Geopolymers as infill material for conservation of azulejos

The restoration materials currently used to fill gaps in architectural historical azulejos (e.g. lime or organic resin pastes) usually show serious drawbacks in terms of compatibility, effectiveness and durability. The existing solutions do not fully protect azulejos in outdoor conditions and frequently result in further deterioration. Geopolymers can be a potential solution for azulejo lacunae infill given the chemical-mineralogical similitude to the ceramic body, and also the durability and versatile range of physical properties that can be obtained through the manipulation of their formulation and curing conditions. This work presents and discusses the viability of the use of geopolymeric pastes to fill lacunae in azulejos or to act as “cold” cast ceramic tile surrogates reproducing missing azulejo fragments. The formulation of geopolymers, namely the type of activators, the aluminosilicate source, the amount of water (to meet adequate workability requirements) and curing conditions were studied. The need for post-curing desalination was also considered envisaging their application in the restoration of outdoor architectural historical azulejos frequently exposed to adverse environmental conditions. The possible advantages and disadvantages of the use of geopolymers in the conservation of azulejos are also discussed. Several techniques were used to study the chemical and physical behavior of geopolymers, namely FT-IR, XRD, MIP, SEM-EDS, WDXRF, electrical conductivity, open porosity, bending strength, adhesion strength, water vapour permeability, thermal expansion and hydric expansion. The results indicate that geopolymers are a promising material for restoration of azulejos, exhibiting some properties, such as adhesion to the ceramic substrate, higher than inorganic materials used nowadays, such as aerial lime based pastes.

Improvement of a photoautotrophic chassis robustness for synthetic biology applications

Cyanobacteria are photoautotrophic microorganisms with great potential for the biotechnological industry due to their low nutrient requirements, photosynthetic capacities and metabolic plasticity. In biotechnology, the energy sector is one of the main targets for their utilization, especially to produce the so called third generation biofuels, which are regarded as one of the best replacements for petroleum-based fuels. Although, several issues could be solved, others arise from the use of cyanobacteria, namely the need for high amounts of freshwater and contamination/predation by other microorganisms that affect cultivation efficiencies. The cultivation of cyanobacteria in seawater could solve this issue, since it has a very stable and rich chemical composition. Among cyanobacteria, the model microorganism Synechocystis sp. PCC 6803 is one of the most studied with its genome fully sequenced and genomic, transcriptomic and proteomic data available to better predict its phenotypic behaviors/characteristics. Despite suitable for genetic engineering and implementation as a microbial cell factory, Synechocystis’ growth rate is negatively affected by increasing salinity levels. Therefore, it is important to improve. To achieve this, several strategies involving the constitutive overexpression of the native genes encoding the proteins involved in the production of the compatible solute glucosylglycerol were implemented, following synthetic biology principles. A preliminary transcription analysis of selected mutants revealed that the assembled synthetic devices are functional at the transcriptional level. However, under different salinities, the mutants did not show improved robustness to salinity in terms of growth, compared with the wild-type. Nevertheless, some mutants carrying synthetic devices appear to have a better physiological response under seawater’s NaCl concentration than in 0% (w/v) NaCl.

Going Public: Conservation of Contemporary Artworks. Between Backstage and Frontstage in Contemporary Art Museums

In museum studies and history of art, what happens behind the scenes of museums stays relatively unseen and unspoken about. In the arts, generally speaking, what is dismissed as irrelevant (e.g. the realm of practices) is deliberately detached from what is thought to really matter; theory, discourse, content and meaning. Up till recently, backstage activities such as conservation practices are merely discussed among specialists and museum professionals. Only the outcomes of these discussions are sometimes – if at all – explicitly communicated to a larger public. Studies into the practices of contemporary art conservation however show that practices behind the scenes play an important role in the perpetuation of these artworks. What happens behind the scenes in terms of conservation has, in several ways, important effects on the ongoing life of these artworks in a museum context. Conservation practices, I argue, should therefore become a necessary part of museum studies and history of art. How can the working practices of conservators become more visible and transparent to a diversity of audiences, including researchers? And what does this mean in terms of research methodology?

