Estudios en distintos ambientes, como el marino o el hipersalino, han demostrado que la combinación de metodologías y la integración de resultados tienen el potencial de aportar nueva información que las técnicas no pueden proveer de manera individual. En esta tesis se han utilizado tres cepas de S. ruber utilizadas para la descripción de la especie para el aislamiento de virus. Un total de 8 virus de aguas de diferentes ambientes hipersalinos se han aislado y seleccionado para realizar una descripción fenotípica, genética y ecogenómica en profundidad. La descripción se ha combinado con un estudio mediante metagenómica vírica dirigida para obtener información acerca de la dinámica poblacional de los virus de S. ruber. Para ahondar en el estudio de la virosfera de Haloquadratum hemos usado diferentes estrategias combinando metodologías dependientes e independientes cultivo. Se realizó un experimento de metagenómica dirigida mediante la incubación de los virus de una muestra de agua hipersalina con dos cepas de Hqr. walsbyi. El metaviroma obtenido se estudió mediante ensamblaje de secuencias y se utilizó para hibridación con un microarray vírico previamente construido. Los genomas halovíricos con señal positiva a la hibridación fueron también analizados y sus secuencias sirvieron como plantilla para la realización de experimentos de phageFISH, en un trabajo independiente a esta tesis, que confirmó su hospedador. Por último se combinó la técnica de single-cell genomics con la hibridación en virochips para detectar células de Haloquadratum infectadas. / Tesis financiada por los proyectos del Plan Nacional CGL2012-39627-C03-01 y CGL2015-66686-C3-3-P, del Ministerio de Economía y Competitividad.

Nature uses inorganic elements for biological processes based on the useful chemistry, abundance, and availability of each metal. Transition metals are critical in the biogeochemical cycling of essential elements and the bioinorganic chemistry of organisms. Hydrolysis-prone metals such as iron and titanium are abundant on Earth but are mostly insoluble in oxic aqueous environments. Nearly every organism requires iron for survival, therefore Nature evolved to stabilize iron from hydrolysis and hydrolytic precipitation through protein and small molecule mechanisms. Like iron, titanium primarily exists as insoluble mineral oxides and is second only to iron as the most abundant transition metal in the Earth’s crust. Despite the reputation as an inert and insoluble metal, titanium can be solubilized and made bioavailable through by chemical and biological weathering. Currently there is no known native role for titanium, however it is quite bioactive. As a stronger Lewis acid, titanium can compete with iron in binding to biomolecules and proteins. It is of interest to investigate the interactions between hydrolysis-prone metals and biological systems, from whole cell organisms to proteins and small molecules. The non-pathogenic bacterium Rhodococcus ruber GIN-1 was isolated for its ability to strongly adhere to titanium dioxide (TiO2) over other metal oxides, providing an opportunity to study the interactions between whole bacterial cells and metal oxides. The GIN-1 strain incorporates Ti(IV) ions into its biomass after adherence to anatase, rutile, and a mixture of the two morphologies. Six metals were quantitated in TiO2-exposed and control (unexposed) cells by inductively coupled plasma optical emission spectroscopy. The exposure to TiO2 caused a significant uptake of titanium with concomitant loss of iron, zinc, and possibly manganese. A collaborative project with the Strongin laboratory at Temple University works to develop stable, biomaterial photocatalysts for environment remediation of toxic inorganic contaminants. Ferritins are a class of proteins that mineralizes and stores iron as a non- toxic ferrihydrite nanoparticle. These proteins can be photoactivated with ultraviolet light to release iron from its core to remediate environmental contaminants. Ferritin can be sensitized with plasmonic gold nanoparticles to extend the photoactivity of the catalyst to the visible spectrum. Work in this thesis highlights the contribution to this collaboration from the Valentine laboratory, included the expression and purification of proteins in E. coli (human H-chain ferritin, human L-chain ferritin, and bacterial DNA protection from starved cells protein), mutation of proteins to improve sensitization of catalyst, and biomineralization with iron and titanium. The trafficking of hydrolysis prone metals is vital for the survival of nearly every organism. Iron transport proteins such as