Assessing the balance between biodiversity conservation and coastal and estuarine management of the Sado Estuary Natural Reserve, Portugal, using BIO-SAFE

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em Engenharia do Ambiente, perfil de Engenharia Ecológica

Modulation of inflammatory mediators by Opuntia ficus-indica and Prunus avium bioproducts using an in vitro cell-based model of intestinal inflammation

Dissertation to obtain a Master Degree in Biotechnology

Adaptation from standing genetic variation and from mutation

Dissertation presented to obtain the Ph.D degree in Evolutionary Biology

Visitor behaviours and visitor effects: a case study on the white-crowned mangabey (cercocebus atys lunulatus) of ZSL London Zoo

Tese de mestrado em Antropologia, especialização natureza e conservação

Predicting the impacts of climate change on the distribution and conservation of endemic forest land snails of Madeira Island

Climate change is emerging as one of the major threats to natural communities of the world’s ecosystems; and biodiversity hotspots, such as Madeira Island, might face a challenging future in the conservation of endangered land snails’ species. With this thesis, progresses have been made in order to properly understand the impact of climate on these vulnerable taxa; and species distribution models coupled with GIS and climate change scenarios have become crucial to understand the relations between species distribution and environmental conditions, identifying threats and determining biodiversity vulnerability. With the use of MaxEnt, important changes in the species suitable areas were obtained. Laurel forest species, highly dependent on precipitation and relative humidity, may face major losses on their future suitable areas, leading to the possible extinction of several endangered species, such as Leiostyla heterodon. Despite the complexity of the biological systems, the intrinsic uncertainty of species distribution models and the lack of information about land snails’ functional traits, this analysis contributed to a pioneer study on the impacts of climate change on endemic species of Madeira Island. The future inclusion of predictions of the effect of climate change on species distribution as part of IUCN assessments could contribute to species prioritizing, promoting specific management actions and maximizing conservation investment.

Isolation and encapsulation of a natural colourant using green technologies

The use of natural pigments instead of synthetic colourants is receiving growing interest in the food industry. In this field, cactus pears (Opuntia spp.) have been identified to be a promising betalainic crops covering a wide coloured spectrum. The aim of this work was to develop adequate clean and mild methodologies for the isolation and encapsulation of betacyanins, from cactus pear fruits (Opuntia spp.). Firstly, two different emerging technologies, namely PLE (Pressurized Liquid Extraction) and HPCDAE (High Pressure Carbon Dioxide-Assisted Extraction), were exploited to isolation of betacyanins form cactus pear fruits. Different process conditions were tested for the maximum recovery of betacyanins. Results showed that highest extraction yields were achieved for HPCDAE and mass ratio of pressurized carbon dioxide vs. acidified water was the parameter that most affected the betacyanins extraction. At optimum conditions of HPCDAE, Opuntia spp. extract presented a total betacyanin content of 211 ± 10 mg/100 g whereas extracts obtained using conventional extraction, PLE in static and in dynamic mode presented a total betacyanin content of 85 ± 3, 191 ± 2 and 153 ± 5 mg/100 g, respectively. HPCDAE has proven to be a successful technology to extract betacyanins from Opuntia spp. fruits. Afterward, Supercritical Fluid Technology was exploited to develop lipidic particles of betalain-rich extract. A betacyanin-rich conventional extract was encapsulated by PGSS® (Particles from Gas Saturated Solutions) technique. Different process conditions were tested in order to model the encapsulation of betacyanins. The pressure had a negative effect on betacyanin encapsulation. Lower pressures leads to an increase in the betacyanin encapsulation. This effect was more pronounced at higher temperatures and lower equilibrium time. At these conditions, Opuntia spp. particles presented 64.4 ± 4.5 mg/100 g and high antioxidant capacity. When compared with the Opuntia spp. dried extract, lipidic particles contributed to a better homogenization of the pink colour after incorporation in ice cream.

