O Turismo e o Recreio Náutico na Região Autónoma da Madeira

Dissertação apresentada para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Gestão do Território

Genetic parameters and mapping quantitative trait loci associated with tibia traits in broilers.

Abstract: Selection among broilers for performance traits is resulting in locomotion problems and bone disorders, once skeletal structure is not strong enough to support body weight in broilers with high growth rates. In this study, genetic parameters were estimated for body weight at 42 days of age (BW42), and tibia traits (length, width, and weight) in a population of broiler chickens. Quantitative trait loci (QTL) were identified for tibia traits to expand our knowledge of the genetic architecture of the broiler population. Genetic correlations ranged from 0.56 +/- 0.18 (between tibia length and BW42) to 0.89 +/- 0.06 (between tibia width and weight), suggesting that these traits are either controlled by pleiotropic genes or by genes that are in linkage disequilibrium. For QTL mapping, the genome was scanned with 127 microsatellites, representing a coverage of 2630 cM. Eight QTL were mapped on Gallus gallus chromosomes (GGA): GGA1, GGA4, GGA6, GGA13, and GGA24. The QTL regions for tibia length and weight were mapped on GGA1, between LEI0079 and MCW145 markers. The gene DACH1 is located in this region; this gene acts to form the apical ectodermal ridge, responsible for limb development. Body weight at 42 days of age was included in the model as a covariate for selection effect of bone traits. Two QTL were found for tibia weight on GGA2 and GGA4, and one for tibia width on GGA3. Information originating from these QTL will assist in the search for candidate genes for these bone traits in future studies.

Assessment of hydrogen sulfide removal from biogas using a field scale anoxic biotrickling filter.

Hydrogen sulphide is one of the most toxic and corrosive compound present in swine-derived biogas streams.In this study, afield scale biotrickling filter for the removal of hydrogen sulfide was investigated.A Biofilter packed with supporting biofilm materials was fed continuously with a proprietary nutrient solution and operatedfor over 73days. The system has been operating with a H2S inlet concentrations ranging from 1,000to 3,000 ppm.Significant removal efficiencies >95% was demonstrated. pH of the stock feeding solution decreased from 6.2 to as low as 3.5within couple days.The resulting drop in pH provided circumstantial evidence to support biological H2 Soxidation to sulphuric acid by sulfide-oxidizers. Sulfur precipitation was also observed to occur. The results suggested that H2S removal from biogas stream can be efficiently achieved using portable, low cost and maintenance free biotrickling filters.

Use of swine waste-derived biogas to enhance microalgae productivity.

The effects of swine wastewater-derived biogas on microalgae productivity were determined. Experiments were conducted in a closed photobioreactor containing digestate effluent as culturing media and biogas in the headspaceas source of CO2. Experiments were carried out under mixothrophic and autothrophic conditions. Results showed that autotrophic growth rate (0.6 d-1)was twofoldfaster than mixotrophic. Frequent reinjections of biogas containing up to 2,000 ppm of hydrogen sulfide was not inhibitory to microalgae growth. The rapid removal of H2S in the system suggests photobioreactors can be an interesting alternative to biogas purification. A model to estimate microalgae productivity based on the amount of available CO2, inorganic and organic carbon was developedand showed good data fit correlation (r²= 0.99).

Determination of microalgae cellular composition after phycoremediation of swine wastewaters.

Phycoremediation of swine wastewaters has been widely reported as an attractive tertiary treatment system, that effectively removes the excessive nutrient loadswhilst offering a valuable source of feedstock biomass. Digestate from an upflow anaerobic sludge blanket (UASB, 6%v/v) and a nitrification reactor (NR; 50% v/v) were used as culturing media to microalgae. Experiments were carried out in lab scale photobioreactors (PBRs) using a consortia of Chlorella and Scenedesmus. Ammonia (44 to 90%) and phosphorus (77%) were efficiently removed from both effluents tested after 4 days. Microalgae biomass harvested from the UASB effluent showed 57, 34 and 1% of proteins, carbohydrates and lipids, respectively. Comparatively, the cellular composition of microalgae grown on NR effluent had lower protein (43%) but higher carbohydrate (42%) contents. Negligible difference in lipid fraction was observed independently of the effluents tested. The results suggest that the biomass harvested from phycoremediation of swine wastewaters can offer a valuable protein and carbohydrate feedstock for nutritional and biotechnological applications.

Biodiversity and quantification of functional bacteria in deammonification process.