transferrins are studied to understand how nature utilizes a difficult essential metal across the domains of life. Most transferrins have two homologous lobes and are believed to have evolved from a gene duplication of a monolobal transferrin. The ascidian Ciona intestinalis has genes for both a bilobal and monolobal transferrin. Nicatransferrin (nicaTf), the monolobal transferrin from C. intestinalis, is a primitive protein that may provide insight on the evolution of transferrins in higher organisms. It is advantageous to use E. coli expression systems to produce recombinant proteins, however protein misfolding and aggregation can be a concern. To improve expression of nicaTf in E. coli, codon optimization and disulfide bonded protein expression were used. Finally, siderophores are small, high affinity iron-chelating molecules secreted from lower organisms that scavenge iron in iron-limiting conditions. R. ruber GIN-1 and R. ruber DSM 43338 strains both secrete siderophores in artificial seawater media. There are several siderophores identified from Rhodococcus species, however none have been reported from any R. ruber strain. A new siderophore was isolated and preliminary work has been done to purify and characterize the molecule. Understanding the siderophore- metal ion interactions may help elucidate the mechanism of how R. ruber cells obtain titanium from the metal-oxide particles. / Chemistry

O objetivo do presente estudo foi investigar a resposta da assembleia de foraminíferos planctônicos frente às variações paleoceanográficas superficiais na porção oeste do Atlântico Sul, o qual desempenha um papel fundamental na Célula de Revolvimento Meridional do Atlântico (AMOC, Meridional Overturning Circulation). Para esta finalidade, foi analisado um registro sedimentar marinho contínuo desde o Último Máximo Glacial até o presente, com base na assembleia de foraminíferos planctônicos, isótopos de oxigênio e na razão Mg/Ca. Adicionalmente, foram avaliadas as variações na ocorrência e na geoquímica dos dois morfotipos de G. ruber (branca), principal espécie utilizada em estudos paleoceanográficos de regiões tropicais e subtropicais. Os resultados mostraram que as diferenças geoquímicas e de abundância relativa dos morfotipos de G. ruber branca evidenciam dois grupos de espécimes ecologicamente distintos, confirmando que o morfotipo sensu stricto calcifica em menores profundidades com relação ao morfotipo sensu lato e que reconstituições paleoceanográficas baseadas no uso não-seletivo dos morfotipos poderiam ser tendenciosas. A Análise Fatorial realizada nos dados de abundância relativa dos foraminíferos planctônicos identificou 4 fatores principais: o Fator 1, representado pelas espécies G. tenella, G. calida, e G. rubescens e relacionado com a temperatura subsuperficial; o Fator 2, representado pelas espécies N. incompta, N. dutertrei e G. truncatulinoides, relacionado com a profundidade da termoclina; o Fator 3, representado pela espécie G. bulloides, associado com o processo de ressurgência e o Fator 4, representado pelas espécies G. ruber e G. glutinata, associado com a intensidade da Corrente do Brasil. As variações mais expressivas de paleotemperatura e paleosalinidade da superfície do mar e da fauna de foraminíferos planctônicos estão relacionadas com os eventos climáticos abruptos do Hemisfério Norte ocorridos durante a deglaciação, tendo sido moduladas pelas variações de arranjo e intensidade da AMOC. Durante os eventos frios Heinrich 1 e Younger Dryas, relacionados a um enfraquecimento da AMOC, foi observado um aumento das paleotemperatura e paleosalinidade e da intensidade da Corrente do Brasil (Fator 4). Contrariamente, durante o evento quente Bolling-Allerod, a paleotemperatura e a paleosalinidade diminuíram expressivamente, como resposta à retomada da AMOC, e houve aumento da produtividade e presença de uma termoclina mais rasa (Fatores 2 e 3). Sugere-se ainda, que entre 26 e 15 ka houve um deslocamento para norte da Confluência Brasil-Malvinas, possivelmente alcançando a latitude da área de estudo (∼23°S), inferido pela presença da espécie G. inflata. Durante o Holoceno, também foi registrada variação significativa na intensidade da Corrente do Brasil, indicada pelos valores máximos do Fator 4 em torno de 7 ka. O reaparecimento das espécies do plexo G. menardii se deu em 8 ka, assim como o aumento das espécies de foraminíferos planctônicos subsuperficiais, indicado pelo Fator 1. Essas mudanças no Holoceno foram atribuídas à entrada efetiva das águas quentes e salinas do Oceano Índico via vazamento das Agulhas, as quais foram essenciais para o restabelecimento