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dc.contributor.authorSantos, Priscilla Nunes dos
dc.date.accessioned2023-12-22T01:57:27Z-
dc.date.available2023-12-22T01:57:27Z-
dc.date.issued2016-02-24
dc.identifier.citationSANTOS, Priscilla Nunes dos. Caracterização molecular de msp1 alfa da cepa AmRio1 de Anaplasma marginale em bezerro e carrapatos experimentalmente infectados. 2016. 31 f. Dissertação (Mestrado em Ciências Veterinárias) - Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, 2016.por
dc.identifier.urihttps://rima.ufrrj.br/jspui/handle/20.500.14407/11818-
dc.description.abstractA anaplasmose bovina, causada pela rickettsia intraeritrocítica Anaplasma marginale (ordem Rickettsiales, família Anaplasmataceae), é uma doença economicamente importante de bovinos e é endêmica em regiões tropicais e subtropicais do mundo. As proteínas principais de superfície (MSPs) têm sido muito utilizadas em estudos envolvendo A. marginale, sendo MSP1a uma das mais estudadas, pois esta é uma adesina de eritrócitos bovinos e células de carrapato, possuindo assim um papel importante no fenótipo de transmissão do agente. Ao mesmo tempo, é considerada um marcador estável para cepas de A. marginale, uma vez que possui uma região C-terminal altamente conservada e uma região N- terminal, altamente variável, porém, constante para cepas. Acredita-se que esta proteína pode sofrer pressão seletiva pois possui um epítopo sensível à neutralização em sua região variável. Assim, em alterações casuais da estrutura das sequências repetidas devido à mutação, cepas bem adaptadas poderiam prevalecer no ambiente. Desta forma, o objetivo deste estudo foi avaliar a diversidade do gene msp1 α de A. marginale na infecção aguda e crônica em um bezerro imunocompetente experimentalmente infectado e em carrapatos. No estudo, foi utilizado um bezerro, que foi infectado experimentalmente com a cepa AmRio 1 de A. marginale por via endovenosa. Este bezerro foi acompanhado através da observação de sinais clínicos, temperatura retal, esfregaços sanguíneos e hematócrito do dia 2 após infecção experimental até o dia 28 após infecção. O bovino posteriormente foi infestado com 1g de larvas de carrapatos Rhipicephalus (Boophilus) microplus. O sangue do bezerro foi colhido entre o dia 2 ao dia 119 pós infecção. Os carrapatos foram recuperados na fase de partenógina (n=61) sendo 43 submetidos à dissecção para remoção de intestino e glândula salivar, e 18 para a coleta de saliva utilizando pilocarpina 2%. As amostras foram submetidas à Reação em Cadeia da Polimerase (PCR), possuindo como gene alvo msp1 α. O produto desta reação foi enviado para sequenciamento em ambas as direções. Após a inoculação da cepa AmRio 1 de A. marginale, o bovino apresentou sinais clínicos e alterações em hematócrito e temperatura, havendo necessidade de tratamento de suporte para a recuperação do animal. Isto demonstra a patogenicidade da cepa. Todas as amostras do bezerro e dos carrapatos foram positivas na PCR. As sequências revelaram uma estabilidade nas sequências repetidas de MSP1a, tanto na infecção aguda quanto na crônica no bezerro e em todos os órgãos e salivas de carrapato. A positividade das salivas de carrapato sugere que a cepa AmRio 1 de A. marginale é transmitida biologicamente por R. (Boophilus) microplus. Além disto, nos órgãos de carrapato havia grandes quantidades de DNA de A. marginale, indicando a replicação do agente.por
dc.description.sponsorshipCNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológicopor
dc.formatapplication/pdf*
dc.languageporpor
dc.publisherUniversidade Federal Rural do Rio de Janeiropor
dc.rightsAcesso Abertopor
dc.subjectAnaplasmosepor
dc.subjectAmRio 1por
dc.subjectMSP1apor
dc.subjectBovinopor
dc.subjectRhipicephalus (Boophilus) micropluseng
dc.subjectAnaplasmosiseng
dc.subjectBovineeng
dc.titleCaracterização molecular de msp1 alfa da cepa AmRio1 de Anaplasma marginale em bezerro e carrapatos experimentalmente infectadospor
dc.title.alternativeMolecular characterization of msp1 alpha from Anaplasma marginale AmRio1 strain in calf and experimentally infected tickseng
dc.typeDissertaçãopor
dc.description.abstractOtherBovine anaplasmosis, caused by intraerythrocytic rickettsia Anaplasma marginale (Rickettsiales: Anaplasmataceae), is an economically important disease of cattle which is endemic in tropical and subtropical regions of the world. Major surface proteins (MSP) have been widely used in studies involving A. marginale, MSP1a being one of the most studied because it is an adhesin for bovine erythrocytes and tick cells, thus having an important role in the agent's transmission phenotype. At the same time, it is regarded as a stable marker for strains of A. marginale, since it has a highly conserved C-terminal region and a highly variable N-terminal region, but constant for strains. It is believed that this protein can undergo selective pressure because it has a neutralization-sensitive epitope in its variable region. Therefore, in random changes in the structure of tandem repeats due to mutation, well-adapted strains could prevail in the environment. Thus, the aim of this study was to evaluate the diversity of msp1 α gene of A. marginale in acute and chronic infection in one experimentally infected immunocompetent calf and in ticks. In the study, a calf, which was experimentally infected intravenously with AmRio 1 strain of A. marginale, was used. This calf was monitored by observing clinical signs, rectal temperature, blood smears and hematocrit of day-2 after experimental infection until the day 28 after infection. The bovine was subsequently infested with 1g of Rhipicephalus (Boophilus) microplus larvae. The calf's blood was collected from day 2 to day 119 after infection. Ticks were recovered in partenogenetic phase (n=61) and 43 underwent dissection to remove the gut and salivary gland, and 18 for saliva collection using pilocarpine 2%. The samples were submitted to Polymerase Chain Reaction (PCR), having msp1 α as target gene. The product of this reaction was submitted for sequencing in both directions. After inoculation of the strain AmRio 1 of A. marginale, the bovine showed clinical signs and changes in hematocrit and temperature, with supportive treatment necessary for the animal's recovery. This demonstrates the pathogenicity of the strain. All samples of the calf and the ticks were positive in PCR. The sequences revealed a stability in repeated sequences of MSP1a in both acute and chronic infection in the calf and all tick organs and saliva. Positivity of tick saliva suggests that the strain AmRio 1 of A. marginale is transmitted biologically by R. (Boophilus) microplus. In addition, tick organs had large amounts of DNA of A. marginale, indicating the agent's replication.eng
dc.contributor.advisor1Fonseca, Adivaldo Henrique da
dc.contributor.advisor1IDCPF: 475.018.557-49por
dc.contributor.advisor1Latteshttp://lattes.cnpq.br/441144116286260por
dc.contributor.referee1Cunha, Nathalie Costa da
dc.contributor.referee2Guedes Junior, Daniel da Silva
dc.creator.IDCPF: 143.225.707-20por
dc.creator.Latteshttp://lattes.cnpq.br/9669827127440973por
dc.publisher.countryBrasilpor
dc.publisher.departmentInstituto de Veterináriapor
dc.publisher.initialsUFRRJpor
dc.publisher.programPrograma de Pós-Graduação em Ciências Veterináriaspor
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