O vírus da mancha das orquídeas (“Orchid fleck virus” - OFV), transmitido pelo ácaro Brevipalpus californicus, causa manchas cloróticas e necróticas em orquídeas de vários gêneros e foi relatado em diversos países. O diagnóstico de "orchid fleck", doença causada pelo OFV, tem sido feito através da análise dos sintomas, sorologia, observação de cortes ultrafinos de tecido infectado em microscópio eletrônico de transmissão ou RT-PCR. No entanto, apesar de testes moleculares serem freqüentemente mais eficientes e específicos que outros métodos, os "primers" disponíveis na literatura nem sempre detectam o vírus em baixas concentrações no tecido vegetal, ou amplificam regiões da planta sadia. Com base nas seqüências nucleotídicas da capa protéica viral depositadas no GenBank foram desenhados novos “primers”, que amplificam um fragmento de 326 pb. Esses “primers” foram utilizados para a detecção do OFV por RT-PCR e para a marcação com digoxigenina de sondas para hibridização. A variabilidade de um fragmento do gene da capa protéica deste vírus foi estudada por polimorfismo conformacional de fita simples ("Single strand conformational polymorphism" – SSCP) e seqüenciamento de nucleotídeos. Quarenta e oito amostras de 18 gêneros de orquídeas foram coletadas no Brasil, Costa Rica e Austrália. As análises dos padrões de SSCP resultaram em seis haplótipos diferentes e em agrupamentos baseados na origem geográfica das amostras. Amostras representando cada um desses padrões foram seqüenciadas e comparadas com aquelas disponíveis no GenBank. A análise de SSCP provou ser eficiente para fornecer informações preliminares sobre a variabilidade do OFV. No entanto, apenas através do seqüenciamento de nucleotídeos das amostras foi possível determinar a real variabilidade das mesmas. / Orchid fleck virus (OFV), transmitted by the mite Brevipalpus californicus, causes chlorotic and necrotic ringspots in many orchid genera and was reported in several countries. The diagnosis of the Orchid fleck disease has been performed by symptomatology, transmission electon microscopy, serology or RT-PCR. Even though the molecular tests are usually more efficient and specific than other methods, the available primers did not always detect the OFV in low concentrations or sometimes amplified healthy plant samples. Based on the nucleotide sequences of the coat protein gene (cp) available in the GenBank, new primers were designed. These primers amplified a 326 pb specific OFV fragment and were used for RT-PCR and as hybridization probes. The variability of a fragment of the cp of this virus was investigated by “single strad conformational polymorphism (SSCP)” and nucleotide sequencing. Forty eight samples of 18 genera of orchids were collected from Brazil, Costa Rica and Australia. The SSCP analysis resulted in six different haplotypes and demonstrated a clustering in samples based on geographical origin. Samples representing the different SSCP patterns were sequenced and compared with those available in the GenBank. The SSCP analysis proved to be efficient to provide preliminary information about OFV variability. However, only through nucleotide sequencing it was possible to determine the actual variability amongst the samples.

Submitted by Maria Dulce (mdulce@ndc.uff.br) on 2014-01-30T17:46:13Z No. of bitstreams: 1 Passos, Pamella-Tese-2013.pdf: 3216374 bytes, checksum: 476a2aa1e9ebc02e7b439404d314731a (MD5) / Made available in DSpace on 2014-01-30T17:46:13Z (GMT). No. of bitstreams: 1 Passos, Pamella-Tese-2013.pdf: 3216374 bytes, checksum: 476a2aa1e9ebc02e7b439404d314731a (MD5) Previous issue date: 2013 / Tendo sido apontada como responsável por 49% dos acessos à internet no país, em 2007, as lan houses assumiram importante papel no debate sobre inclusão digital no Brasil. Equipadas com computadores conectados à internet e cobrando por suas horas de uso, esses estabelecimentos espalharam-se rapidamente, sobretudo, nos espaços populares. Partindo do acompanhamento de duas lan houses situadas em favelas cariocas: Acari e Santa Marta, analisamos seus impactos sociais nos territórios em que estão inseridas. Elegendo três pontos analisadores: o Estado, os donos de lan house e seus frequentadores, refletimos acerca dos usos e mediações que observamos em nosso trabalho de campo. Nesse contexto, escolhemos a lan house como dispositivo para reflexão das Políticas Públicas no campo da segurança e da educação. Compartilhando dos referenciais da pesquisa-intervenção, recorremos à realização de oficinas que serviram como grupos focais, entrevistas semiestruturadas e observação participante. Com isso, buscamos tecer encontros entre uma pesquisa etnográfica e as concepções da cartografia na produção de uma análise histórica do tempo presente. Priorizando a dimensão qualitativa, o trabalho orienta-se pela valorização da experiência e do cotidiano, para compreender a cultura popular a partir de suas significações no seio dos embates da indústria cultural. / Having been singled out as responsible for 49% of the internet access in the country in 2007, lan houses have taken on an important role in the debate about digital inclusion in Brazil. Equipped with computers connected to the internet and charging for their hours of use, these establishments have spread rapidly, especially in popular spaces. Based on the monitoring of two lan houses located in favelas in Rio: Acari and Santa Marta, we analyzed their social impacts in the territoriesin which they operate. Three analyzing points were chosen: The State, the owners of lan houses and its regulars, we reflected about the uses and mediations which we had observed in our field work. In this context, we elected the lan house as a device for reflection of public policies in the field of security and education. Sharing of research-intervention reference, we used workshops which served as focus groups, semi-structured interviews and participants observation. This way, we tried to make meetings between an ethnographic research and concepts of cartography in the production of a historical analysis of the present time. Prioritizing the qualitative dimension, the work is guided by the appreciation of everyday experience to understand popular culture from their meanings within the cultural industry ties.