2015

The influence of use additives on nitrous oxide emission during swine slurry composting.

use of additives (Mg/P and nitrification inhibitor dicyandiamide - DCD), on nitrous oxide emission during swine slurry composting. The experiment was run in duplicate; the gas was monitored for 30 days in different treatments (control, DCD, Mg/P and DCD + Mg/P). Nitrous oxide emissions rate (mg of N2O-N.day-1) and the accumulated emissions were calculated to compare the treatments. Results has shown that emissions of N-N2O were reduced by approximately 70, 46 and 96% through the additions of DCD, MgCl2.6H2O + H3PO4 and both additives, respectively, compared to the control. Keywords Composting; swine slurry; additives; nitrous

Avian infectious bronchitis virus (IBV): differential pathological and immunological profiles of two Brazilian variants.

2015

Apparent metabolizable energy from microalgae spirulina maxima for broiler chickens.

2016

Broiler chicken ileal apparent digestibility of calcium and phosphorus from microalgae spirulina maxima.

2016

Genome-wide association studies reveal genomic windows and candidate genes related to fat deposition in chickens.

2016

Identification of CNVs on the genome of Brazilian chicken lines.

2016

Haida Totem Poles: Reflections of a Society

The Haida were a First Nations group located on the Northwest Coast of Canada. They were exceptional wood carvers that produced various types of works, the most monumental of which were their totem poles. This dissertation analyses 26 of these open architectural structures, each of which belonged to one of the following five types: frontal, mortuary, memorial, house post, or corner post. Of the representations found on poles, 28 different figures were identified. However, individual poles were found to contain between one and fourteen different figures with frontal poles generally featuring the most. The predominant figures on the inventoried poles proved to be birds, humans, and bears. An iconographic structure of a tripartite character was detected that reflects the religious ideology of the Haida population with birds being featured at the top of the poles, humans in the middle, and bears at the lowest point. It also suggests the possible transition from a hunter-gatherer economy to a food producing economy.***********************************************************************************Os Haida foram um grupo das Primeiras Nações que habitaram a costa Noroeste do Canadá. Eram excepcionais escultores de madeira, tendo produzido variados tipos de artefactos, dos quais os totem poles eram os mais monumentais. Na presente dissertação são analisadas 26 dessas estruturas arquitectónicas de exterior, sendo que cada uma delas pertenceria a um dos seguintes cinco tipos: frontal, funerário, memorial, travemestra, ou postes de esquina. Das representações encontradas nos postes, foram individualizadas 28 figuras. No entanto, em postes isolados foi possível identificar entre uma e catorze figuras distintas, sendo os postes frontais os que apresentavam, geralmente, o maior número. A análise destes postes permitiu detectar uma estrutura iconográfica tripartida que reflectia a ideologia religiosa das populações Haida, representando-se as aves no topo, os humanos a meio, e os ursos na base dos postes. Por outro lado, foi possível sugerir, para estas populações, a transição de uma economia baseada na caça-recolecção para uma economia de produção alimentar.

Primary cilia and the regulation of hedgehog signaling: link to cardiac development

