Suchergebnisse

Natural compounds from agro-food by-products have fostered interest in food industries. The aim of this study was to unravel potential uses for Pinus pinaster bark extracts (PBE). As functional features of this type of extracts are usually attributed to phenolic compounds, the extraction process was studied. Different PBEs were achieved, with high content in phenolic compounds, using different water/ethanol combinations as a solvent. These PBEs were chemically characterized, and their bioactivity and in vitro cell viability were evaluated. Extracts obtained with hydroethanolic solvents had higher content in phenolic and flavonoid compounds. All the PBEs presented high antioxidant, antibacterial and antihyperglycemic activities. Moreover, PBEs have low cytotoxicity and a selective activity against cancer cells as these were negatively affected. These features may allow the extracts to be used in food formulation and processing (as preservatives, antioxidants or bioactive ingredients), but they showed also potential for the pharmaceutical or nutraceutical sectors. ; This research was funded by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund (FEDER) under the scope of Norte2020—Programa Operacional Regional do Norte, by program INTERREG V-B Sudoe (REDVALUE, SOE1/P1/E0123) and by project MOBFOOD (POCI-01-0247-FEDER-024524), cofounded by PORTUGAL2020, Lisb@a2020, COMPETE 2020 and the European Union. Zlatina Genisheva is supported by the project OH2O (POCI-01-0145-FEDER-029145) funded by FCT and FEDER under the scope of Programa Operacional de Competividade e Internacionalizaçao (POCI)-COMPETE 2020 and PORTUGAL2020. Pedro Santos is recipient of a fellowship supported by a doctoral advanced training (call NORTE-69-2015-15), funded by the European Social Fund under the scope of Norte2020 (NORTE-08-5369-FSE-000036). ...

Producción Científica ; Carbon isotope ratios, recorded as 13C/12C variations in tree rings of woody species, are the result of physiological changes related to environmental conditions. The objective of this work was to analyze the association among carbon thirteen variability (δ13C), climate variables and tree-ring growth of Pinus pinaster and Pinus sylvestris in central Spain. Pulverized woody material from the period 1975-1999 from four trees for each pine species was analyzed. To detect common patterns in δ13C within each species and between δ13C and growth indices, a principal component analysis (PCA) was performed. δ13C of trees and the residual tree-ring chronologies were used at the PCA. Multilevel mixed linear models were applied between intrinsic water use efficiency (iWUE) and climate variables. Our results show an inverse significant correlation between δ13C and tree-ring growth of both species. Winter and spring air moisture was negatively correlated with iWUE of Pinus pinaster. July maximum temperature was positively correlated with iWUE of Pinus sylvestris. As δ13C is significantly related to climate and growth and it may be recommended as a valuable tool for tree growth dynamic analysis to withstand increasingly stressful climate conditions. ; Junta de Castilla y León (project VA096A05) ; Ministerio de Educación y Ciencia (project AGL2007-65795-C02-01) ; Alβan Programme, the European Union Programme of High Level Scholarships for Latin America (Scholarship # E05D049920AR)

Integrating fungal-based ecosystem services into forest management planning and policy-making requires quantitative knowledge of the yields of fungal sporocarps and their environmental drivers. The aim of this study was to predict edible mushroom yield in Pinus pinaster forests of Central Spain, based on a 17-year data series. Two-stage mixed-effects models were used to examine the effect of predictors on mushroom occurrence and yield separately with the aim of providing further insight into the ecological system. Changes in seasonal precipitation represented the main weather-related driver affecting sporocarp emergence and production, since they were both positively influenced by late summer and early autumn precipitation. Soil acidity positively influenced Lactarius yield. Stand age and sandy soils showed a negative influence on mushroom production. The diversity of drivers became more apparent at the fungal species level. The models can be used for predicting the production of edible fungi under different meteorological and site conditions. ; This study was partially funded by the research projects AGL2012-40035-C03-01 and AGL2012-40035-C03-03 (Ministerio de Economía y Competitividad of Spain, Secretaría de Estado de Investigaci on, Desarrollo e Innovaci on), by the Micosylva þ project (Interreg IVB SUDOE SOE3/P2/E533), and by the Mycological Programme of Castilla y Leon (www.micocyl.es). This study also received funding from the European Union's Horizon 2020 research and innovation programme within the framework of the Multi- FUNGtionality Marie Skłodowska-Curie Individual Fellowship (IFEF) under grant agreement No 655815. This work further benefited from the Erasmus Mundus Master Course MEDFOR (Mediterranean Forestry and Natural Resources Management) (520137-1-2011-1- PT-ERA MUNDUS-EMMC), which provided one scholarship to Mr. Zelalem Mengiste Taye. The inventory and monitoring of the sample plots was funded by Junta de Castilla y Leon and ADEMA from 1997 to 2007, and by Junta de Castilla y Leon and CESEFOR from 2007 to 2014.

