Desvendando o potencial bioativo de bactérias marinhas

Besides a primary metabolism, living organisms possess a secondary metabolism which allows for the production of secondary metabolites that, normally, are not fundamental to the organism’s growth and survival. These metabolites allow for a better adaptability to their environment as they act as defence mechanisms, can be bioactive against several pathogenic agents and can be used as drugs to efficiently eliminate them. At first, the search for new natural compounds by the scientific community was based on terrestrial environments. However, more recently, more and more attention has been paid to the oceans. Since there has been a general abuse in the use of antibiotics, the numbers of antibiotic resistant bacteria have been rising rapidly. Therefore, it is essential to invest in the discovery new compounds that can be used as alternatives to the traditional antibiotics. The main aim of this work was the analysis of the antimicrobial potential of a collection of marine bacteria by molecular and screening assays. Aiming at obtaining a higher yield of secondary metabolites, in this work a transformation of the planctomycete Roseimaritima ulvae UC8T was performed using the triparental mating technique. This bacterium was chosen since Planctomycetes have shown promising bioactive potential. However, this strain exhibited the production of some kind of bioactive compounds that inhibited the growth of Escherichia coli ATCC 25922 and possibly affected the growth of the donor and helper E. coli DH5α involved in the transformation process. In parallel, a molecular screening for the amplification of polyketide synthases type I (PKS-I) and non-ribosomal peptide synthetases (NRPS) was performed in 329 newly isolated strains which belong to Laboratório de Ecofisiologia Microbiana da Universidade do Porto (LEMUP) collection. From these strains 36% generated amplicons for PKS-I and 24% for NRPS. The rest did not amplify for either one of these genes. Then, a selection was made based on the amplification for these genes and the promising strains were chosen to be tested against E. coli ATCC 25922 and Staphylococcus aureus ATCC 29213. Furthermore, a one strain many compounds (OSMAC) approach was performed to the strains that showed to be bioactive in the preliminary antimicrobial assay in 5 different culture media (1:10 M607, M607, M600, MA and CGY). From all the bacteria that were selected only 16 strains were considered bioactive, and mostly against E. coli ATCC 25922, unlike Streptomyces flavoviridis PMIC_1A8B which was highly bioactive against S. aureus ATCC 29213. Most of the bioactive strains belong to the Actinobacteria phylum except for Arenibacter aquaticus PMIC_1E11B.2, Aquimarina algiphila PMO90_19.1 and the new species PMO138_17 closely affiliated with Methylotenera mobilis. In the OSMAC assay, the highest values of bioactivity were obtained in extracts from strains grown in 1:10 M607, the lowest nutrient rich culture medium. Also, high activities were displayed in the more nutrient rich CGY culture medium. In conclusion, the OSMAC approach showed the importance of the culture medium composition for the production of bioactive molecules.Para além de um metabolismo primário, os organismos vivos possuem um metabolismo secundário que permite a produção de metabolitos secundários que, normalmente, não são fundamentais para o crescimento e sobrevivência do organismo. Estes metabolitos permitem uma melhor adaptação ao ambiente envolvente, uma vez que atuam como mecanismos de defesa e podem ser bioativos contra vários agentes patogénicos, podendo ser utilizados como medicamentos para os eliminar eficazmente. Inicialmente, a procura por novos compostos naturais pela comunidade científica era focada em ambientes terrestres. No entanto, mais recentemente, cada vez mais atenção é dada aos oceanos. Uma vez que tem existido um abuso geral na utilização de antibióticos, o número de bactérias resistentes a antibióticos tem vindo a aumentar rapidamente. Por isso, é essencial investir na descoberta de novos compostos que podem ser usados como alternativas aos antibióticos tradicionais. Este trabalho teve como principal objetivo a análise do potencial antimicrobiano de uma coleção de bactérias marinhas através de ensaios moleculares e de despiste. Com o objetivo de aumentar a produção de metabolitos secundários, neste trabalho, uma transformação do planctomycete Roseimaritima ulvae UC8T foi realizada utilizando a técnica de conjugação triparental. Esta bactéria foi escolhida uma vez que Planctomycetes mostraram serem promissores a nível de potencial bioativo. No entanto, esta estirpe exibiu a produção de algum tipo de composto bioativo capaz de inibir o crescimento de Escherichia coli ATCC 25922 e que, possivelmente, afetou o crescimento da E. coli DH5α dadora e ajudante envolvida no processo de transformação. Em paralelo, um ensaio molecular para a amplificação de sintases de policétidos tipo I (PKS-I) e sintetases de péptidos não ribossomais foi realizado com 329 estirpes recentemente isoladas e pertencentes ao Laboratório de Ecofisiologia Microbiana da Universidade do Porto. Destas estirpes 36% deram origem a amplicões para PKS-I e 24% para NRPS. As restantes não amplificaram para nenhum destes genes. Posteriormente, uma seleção foi realizada baseada na amplificação destes genes e as estirpes promissoras foram escolhidas para serem testadas contra E. coli ATCC 25922 e Staphylococcus aureus ATCC 29213. Para além disso, uma abordagem de uma estirpe muitos compostos (OSMAC) foi realizada com as estirpes que mostraram ser bioativas no ensaio preliminar em 5 meios de cultura diferentes (1:10 M607, M607, M600, MA e CGY). Das bactérias selecionadas apenas 16 estirpes foram consideradas bioativas e principalmente contra E. coli ATCC 25922, ao contrário de Streptomyces flavoviridis PMIC_1A8B que foi altamente bioativa contra S. aureus ATCC 29213. A maior parte das estirpes bioativas pertencem ao filo Actinobacteria exceto Arenibacter aquaticus PMIC_1E11B.2, Aquimarina algiphila PMO90_19.1 e uma nova espécie PMO138_17 relacionada com Methylotenera mobilis. No ensaio OSMAC, os valores mais altos de bioatividade foram obtidos nos extratos de estirpes crescidas em 1:10 M607, o meio de cultura menos rico em nutrientes. Para além disso, também foram obtidas bioatividades altas para o meio de cultura mais rico em nutrientes CGY. Em conclusão, a abordagem OSMAC mostrou a importância da composição do meio de cultura para a produção de compostos bioativos.Mestrado em Microbiologi