The perfect paint in modern art conservation: a comparative study of 21st century vinyl emulsions

Contemporary painting places, and will continue to place, several questions about its meaning, its chemical nature, its durability and the best way to preserve it. This research aims at putting together comprehensive data on vinyl based paints, including their components, their properties, their aging behavior and their response to selected cleaning products. In this project degradation mechanisms of vinyl binders and formulations used in the 20th and 21st century were studied. Stability over time of selected vinyl polymers was assessed through natural indoor and artificially aging. The objective was to enhance knowledge and understanding of vinyl emulsion formulations and their performance over time. Overall conservation state of pictorial layers namely, adhesion, cohesion and discoloration of selected case studies from the Portuguese artist Julião Sarmento (b.1948) was correlated with the observed molecular level changes studied in laboratory experiments. Sarmento’s paintings were chosen due to conservation concerns (discoloration) on some of his works from the 90’s. Besides, research was carried out to start increasing the knowledge of what can be expected of PVAc based paints in terms of response to conservation treatments namely, surface cleaning. Artificial aging showed that the most recent formulations which are based on a poly(vinyl acetate), poly(vinyl chloride) and polyethylene terpolymer are less stable when compared to some homopolymer formulations. From the four pigments studied, titanium dioxide rutile and a carbon based black proved to be stabilizers for both types of polymer. The mixture lithopone plus calcium carbonate has showed to have a photocatalytic effect on the binders. The studied paintings showed to be in an overall good state of conservation except for the paintings created in the 90’s with white glue and a mixture of white lithoponeand calcium carbonate. Discoloration of this white paint seems to be irreversible and ongoing and is still a major concern. The disapearance of the plasticizer was the only change detected. The current works created by Sarmento are expected to be more stable as they were painted using the rutile titanium dioxide. Immersion/cleaning tests showed that vinyl based paints can be susceptible to water and organic solvents like ethanol as some evidences point to the removal/diffusion of additives from the paint. The observations made point to the need to further proceed in this research field.

Biology of amazonian mosquitoes. III. Esterase isozymes in Anopheles darlingi.

Six esterase isozymes were studied during the development of Anopheles darlingi by using polyacrylamide gel electrophoresis and two different substrates, a-naphthylcelate and a-naphthylpropionate. Esterases 5 and 6'were detected in all developmental stages esterases 1 and 2 were more intensively stained if larvae, while esterases 3 and 4 were better visualized in pupae and adults. Strong differences in intensity of some of the isozymes were observed during the pupal stage.Four out of the six isozymes showed variation in the electrophoretic mobility. Esterase-2 was choosed for genetic studies, because was the best stained isozyme in the gels. Two codominant alleles {Est2*S and Est2*F) code for this polymorphic system, with the Est*S frequency equal to 0.521. Phenotypic distribution is in agreement with hardy-Weinberg expectations.

Reproductive biology of Bellucia (MELASTOMATACEAE)

The reproductive biology of five of the seven species of Bellucia (Melastomataceae), a genus of shrubs and small trees, was investigated in Amazonia. Sucessful fruit-set by Bellucia requires floral visitation by bees. The flowers are produced continuously all year, and are visited by a wide variety of female bees, the principal pollinators being Xylocopa, Centris, Ptilotopus, Epicharis, Eulaema, Bombus, and Oxaea. The floral attractants are color and the odor produced by the pollen, stamens, and petals; the reqard is pollen. Three species of Bellucia are self-incompatible. Indiscriminate visitor behavior and lack of phenological, morphological, or genetic barriers lead to hybridization between sympatric species of Bellucia, and no more than two species occupy the same habitat at anu one locality. Bellucia produces berries with numerous small seeds, and is dispersed by birds, bats, monkeys, tapirs, turtles, and ants. Seedling establisment requires full sunlight, and occurs on a variety of soil types. The reproductive strategy is interpreted as that of a pioneer species.