da AMOC moderna. / The objective of the present study was to investigate the response of the planktonic foraminiferaassemblage to the superficial paleoceanographic variations in the western South Atlantic, which plays a key role in the AMOC. For this purpose, a continuous marine sedimentary record was analyzed from the Late Glacial Maximum to the present, based on the planktonic foraminifera assemblage, oxygen isotopes and the Mg/Ca ratio. In addition, variations in the occurrence and geochemistry of the two morphotypes of G. ruber (white) were evaluated. This is the main species used in paleoceanographic studies of tropical and subtropical regions. The results showed that the geochemical and relative abundance differences between the G. ruber morphotypes show two ecologically distinct groups of specimens, confirming that the morphotype sensu stricto calcifies at lower depths with respect to the morphotype and sensu lato, and paleoceanographic and paleoceanographic reconstructions using non-selective mixture of morphotypescould potentially be biased. The Factorial Analysis identified four factors: Factor 1, represented by G. tenella, G. calida, and G. rubescens, is related to the subsurface temperature; Factor 2, represented by N. incompta, N. dutertrei and G. truncatulinoides is related to the depth of the thermocline; Factor 3, represented by G. bulloides, is related toupwelling; and Factor 4, represented by G. ruber and G. glutinata, is related to the intensity of the Brazil Current. The results showed that the most significant variations of paleotemperature and paleosalinity and the planktonic foraminifera assemblage are related to the abrupt climatic events of the Northern Hemisphere occurred during deglaciation and were modulated by variations in AMOC arrangement and intensity. During the cold events Heinrich 1 and Younger Dryas, related to a weakening of the AMOC, an increase of paleotemperature and paleosalinity and intensity of the Brazil Current (Factor 4) in the western portion of the South Atlantic was observed. Conversely, during the hot event Bolli-Allerod, paleotemperature and paleosalinity decreased expressively, as a response to AMOC resumption, with increased productivity and presence of a shallower thermocline (Factors 2 and 3). It is also suggested that between 26 and 15 ka there was a displacement to the north of the Brazil-Malvinas Confluence, possibly reaching the latitude of the study area (∼23°S), inferred by the presence of the G. inflata. Along the Holocene, there was also a significant variation in the intensity of the Brazil Current, indicated by the maximum values of Factor 4 around 7ka. The reappearance of G. menardii plexus occurred in 8ka, as well as the increase of the species of subsurface planktonic foraminifera, indicated by Factor 1. These changes in the Holocene were attributed to the effective entrance of the hot and saline waters from the Indian Ocean via the Agulhas Leakage, which were essential for the reestablishment of the modern AMOC.

O objetivo do presente estudo foi investigar a resposta da assembleia de foraminíferos planctônicos frente às variações paleoceanográficas superficiais na porção oeste do Atlântico Sul, o qual desempenha um papel fundamental na Célula de Revolvimento Meridional do Atlântico (AMOC, Meridional Overturning Circulation). Para esta finalidade, foi analisado um registro sedimentar marinho contínuo desde o Último Máximo Glacial até o presente, com base na assembleia de foraminíferos planctônicos, isótopos de oxigênio e na razão Mg/Ca. Adicionalmente, foram avaliadas as variações na ocorrência e na geoquímica dos dois morfotipos de G. ruber (branca), principal espécie utilizada em estudos paleoceanográficos de regiões tropicais e subtropicais. Os resultados mostraram que as diferenças geoquímicas e de abundância relativa dos morfotipos de G. ruber branca evidenciam dois grupos de espécimes ecologicamente distintos, confirmando que o morfotipo sensu stricto calcifica em menores profundidades com relação ao morfotipo sensu lato e que reconstituições paleoceanográficas baseadas no uso não-seletivo dos morfotipos poderiam ser tendenciosas. A Análise Fatorial realizada nos dados de abundância relativa dos foraminíferos planctônicos identificou 4 fatores principais: o Fator 1, representado pelas espécies G. tenella, G. calida, e G. rubescens e relacionado com a temperatura subsuperficial; o Fator 2, representado pelas espécies N. incompta, N. dutertrei e G. truncatulinoides, relacionado com a profundidade da termoclina; o Fator 3, representado pela espécie G. bulloides, associado com o processo de