Evaluation of neuroprotective effects of natural extracts obtained from portuguese agro-food residues

Countries are currently faced with problems derived from changes in lifespan and an increase in lifestyle-related diseases. Neurodegenerative disorders such Parkinson’s (PD) and Alzheimer’s (AD) diseases are an increasing problem in aged societies. Data from World Alzheimer Report 2011 indicate that 36 million people worldwide are living with dementia. Oxidative stress has been associated with the development of AD and PD. Therefore there is interest to search for effective compounds or therapies to combat the oxidative damage in these diseases. Current evidence strongly supports a contribution of phenolic compounds present in fruits and vegetables to the prevention of neurodegenerative diseases such AD and PD. The industrial processing of a wide variety of fruits results in the accumulation of by-products without commercial value. Opuntia ficus-indica (cactus pear) is consumed fresh and processed like in juice. Prunnus avium (sweet cherry) is consumed fresh but the organoleptics characteristics of the fruits leads to the smaller and ragged fruits have no commercial value. Fruit extracts of both species has described to be rich in phenolic compounds and to have high antioxidant activities due to its composition. The aim of this work was assessing the efficacy of O. ficus-indica and P. avium by-products extracts obtained with conventional solvent extraction and pressurized liquid extraction in a neurodegeneration cell model. All extracts have protected neuroblastoma cells from H2O2-induced death at low, non-toxic levels, which approach to physiologically-relevant serum concentration. However, cherry extract has a slighter neuroprotective activity. The protective effect of Opuntia extracts are not conducted by a direct antioxidant activity since there are not decreases in intracellular ROS levels in cell treated with extracts and challenged with H2O2, while cherry extract neuroprotection seems to be due to a direct scavenging activity. Extracts from different biological matrixes seems to protect neuronal cells trough different cellular mechanisms.

Seasonal effects and antifungal activity from bark chemical constituents of Sterculia apetala (Malvaceae) at Pantanal of Miranda, Mato Grosso do Sul, Brazil

In the Southern Pantanal, the hyacinth macaw (Anodorhynchus hyacinthinus), an endangered species, often chooses the manduvi tree (Sterculia apetala) as a nesting site, because of its physical properties. In addition, the chemical composition of the wood may also contribute to a nesting selection by the hyacinth macaws. The objective of this study was to determine the main chemical components of S. apetala bark for two seasons, and evaluate its fungicidal potential. Bark samples from S. apetala trees with and without nests of A. hyacinthinus were collected in January (wet season) and August (dry season) of 2012. The inhibition of mycelium growth (MGI) from tree samples with and without nests were assessed using a phytochemical analysis to evaluate their antifungal activity against Trichoderma sp. Phytochemical analysis confirmed the presence of phenolic compounds and flavonoids. In both seasons, samples obtained from nested trees had higher content of total phenols than those collected from non-nested trees. The average content of total flavonoids was higher in January for samples with nest and in August for samples without nest. All selected samples showed antifungal activity, and those with nest collected in August (peak of hyacinth macaw breeding) resulted in an MGI of 51.3%. Therefore, this percentage, related to the content of flavonoids and the presence of coumarins, may influence the reproductive success of hyacinth macaws and other species of birds, in this region. This is the first chemical study report with the stem bark of S. apetala.