The diversity of the Eriophyoidea is largely unknown and their systematic study mostly entails alpha-taxonomy which is critically important for these mites. Eriophyoid morphology is almost exclusively studied on slide-mounted specimens, and truly permanent specimen slides cannot be prepared and are eventually lost. Shortcomings in taxon descriptions are persistent, and too few morphological characters are available for systematic use, particularly for phylogenetic studies. The fragile, simplified and minute eriophyoid bodies, and the inadequacy of study methods and technology, including preparation and light microscopy, contribute to these problems. The present eriophyoid classification is widely accepted, relatively stable and useful. The major part of the classification, however, is probably artificial, and some taxon delimitations and identifications are becoming increasingly difficult. Scanning electron microscopy (SEM) is only sporadically used to supplement conventional descriptions of eriophyoid mites, and their phylogeny has hardly been studied. In the present study some aspects of eriophyoid systematics and its improvements by incorporating SEM for morphological study and phylogenetic analyses for testing and improving the naturalness of the present eriophyoid classification, are used and appraised. The morphology of about 64 species, mostly from South Africa, was studied with low-temperature (cryo) SEM. The specimens remained turgid and the shape of the mites largely unaltered. A general overview of the contribution of the SEM study towards systematic morphology of the Eriophyoidea is presented. Discrepancies between species descriptions from slide-mounted specimens and the SEM images were found. These include body form, interpretation of structures, resolution and information on minute morphology, and the presence of secretions. Some of these differences were caused by artefacts introduced with slide-mounting of specimens. The SEM study includes a comparative morphological study of the gnathosoma, including a review and appraisal of characters presently used in eriophyoid systematics. New morphological information was found, including new characters that may be of systematic use. Morphology studied with SEM should be routinely incorporated into eriophyoid descriptions, which is not presently the case. The phylogeny of the Eriophyoidea was studied at genus level, using morphological data, to test the monophyly of the present suprageneric taxa. Three data matrices with 66, 60 and 27 informative characters of 316 (including most Diptilomiopus spp.), 64 and 17 eriophyoid ingroup species respectively were analyzed with parsimony analyses, and trees were searched under different parameters. This was done to find different hypotheses regarding the taxon relationships, to roughly assess the robustness of the tree groups, and to use different approaches: a very comprehensive taxon sample, but with low ratio of characters to taxa; an exemplar species sample to improve the ratio between characters to taxa; and a very small taxon sample with a good ratio between characters and taxa, but very little inclusion of variation found in the Eriophyoidea. Most groups found were supported only by homoplasy, but many made biological sense and various potentially monophyletic groups, additional to taxa in the present classification, are proposed for further study. The robustness and convergence of these groups on monophyly are discussed. The Phytoptidae was found to be polyphyletic. Part of the Nalepellinae is probably positioned outside the remainder of the Eriophyoidea, while the rest of the Phytoptidae were positioned in smaller subgroups among the Eriophyidae. The Phytoptinae and Sierraphytoptinae, including Pentasetacus, may group together. The Eriophyidae never grouped together with much support, and the family is both polyphyletic and paraphyletic. The Diptilomiopidae was largely found to be monophyletic, with a relatively strong phylogenetic structure. The Rhyncaphytoptinae is mainly paraphyletic, and the Diptilomiopinae polyphyletic, but part of the Diptilomiopinae may be monophyletic. Three new Diptilomiopus spp. from South Africa are described as part of the study: D. faurius sp. nov. from Faurea rochetiana (A. Rich.) Pic. Serm. (Proteaceae); D. apobrevus sp. nov. and D. apolongus sp. nov. from Apodytes dimidiata E. Mey. ex Arn. (Icacinaceae). They were leaf vagrants not causing any observable symptoms. / Thesis (PhD)--University of Pretoria, 2011. / Zoology and Entomology / unrestricted