ressurgência e o Fator 4, representado pelas espécies G. ruber e G. glutinata, associado com a intensidade da Corrente do Brasil. As variações mais expressivas de paleotemperatura e paleosalinidade da superfície do mar e da fauna de foraminíferos planctônicos estão relacionadas com os eventos climáticos abruptos do Hemisfério Norte ocorridos durante a deglaciação, tendo sido moduladas pelas variações de arranjo e intensidade da AMOC. Durante os eventos frios Heinrich 1 e Younger Dryas, relacionados a um enfraquecimento da AMOC, foi observado um aumento das paleotemperatura e paleosalinidade e da intensidade da Corrente do Brasil (Fator 4). Contrariamente, durante o evento quente Bolling-Allerod, a paleotemperatura e a paleosalinidade diminuíram expressivamente, como resposta à retomada da AMOC, e houve aumento da produtividade e presença de uma termoclina mais rasa (Fatores 2 e 3). Sugere-se ainda, que entre 26 e 15 ka houve um deslocamento para norte da Confluência Brasil-Malvinas, possivelmente alcançando a latitude da área de estudo (∼23°S), inferido pela presença da espécie G. inflata. Durante o Holoceno, também foi registrada variação significativa na intensidade da Corrente do Brasil, indicada pelos valores máximos do Fator 4 em torno de 7 ka. O reaparecimento das espécies do plexo G. menardii se deu em 8 ka, assim como o aumento das espécies de foraminíferos planctônicos subsuperficiais, indicado pelo Fator 1. Essas mudanças no Holoceno foram atribuídas à entrada efetiva das águas quentes e salinas do Oceano Índico via vazamento das Agulhas, as quais foram essenciais para o restabelecimento da AMOC moderna. / The objective of the present study was to investigate the response of the planktonic foraminiferaassemblage to the superficial paleoceanographic variations in the western South Atlantic, which plays a key role in the AMOC. For this purpose, a continuous marine sedimentary record was analyzed from the Late Glacial Maximum to the present, based on the planktonic foraminifera assemblage, oxygen isotopes and the Mg/Ca ratio. In addition, variations in the occurrence and geochemistry of the two morphotypes of G. ruber (white) were evaluated. This is the main species used in paleoceanographic studies of tropical and subtropical regions. The results showed that the geochemical and relative abundance differences between the G. ruber morphotypes show two ecologically distinct groups of specimens, confirming that the morphotype sensu stricto calcifies at lower depths with respect to the morphotype and sensu lato, and paleoceanographic and paleoceanographic reconstructions using non-selective mixture of morphotypescould potentially be biased. The Factorial Analysis identified four factors: Factor 1, represented by G. tenella, G. calida, and G. rubescens, is related to the subsurface temperature; Factor 2, represented by N. incompta, N. dutertrei and G. truncatulinoides is related to the depth of the thermocline; Factor 3, represented by G. bulloides, is related toupwelling; and Factor 4, represented by G. ruber and G. glutinata, is related to the intensity of the Brazil Current. The results showed that the most significant variations of paleotemperature and paleosalinity and the planktonic foraminifera assemblage are related to the abrupt climatic events of the Northern Hemisphere occurred during deglaciation and were modulated by variations in AMOC arrangement and intensity. During the cold events Heinrich 1 and Younger Dryas, related to a weakening of the AMOC, an increase of paleotemperature and paleosalinity and intensity of the Brazil Current (Factor 4) in the western portion of the South Atlantic was observed. Conversely, during the hot event Bolli-Allerod, paleotemperature and paleosalinity decreased expressively, as a response to AMOC resumption, with increased productivity and presence of a shallower thermocline (Factors 2 and 3). It is also suggested that between 26 and 15 ka there was a displacement to the north of the Brazil-Malvinas Confluence, possibly reaching the latitude of the study area (∼23°S), inferred by the presence of the G. inflata. Along the Holocene, there was also a significant variation in the intensity of the Brazil Current, indicated by the maximum values of Factor 4 around 7ka. The reappearance of G. menardii plexus occurred in 8ka, as well as the increase of the species of subsurface planktonic foraminifera, indicated by Factor 1. These changes in the Holocene were attributed to the effective entrance of the hot and saline waters from the Indian Ocean via the Agulhas Leakage, which were essential for the reestablishment of the modern AMOC.