RESUMO: As células eucarióticas evoluíram um sistema de sinalização complexo que lhes permite responder aos sinais extracelulares e intracelulares. Desta forma, as vias de sinalização são essenciais para a sobrevivência da célula e do organismo, uma vez que regulam processos fundamentais, tais como o desenvolvimento, o crescimento, a imunidade, e a homeostase dos tecidos. A via de transdução de sinal Hedgehog (Hh) envolve o receptor Patched1 (Ptch1), que tem um efeito inibidor sobre a proteína Smoothened (Smo) na ausência dos seus ligandos, as proteínas Sonic hedgehog (Shh). Estas proteínas são reguladores fundamentais do desenvolvimento embrionário, como ilustrado pelas malformações drásticas observadas em embriões humanos e de murganho com perturbações da transdução de sinal da via Hh e que incluem polidactilia, defeitos craniofaciais e malformações ósseas. Igualmente importantes são as consequências da ativação inapropriada da via de sinalização Hh na formação de tumores. Curiosamente, os componentes desta via localizam-se nos cílios primários. Além disso, demonstrou-se que esta localização é crucial para a sinalização através da via Hh. Na presença dos ligandos, Ptch1 é internalizado e destinado a degradação ou sequestrado num compartimento da célula de onde não pode desempenhar o seu papel inibitório. A proteína Arl13b é uma pequena GTPase pertencente à família Arf/Arl da superfamília Ras de pequenas GTPases e foi implicada no síndrome de Joubert, uma ciliopatia caracterizada por ataxia congénita cerebelar, hipotonia, atrso mental e cardiopatia congénita. Murganhos deficientes para Arl13b, chamado hennin (hnn) morrem morrem prematuramente ao dia 13,5 de gestação (E13,5) e exibem anomalias morfológicas nos cílios que levam à interrupção da sinalização Hh. Além disso, a Arl13b está diretamente envolvida na regulação da via Hh, controlando a localização de vários componentes desta via nos cílios primários. Neste trabalho, mostramos que a Arl13b se localiza em circular dorsal ruffles (CDRs), que são estruturas de actina envolvidas em macropinocitose e internalização de recetores, e que regula a sua formação. Além disso, aprofundámos o conhecimento do processo de ativação da via de sinalização Hh, mostrando que as CDRs sequestram seletivamente e internalizam o recetor Ptch1. As CDRs formam-se minutos após ativação da via por ligandos Shh ou pelo agonista de Smo SAG e continuam a ser formadas a partir daí, sugerindo uma indução contínua da reorganização do citoesqueleto de actina quando a via está ativada. Observámos ainda que a inibição da formação de CDRs através do silenciamento de WAVE1, uma proteína necessária para a formação destas estruturas, resulta na diminuição da ativação da via de sinalização Hh. Além disso, o bloqueio da macropinocitose, que se segue ao fecho das CDRs, através do silenciamento de uma proteína necessária para a cisão de macropinossomas, nomeadamente a proteína BARS, tem um efeito semelhante. Estes resultados sugerem que as CDRs e a macropinocitose são necessárias para a ativação da via de sinalização Hh e indicam que esta via de internalização controla os níveis de sinal Hh. Durante o desenvolvimento, as células proliferativas dependem do cílio primário para a transdução de várias vias de sinalização. A via Hh induz a diferenciação do músculo cardíaco. Por conseguinte, os murganhos deficientes na via de sinalização Hh exibem uma variedade de defeitos de lateralidade, incluindo alteração do looping do coração, como pode ser visto em murganhos deficientes para Arl13b. Por conseguinte, investigámos o papel da Arl13b no desenvolvimento do coração. Mostramos que a Arl13b é altamente expressa no coração de embriões de murganho e de murganhos adultos ao nível do mRNA e da proteína. Além disso, o perfil de distribuição da Arl13b no coração segue o dos cílios primários, que são essenciais para o desenvolvimento cardíaco. Corações de murganhos hnn no estadio E12,5 mostram um canal átrio-ventricular aberto, espessamento da camada compacta ventricular e aumento do índice mitótico no ventrículo esquerdo. Além disso, um atraso de 1 a 2 dias no desenvolvimento é observado em corações de murganhos hnn, quando comparados com controlos selvagens no estadio E13,5. Assim, estes resultados sugerem que a Arl13b é necessária para o desenvolvimento embrionário do coração e que defeitos cardíacos podem contribuir para a letalidade embrionária de murganhos hnn. Em suma, foi estabelecido um novo mecanismo para a regulação dos níveis de superfície do recetor Ptch1, que envolve a remodelação do citoesqueleto de actina e a formação de CDRs após a ativação da via de sinalização Hh. Este mecanismo permite um feedback negativo que evita a repressão excessiva da via através da remoção de Ptch1 da superfície da célula. Além disso, determinou-se que uma mutação de perda de função na Arl13b causa defeitos cardíacos durante o desenvolvimento, possivelmente relacionados com a associação dos defeitos em cílios primários e na sinalização Hh, existentes em murganhos deficientes para Arl13b. A via de sinalização Hh tem tido um papel central entre as vias de sinalização, uma vez que a sua regulação é crucial para o funcionamento apropriada da célula. Assim, a descoberta de um novo mecanismo de tráfego através de macropinocitose e CDRs que controla a ativação e repressão da via de sinalização Hh traz novas perspetivas de como esta via pode ser regulada e pode ainda conduzir à identificação de novos alvos e estratégias