Por favor, use este identificador para citar o enlazar este ítem: https://rima.ufrrj.br/jspui/handle/20.500.14407/19943
Tipo do documento: Tese
Título: Diversidade do Gene Msp1α de Anaplasma Marginale e Isolamento e Propagação de um Novo Genótipo de Anaplasma Sp. em Células Ide8
Otros títulos: Diversity of the Anaplasma marginale msp1α gene and isolation and propagation of a novel genotype of Anaplasma sp. in IDE8 cells
Autor: Santos, Priscilla Nunes dos
Orientador(a): Fonseca, Adivaldo Henrique da
Primeiro coorientador: Silva, Claudia Bezerra da
Segundo coorientador: Silva, Jenevaldo Barbosa da
Primeiro membro da banca: Fonseca, Adivaldo Henrique da
Segundo membro da banca: Santos, Huarrisson Azevedo
Terceiro membro da banca: Peixoto, Maristela Peckle
Quarto membro da banca: Baêta, Bruna de Azevedo
Quinto membro da banca: Zweygarth, Erich Peter
Palabras clave: Diversity;Rhipicephalus microplus;IDE8;msp1α
Área(s) do CNPq: Medicina Veterinária
Medicina Veterinária
Idioma: por
Fecha de publicación: 23-jun-2020
Editorial: Universidade Federal Rural do Rio de Janeiro
Sigla da instituição: UFRRJ
Departamento: Instituto de Veterinária
Programa: Programa de Pós-Graduação em Ciências Veterinárias
Citación: SANTOS, Priscilla Nunes dos. Diversidade do Gene Msp1α de Anaplasma Marginale e Isolamento e Propagação de um Novo Genótipo de Anaplasma Sp. em Células Ide8. 2020. 127 f. Tese (Doutorado em Ciências Veterinárias) - Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, 2020.
Resumen: -
Abstract: This thesis consists of two chapters: The purpose of the first chapter was to evaluate the tandem repeat diversity in the major surface protein MSP1a of Anaplasma marginale in experimentally infected cattle and in Rhipicephalus microplus over the span of 1 year, and the re-isolation of AmRio1 strain of A. marginale, using the buffy coat separation protocol, was performed. The second chapter was aimed at isolating, propagating, and characterizing a novel Anaplasma sp. genotype from Portugal in IDE8 cells. To evaluate A. marginale genetic diversity, cattle were infested with R. microplus larvae and subsequently infected by A. marginale from tick cell cultures. The strains AmRio1 and AmRio2 were inoculated in bovine 1 and bovine 2, respectively. Subsequently, blood samples were collected over 1 year for blood smears, hematocrit, plasma proteins, and semi-nested polymerase chain reaction (PCR) for the msp5 and msp1α genes. The buffy coat separation protocol was used to isolate Anaplasma sp. Ticks were collected and their salivary glands and guts DNA were tested using semi-nested PCR for the msp5 and msp1α genes. Both animals showed stability in the evaluated parameters and absence of clinical signs, showing that they had persistent infection. A blood smear from Animal 1 showed inclusions inside the leukocytes, and four attempts were made to isolate Anaplasma sp. using the buffy-coat as inoculum. Animal 1 became positive for the msp5 gene on day 12 after infection, while positivity was detected in Animal 2 only 105 days after first exposure to the agent. The tandem repeats were stable when the blood and tick samples of both animals were evaluated, demonstrating the presence of the strains AmRio1 in Animal 1 and AmRio2 in Animal 2. In the second chapter, erythrocytes from a naturally infected bovine from Portugal were used for isolating Anaplasma sp. in IDE8 cells. Suggestive inclusions of infection were observed in the IDE8 cells 35 days after inoculation. The Anaplasmataceae family 16S rRNA gene from the cultures was sequenced and showed that the isolated rickettsia was close to the A. platys species in the phylogenetic tree.
URI: https://rima.ufrrj.br/jspui/handle/20.500.14407/19943
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