Guia dos peixes de água doce de Portugal Continental

O trabalho desenvolvido insere-se num projecto do FFishGUL, Freshwater Fish Group of the University of Lisbon, que consiste na publicação de um guia ilustrado que reúna e disponibilize informação sobre a fauna ictiológica dulciaquícola de Portugal Continental, dirigido a diversos públicos-alvo. Este guia procura promover o conhecimento sobre as espécies e comunidades piscícolas e ser uma ferramenta técnica de resposta a questões prioritárias em termos de conservação do património natural, gestão dos recursos hídricos e ordenamento do território. O âmbito deste trabalho de mestrado centra-se na ilustração de algumas das espécies de peixes tratadas, investigando e criando pela primeira vez uma representação rigorosa que promova a documentação e identificação destas, como ponto de partida para o trabalho a realizar para a totalidade das espécies a tratar no guia. O trabalho desenvolvido é um trabalho que se pode considerar inédito, na medida em que não existe, tanto quanto sabemos e após extensiva pesquisa bibliográfica neste domínio, nada publicado que se assemelhe aos níveis de rigor, detalhe e coerência entre todo o conjunto apresentado. O resultado pretendido para este guia é o de uma publicação de referência, de grande utilidade técnica e de cuidada comunicação visual. Nesse sentido foi também desenvolvido um layout gráfico, visando uma apresentação cuidada e apelativa dos conteúdos do guia, pretendendo contribuir para uma maior sensibilização dos vários públicos a que se destina; ABSTRACT: This Masters thesis is part of a larger project, developed by FFishGUL, the Freshwater Fish Group of the University of Lisbon, consisting of the edition of an illustrated guide to the freshwater fish of mainland Portugal. This guide aims to create a hitherto nonexistent tool to promote knowledge on Portuguese freshwater fish communities and species and help on the management of water resources and land-use planning. This Masters thesis is centered on the illustration of some of the guide’s fish species, aiming to study and create an accurate representation that will facilitate their correct identification. These illustrations serve as an example of the process that will be followed for the total guide’s species range. The developed work can be regarded as unprecedented, to the extent that there is not, as far as we know, and after extensive bibliographic research in this field, any published work resembling the levels of accuracy, detail and coherence of what is here presented. This guide intends to be a reference publication, of major technical utility and careful visual communication. In this sense a graphic layout was also developed, aiming a careful and attractive presentation of the guide’s contents and contributing to greater awareness of the various target audiences.

RNA-FISH Technique: Probe Cocktails vs Single Probes

RNA-Fluorescence In Situ Hybridization (RNA-FISH) enables to analyze and visualize the microorganisms of interest within microbial communities in their natural environments by fluorescent labelling of specific RNA sequences. Poor accessibility or low content of the RNA target region can cause false positives/negatives due to low fluorescence of the cells. To reduce the chances of this occurring, probe cocktails – i.e. mixture of several probes that hybridize to different regions of the target RNA- has been proposed as an alternative to single probes use for increasing the Fluorescence Intensities (FI). However, is this really a good solution? The key finding of this work was that the use of probe cocktails is not always a good solution for maximizing the FI as at high concentrations the single probe EUK516-6 FAM yielded higher FI than the probe cocktails.

A SHORTENED RNA-FISH PROTOCOL FOR ANALYZING ARTWORKS’ MICROCOLONIZERS

Microorganisms are involved in the deterioration of Cultural Heritage. Thus, there is a need to enhance the techniques used for their detection and identification. RNA Fluorescent In Situ Hybridization (RNA-FISH) has been successfully applied for phylogenetic identification of the viable components of the microbial communities colonizing artworks both in situ and ex situ. Until recently, it was time-consuming, taking not less than 6 h for the analysis. We have developed an RNA-FISH in suspension protocol that allowed ex situ analysis of microorganisms involved in artworks’ biodeterioration in 5 h. In this work, three modified protocols, involving microwave heating, were evaluated for further shortening two of the four main critical steps in RNA-FISH: hybridization and washing. The original and modified protocols were applied in cellular suspensions of bacteria and yeast isolates. The results obtained were evaluated and compared in terms of detectability and specificity of the signals detected by epifluorescence microscopy. One of the methods tested showed good and specific FISH signals for all the microorganisms selected and did not produce signals evidencing non-specific or fixation-induced fluorescence. This 3 h protocol allows a remarkable reduction of the time usually required for performing RNA-FISH analysis in Cultural Heritage samples. Thus, a rapid alternative for analyzing yeast and bacteria cells colonizing artworks’ surfaces by RNA-FISH is presented in this work.