Linguistic innovations in the immigration context as initial stages of a partially restructured variety: Evidence from SE constructions in the Portuguese of the East Timorese diaspora in Portugal

Portuguese became one of the official languages of independent East Timor after ca. 25 years of Indonesian rule; this prevented the partial restructuring of an East Timorese variety of Portuguese in a similar way to that undergone by other Portuguese varieties (e.g., Mozambican, Angolan and Vernacular Brazilian Portuguese). We will discuss the idiosyncratic use of SE constructions in the speech of literate Portuguese-speaking East Timorese immigrants in Portugal, who will go back to East Timor and will be likely to lead language change. Given this particular link between East Timor and its diasporas, linguistic innovations in the immigration context can shed light on the initial stages of a future partially-restructured East Timorese Portuguese variety. SE constructions are highly polysemous and marked and the data show that innovative patterns are emerging, comprising deletion and generalization of the clitic as well as creative uses of these constructions, mainly observed in impersonal and spontaneous situation types. These innovative patterns can be attributed to L2 acquisition and to the interference of Tetum Dili.We gratefully acknowledge the support of the British Academy (Small Grant SG100616) for the data collection, which took place from June 2010 to March 2011

Improving by sustainability in sport facilities

Comunicação apresentada em Energy for Sustainability 2013 - Sustainable Cities: Designing for People and the Planet, Coimbra, Portugal, 8-10 September 2013Sporting structures particularly in Portugal, which were built for Euro 2004, in addition to belonging to clubs, the rest, have a very low utilization rate. But all in general have high operating costs and little control in the use of resources available. From the cases of stadiums Letzigrund, Zurich, Switzerland and Amsterdam Arena, the Netherlands, there may be a new vision for the improvement of Portuguese stadiums. In the case of the stadium was the Letzigrund stadium in its construction was dubbed the Green Stadium, built for Euro 2008. Among the recommended measures were implemented immediately: Green roof: with a photovoltaic system with an area of 2500 m2 and Solar Thermal Collectors 90 m2, heating by pellets; Promoting green transport: public transport; Waste management by implementing reusable cup; Lighting System: Minergie. The Amsterdam Arena, even having been built in 1998, has now the objective for 2015, be the first zero carbon stadium. The Amsterdam ArenA strives: to be a platform for sustainable innovations; to reduce our own environmental impact, and that of our events; to be a good neighbor and a positive boost for the economy; to increase our sustainable impact together with others. It presents an analysis of the measures advocated by these two football stadiums for sustainability, and measures can be defined and advocated an example and be applicable in Euro 2004 stadiums. It is considered that the implementation of measures for sustainability in sports facilities can be made throughout his life in order to have lower cost and operating efficiency

Sustainability as architecture for people: the case of football stadiums

Comunicação apresentada em Energy for Sustainability 2013 - Sustainable Cities: Designing for People and the Planet, Coimbra, Portugal, 8-10 September 2013The football stadiums are a center of emotions. In use the assembly is of thousands of people in a short period of time, a few hours. The sustainability of these sports facilities can not only be associated with its construction and operation. In 2011 the Quality Manual of Stadiums developed by UEFA appears the concept of Sustainable Architecture - Architecture for people. The concept of "architecture for people," emphasizes the need for well-being, comfort, and both psychological and physical, which should be an integral feature of any construction project sustainable. This can be defined as sustainable architecture for the planet and for people. Sustainable architecture can be considered as a simple, but important to the human scale, psychology, culture and ergonomics. It also encourages the design sensibility and interpretation that seeks to go beyond the basic requirements of the customer and therefore aims to give added value to the project. Sustainable architecture also focuses on the importance of creating a sense of place and encourage social interaction, which is particularly significant buildings, such as stadiums, where the idea of promoting the community is very important. This can be promoted through a variety of additional activities and facilities within the stadium complex that can offer the community recreation options as needed, without forgetting the business benefits they will bring to the development of the stadium itself. Thus presents the current situation of the Euro 2004 stadiums in relation to the parameters of Sustainable Architecture defined by UEFA as well as what they can improve, given that facilities that are still present only one fifth of its useful life. Football stadiums can thus leverage new forms of interaction, connection and even change their mindsets thousands of users