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-05-05T15:56:37Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_IncubacaoOvosCriacao.pdf: 1570019 bytes, checksum: 00e3d234ee38b3c294a675c5d631d88a (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-05-11T13:48:49Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_IncubacaoOvosCriacao.pdf: 1570019 bytes, checksum: 00e3d234ee38b3c294a675c5d631d88a (MD5) / Made available in DSpace on 2017-05-11T13:48:49Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_IncubacaoOvosCriacao.pdf: 1570019 bytes, checksum: 00e3d234ee38b3c294a675c5d631d88a (MD5) Previous issue date: 2015-06-12 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A população de guará é considerada extinta em diversas regiões. A reprodução em cativeiro pode ser uma ferramenta importante para a conservação. Os objetivos foram desenvolver um protocolo alimentar e ambientação para a criação de filhotes de guará, intensificar a reprodução e aumentar o número de indivíduos em cativeiro, descrever as fases de desenvolvimento dos filhotes e desenvolver equações para estimar o crescimento. Os filhotes receberam três tipos de dietas: Dieta R (ração comercial), C (ração comercial e camarão) e P (ração comercial e peixe). A condição corporal e o peso dos filhotes foram obtidos diariamente e as medidas corporais a cada sete dias. A taxa de mortalidade na Dieta R foi 100% e nas Dietas C e P foram 0%. As médias dos pesos corporais e comprimentos dos ossos foram maiores com a Dieta P e as médias do comprimento da despigmentação do bico não foram diferentes com as Dietas C e P (P<0,05). A Dieta P apresentou maiores teores nutricionais. No Ano I a criação foi 100% artificial e no Ano II a criação em uma área do parque foi natural. Na criação artificial, o desenvolvimento dos membros e a condição corporal dos filhotes foram avaliados diariamente, e a pesagem a cada sete dias. A taxa de mortalidade foi menor e o número de nascimentos e a taxa de sobrevivência foram maiores durante a criação artificial. Todos os filhotes apresentaram ganho de peso (P<0,05), condição corporal boa e não foram observadas distrofias. As fases de desenvolvimento dos filhotes e o peso foram monitorados diariamente, e as mensurções da despigmentação do bico e ossos foram realizadas a cada sete dias. Os olhos abriram no dia 4,73 ± 0,12, no dia 6,31 ± 0,18 os filhotes se moveram dentro do ninho e no dia 15,3 ± 0,68 sairam do ninho. As médias do peso e das medidas dos ossos aumentaram (P<0,05). As médias da despigmentação do bico diminuiram (P<0,05). No 7º dia o bico começou a pigmentar e no dia 35º ficou completamente pigmentado. A correlação entre todos os parâmetros e os coeficientes de determinação das equações de regressão foram altos. A Dieta P foi considerada a melhor para a criação artificial. O protocolo proporcionou ambientação e alimentação adequadas, intensificou a reprodução e aumentou o número de guarás. Foi possível estabelecer as equações de estimativa de crescimento e descrever as fases de desenvolvimento dos filhotes. A pigmentação do bico foi considerada um parâmetro eficiente para estimar a idade. / The population of scarlet ibis is considered to be extinct in several regions. Captive breeding techniques are complementary tools for species conservation. The objectives were to develop a feeding protocol and an ambient environment for the rearing of scarlet ibis chicks, to intensify reproduction in captivity and increase the number of ibises in the colony, describe the developmental stages of chicks and develop growth estimation equations. The chicks were fed with three diets: Diet C (commercial feed), S (commercial ration and shrimp) and F (commercial ration and fish). The body condition and weight were taken daily, and the body measurements every seven days. The mortality rate under Diet C was 100%, and 0% under Diets S and F. The mean body weight and bones lengths were higher with Diet F and the mean length of the unpigmented portion of the beak did not differ among the diets (P<0.05). Diet F had higher nutritional contents. Rearing was artificial during year I and natural during year II at one of the areas of the park. The limbs development and body condition were evaluated daily, and the weight every seven days during artificial rearing. The mortality rate was lower and the number of births and survival rate were higher during artificial rearing. All of the young exhibited weight gain (P<0.05), good body condition and no dystrophies were observed. The development stages of chicks and the weight were monitored daily, and the measurements of the beak and bones were taken every seven days. The eyes opened on day 4.73±0.12, the nestlings moved within the nest on day 6.31±0.18 and left the nest on day 15.3±0.68. The means weight and measurements of the bones increased (P<0.05). The means of beak despigmentation decreased (P<0.05). The beak pigmentation started on day 7º and on day 35º was completely pigmented. The correlation between all parameters and the determination coefficients of regression equations were high. Diet F was found to be best for the artificial rearing. The artificial rearing protocol provided an adequate environment and feeding, intensified the reproduction and increased the number of scarlet ibises. Growth estimation equations were developed to assess the chicks growth and was possible to describe the developmental stages. Beak pigmentation was found to be a useful parameter for estimating the age.