Description of behavioral patterns of Podocnemis erythrocephala (Spix, 1824) (Testudines: Podocnemididae) (Red-headed river turtle) in captivity, Manaus, Amazonas, Brazil

The biology and ecology of South American turtles is still poorly known, particularly, for the Brazilian species. Laboratory studies are essential to understand the life cycles of aquatic turtles species and to help in formulating management plans for their conservation. As a contribution to the knowledge of Podocnemis erythrocephala species, we give a description of its species-typical behaviors, categorized as: maintenance, locomotion, feeding, agonistic and reproduction, based on captives observations of four pairs of turtles in an aquarium in Manaus, Brazil. Similarities and differences with the repertoires of other turtle species are discussed, concluding that turtles have much more complex adaptative strategies and social life than was believed.

The influence of the conservation status and changes in the rainfall regime on forest-savanna mosaic dynamics in Northern Brazilian Amazonia

Coupled carbon/climate models are predicting changes in Amazon carbon and water cycles for the near future, with conversion of forest into savanna-like vegetation. However, empirical data to support these models are still scarce for Amazon. Facing this scenario, we investigated whether conservation status and changes in rainfall regime have influenced the forest-savanna mosaic over 20 years, from 1986 to 2006, in a transitional area in Northern Amazonia. By applying a spectral linear mixture model to a Landsat-5-TM time series, we identified protected savanna enclaves within a strictly protected nature reserve (Maracá Ecological Station - MES) and non-protected forest islands at its outskirts and compared their areas among 1986/1994/2006. The protected savanna enclaves decreased 26% in the 20-years period at an average rate of 0.131 ha year-1, with a greater reduction rate observed during times of higher precipitation, whereas the non-protected forest islands remained stable throughout the period of study, balancing the encroachment of forests into the savanna during humid periods and savannization during reduced rainfall periods. Thus, keeping favorable climate conditions, the MES conservation status would continue to favor the forest encroachment upon savanna, while the non-protected outskirt areas would remain resilient to disturbance regimes. However, if the increases in the frequency of dry periods predicted by climate models for this region are confirmed, future changes in extension and directions of forest limits will be affected, disrupting ecological services as carbon storage and the maintenance of local biodiversity.

Reproductive biology of Amasonia obovata Gleason (Laminaceae)

Floral mechanisms that ensure seed production via autogamy are more likely to occur in species growing in environments where pollination is scarce. Amasonia obovata was studied in the State of Mato Grosso-Brazil, from 2009 to 2012, to analyze the morphological and reproductive characteristics, aside from investigating the association of the reproductive success with the pollinator frequency and identity. The flowering and fruiting of A. obovata was concentrated in a period of five months during the rainy season. The dichogamy in flowers of A. obovata is not clearly defined, since the sexual functions were overlapped in the male and female phases. The species is self-compatible and not apomictic. The fruiting percentage obtained by hand self-pollination did not differ from cross-breeding (F = 0.74, P =0.39). In the observations from 2010 to 2012, a hummingbird (Thalurania furcata) legitimate visited 20-100% of the flowers in the male and female phases on different A. obovata plants. Due to the high frequency, this hummingbird was considered the single potential pollinator of the species. These findings show that a limited availability of pollinators may select for floral traits and plant mating strategies that lead to a system of self-fertilization.

Reproductive biology of Ameerega trivittata (Anura: Dendrobatidae) in an area of terra firme forest in eastern Amazonia