Abstract - The genus Bursaphelenchus comprises almost 100 species mainly from the northern hemisphere, with conifers as the most important hosts. Among the various nematode species, the pine wood nematode (PWN), Bursaphelenchus xylophilus, is the casual agent of pine wilt disease (PWD), and the most important forest pest for pines worldwide, classified as an A1 quarantine organism within the European Union. In 1999 this nematode was detected for the first time in Portugal and Europa associated with maritime pine, Pinus pinaster. Following detection, a national program denominated "Programa Nacional de Luta contra o Nemátodo da Madeira do Pinheiro" (PROLUNP) was created to, among other objectives, determine the distribution of the PWN and its associated vector(s) and host(s), and therefore intensive surveys covering the entire country were conducted with thousands of wood samples and suspected insects being analyzed. This thesis presents the listing, distribution, frequency and the insects associated with Bursaphelenchus species found associated with maritime pine in Portugal, identifying and characterizing the various species by morphological, biometrical and molecular biology (ITS-RFLP and rDNA sequencing analysis) techniques. To achieve the objectives, a total of 4813 maritime pine wood samples and 3294 insects from 22 species and six families were individually analyzed. A total of nine Bursaphelenchus species were found, namely: B. antoniae, B. hellenicus, B. leoni, B. mucronatus, B. pinasteri, B. sexdentati, B. teratospicularis, B. tusciae and B. xylophilus, all of them (with the exception of B. xylophilus) being new records for Portugal. Some of the species appear to have a widespread distribution, such as B. leoni, B. teratospicularis and B. tusciae while others were very rarely found and apparently have a localized distribution range within the country, namely B. antoniae and B. mucronatus. The majority of the species is characteristic of the Mediterranean region and can also be found in countries such as Spain, Italy and Greece, reflecting the affinity of our fauna with those locations. The association of B. hellenicus and B. tusciae with maritime pine is here reported for the first time. Six of the Bursaphelenchus species were also found associated with insects, mainly from the family Scolytidae (Coleoptera). Some of these interactions were described for the first time, namely: B. hellenicus with both Ips sexdentatus and Hylurgus ligniperda, B. sexdentati with both H. ligniperda and Orthotomicus erosus and B. tusciae with H. ligniperda. The exclusive association of B. xylophilus with the cerambycid Monochamus galloprovincialis was also confirmed. The nematode's dauer juveniles were usually found in low numbers in the insect vectors (ca 10-100 per insect), although for B. xylophilus a few thousand specimens per insect were sometimes found. The location of the dauer juveniles differed according to the species, although they were more common under the elytra and wings of the adult insects. A species new to science was detected and formally described as B. antoniae, associated with Hylobius sp. (Coleoptera; Curculionidae) beetles. Morphologically, this new species is very similar to B. hylobianum, although it's distinct ITS-RFLP molecular pattern (with only the enzyme Haelll producing comparable restriction bands) and the failure of hybridization supported the two species as distinct entities. Additional phylogenetic analysis of the 18S rDNA sequence further supported the taxonomical proximity of B. antoniae with B. hylobianum. Concerning the PWN, detailed studies on the development and morphology of B. xylophilus were conducted, and comparative measurements of field-collected and laboratory-maintained populations demonstrated that nematodes from the second group displayed larger size in all morphometric parameters, which could derive from more adequate conditions of nourishment and/or temperature. Taxonomical studies on the development stages of B. xylophilus confirmed the existence of four propagative juvenile stages (J1,J2,J3 and J4), an adult stage with both sexes and two dispersal stages (jIII e jIV), with the measurements of the gonad length allowing the separation of the propagative stages. It is hoped that the acquired knowledge will be useful on future surveys of nematodes of the Bursaphelenchus genus collected from either wood material or insect vectors, and facilitate the correct distinction and identification of the various species which are now known to occur. ### Resumo - 0 género Bursaphelenchus compreende quase 100 espécies, distribuídas sobretudo nos países do hemisfério norte do globo terrestre. Embora algumas espécies já tenham sido detectadas em plantas herbáceas, os hospedeiros vegetais mais comuns deste género são as coníferas, particularmente pinheiros. 