• JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
 
  Bookmark and Share
 
 
Doctoral Thesis
DOI
https://doi.org/10.11606/T.74.2022.tde-02022023-120346
Document
Author
Full name
Samara Rita de Lucca Maganha
Institute/School/College
Knowledge Area
Date of Defense
Published
Pirassununga, 2022
Supervisor
Committee
Sousa, Ricardo Luiz Moro de (President)
Carrasco, Adriano de Oliveira Torres
Keid, Lara Borges
Moreli, Marcos Lazaro
Pandolfi, José Rodrigo Cláudio
Title in Portuguese
Estudos in vitro com estirpe brasileira de Megalocytivirus e Lymphocystivirus: isolamento e caracterização fenogenotípica
Keywords in Portuguese
Iridoviridae
Cultivo celular
Propriedades físico-químicas
Abstract in Portuguese
O gênero Megalocytivirus está associado a ocorrência de surtos com altas taxas de mortalidade em peixes ao redor do mundo, causando graves perdas econômicas na produção aquícola mundial e limitando a maior expansão do setor. Já o gênero Lymphocystivirus, embora não esteja associado a altas taxas de mortalidade, causa gaves perdas econômicas para a aquicultura mundial ao prejudicar a aparência do animal, alterando assim o seu valor comercial. Desse modo, o presente estudo objetivou o isolamento e a caracterização fenogenotípica das estirpes brasileiras de Megalocytivirus e Lymphocystivirus. Para tanto, pools de tecidos de peixes ornamentais sabidamente positivos para os dois vírus foram inoculados em células BF-2, posteriormente foi realizada a reconstrução filogenética para ambos os isolados com base em um fragmento do gene MCP, caracterização viral através da realização dos testes de sensibilidade a solvente lipídico, pH, temperatura e descongelamento bem como a construção da curva de replicação viral e para Megalocytivirus incluiu-se a caracterização morfológica por microscopia eletrônica de transmissão. Os resultados indicaram que a estirpe brasileira de Megalocytivirus apresenta uma população de vírions majoritariamente envelopados, é sensível a temperatura de 56°C e aos valores de pHs de 2,0 e 10,0. A técnica de microscopia eletrônica de transmissão revelou a presença de partículas hexagonais no citoplasma celular compatíveis com o formato icosaédrico de Megalocytivirus. O isolado brasileiro de Megalocytivirus agrupou-se com outros isolados descritos como pertencentes ao genótipo I. Os resultados para a estirpe brasileira de Lymphocystivirus revelaram uma população de vírions majoritariamente envelopados, sensível a temperatura de 56°C, aos valores de pHs de 2,0 e 12,0, sendo susceptível a alterações no título viral quando descongelado por sucessivas vezes. O isolado brasileiro de Lymphocystivirus agrupou-se com outros isolados descritos como pertencentes ao genótipo V. A curva de replicação viral para os isolados demonstrou que o aumento no título viral acompanha a progressão do efeito citopático, alcançando o maior valor de TCID50.ml-1 para Megalocytivirus no quarto dia pós inoculação e, para Lymphocystivirus, no quinto dia pós inoculação. Os resultados obtidos fornecem subsídios importantes para estudos posteriores nas áreas de controle e prevenção das infecções causadas por vírus desses grupos.
Title in English
In vitro studies with Brazilian strain of Megalocytivirus and Lymphocystivirus: isolation and phenogenotypic characterization
Keywords in English
Iridoviridae
Cell culture
Physicochemical properties
Abstract in English
The genus Megalocytivirus is associated with the occurrence of outbreaks with high mortality rates in fish around the world, causing serious economic losses in the world aquaculture and limiting the greater expansion of the sector. Although the genus Lymphocystivirus isn´t associated with high mortality rates, it causes great economic losses for the world aquaculture by impairing the animal's appearance and altering its commercial value. Thus, the present study aims the isolation and phenogenotypic characterization of the Brazilian strain of Megalocytivirus and Lymphocystivirus. For this purpose, pools of ornamental fish tissues known to be positive for Megalocytivirus and Lymphocystivirus were used for isolation in BF-2 cells. After confirmation of the isolation, viral characterization was carried out in the same cell line including morphological characterization by transmission electronic microscopy for Megalocytivirus and the tests of sensitivity to lipid solvent, pH, temperature and thawing as well as the construction of viral growth curve for Megalocytivirus and Lymphocystivirus. The results indicated that the Brazilian Megalocytivirus strain has a population of enveloped virions, is heat labile (56°C), sensible to pH values of 2,0 and 10,0 and clustered with other isolates described as belonging to the genotype I. Transmission electron microscopy revealed the presence of hexagonal particles in the cell cytoplasm designed with the icosahedral shape of Megalocytivirus. The results for Brazilian Lymphocystivirus strain revealed a population of enveloped virions, heat labile (56°C), sensible to pH values of 2.0 and 12.0, being susceptible to changes in viral titer when thawed successively, and clustered with other isolates described as belonging to the genotype V. The growth curve demonstrated that the increase in viral titres accompanies the progression of the cytopathic effect, reaching the highest value of TCID50.ml-1 for Megalocytivirus on the fourth day post inoculation and for Lymphocystivirus on the fifth day post inoculation. The results obtained provide important subsidies for further studies in the areas of control and prevention of infections caused by viruses from these groups.
 
WARNING - Viewing this document is conditioned on your acceptance of the following terms of use:
This document is only for private use for research and teaching activities. Reproduction for commercial use is forbidden. This rights cover the whole data about this document as well as its contents. Any uses or copies of this document in whole or in part must include the author's name.
DO6787490COR.pdf (4.96 Mbytes)
Publishing Date
2023-02-03
 
WARNING: Learn what derived works are clicking here.
All rights of the thesis/dissertation are from the authors
CeTI-SC/STI
Digital Library of Theses and Dissertations of USP. Copyright © 2001-2024. All rights reserved.