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.

Controlo e monitorização do efluente de uma indústria vitivinícola

O tema deste trabalho é "Controlo e monitorização do efluente de uma indústria vitivinícola" e teve lugar na Adega Cooperativa de Reguengos de Monsaraz - CARMIM. O controlo e monitorização, bem como a caracterização físico-química do efluente deste tipo de indústrias, especialmente na CARMIM, pretendem representar um instrumento importante no sentido da necessidade de se construir uma estação de pré-tratamento de águas residuais, antes destas fazerem parte do sistema de águas residuais municipais. A monitorização do efluente ocorreu em duas épocas distintas, a Época Baixa e a Época de Campanha. Em cada uma das épocas foram monitorizadas operações distintas, como a lavagem dos filtros de terras de diatomáceas, a higienização do equipamento de filtração tangencial, a higienização das cubas de inox, lavagens de cubas de vinho tinto e lavagens de depósitos de vinho branco. Foi também monitorizado o efluente resultante do edifício de engarrafamento, da Adega Velha e da caixa colectora global. Acompanhou-se em cada operação e caixas colectoras, o pH, temperatura e caudal, sempre que possível. Para cada amostra foi efectuada uma caracterização físico-química, que inclui os seguintes parâmetros: carência química em oxigénio, azoto total, fósforo total, nitratos, sólidos suspensos totais e metais como ferro, cobre e manganês. ABSTRACT: "Winery wastewater control and monitoring" is the main theme of all this work. All this occurred in CARMIM - Cooperativa Agrícola de Reguengos de Monsaraz. The control and monitoring, as well as physical and chemistry features of this type of wastewater, particularly related with CARMIM, intend to represent an important instrument towards the necessity of building a pre-treatment wastewater station, before they are part of the municipal wastewater system. The wastewater monitoring took place in two different seasons, the non-vintage and vintage season. ln both seasons there were checked different operations such as, the washing of earth filtering equipment, red and white wine tanks and finally inox tank caustic washing. At the same time, there was also possible monitoring the final wastewater resulting from the bottling line facility, "Old “Cellar (Adega Velha) and global drain boxes. There was preformed a physical chemistry feature for each sample. These features comprehend several parameters: Chemical oxygen demand, total nitrogen, total phosphorous, nitrates, total suspended solids and metals such as iron, cupper and manganese.