0 nemátode da madeira do pinheiro (NMP), Bursaphelenchus xylophilus, é considerado a espécie mais importante deste género uma vez que é o agente causal da doença da murchidão dos pinheiros ("pine wilt disease"). Originário dos Estados Unidos, onde não causa grande impacte, o NMP foi introduzido em alguns países da Ásia (China, Japão, Coreia e Taiwan) e mais recentemente na Europa (Portugal). Nestas regiões é responsável pela destruição de milhares de hectares de coníferas, assumindo uma elevada importância económica. Em Portugal, depois da sua detecção em 1999, associado a Pinus pinaster, foi implementado um programa nacional "Programa Nacional de Luta contra o Nemátodo da Madeira do Pinheiro" (PROLUNP) que permitiu determinar a área afectada pela praga (a sul do rio Tejo, península de Setúbal) bem como definir e implementar estratégias de controlo e prevenção da disseminação do NMP a outras zonas de Portugal. Recentemente, em Junho de 2008, foi confirmada a presença de B. xylophilus em outras regiões de Portugal levando as autoridades oficiais a definir todo o território continental como zona afectada e de restrição. As prospecções intensivas realizadas nos últimos anos incluíram a recolha e análise de milhares de amostras de madeira de pinheiro bem como de insectos associados ao pinheiro bravo conduzindo à identificação de várias espécies de Bursaphelenchus. Assim, os estudos conduzidos neste trabalho tiveram como objectivos efectuar uma caracterização morfológica, biométrica e molecular das espécies associadas a P. pinaster em Portugal bem como a sua distribuição geográfica e abundância. Os estudos biométricos foram realizados com populações extraídas directamente do meio natural. Foi ainda realizada uma pesquisa que permitiu identificar os insectos a que estão associadas essas espécies, os seus possíveis vectores. Foram analisadas no total 4813 amostras de P. pinaster e 3294 insectos (22 espécies pertencentes a seis famílias diferentes). Foram identificadas um total de nove espécies: B. antoniae n. sp., B. hellenicus, B. leoni, B. mucronatus, B. pinasteri, B. sexdentati, B. teratospicularis, B. tusciae e B. xylophilus. Foram realizados estudos morfológicos e biométricos de todas as espécies com excepção de B. mucronatus; o reduzido número de exemplares encontrados em apenas uma amostra foram utilizados para efectuar o diagnóstico molecular desta espécie (ITS-RFLP). Apesar de ter havido, sempre que possível, a confirmação molecular, na maioria dos casos a caracterização morfológica e biométrica permitiu a correcta identificação das espécies. Contudo, foi imprescindível a análise molecular em algumas amostras, nomeadamente para a identificação de B. xylophilus e B. sexdentati; dada a grande semelhança entre B. xylophilus e B. mucronatus e tendo sido encontradas algumas populações de B. xylophilus que possuíam fêmeas com cauda mucronada, foi necessária a realização da confirmação molecular. Com excepção de B. xylophilus, todas as outras espécies foram reportadas pela primeira vez em Portugal. Juntamente com B. xylophilus, B. pinasteri foi a espécie encontrada nas amostras de madeira de pinheiro com maior frequência. Algumas destas espécies como B. leoni, B. teratospicularis e B. tusciae foram reportadas em diferentes localidades do norte, centro e sul de Portugal, apresentando uma vasta distribuição geográfica; este resultado está em consonância com a forte associação destas espécies a climas mediterrânicos tal como acontece em Espanha, França, Itália e Grécia. Em oposição, espécies como B. antoniae e B. mucronatus foram encontradas apenas numa ocasião na região centro (Leiria) e norte (Figueira da Foz) do país, respectivamente. Bursaphelenchus mucronatus é igualmente pouco frequente em Espanha onde ocorre sobretudo na região norte, na Galiza. Esta espécie preferirá climas mais frios, ocorrendo com uma maior frequência nas regiões de latitude norte; esta análise é corroborada pela presença constante em países como Alemanha, Finlândia, França, Noruega, Rússia e Suécia. A nível mundial são descritas neste trabalho pela primeira vez as associações das espécies B. hellenicus e B. tusciae ao hospedeiro vegetal P. pinaster.