My thesis is engaged in description of ineligibility original version of the elbow draft tube new constructed small hydro power plant Doudlevce. Other main purpose was construction of better hydraulic thus more efficient this draft tube. First chapter meets project small hydro power plant Doudlevce and it specifies this interesting contstruction of the power plant (for example position of the turbine in vacuum fontain) and many others information about it. Second chapter and its particular parts there deal with functions of this draft tube, for example out-flow of water from turbine and a principle of water-energy utilization placing behind turbine runner. There is also described energy profit and how to get it throught draft tube. There is an analysis of important quantities, which are used during the scoring of flow in the draft tube in third chapter. This part of my thesis is aimed to meeting their principles. There is one of the main purposes of this thesis, where is a description of ineligibility original version of the elbow draft tube in fourth chapter. It results sizable from Kaplan´s attemps and analysis of flowing based on computations in computer programme CFD Fluent. Fifth chapter is about general principles, which should be looked out during the construction of the draft tube. There is also project of new hydraulic better draft tube and its another parts. Last chapter compares original version with new better project based on computations in computer programme CFD Fluent. Comparison is made of specific criterion flowing of water, where are described quantites in third chapter, and efficiency of this draft tube.

The possibilities of double layer porous asphalt usage and rules for design of porous asphalt mixtures are introduced in this work. The porous asphalt mixtures PA 8, PA 11 and PA 16 are designed with different content of asphalt-rubber according to Marshall Test method. The water sensitivity, drainability, particle loss of these PA are determined. The permanent deformation and drainability of double layer PA 8 and PA 11 were also determined. The possibility of usage of this double layer porous asphalt with expected decreasing of traffic noise was proved.

Submitted by Irvana Coutinho (irvana@ufpa.br) on 2017-05-04T16:35:56Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_EcologiaTroficaEspacial.pdf: 871595 bytes, checksum: 37dd504bab4b7c8a0049cca1218bf0da (MD5) / Approved for entry into archive by Irvana Coutinho (irvana@ufpa.br) on 2017-05-04T16:38:19Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_EcologiaTroficaEspacial.pdf: 871595 bytes, checksum: 37dd504bab4b7c8a0049cca1218bf0da (MD5) / Made available in DSpace on 2017-05-04T16:38:19Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_EcologiaTroficaEspacial.pdf: 871595 bytes, checksum: 37dd504bab4b7c8a0049cca1218bf0da (MD5) Previous issue date: 2016-04-20 / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / Três fatores essenciais determinam o nicho principal das espécies: dieta, uso do espaço e tempo. Os nichos trófico e espacial desempenham um importante papel para o sucesso reprodutivo das espécies de anuros que coabitam ambientes restritos como as poças temporárias. As espécies alvo deste estudo são comuns em poças temporárias em áreas de fragmentos urbanos localizado na região leste da Amazônia. Com isso, nosso objetivo principal foi analisar como três espécies de hilídeos sintópicos, partilham os recursos tróficos e espaciais entre si, e que mecanismos utilizam para evitar uma possível competição. Para isso, utilizamos diversas medidas e análises de dieta e uso de hábitat, além de testar se as espécies são sensíveis a fatores ambientais e estruturais dos corpos d’água onde elas ocorrem. Nenhum dos fatores analisados (dieta, uso do espaço, fatores ambientais) foi limitante para a coexistência das espécies, sendo que não houve qualquer interação negativa entre elas. Os hilídeos estudados utilizam diversas estratégias as quais contribuem para diminuir a sobreposição entre si, como o uso de micro-habitats diferenciados, poleiros em diferentes alturas, poças com profundidades distintas, utilização de diferentes graus de importância alimentar para itens compartilhados pelas espécies, bem