Desenvolvimento de materiais para isolamento térmico com base em biomimética

Dissertação de mestrado Internacional em Sustentabilidade do Ambiente Construído

Estudos experimentais sôbre a origem do milho

1) It may seem rather strange that, in spite of the efforts of a considerable number of scientists, the problem of the origin of indian corn or maize still has remained an open question. There are no fossil remains or archaeological relics except those which are quite identical with types still existing. (Fig. 1). The main difficulty in finding the wild ancestor- which may still exist - results from the fact that it has been somewhat difficult to decide what it should be like and also where to look for it. 2) There is no need to discuss the literature since an excellent review has recently been published by MANGELSDORF and REEVES (1939). It may be sufficient to state that there are basically two hypotheses, that of ST. HILAIRE (1829) who considered Brazilian pod corn as the nearest relative of wild corn still existing, and that of ASCHERSON (1875) who considered Euchlaena from Central America as the wild ancestor of corn. Later hypotheses represent or variants of these two hypotheses or of other concepts, howewer generally with neither disproving their predecessors nor showing why the new hypotheses were better than the older ones. Since nearly all possible combinations of ideas have thus been put forward, it har- dly seems possible to find something theoretically new, while it is essential first to produce new facts. 3) The studies about the origin of maize received a new impulse from MANGELSDORF and REEVES'S experimental work on both Zea-Tripsacum and Zea-Euchlaena hybrids. Independently I started experiments in 1937 with the hope that new results might be obtained when using South American material. Having lost priority in some respects I decided to withold publication untill now, when I can put forward more concise ideas about the origin of maize, based on a new experimental reconstruction of the "wild type". 4) The two main aspects of MANGELSDORF and REEVES hypothesis are discussed. We agree with the authors that ST. HILAIRE's theory is probably correct in so far as the tunicata gene is a wild type relic gene, but cannot accept the reconstruction of wild corn as a homozygous pod corn with a hermaphroditic tassel. As shown experimentally (Fig. 2-3) these tassels have their central spike transformed into a terminal, many rowed ear with a flexible rachis, while possessing at the same time the lateral ear. Thus no explanation is given of the origin of the corn ear, which is the main feature of cultivated corn (BRIEGER, 1943). The second part of the hypothesis referring to the origin of Euchlaena from corn, inverting thus ASCHERSON's theory, cannot be accepted for several reasons, stated in some detail. The data at hand justify only the conclusion that both genera, Euchlaena and Zea, are related, and there is as little proof for considering the former as ancestor of the latter as there is for the new inverse theory. 5) The analysis of indigenous corn, which will be published in detail by BRIEGER and CUTLER, showed several very primitive characters, but no type was found which was in all characters sufficiently primitive. A genetical analysis of Paulista Pod Corn showed that it contains the same gene as other tunicates, in the IV chromosome, the segregation being complicated by a new gametophyte factor Ga3. The full results of this analysis shall be published elsewhere. (BRIEGER). Selection experiments with Paulista Pod Corn showed that no approximation to a wild ancestor may be obtained when limiting the studies to pure corn. Thus it seemed necessary to substitute "domesticated" by "wild type" modifiers, and the only means for achieving this substitution are hybridizations with Euchlaena. These hybrids have now been analysed init fourth generation, including backcrosses, and, again, the full data will be published elsewhere, by BRIEGER and ADDISON. In one present publication three forms obtained will be described only, which represent an approximation to wild type corn. 6) Before entering howewer into detail, some arguments against ST. HILAIRE's theory must be mentioned. The premendelian argument, referring to the instability of this character, is explained by the fact that all fertile pod corn plants are heterozygous for the dominant Tu factor. But the sterility of the homozygous TuTu, which phenotypically cannot be identified, is still unexplained. The most important argument against the acceptance of the Tunicata faetor as wild type relic gene was removed recently by CUTLER (not yet published) who showed that this type has been preserved for centuries by the Bolivian indians as a mystical "medicine". 7) The main botanical requirements for transforming the corn ear into a wild type structure are stated, and alternative solutions given. One series of these characters are found in Tripsacum and Euchlaena : 2 rows on opposite sides of the rachis, protection of the grains by scales, fragility of the rachis. There remains the other alternative : 4 rows, possibly forming double rows of female and male spikelets, protection of kernels by their glumes, separation of grains at their base from the cob which is thin and flexible. 