Tese de Doutoramento em Engenharia Química (Processos Químicos) apresentada à Faculdade de Ciências e Tecnologia da Universidade de Coimbra ; Agricultural processing inevitably goes along with the production of large amounts of agro-residues, which may represent a major waste disposal problem. Legislation regulating the management of waste materials has appeared throughout the European Union and has significantly contributed to the introduction of sustainable waste management procedures. The reuse and recycling of agro-residues has been highly encouraged and new technologies applying environmentally clean processes have been playing a central role within this context. These strategies may boost both the environmental and economic profiles of the implied industries, since they may create value in the entire chain-production pipeline. Typically, a substantial part of the agro-residues produced during the handling and processing of fruits, vegetables and forest resources still comprises important amounts of the original plant materials, such as fruit skins, fruit seeds, flowers, leaves, stems, barks, and roots. High-value natural compounds can be found in most of these vegetable residues, many of them having health-promoting characteristics. It is the case of, for example, phenolic compounds (which have been associated to the alleged health-promoting effects related to the consumption of fruits and vegetables), since they are preferentially biosynthesised in the external vegetable tissues. Usually, and despite their significant potential value, these residues are often underexploited and thus their potential value as natural products is frequently lost. Efficient techniques can thus be explored for separation and isolation of oils, antioxidants, fragrances, colorants, biocides and other bioactive substances of natural origin. Therefore, with some additional and adequate processing steps, these materials can be easily transformed, from a residual low-value status into a very interesting high-value status, in terms of well-known and consumer high-value and accepted uses in the food, cosmetic and pharmaceutical industries, among others. This Thesis is focused on the extraction of potentially valuable compounds from agroresidues of elder and maritime pine (Portuguese native plants) and also from tara (a Peruvian native plant). Solvents and techniques generally regarded as "environmental friendly" were applied. Despite conventional methods such as hydrodistillation and Soxhlet extraction had been employed, special attention was given to high pressure extraction procedures, namely supercritical and pressurized solvent extractions using, respectively, carbon dioxide and mixtures of carbon dioxide, ethanol and water as solvents. In general terms and besides other particular aspects, the effect of several process variables/conditions on the yields and on the characteristics of the obtained extracts, such as composition and antioxidant and antiinflammatory activities, was analyzed. Extract fractionation, solvent flow rate and composition, in addition to extraction time, temperature and pressure, were among these studied process variables/conditions. Characterization of extracts involved the use of diverse analytical techniques including spectrophotometric and chromatographic ones. Elderberry pomace and elderflower were evaluated as sources of potentially useful natural compounds. Elderberry pomace is a juice-processing industry residue which comprises the fruit skins and seeds, and is known to be rich in anthocyanins and other phenolic compounds. It was subjected to high pressure extractions at 313 K and ~20 MPa, applying supercritical carbon dioxide in a first step and pressurized solvent mixtures of carbon dioxide, ethanol and water in a second step. Extract fractionation was achieved by such a methodology. While the first step led to extracts rich in lipids and other apolar substances, obtained second step extracts revealed high contents of anthocyanins and presented high antioxidant activities, which were strongly influenced by the employed solvent mixture composition. The results of this study highlight the potential of elderberry pomace as a source of valuable compounds, and may assist in the development of strategies for integral exploitation of elderberries, reducing the environmental impact of elderberry juice processing industries. Elderflower was, in its turn, subjected to supercritical carbon dioxide extraction. The effect of solvent density (300-900 kg/m3) and of extraction temperature (308-328 K) on extracts characteristics was analyzed. Most of the obtained extracts revealed high contents of apolar compounds. Their phenolic compounds contents and the resulting antioxidant activities were relatively low, but they may possibly be increased if ethanol and water mixtures are added as cosolvents to carbon dioxide (which is planned to be performed in the near future). Nevertheless, obtained results revealed that elderflower extracts achieved by supercritical fluid technology may be added as natural aromatic additives to cosmetic and food systems. Maritime pine bark is an abundant residue from pine wood processing industries which is rich in condensed tannins. This residue was subjected to conventional solid-liquid aqueousbased extractions as well as to supercritical and pressurized solvent extraction procedures in order to obtain efficient vegetable tanning agents and bioactive phenolic-based extracts, respectively. Conventional aqueous extractions were optimized by adding small amounts (up to 15%, v/v) of ethanol. This