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Master's Dissertation
DOI
https://doi.org/10.11606/D.10.2016.tde-09052016-102446
Document
Author
Full name
Renata Khodair Silva
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2016
Supervisor
Committee
Ferreira, Helena Lage (President)
Carregaro, Valéria Maria Lara
Keid, Lara Borges
Title in Portuguese
Detecção e caracterização moleculares dos paramixovírus aviários tipo 1 em materiais provenientes de aves silvestres utilizando testes para a detecção dos vírus da família viral Paramyxoviridae
Keywords in Portuguese
Aves silvestres
Detecção
Molecular
Vírus
Abstract in Portuguese
As aves silvestres são importantes reservatórios de vírus que podem acometer as aves domésticas. O monitoramento da circulação viral em aves silvestres é de extrema importância para garantir a sanidade dos plantéis avícolas. O presente estudo teve como objetivo 1) comparar dois testes moleculares de RT-PCR para a detecção dos vírus da família Paramyxoviridae em aves silvestres e sinantrópicas; 2) caracterizar os vírus detectados nestas amostras. Dois testes de RT-PCR e testes específicos de RT-PCR em tempo real (RRT-PCR) para o vírus da doença de Newcastle (NDV) e o metapneumovírus aviário (aMPV) foram utilizados para comparar o limite de detecção entre as amostras. As amostras de aves silvestres foram testadas por dois testes de RT-PCR. Um pequeno fragmento da região do sítio de clivagem do gene F das amostras positivas foi sequenciado. Os testes de RT-PCR foram validados com sucesso, mas apresentaram diferenças entre os limites de detecção quando comparados aos testes específicos de RRT-PCR utilizando diferentes vírus. No total, 100 amostras de aves (suabes) foram testados pelo teste RT-PCR que apresentou um limite de detecção similar entre os diferentes agentes virais. O teste selecionado foi capaz de detectar duas amostras de aves silvestres que foram também detectadas pelo testes específico para NDV e relacionadas às amostras de NDV vacinais do genótipo II da classe II referentes aos vírus de NDV lentogênico (113RQGR ↓ L117). Nosso estudo demonstra a deficiência na biosseguridade adotada pelos sistemas avícolas por permitir a saída dos vírus vacinais para as aves silvestres
Title in English
Molecular detection and characterization of avian paramyxovirus type I in samples from wild birds by using molecular tests for detection of viruses from Paramyxoviridae family
Keywords in English
Detection
Molecular
Virus
Wild birds
Abstract in English
Wild birds are important reservoirs of viruses can affect poultry. Surveillance of circulating viruses in wild birds is of the most importance tool to ensure the poultry health. The present study aimed to 1) compare two molecular tests for detection of family Paramyxoviridae viruses in wild and feral birds; 2) characterize the detected viruses in those samples. Two RT-PCR techniques and specific real time RT-PCR (RRT-PCR) assays for detection of Newcastle disease virus (NDV) and avian metapneumovirus (aMPV) were used to compare the limit of detection among them. Wild bird samples were tested by using two RT-PCR tests. A small fragment of cleavage site region in F gene from positive samples was sequenced. The RT-PCR were successfully validated, however they had differences in the limit of detection when compared to specific RRT-PCR assays using different viruses. In total, 100 wild bird samples (swabs) were tested by the selected RT-PCR with similar limit of detection between tested viruses. Two samples were positive by this test and they were also detected by the specific RRT-PCR for NDV. These samples were closed related to vaccinal NDV strains belonging to genotype II class II. Deduced amino acid sequences of cleavage site region from detected samples were characterized as lentogenic NDV strains (113RQGR ↓ L117). Our study demonstrates the poor biosafety used by poultry industry allowing the vaccinal escape to wild bird species
 
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Publishing Date
2016-06-01
 
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