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Digital Annex for the following thesis: Vila-Viçosa, C. (2023). Natural History, Biogeography and Evolution of the Iberian white oak syngameon (Quercus L. Sect. Quercus). Ph.D. Thesis, Faculdade de Ciências da Universidade do Porto, Portugal Abstract: The genus Quercus L. is one of the most diverse and important group of woody plants, particularly when considering that they are the trees that rule the Northern Hemisphere forests. Oaks have an intricate Biogeography that criss-crosses diverse climatic and edaphic gradients, encompassing a huge ambiguity in terms of species delimitation. Frequently, the taxonomic proposals brought by traditional Linnaean Botany are either insufficient or rather inflate the number of species and nomenclatural assignments, which are further diluted into inconsistent taxonomic ranks, varying from species to subspecies and varieties. The supremacy given to morphological characters that are inherently fragile and plastic, spread across the distribution areas of distinct lineages, may carry ambiguity on the identification and proper species delimitation. From the oaks that are distributed across the Western Palearctic region, the ones that are deciduous or brevi-deciduous present higher levels of ambiguity in terms of species number and their delimitation. This ambiguity is particularly strong in the circummediterranean region and in the transitional areas between the two major biogeographic Regions of the western Palearctic region, the Euro-Siberian and Mediterranean. This degree of uncertainty, which increases towards the Southern European Peninsulas, is amplified by the ease that the different species of oaks tend to hybridize among them. The present work provides a holistic framework that covers multiple areas, from the taxonomic and evolutive study of this genus, to biogeography and molecular characterization. Its major objective was to resolve the species delimitation of the Iberian deciduous and marcescent oaks and putative introgression among them, enhancing the available knowledge about species diversity, which can foster suitable species and forest conservation. A specific objective was to cross-reference the natural history revision and the different taxonomic treatments brought by distinct authors, with personal observations. These data were then incorporated into ecological modelling and molecular characterization, which in the end fed a newly updated taxonomic proposal. In Section A we obtained results from extensive field, herbaria, and literature review, updating the nomenclature of the Portuguese and western Mediterranean oaks. Section B was supported by Section A’s in-depth review and enabled finer species distribution models, nurturing both hindcast (since ca. 20 Kyr) and forecast (2070-2100) exercises of the range dynamics of Mediterranean oaks species. The study of past and future range shifts solved important pending biogeographic questions, especially related to past range-shifts. Such past-range shifts improved our knowledge on species responses to climate dynamics and allowed a better anticipation of future responses of range shifts driven by climate change. Section C encompassed the molecular characterization of Iberian white oak species and their hybrids, whose delimitation is often faltering when one intends to infer about species rank, or hypothesize about the participation of parent taxon in natural hybrid swarms. This work allowed us to solve the phylogenetic backbone of western Palearctic white oaks, suggesting a significant segregation of the Iberian pedunculate oaks and unveiling two subsections inside Section Quercus. These subsections are biogeographically well-segregated and present diverse levels of introgression among species. Results demonstrated the efficiency of RADSeq for rebuilding the reticulate phylogeny of the Eurasian white oaks, showcasing the significance of the Iberian Peninsula as a major hotspot for oak diversity. We implemented a circular approach to these methods, which retro-fed themselves in terms of insight generation, enabling a powerful strategy to solve the evolutionary history of this difficult groups of plants. We estimate that the reticulate historical biogeography of the western Palearctic white oaks deserves further scrutiny by adding vicariant oak populations from northern Africa, the Near East and southern European Peninsulas. Methods should again follow this similar additive and sequential process of adjoining deep Natural History examination, with extensive fieldwork in type populations and genome-wide molecular surveys, in order to solve this group of plants. With the present work, we were able to significantly improve on the depiction of the basic unit of Biodiversity (the Species), in the complex Quercus genus. We provided tools to enable further efforts for the conservation of the Mediterranean oak forests, which overwhelm one of the most important (and one of the most threatened) Biomes for plant conservation at the global scale.

