Document details

Red-legged partridge (Alectoris rufa):looking for the circulating viruses in th...

Author(s): Dias, Maryline Marques cv logo 1

Date: 2009

Persistent ID: http://hdl.handle.net/10451/1472

Origin: Repositório da Universidade de Lisboa

Subject(s): Aves; Perdiz-vermelha; Vírus; Teses de mestrado


Description
Tese de mestrado, Biologia (Biologia Humana e Ambiente), 2009, Universidade de Lisboa, Faculdade de Ciências Resumo alargado em português disponível no documento Partridges of the Alectoris genus are birds with economic and game relevance for some of the southern European countries, like Portugal. Alectoris rufa is the only indigenous species present throughout Portuguese territory where populations in captivity are also to be considered since they represent a significant proportion of the local and national economy. Relatively little is known about A. rufa pathogenic agents like parasites and viruses. This study aims the optimization and application of PCR and RTPCR methodologies to look for viruses in partridges in captivity. Animal samples were collected and carried to the laboratory by veterinaries from DirecçãoGeral dos Recursos Florestais. From the different methodologies assayed, direct PCR with Phire Hot Start DNA polymerase (Finnzymes) seemed to be the best one to amplify DNA sequences and RNA extraction with Trisure (Bioline) rendered high quality and amplifiable RNA; small amounts of DNA that remains in these samples turned out to be useful because, under specific conditions, these samples could also amplify DNA. The chosen methodology to amplify RNA was the one with OneStep RTPCR kit (Qiagen). Because DNA and RNA sequences from partridges could be used to validate the nucleic acid samples as well as PCR and RTPCR experiments, beta actin was chosen; the primers for its amplification were selected from Gallus gallus published sequences. A. rufa PCR and RTPCR products were sequenced and showed respectively 85% and 97% of identity with citoplasmatic beta actin and beta actin mRNA of Gallus gallus. Positive results were achieved when searching for Poxvirus, Paramyxovirus and Coronavirus sequences. Positive results for Poxvirus and Paramyxovirus were not subjected to sequencing. One of the 15 in 20 PCR products obtained with Coronavirus primers was sequenced and matched with sequences from other avian Coronaviruses. This result allowed concluding that Coronaviruses are probably circulating among partridges in captivity.
Document Type Master Thesis
Advisor(s) Caeiro, Filomena; Dias, Deodália Maria Antunes, 1952-
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