methodology was selective for condensed tannins and the obtained extracts revealed adequate characteristics for leather tanning applications. Concerning supercritical fluid extraction experiments, carbon dioxide and a mixture of carbon dioxide and ethanol (10%) were used in two consecutive steps, varying extraction pressure in the range 10-30 MPa, and extraction temperature in the range 303-323 K. The employed fractionated extraction methodology showed that it was possible to obtain different extract fractions having diverse antioxidant activities from maritime pine bark. Moreover, it showed that the process could be further optimized through the usage of different solvent mixtures as well as other operational conditions. Therefore, pressurized solvent extractions were consecutively performed varying some process variables such as solvent mixture composition (carbon dioxide and ethanol, with up to 90% of ethanol) and flow rate. Extraction kinetics and extracts composition (in terms of phenolic compounds in general, and of condensed tannins in particular) were considerably affected by the solvent mixture composition. The results of these studies showed that both conventional and high pressure extraction methodologies may be feasible strategies for the valorization of maritime pine bark, therefore reducing the environmental impact and increasing the profit of the involved industries. Finally, tara seed coat, which is a residue of tara powder and of tara gum processing industries, was evaluated as a source of phenolic compounds in general, and of hydrolysable tannins, in particular. Pressurized solvent extractions using homogeneous solvent mixtures of carbon dioxide, ethanol and water were performed at 313 K and 20 MPa. An experimental mixture design was applied in order to optimize the solvent mixture composition in terms of selectivity towards the above referred compounds, as well as in terms of antioxidant activities and lipoxygenase inhibition activities (indicative of their anti-inflammatory capacities) of the whole extracts. The achieved mixture models permitted to discern the contribution of the three individual solvents used: while water revealed to be the most effective solvent to obtain high extraction yields, ethanol-rich mixtures originated the maximum extracts´ phenolic contents. Moreover, the obtained extracts presented quite relevant antioxidant and lipoxygenase inhibition activities. Tara seed coat is, therefore, a rich source of bioactive compounds, which may be applied as natural additives in food, cosmetic or pharmaceutical goods, increasing their shelf-lives and/or acting as human health promoters. In conclusion, instead of being disposed as waste of environmental concern or utilized in low value applications, agro-residues of elder, maritime pine and tara are promising sources of important constituents and should be considered for valorization. Environmentally clean methodologies and solvents may be applied for the extraction of natural colorants, flavors, aromas, preservatives, among others, some of them presenting bioactive properties. The choice of the appropriate extraction methodology and extraction conditions is of extreme importance for the selectivity of the extraction towards the intended compounds and for the resulting properties of the obtained natural extracts, defining their future potential applications. The successful exploitation of the studied agro-residues still requires further research and development, besides the one presented in this Thesis. A more precise identification of the natural compounds present in the obtained extracts is still required and will be addressed in future works. Moreover, other issues like the toxicity, metabolism, bioavailability and physiological activities of the extracted compounds also need to be considered. Nevertheless, a contribution has been given to the sustainability of elder, maritime pine and tara processing industries. ; O processamento industrial de produtos agrícolas e florestais origina enormes quantidades de resíduos que podem constituir um sério problema ambiental, se forem sujeitos a procedimentos incorrectos de aproveitamento e/ou eliminação. Frequentemente, estes resíduos são apenas parcialmente valorizados, sendo utilizados para alimentação animal, como fertilizantes ou transformados em biocombustível. Contudo, nos últimos anos tem-se verificado uma tendência crescente para a sua valorização, acompanhando a evolução de uma legislação ambiental cada vez mais restritiva. Neste contexto, a reutilização e reciclagem destes resíduos naturais têm sido encorajadas, contribuindo para agregar valor económico na linha de produção das empresas do sector, podendo mesmo constituir uma excelente oportunidade de negócio. Tipicamente, uma parte substancial dos resíduos produzidos durante o processamento de frutos, vegetais e recursos florestais consiste da matéria-prima original, incluindo a pele e as sementes dos frutos, flores, folhas, caules, casca de árvores e raízes. Na maior parte destes resíduos existem quantidades substanciais de compostos com elevado valor acrescentado, muitos deles possuindo propriedades benéficas para a saúde humana. De entre eles, destacamse os compostos fenólicos (que têm sido associados aos benefícios relacionados com o consumo de frutos e vegetais), uma vez que são preferencialmente sintetizados nos tecidos vegetais exteriores. Usualmente, e apesar do seu valor significativo, estes resíduos são comummente sub-explorados, e o seu potencial valor como produto natural é frequentemente perdido. Sendo assim, uma das áreas com