ScopeThis study was designed to evaluate the influence of ethanol on the bioavailability of blackberry anthocyanins.Methods and resultsA total of 18 participants were recruited to consume 250 mL of a blackberry puree (650 mg of anthocyanins) without (BBP) or with 12% ethanol (BBP 12%). Venous blood was collected from participants at baseline and at 15, 30, 60, and 120 min after puree ingestion. Urine samples were collected at baseline and at 120 min. Plasma and urine concentration of anthocyanins and anthocyanin conjugates were quantified by HPLC‐DAD. Methyl‐cyanidin‐glucuronide (Me‐Cy‐Glucr) and 3′‐methyl‐cyanidin‐3‐glucoside (3′‐Me‐Cy3glc) were the main anthocyanin conjugates detected in all plasma and urine samples. Urinary concentration of these anthocyanin conjugates were positively correlated with their plasma concentration. Ethanol increased plasma Cmax of Me‐Cy‐Glucr and 3′‐Me‐Cy3glc. Participants were then stratified according to their body mass index (BMI) and body fat mass. After BBP consumption, plasma Cmax of Me‐Cy‐Glucr and 3′‐Me‐Cy3glc tended to be decreased in overweight/obese participants, in comparison to normal weight participants. The increase on plasma Cmax of Me‐Cy‐Glucr and 3′‐Me‐Cy3glc induced by ethanol was more pronounced in the group of overweight/obese participants.ConclusionsEthanol seems to enhance Cy3glc metabolism that appears to be compromised in overweight and obese individuals.

In this study, air concentrations of BaP in two different seasons (winter 2015 and summer 2016) and BaP levels in ground vegetation from Tarragona County were used as control simulations performed with the WRF-CHIMERE air quality modelling system, in order to reproduce the incidence of that hazardous chemical in air and soils. The CTM was validated for the present climatology, showing a good ability to represent air and soil concentrations of BaP over the target domain (petrochemical, chemical, urban and background sites), particularly in the winter. Then, the variation of the BaP concentrations in air and soils were simulated for the time series 1996-2015 and for the climate change scenario RCP8.5 (2031-2050). While an increase is projected for the levels in air, particularly in chemical and remote sites where the variation can go up to 10%, in terms of soil deposition the findings are the opposite, with an evident decrease in soil BaP concentrations, particularly for background sites. Finally, a potential health effect of BaP for the local population (lung cancer) was assessed. Although according to the projections the EU threshold for BaP atmospheric incidence (1 ng m-3) will not be reached by 2050, there will be an increase in the life-time risk of lung cancer, particularly in the most populated areas within the simulation domain.

Abstract Portugal’s performance concerning the Refuse Derived Fuel (RDF) strategy, with particular focus on perspectives for the Northern Region based on the current status of integrated municipal waste management systems in that region, was evaluated. Legal documents, official reports and the market were analyzed. The 2015 national installed capacity of waste treatment technologies, from which the high calorific value fraction (HCVF) for RDF production is obtained, is not far from what was predicted for 2013. However, only around 11% of the HCVF had such destiny and the amount of effectively produced RDF was only 30% of the material sent to RDF production (around 30 kt). In Northern Portugal, no RDF production plant was in operation in 2015; the amount of HCVF was estimated to range from 105 to 204 kt for 2015 and around 2.4 times more is expected in 2020. The strategy laydown for RDF failed so far, and thus, low diversion of waste from landfills is observed, mostly due to low landfill taxes and low HCVF quality, resulting in RDF being non-competitive with the imported higher quality product from countries with very high landfilling fees. Moreover, co-incineration and dedicated recovery solutions are scarce. In the Northern region, the HCVF may be treated in the existing municipal waste incinerator or the RDF might be recovered in new dedicated plants, receiving also non-hazardous waste fractions from other origin. Overall, to ensure a sustainable municipal waste management, the intensification of the reuse, preparation for reuse and recycling of waste components from source segregated collection must be significantly promoted.

