Satellite DNA as a target for TaqMan real-time PCR detection of the pinewood nematode, Bursaphelenchus xylophilus

The pinewood nematode (PWN), Bursaphelenchus xylophilus , is a major pathogen of conifers, which impacts on forest health, natural ecosystem stability and international trade. As a consequence, it has been listed as a quarantine organism in Europe. A real-time PCR approach based on TaqMan chemistry was developed to detect this organism. Specific probe and primers were designed based on the sequence of the Msp I satellite DNA family previously characterized in the genome of the nematode. The method proved to be specific in tests with target DNA from PWN isolates from worldwide origin. From a practical point of view, detection limit was 1 pg of target DNA or one individual nematode. In addition, PWN genomic DNA or single individuals were positively detected in mixed samples in which B. xylophilius was associated with the closely related non-pathogenic species B. mucronatus , up to the limit of 0.01% or 1% of the mixture, respectively. The real-time PCR assay was also used in conjunction with a simple DNA extraction method to detect PWN directly in artificially infested wood samples. These results demonstrate the potential of this assay to provide rapid, accurate and sensitive molecular identification of the PWN in relation to pest risk assessment in the field and quarantine regulation.

"Arqueologias" das aprendizagens em Capelins (Santo António) - Alandroal (1997-2007)

Esta dissertação de mestrado desenvolveu-se no âmbito do projecto “Arqueologia” das Aprendizagens no concelho de Alandroal, que teve, como objectivo primordial, a identificação e caracterização do universo de aprendizagens institucionais disponíveis e concretizadas pela comunidade do território em estudo (Freguesia de Capelins (Santo António) /Alandroal), na década 1997 – 2007. Foram inquiridas, nesta freguesia, 16 instituições e identificadas 30 aprendizagens institucionais, que tiveram maior ocorrência nos anos de 2004, 2005 e 2006. Embora seja uma freguesia essencialmente rural, a área de actividade da maioria das instituições identificadas inscreve-se no sector do comércio por grosso e a retalho e de reparação de veículos automóveis e motociclos. São, na sua maioria, instituições de natureza privada e reveladoras de organização, no que respeita à sua organização. No entanto, o relacionamento interinstitucional foi pouco dinâmico, existindo um fraco trabalho em rede. Quando existiram parcerias, estas ocorreram extra freguesia e, até mesmo, extra concelho. Seguindo uma análise quantitativa dos dados extraídos da aplicação dos questionários, com recurso a análise estatística, conclui-se que os processos de aprendizagem das instituições inquiridas da freguesia de Capelins apresentam características centradas nas próprias instituições promotoras dessas mesmas aprendizagens, assentes na formação do pessoal, com carácter formal e não formal e relacionaram-se com as áreas de administração pública e defesa, alojamento e restauração e ainda com a área do comércio por grosso e a retalho; ABSTRACT: This dissertation developed under the project "Archaeology" of Learning in the county of Alandroal, which had as its primary objective, the identification and characterization of the universe of available institutional learning and implemented by the community of the area under study (Parish of capelin (San Antonio) / Alandroal), in the decade from 1997 to 2007. Were surveyed in this parish, and 16 institutions identified 30 institutional learning, which have a higher occurrence in the years 2004, 2005 and 2006. Although it is an essentially rural, the parish area of activity of most institutions identified in the sector of wholesale and retail trade and repair of motor vehicles and motorcycles. Are, for the most part, private institutions and revealing of organization, with regard to its organization. However, the interinstitutional relationship was weak and there is a little dynamic networking. When partnered, this extra parish occurred and even extra municipality. Following a quantitative analysis of data taken from the application of questionnaires, using statistical analysis, it is concluded that the learning processes of the institutions surveyed in the parish of Capelins characteristics focused on promoting institutions of those learnings, based on staff training, formal and non-formal nature and related to the areas of public administration and defense, hotels and restaurants and also the area of wholesale and retail trade.