maior potencial de exploração é o aproveitamento de substâncias com elevado valor acrescentado. De facto, após o tratamento adequado das matérias-primas, seguido de técnicas de separação adequadas, a maior parte dos resíduos agro-industriais pode constituir uma fonte rica de antioxidantes, corantes, fragrâncias e biocidas naturais, entre outros, com enorme interesse para as indústrias farmacêutica, cosmética ou alimentar. Neste trabalho foi estudada a extracção de compostos com potencial valor acrescentado a partir de resíduos de três plantas: o sabugueiro e o pinheiro-bravo (nativas de Portugal), e a tara (nativa do Peru). Foram usadas técnicas e solventes de extracção considerados "amigos do ambiente". Apesar de terem sido usadas técnicas convencionais de extracção, como a hidrodestilação e a extracção por Soxhlet, foi dado especial destaque à extracção supercrítica e à extracção com solvente pressurizado, usando como solventes, respectivamente, o dióxido de carbono supercrítico e misturas pressurizadas de dióxido de carbono, etanol e água. Em termos gerais, e para além de outros aspectos particulares, foi estudado o efeito de diversas variáveis/condições de processo (fraccionamento de extracção, composição do solvente, temperatura e pressão de operação, caudal de solvente, entre outros) nos rendimentos e nas características dos extractos obtidos, nomeadamente em termos de composição e actividades antioxidante e anti-inflamatória. Para a caracterização analítica dos extractos foram usadas diversas técnicas analíticas, incluindo espectrofotométricas e cromatográficas. O bagaço e a flor de sabugueiro foram avaliados como possíveis fontes de compostos naturais de interesse. O bagaço de sabugueiro, um resíduo proveniente da indústria de processamento de sumo, é constituído essencialmente pelas peles e grainhas das bagas e é uma fonte rica de antocianinas e outros compostos fenólicos. Este resíduo foi sujeito a extracção a alta pressão a 313 K e a ~20 MPa, usando dióxido de carbono supercrítico numa primeira etapa, e misturas pressurizadas de dióxido de carbono, etanol e água numa segunda etapa. Esta metodologia de extracção permitiu obter o fraccionamento do extracto. A primeira etapa de extracção originou extractos contendo elevados teores de lípidos e outras substâncias apolares. Os extractos obtidos na segunda etapa revelaram ter elevados teores de antocianinas e apresentaram uma elevada actividade antioxidante, com uma acentuada influência da composição do solvente. Os resultados deste estudo evidenciaram o potencial do bagaço do sabugueiro como fonte de compostos com elevado valor acrescentado, podendo contribuir para o desenvolvimento de estratégias de utilização integral das bagas, reduzindo o impacto ambiental das indústrias do sector. Por outro lado, a flor foi sujeita a extracção supercrítica usando dióxido de carbono como solvente. Foi avaliado o efeito da massa específica do solvente (300-900 kg/m3) e da temperatura de extracção (308-328 K) nos rendimentos e nas características dos extractos obtidos. A maior parte dos extractos obtidos revelaram ter um elevado teor de compostos apolares. No entanto, os seus teores de compostos fenólicos e as suas actividades antioxidantes foram relativamente baixos, podendo eventualmente ser melhorados se misturas de etanol e água forem usadas como co-solventes (o que se prevê que venha a ser feito em trabalhos futuros). Os resultados obtidos revelaram que os extractos de flor de sabugueiro obtidos recorrendo à tecnologia de extracção supercrítica podem, eventualmente, ser usados como aditivos aromáticos naturais em produtos alimentares e cosméticos. A casca do pinheiro-bravo, um resíduo abundante da indústria madeireira e rico em taninos condensados, foi sujeita a extracções convencionais (com o objectivo de obter extractos eficazes na curtimenta de peles de animais) e a extracções supercríticas e com solvente pressurizado (com o objectivo de obter extractos bioactivos). Nas extracções convencionais, foi utilizada água como solvente e foi optimizado o aditivo usado. Verificouse que uma pequena percentagem de etanol (até 15%, v/v) foi selectiva para os taninos condensados e que os extractos obtidos tinham condições adequadas para serem usados na curtimenta de pele de animais. Nas extracções supercríticas foram estudadas as condições de pressão (10-30 MPa) e temperatura (303-323 K), usando o dióxido de carbono e uma mistura de dióxido de carbono e etanol (10%) como solventes, em dois passos consecutivos. A metodologia de extracção aplicada mostrou que é possível obter fracções de extracto com actividades diversas a partir da casca de pinheiro-bravo. Para além disso, mostrou que o processo podia, ainda, ser optimizado utilizando diferentes condições experimentais, entre as quais a composição do solvente. Sendo assim, foram efectuadas extracções com solvente pressurizado variando algumas condições de processo, tais como a composição do solvente (misturas de dióxido de carbono e etanol, contendo até 90% de etanol) e o caudal de solvente. Tanto as cinéticas de extracção como a composição dos extractos (em termos de compostos fenólicos, em geral, e de taninos condensados, em particular) foram consideravelmente afectados pela composição da mistura do solvente. Este estudo mostrou que, tanto a metodologia de extracção convencional como a de alta pressão, podem ser estratégias viáveis de valorização da casca do pinheiro-bravo, contribuindo para a redução do impacto ambiental e para o aumento do lucro das indústrias do sector. Finalmente, foi avaliada a possibilidade de aproveitamento