The olive oil mill wastewater (OMW) is a by-product (with a high organic load) derived from the production of the olive oil. The OMW steam reforming (OMWSR) process was studied herein, aiming to decrease the environmental damage of such effluents; simultaneously, the waste is economically and energetically valorized with the H-2 production. Several Ni-containing catalysts were prepared and tested to compare their performances for the OMWSR using a synthetic OMW effluent; still, stability tests were also carried out. The materials were extensively characterized: thermogravimetric analysis, temperature programmed oxidation/reduction, temperature-programmed desorption of CO2/NH3, chemisorption of H-2, inductively coupled plasma optical emission spectrometry and physical adsorption-desorption of N-2 at-196 & DEG;C. Amongst the materials tested, the Ni-Ru/SiO2 sample stood out, exhibiting high catalytic performance: at 400 & DEG;C, the H-2 yield (> 8 mol(H2)& BULL;mol(OMW)(-1)) and conversion of total organic carbon (asymptotic to 75%) were high during all the 24 h of the long-term test, with only a small deactivation being noticed.

The objective of the present work was: (i) to enable biodiesel production from acid waste lard; (ii) to study the esterification reaction as possible pre-treatment at different temperatures, catalyst amount and reaction times; (iii) to evaluate biodiesel quality according to EN 14214 after basic transesterification of the pre-treated fat; and (iv) to predict the impact of using such waste as raw material in mixture with soybean oil. Temperature and catalyst amount were the most important reaction conditions which mostly affected biodiesel quality, namely viscosity and purity. The selected pre-treatment conditions were 65 degrees C, 2.0 wt% H(2)SO(4) and 5 h, which allowed obtaining a product with a viscosity of 4.81 mm(2) s(-1) and a purity of 99.6 wt%. The proposed pre-treatment was effective to enable acid wastes as single raw materials for biodiesel production with acceptable quality; however, low yields were obtained (65 wt%). Alkali transesterification of a mixture of waste lard and soybean oil resulted in a product with a purity of 99.8 wt% and a yield of 77.8 wt%, showing that blending might be an interesting alternative to recycle such wastes. Also, because in addition to using conventional and relatively economical processes, some biodiesel properties depending on the raw material composition (such as the iodine value) might even be improved.

The catalytic oxidation of binary VOC mixtures, ethyl acetate, and butyl acetate, and the effect of water vapor on their oxidation, was studied over manganese oxide–cryptomelane‐type catalyst. This catalyst, prepared by a novel solid‐state reaction with ball milling, was shown to be very active for the oxidation of ethyl acetate and butyl acetate, promoting their complete oxidation at relatively low temperatures, the CO2 yield being influenced by the type of VOC. The mixture with equal amounts of each ester shows a light‐off curve similar to that observed for butyl acetate alone, but the light‐off curves are shifted to higher temperatures when different proportions of these VOCs are present in the mixture. When processing VOC mixtures, the temperature required for 100% CO2 yield depends on the VOC, which is in larger amount. Water vapor also has a significant effect on the oxidation of these VOCs, particularly in the case of ethyl acetate. © 2016 American Institute of Chemical Engineers Environ Prog, 35: 1324–1329, 2016

Molinate (S-ethyl-azepane-1-carbothioate) is a thiocarbamate herbicide used in rice cultivation for the control of grass weeds. Environmental contamination with molinate is of major concern due to the adverse effects described both for humans and animals. Molinate hydrolase, a novel amidohydrolase previously characterized, is responsible for the initial breakdown of molinate, cleaving the thioester bond of molinate, releasing ethanethiol and azepane-1-carboxylate (ACA). Biotechnology is the key for sustainable farming. With advances in biotechnology, bioremediation has become one of the most rapidly developing fields of environmental restoration. Through the microencapsulation of molinate hydrolase, we are aiming to develop a bioremediation process for the effective molinate degradation in rice paddies. The purpose of this work was to develop and validate an UV method to effectively quantify the substrate (molinate) in further assays with free and microencapsulated molinate hydrolase. The analytical method was validated and the main parameters, as limit of detection, linearity range, precision and accuracy were determined, and compared to those obtained by HPLC (regarding free enzyme kinetics). Both methods show to be linear (r > 0.999) over the concentration range of 0.005-0.150 mM molinate. The global uncertainty, estimated accordingly to the bottom-up approach used by Eurachem, was estimated for both methods. The UV analytical method is effective and seems that it can be applied in future for the quantification of molinate breakdown by free and encapsulated molinate hydrolase.