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Master's Dissertation
DOI
10.11606/D.87.2014.tde-10122014-094539
Document
Author
Full name
Rodrigo Pinheiro Araldi
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2014
Supervisor
Committee
Stocco, Rita de Cassia (President)
Kfoury Junior, José Roberto
Sant'Anna, Osvaldo Augusto Brazil Esteves
Title in Portuguese
Isolamento e identificação do papilomavírus bovino em grupo experimental de bovinos para obtenção de um banco de vírus.
Keywords in Portuguese
Papovaviridae
Histopatologia animal
Imunohistoquímica
Mutagênese
Oncologia veterinária
Vírus oncogênico
Abstract in Portuguese
O Papilomavírus bovino (BPV) gera prejuízos à pecuária. O trabalho buscou isolar vírions de BPV de papilomas cutâneos previamente diagnosticados e avaliar o potencial clastogênico do vírus através do ensaio cometa (EC). O DNA tecidual foi extraído e submetido a PCR. As bandas foram purificadas e sequenciadas. As sequências foram analisadas através de bioinformática. A tipagem mostrou a prevalência de BPV-2. A análise histopatológica revelou acantose, coilocitose e hiperqueratose. Foi possível detectar a presença de fibroblastos transformados, sugerindo uma via de infecção através do sangue. A imuno-detecção das proteínas L1 e E7 no estroma sugere atividade transformadora do BPV em fibroblastos. O EC mostrou a ação clastogênica estatisticamente igual entre os tipos virais BPV-2, 5 e 9. O isolamento viral foi realizado através de ultracentrifugação em densidade única de cloreto de césio, empregando uma nova metodologia, que se mostrou menos laboriosa do que as já descritas, permitindo o isolamento de vírions de BPV-2, iniciando o banco de vírus proposto.
Title in English
Isolation and identification of bovine papillomavirus in experimental group of cattle in order to obtain a virus bank.
Keywords in English
Papovaviridae
Animal histopathology
Immunohistochemistry
Mutagenesis
Oncogenic vírus
Veterinary oncology
Abstract in English
The Bovine papillomavirus (BPV) generates losses to livestock. The study sought to isolate BPV virions from cutaneous papillomas previously diagnosed and evaluate the clastogenic potential of the virus through the comet assay (EC). The tissue DNA was extracted and subjected to PCR. The bands were purified and sequenced. Sequences were analyzed using bioinformatics. The typing showed the prevalence of BPV-2. Histopathology showed acanthosis, hyperkeratosis and koilocytosis. It was possible to detect the presence of transformed fibroblasts, suggesting a route of infection through the blood. Immunohistochemical detection of L1 and E7 proteins in the stroma suggests transforming activity of BPV in fibroblasts. The EC clastogenic action showed statistically equal among the virus types BPV-2, 5 and 9. Viral isolation was performed by ultracentrifugation in a single cesium chloride density, using a new method that was less laborious than those already described, allowing the isolation of BPV-2 virions, starting the proposed stock virus.
 
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Publishing Date
2014-12-11
 
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