da casca da semente de tara (um resíduo proveniente do processamento do fruto para a obtenção de pó e goma de tara) para a recuperação de compostos fenólicos, em geral, e de taninos hidrolisáveis, em particular. Foram efectuadas extracções pressurizadas usando misturas homogéneas de dióxido de carbono, etanol e água, a 313 K e a 20 MPa. Foi utilizado um desenho experimental para optimizar a composição do solvente de extracção, em termos de selectividade para com os compostos referidos, assim como em termos de actividade antioxidante e de inibição da lipoxigenase (indicativa da actividade anti-inflamatória). Os modelos de mistura obtidos permitiram avaliar a contribuição de cada um dos solventes individuais usados: enquanto que a água foi eficaz na promoção de elevados rendimentos de extracção, o etanol foi selectivo para os compostos fenólicos. Os extractos obtidos mostraram ter um elevado teor de compostos fenólicos e apresentaram relevantes actividades antioxidante e anti-inflamatória. Sendo assim, a casca da semente de tara é uma fonte rica de compostos bioactivos que podem ser usados como aditivos naturais em produtos alimentares, cosméticos e farmacêuticos, aumentando a sua vida útil e/ou actuando como substâncias promotoras da saúde humana. Em conclusão, os resíduos obtidos a partir de sabugueiro, pinheiro-bravo e tara são fontes promissoras de substâncias importantes devendo, por isso, ser considerados para valorização, em vez de serem encarados como causadores de problemas ambientais, ou serem usados em aplicações de baixo valor acrescentado. Neste estudo foram aplicadas metodologias e solventes "amigos do ambiente" para a extracção de corantes, aromas e antioxidantes, entre outros, de origem natural, alguns deles apresentando potenciais propriedades biológicas. A escolha da metodologia adequada e das condições particulares de extracção é de extrema importância para a selectividade da extracção para com os compostos de interesse, assim como para as propriedades evidenciadas pelos extractos naturais obtidos, definindo as suas potenciais aplicações futuras. O sucesso da exploração dos resíduos agro-industriais considerados requer mais investigação e desenvolvimento, para além dos apresentados neste estudo. Nomeadamente, requer uma identificação mais precisa dos compostos naturais presentes nos extractos obtidos, que irá ser abordada em trabalhos futuros. Outras questões tais como o metabolismo, a bio-disponibilidade e a actividade fisiológica dos compostos presentes nos extractos também necessitam, eventualmente, de ser considerados. No entanto, neste estudo foi dada uma contribuição para a sustentabilidade das indústrias de processamento de sabugueiro, pinheiro-bravo e tara.

Forestry residual biomass from pruning operations is an important, though little studied, potential resource. Residues normally remain in the stand, since tools for their accurate quantification do not exist and it has no particular end use. Traditional tree biomass estimation models consider the whole-tree, but estimating pruned biomass requires the development of more specific equations. This work provides a modelling approach for assessing biomass along the stem and the corresponding residual biomass from forest pruning, and quantitative results from different pruning intensities in Pinus pinaster Ait. are presented. Two types of models were considered: allometric biomass equations (whole-tree) and biomass ratio equations (tree by height along the stem), and the 2-parameter Weibull distribution function resulted in the best characterization. Diameter at breast height was the best explanatory variable in all equations, and model accuracy increased when models were combined with total tree height for the tree stem and thicker branches, or with crown ratio for the remaining tree crown components. This study provides a powerful tool to estimate residual pruned biomass, enabling its better management as a valuable source of bioenergy, as well as the importance in nutrient balance and fire risk which it plays in a sustainable forestry production ; We thank the Forest Services of the Government of the Principality of Asturias for access to the forests used in this study and for financial support. Thank you to people from CETEMAS (L. González, M. García, P. Vallejo) and SERIDA (J.C. Hernández) for their participation in the fieldwork. Thank you to Ronnie Lendrum for reviewing the English and FORRISK project (Interreg IV B SUDOE 2007–2013) for its support during data analysis. Andrea Hevia was financially supported during fieldwork and data analysis by the Spanish Ministry of Education and Science through the FPU scholarship program (Reference AP2006-03890) ; SI

Producción Científica ; Long-term field-data series were used to fit a mushroom productivity model. Simulations enabled us to predict the consequences of management and climate scenarios on potential mushroom productivity. Mushrooms play an important ecological and economic role in forest ecosystems. Human interest in collecting mushrooms for self-consumption is also increasing, giving forests added value for providing recreational services. Pinus pinaster Ait. is a western Mediterranean species of great economic and ecological value. Over 7.5% of the total European distribution of the species is found on the Castilian Plateau in central Spain, where a great variety of mushrooms can be harvested. The aim of this study was to model and simulate mushroom productivity in Maritime pine (Pinus pinaster Ait.) ecosystems in northern Spain under different silvicultural and climatic scenarios. A