; (6)—Cave surroundings.</p

Species accumulation curve, abundance, and species richness of the sand fly fauna of PES from September 2011 to August 2013.

<p>Species accumulation curve, abundance, and species richness of the sand fly fauna of PES from September 2011 to August 2013.</p