8) Three successive stages in the reconstruction of wild corn, obtained experimentally, are discussed and illustrated, all characterized by the presence of the Tu gene. a) The structure of the Fl hybrids has already been described in 1943. The main features of the Tunicata hybrids (Fig. -8), when compared with non-tunicate hybrids (Fig. 5-6), consist in the absence of scaly protections, the fragility of the rachis and finally the differentiation of the double rows into one male and one female spikelet. As has been pointed out, these characters represent new phenotypic effects of the tunicate factor which do not appear in the presence of pure maize modifiers. b) The next step was observed among the first backcross to teosinte (Fig. 9). As shown in the photography, Fig. 9D, the features are essencially those of the Fl plants, except that the rachis is more teosinte like, with longer internodes, irregular four-row-arrangement and a complete fragility on the nodes. c) In the next generation a completely new type appeared (Fig. 10) which resembles neither corn nor teosinte, mainly in consequence of one character: the rachis is thin and flexible and not fragile, while the grains have an abscission layer at the base, The medium sized, pointed, brownish and hard granis are protected by their well developed corneous glumes. This last form may not yet be the nearest approach to a wild grass, and I shall try in further experiments to introduce other changes such as an increase of fertile flowers per spikelet, the reduction of difference between terminal and lateral inflorescences, etc.. But the nature of the atavistic reversion is alveadwy such that it alters considerably our expectation when looking for a still existing wild ancestor of corn. 9) The next step in our deductions must now consist in an reversion of our question. We must now explain how we may obtain domesticated corn, starting from a hypothetical wild plant, similar to type c. Of the several changes which must have been necessary to attract the attention of the Indians, the following two seem to me the most important: the disappearance of all abscission layers and the reduction of the glumes. This may have been brought about by an accumulation of mutations. But it seems much more probable to assume that some crossing with a tripsacoid grass or even with Tripsacum australe may have been responsible. In such a cross, the two types of abscission layer would be counterbalanced as shown by the Flhybrids of corn, Tripsacum and Euchlaena. Furthermore in later generations a.tu-allele of Tripsacum may become homozygous and substitute the wild tunicate factor of corn. The hypothesis of a hybrid origin of cultivated corn is not completely new, but has been discussed already by HARSHBERGER and COLLINS. Our hypothesis differs from that of MANGELSDORF and REEVES who assume that crosses with Tripsacum are responsible only for some features of Central and North American corn. 10) The following arguments give indirects evidence in support of our hypothesis: a) Several characters have been observed in indigenous corn from the central region of South America, which may be interpreted as "tripsacoid". b) Equally "zeoid" characters seem to be present in Tripsacum australe of central South-America. c) A system of unbalanced factors, combined by the in-tergeneric cross, may be responsible for the sterility of the wild type tunicata factor when homozygous, a result of the action of modifiers, brought in from Tripsacum together with the tuallele. d) The hybrid theory may explain satisfactorily the presence of so many lethals and semilethals, responsible for the phenomenon of inbreeding in cultivated corn. It must be emphasized that corn does not possess any efficient mechanism to prevent crossing and which could explain the accumulation of these mutants during the evolutionary process. Teosinte which'has about the same mechanism of sexual reproduction has not accumulated such genes, nor self-sterile plants in spite of their pronounced preference for crossing. 11) The second most important step in domestication must have consisted in transforming a four rowed ear into an ear with many rows. The fusion theory, recently revived byLANGHAM is rejected. What happened evidently, just as in succulent pXants (Cactus) or in cones os Gymnosperms, is that there has been a change in phyllotaxy and a symmetry of longitudinal rows superimposed on the original spiral arrangement. 12) The geographical distribution of indigenous corn in South America has been discussed. So far, we may distinguish three zones. The most primitive corn appears in the central lowlands of what I call the Central Triangle of South America: east of the Andies, south of the Amazone-Basin, Northwest of a line formed by the rivers São Prancisco-Paraná and including the Paraguay-Basin. The uniformity of the types found in this extremely large zone is astonishing (BRIEGER and CUTLER). To the west, there is the well known Andian region, characterized by a large number of extremely diverse types from small pop corn to large Cuszco, from soft starch to modified sweet corn, from large cylindrical ears to small round ears, etc.. The third region extends along the atlantic coast in the east, from the Caribean Sea to the Argentine, and is characterized by Cateto, an orange hard flint corn. The Andean types must have been obtained very early, and undoubtedly are the result of the intense Inca agriculture. The Cateto type may be obtained easily by crosses, for instance, of "São Paulo Pointed Pop" to some orange soft corn of the central region. The relation of these three South American zones to Central and North America are not discussed, and it seems essential first to study the intermediate region of Ecuador, Colombia and Venezuela. The geograprical distribution of chromosome knobs is rapidly discussed; but it seems that no conclusions can be drawn before a large number of Tripsacum species has been analysed.