mixed model was fitted that related total mushroom productivity to stand and weather variables. The model was uploaded to the SiManFor platform to study the effect of different silvicultural and climatic scenarios on mushroom productivity. The selected independent variables in the model were the ratio between stand basal area and density as a stand management indicator, along with precipitation and average temperatures for September and November. The simulation results also showed that silviculture had a positive impact on mushroom productivity, which was higher in scenarios with moderate and high thinning intensities. The impact was highly positive in wetter scenarios, though only slightly positive and negative responses were observed in hotter and drier scenarios, respectively. Silviculture had a positive impact on mushroom productivity, especially in wetter scenarios. Precipitation had greater influence than temperature on total mushroom productivity in Maritime pine stands. The results of this paper will enable forest managers to develop optimal management approaches for P. pinaster forests that integrate Non-Wood Forest Products resources. ; European Union Seventh Framework Programme FP7 under Grant Agreement n 613762 ; Ministerio de Economía y Competitividad under Grant Agreement n 345151982-51982-45-514 ; Ministerio de Economía y Competitividad - Fondo Social Europeo (Grant PTQ-12-05409)

17 Pág. ; A decade of genetic association studies in multiple organisms suggests that most complex traits are polygenic; that is, they have a genetic architecture determined by numerous loci, each with small effect-size. Thus, determining the degree of polygenicity and its variation across traits, environments and time is crucial to understand the genetic basis of phenotypic variation. We applied multilocus approaches to estimate the degree of polygenicity of fitness-related traits in a long-lived plant (Pinus pinaster Ait., maritime pine) and to analyse this variation across environments and years. We evaluated five categories of fitness-related traits (survival, height, phenology, functional, and biotic-stress response) in a clonal common-garden network planted in contrasted environments (over 20,500 trees). Most of the analysed traits showed evidence of local adaptation based on Qst -Fst comparisons. We further observed a remarkably stable degree of polygenicity, averaging 6% (range of 0%-27%), across traits, environments and years. We detected evidence of negative selection, which could explain, at least partially, the high degree of polygenicity. Because polygenic adaptation can occur rapidly, our results suggest that current predictions on the capacity of natural forest tree populations to adapt to new environments should be revised, especially in the current context of climate change. ; This study was funded by the Spanish Ministry of Economy and Competitiveness through projects RTA2010-00120-C02- 02 (CLONAPIN), CGL2011-30182- C02- 01 (AdapCon) and AGL2012-40151- C03- 02 (FENOPIN). The study was also supported by the "Initiative d'Excellence (IdEx) de l'Université de Bordeaux - Chaires d'installation 2015" (EcoGenPin) and the European Union's Horizon 2020 research and innovation programme under grant agreement No 773383 (B4EST) ; Peer reviewed

Herbaceous vegetation is a major source of interference with the regeneration of woody species. This is particularly the case after forest fires, as a dense herbaceous layer usually regenerates naturally. Although the competitive effect of the herbaceous vegetation upon tree seedlings has been widely studied, there are still gaps in knowledge for management related to the effect of tree seedling age and size on the outcome of the interaction. In this study, we seek to determine the response of maritime pine (Pinus pinaster Aiton) seedlings to herbaceous competition at two different seedling ages. For that, two treatments of herbaceous competition were implemented, namely unweeded (no action around pine seedlings) and weeded (herbaceous cover removed around pine seedlings). Treatments were conducted twice (2 and 4 years after the fire), and we monitored seedling survival and growth at the end of each growing season. The treatments were implemented across three adjacent landscape units that differed in the management of burned wood and that are representative of common post-fire scenarios: no intervention, salvage logging, and an intermediate degree of intervention. Weeding increased seedling survival from 44.7% to 67.8% when seedlings were 2 years old, but had no effect for four-year-old seedlings, which showed 99% survival. Seedling growth also increased in the weeding treatment, but only slightly. Moreover, growth (and survival for two-year-old seedlings) was strongly correlated with initial seedling size, particularly in the case of two-year-old seedlings. Initial pine seedling height was strongly and positively correlated with the height of the herbaceous layer, supporting the existence of microsite features that promote plant growth above competitive effects. The results support that management actions conducive to foster post-fire pine forest restoration in this Mediterranean ecosystem should reduce herbaceous competition at early stages after fire (second or third year) and focus on larger seedlings. ; This research was funded by projects 10/2005 from the Organismo Autónomo de Parques Nacionales (Spanish Government), CGL2008-01671 from the Spanish Ministerio de Ciencia e Innovación (Spanish Government), and P12-RNM-2705 from Junta de Andalucía (Andalusian regional Government).