The reproductive success of tropical amphibians is influenced by factors such as body size and the characteristics of breeding sites. Data on reproductive biology are important for the understanding of population dynamics and the maintenance of species. The objectives of the present study were to examine the abundance of Ameerega trivittata, analyze the use of microhabitats by calling males and the snout-vent length (SVL) of breeding males and females, the number of tadpoles carried by the males and mature oocytes in the females, as well as the relationship between the SVL of the female and both the number and mean size of the mature oocytes found in the ovaries. Three field trips were conducted between January and September, 2009. A total of 31 plots, with a mean area of 2.3 ha, were surveyed, resulting in records of 235 individuals, with a mean density of 3.26 individuals per hectare. Overall, 66.1% of the individuals sighted were located in the leaf litter, while 17.4% were perched on decaying tree trunks on the forest floor, 15.7% on the aerial roots of Cecropia trees, and 0.8% on lianas. Males were observed transporting a mean of 10.8 tadpoles on their backs. A significant correlation was found between the size of the females and the mean diameter of the oocytes. New data were collected on the size of oocytes and no pattern was found in the type of perches used by calling males of the different Ameerega species.

Reproductive biology of Macrobrachium surinamicum (Decapoda: Palaemonidae) in the Amazon River mouth

Macrobrachium surinamicum is an indigenous prawn distributed from the lower Amazon and Tocantins river basins to Venezuela in the Orinoco Delta region. It is common bycatch fauna of Macrobrachium amazonicum artisan fishing in the states of Pará and Amapá. The aim of this study was to investigate aspects on reproductive biology (reproductive period, size of sexual maturity population, fecundity, reproductive output and recruitment) of M. surinamicum from four important areas to artisanal prawn fishing located at the Amazon River mouth (Amapá and Pará). The specimens were captured using 20 handcrafted traps called "matapi". A number of 675 prawns were captured, 258 males, 409 females and eight juveniles, resulting in 1:1.6 (Male: Female) sex ratio. The reproductive peak period occurred from March to July, coinciding with the higher rainfall period. The juvenile prawn occurred only in May and July. Total length of egg-bearing females ranged from 12.12 to 38.30 mm, with mean female length at first maturity (L50) of 23.7 mm. Fecundity increased with prawn size and varied between 174 and 1780 eggs per female. Mean egg volume increased gradually from 0.031 (Stage I) to 0.060 mm³ (Stage III) during embryogenesis. Macrobrachium surinamicum depends on brackish water to complete the larval development. Irrespective of female size, reproductive output of M. surinamicum varied between 4.3 % and 35.5 % of their body weight for egg production. The knowledge of the reproductive biology reported in the present study is an important tool to define strategies to preserve M. surinamicum in Amazon River mouth.

Systems biology approaches for the design of novel Saccharomyces cerevisiae winemaking strains for enhanced flavour compounds synthesis

Tese de Doutoramento em Biologia Ambiental e Molecular

Synthetic biology approaches to engineer new pathways for the production of plant secondary metabolites

Secondary metabolites from plants are important sources of high-value chemicals, many of them being pharmacologically active. These metabolites are commonly isolated through inefficient extractions from natural biological sources and are often difficult to synthesize chemically. Therefore, their production using engineered organisms has lately attracted an increased attention. Curcuminoids, an example of such metabolites, are produced in Curcuma longa and exhibit anti-cancer and anti-inflammatory activities. Herein we report the construction of an artificial biosynthetic pathway for the curcuminoids production in Escherichia coli. Different 4-coumaroyl-CoA ligases (4CL) and polyketide synthases (diketide-CoA synthase (DCS), curcumin synthase (CURS) and curcuminoid synthase) were tested. The highest curcumin production (70 mg/L) was obtained by feeding ferulic acid and with the Arabidopsis thaliana 4CL1 and C. longa DCS and CURS enzymes. Other curcuminoids (bisdemethoxy- and demethoxycurcumin) were also produced by feeding coumaric acid or a mixture of coumaric and ferulic acids, respectively. Curcuminoids, including curcumin, were also produced from tyrosine through the caffeic acid pathway. To produce caffeic acid, tyrosine ammonia lyase and 4-coumarate 3-hydroxylase were used. Caffeoyl-CoA O-methyltransferase was used to convert caffeoyl-CoA to feruloyl-CoA. This pathway represents an improvement of the curcuminoids heterologous production. The construction of this pathway in another model organism is being considered, as well as the introduction of alternative enzymes.