Mamíferos carnívoros e sua relação com a diversidade de hábitats no Parque Nacional dos Aparados da Serra, sul do Brasil

A survey of carnivore mammals was accomplished in Aparados da Serra National Park from February 1998 to March 2000. The park has 10,250 ha and is considered a biodiversity core area of the Atlantic Forest Biosphere Reserve in the Rio Grande do Sul State, Brazil. The landscape is characterized by relatively well preserved relicts of Araucaria angustifolia (Bertol.) Kuntze forest, grasslands and Atlantic Forest, which have contributed for the survival of endangered carnivore mammals. The National Park was divided in a grid of 16 km² cells using a 1:50,000 scale map. The animals were recorded using indirect methods, by identifying signs (scats, tracks) and direct observation in 2.5 km long and 5 m wide transects, with 10 replicates in each grid cell. Interviews with local people were also used to confirm the animal presence. A total of 13 species was recorded: Procyon cancrivorus (Cuvier, 1798), Pseudalopex gymnocercus (G. Fischer, 1814), Leopardus pardalis (Linnaeus, 1758) and Cerdocyon thous (Linnaeus, 1766) were the most frequent species registered. Nasua nasua (Linnaeus 1766), Herpailurus yaguarondi (Lacépède, 1809), Chrysocyon brachyurus (Illiger, 1815), Eira barbara (Linnaeus, 1758), Leopardus sp., Puma concolor (Linnaeus, 1771), Galictis cuja (Molina, 1782), Conepatus chinga (Molina, 1892) and Lontra longicaudis (Olfers, 1818) showed lower frequencies. The Park presented areas with significant differences (Mantel Test, P< 0.05) in species richness and composition related to habitat classes. Areas with high habitat richness presented high species richness. The Araucaria forest was the habitat that presented the higher carnivore richness. The border areas of the Park are influenced by several environmental degradation factors that could be affecting the distribution of carnivores.

Conservation status assessment of the amphibians and reptiles of Uruguay

The native species of amphibians and reptiles of Uruguay were categorized according to the IUCN Red List criteria. Out of 47 amphibian species, seven are listed as Critically Endangered (CR), five as Endangered (EN), one as Vulnerable (VU), three as Near Threatened (NT), and two as Data Deficient (DD); the remaining species are considered to be Least Concern (LC). Among the 64 species of reptiles evaluated, one is listed as Critically Endangered (CR), seven as Endangered (EN), two as Vulnerable (VU), one as Near Threatened (NT) and seven as Data Deficient (DD); the rest are considered to be Least Concern (LC). The use of these results as an additional criterion in the definition of protected areas in Uruguay will contribute towards the conservation of the aforementioned threatened species and their associated ecosystems.

Primatology in southern Brazil: a transdisciplinary approach to the conservation of the brown-howler-monkey Alouatta guariba clamitans (Primates, Atelidae)

Human interventions in natural environments are the main cause of biodiversity loss worldwide. The situation is not different in southern Brazil, home of five primate species. Although some earlier studies exist, studies on the primates of this region began to be consistently carried out in the 1980s and have continued since then. In addition to important initiatives to study and protect the highly endangered Leontopithecus caissara Lorrini & Persson, 1990 and Brachyteles arachnoides E. Geoffroy, 1806, other species, including locally threatened ones, have been the focus of research, management, and protection initiatives. Since 1993, the urban monkeys program (PMU, Programa Macacos Urbanos) has surveyed the distribution and assessed threats to populations of Alouatta guariba clamitans (Cabrera, 1940) in Porto Alegre and vicinity. PMU has developed conservation strategies on four fronts: (1) scientific research on biology and ecology, providing basic knowledge to support all other activities of the group; (2) conservation education, which emphasizes educational presentations and long-term projects in schools near howler populations, based on the flagship species approach; (3) management, analyzing conflicts involving howlers and human communities, focusing on mitigating these problems and on appropriate relocation of injured or at-risk individuals; and finally, (4) Public Policies aimed at reducing and/or preventing the impact of urban expansion, contributing to create protected areas and to strengthen environmental laws. These different approaches have contributed to protect howler monkey populations over the short term, indicating that working collectively and acting on diversified and interrelated fronts are essential to achieve conservation goals. The synergistic results of these approaches and their relationship to the prospects for primatology in southern Brazil are presented in this review.

Bird communities in three forest types in the Pernambuco Centre of Endemism, Alagoas, Brazil

The Pernambuco Center of Endemism (PCE) in northeastern Brazil is highly fragmented and degraded. Despite its potential conservation importance the bird fauna in this area is still relatively unknown and there are many remnant fragments that have not been systematically surveyed. Here, we report the results of bird surveys in five forest fragments (one pioneer, two ombrophilous and two seasonal). In total, 162 taxa were recorded, 12 of which are endemic to the PCE. The frequency of endangered species was lower than what has been reported in studies from the same area and most of the taxa considered to be at risk of extinction were sub-species of uncertain taxonomic validity. The comparatively low number of endemic/threatened species may be due to the small size of the fragments in the present study - a consequence of the high levels of habitat loss in this region. Analysis of species richness patterns indicates that ombrophilous forest fragments are acting as refuges for those bird species that are most sensitive to environmental degradation.