terapêuticas. --------------------ABSTRACT: Eukaryotic cells have evolved a complex signaling system that allows them to respond to extracellular and intracellular cues. Signaling pathways are essential for cell and organism survival, since they regulate fundamental processes such as development, growth, immunity, and tissue homeostasis. The Hedgehog (Hh) pathway of signal transduction involves the receptor Patched1 (Ptch1), which has an inhibitory effect on Smoothened (Smo) in the absence of its ligands, the Sonic hedgehog (Shh) proteins. These proteins are fundamental regulators of embryonic development, as illustrated by the dramatic malformations seen in human and mouse embryos with perturbed Hh signal transduction that include polydactyly, craniofacial defects and skeletal malformations. Equally important are the consequences of inappropriate activation of the Hh signaling response in tumor formation. Interestingly, the components of this pathway localize to primary cilia. Moreover, it has been shown that this localization is crucial for Hh signaling. However, in the presence of the ligands, Ptch1 is internalized and destined for degradation or sequestered in a cell compartment where it no longer can play its inhibitory role. ADP-ribosylation factor-like (Arl) 13b, a small GTPase belonging to Arf/Arl family of the Ras superfamily of small GTPases has been implicated in Joubert syndrome, a ciliopathy characterized by congenital cerebellar ataxia, hypotonia, intellectual disability and congenital heart disease. Arl13b-deficient mice, called hennin (hnn) die at embryonic day 13.5 (E13.5) and display morphological abnormalities in primary cilia that lead to the disruption of Hh signaling. Furthermore, Arl13b is directly involved in the regulation of Hh signaling by controlling the localization of several components of this pathway to primary cilia. Here, we show that Arl13b localizes to and regulates the formation of circular dorsal rufles (CDRs), which are actin-basedstructures known to be involved in macropinocytosis and receptor internalization. Additionally, we extended the knowledge of the Hh signaling activation process by showing that CDRs selectively sequester and internalize Ptch1 receptors. CDRs are formed minutes after Hh activation by Shh ligands or the Smo agonist SAG and keep being formed thereafter, suggesting a continuous induction of actin reorganization when the pathway is switched on. Importantly, we observed that disruption of CDRs by silencing WAVE1, a protein required for CDR formation, results in down-regulation of Hh signaling activation. Moreover, the blockade of macropinocytosis, which follows CDR closure, through silencing of a protein necessary for the fission of macropinosomes, namely BARS has a similar effect. These results suggest that CDRs and macropinocytosis are necessary for activation of Hh signaling and indicate that this pathway of internalization controls Hh signal levels. During development, proliferating cells rely on the primary cilium for the transduction of several signaling pathways. Hh induces the differentiation of cardiac muscle. Accordingly, Hh-deficient mice display a variety of laterality defects, including alteration of heart looping, as seen in Arl13b-deficient mice. Therefore, we investigated the role of Arl13b in heart development. We show that Arl13b is highly expressed in the heart of both embryonic and adult mice at mRNA and protein levels. Also, Arl13b localization profile mimics that of primary cilia, which have been shown to be essential to early heart development. E12.5 hnn hearts show an open atrioventricular channel, increased thickening of the ventricular compact layer and increased mitotic index in the left ventricle. Moreover, a delay of 1 to 2 days in development is observed in hnn hearts, when compared to wild-type controls at E13.5. Hence, these results suggest that Arl13b is necessary for embryonic heart development and that cardiac defects might contribute to the embryonic lethality of hnn mice. Altogether, we established a novel mechanism for the regulation of Ptch1 surface levels, involving cytoskeleton remodeling and CDR formation upon Hh signaling activation. This mechanism allows a negative feedback loop that prevents excessive repression of the pathway by removing Ptch1 from the cell surface. Additionally, we determined that the Arl13b loss-offunction mutation causes cardiac defects during development, possibly related to the associated ciliary and Hh signaling defects found in Arl13b-deficient mice. Hh signaling has taken a center stage among the signaling pathways since its regulation is crucial for the appropriate output and function of the cell. Hence, the finding of a novel trafficking mechanism through CDRs and macropinocytosis that controls Hh signaling activation and repression brings new insights to how this pathway can be regulated and can lead to the discovery of novel therapeutic targets and strategies.

Informe preliminar del Crucero de Otoño 1966 (Cabo Blanco - Ilo)

Describe la investigación realizada por el Crucero bio-oceanográfico 6605 del B.A.P. Unanue, correspondiente a la estación Otoño del año 1966, efectuada en dos etapas. El estudio obtuvo información sobre el desove de la anchoveta y otros peces mediante la recolección del plancton con red Hensen, así como se investigó la presencia de cardúmenes de peces por medio de ecosonda y sonar, se efectuaron también observaciones de aves y mamíferos marinos.