como, o uso de itens alimentares exclusivos de cada espécie. Além disso, os fatores ambientais e estruturais dos micro-habitats, influenciam diretamente na abundância de duas das espécies estudadas, regulando seu tamanho populacional e auxiliando no seu período reprodutivo. / Three key factors determine the main niche of the species: diet, use of space and time. The trophic and spatial niche show an important role in the reproductive success of frog species than cohabit restricted environments as temporary ponds. The target species of this study are common in temporary ponds in areas of urban fragments located in the eastern Amazon region. With this, our main objective were analyse how three species of sintopics hylids, share the trophic and spatial resources among themselves, and what mechanisms are used to avoid potential competition. For this, we use different measures and diet analyses and habitat use, and test whether the species are sensitive to environmental and structural factors in ponds where they occur. None of the factors (diet, use of space, influence of environmental factors) was limiting to the coexistence of the species, due no there are negative interaction between them. The studied hylids utilized several strategies than help reduce overlap among the species, how the use of different microhabitats, perches at different heights, ponds with different depths, using different degrees of food importance to items shared by species and the use of unique food items of each specie. In addition, environmental and structural factors of microhabitats, influence directly the abundance of two of the species studied, regulating their population size and auxiliary in their reproductive period.

Unambiguous gender identification (ID) is needed to assess parameters in studies of population dynamics, behavior, and evolutionary biology of Caribbean Flamingo (Phoenicopterus ruber ruber) and other birds. Due to its importance for management and conservation, molecular (DNA-based) avian gender ID assays targeting intron-size differences of the Chromosome Helicase ATPase DNA Binding (CHD) gene of males (CHD-Z) and females (CHD-W) have been developed. Male (ZZ) and female (WZ) genotypes are usually scored as size polymorphisms through agarose or acrylamide gels. For certain species, W-specific restriction sites or multiplex polymerase chain-reaction (PCR) involving CHD-W specific primers are needed. These approaches involve a minimum of three steps following DNA isolation: PCR, gel electrophoresis, and photo-documentation, which limit high throughput scoring and automation potential. In here, a short amplicon (SA) High-resolution Melting Analysis (HRMA) assay for avian gender ID is developed. SA-HRMA of an 81-Base Pair (bp) segment differentiates heteroduplex female (WZ) from homoduplex male (ZZ) genotypes by targeting Single-nucleotide Polymorphisms (SNPs) instead of intron-size differences between CHD-Z and CHD-W genes. To demonstrate the utility of the approach, the gender of Caribbean Flamingo (P. ruber ruber) (17 captive from the Dallas Zoo and 359 wild from Ria Lagartos, Yucatan, Mexico) was determined. The assay was also tested on specimens of Lesser Flamingo (P. minor), Chilean Flamingo (P. chilensis), Saddle-billed Stork (Ephippiorhynchus senegalensis), Scarlet Ibis (Eudocimus ruber), White-bellied Stork (Ciconia abdimii), Roseate Spoonbill (Platalea ajaja), Marabou Stork (Leptoptilos crumeniferus), Greater Roadrunner (Geococcyx californianus), and Attwater's Prairie Chicken (Tympanuchus cupido attwateri). Although the orthologous 81 bp segments of Z and W are highly conserved, sequence alignments with 50 avian species across 15 families revealed mismatches affecting one or more nucleotides within the SA-HRMA forward or reverse primers. Most mismatches were located along the CHD-Z gene that may generate heteroduplex curves and thus gender ID errors. For such cases, taxon and species-specific primer sets were designed. The SA-HRMA gender ID assay can be used in studies of avian ecology and behavior, to assess sex-associated demographics and migratory patterns, and as a proxy to determine the health of the flock and the degree by which conservation and captive breeding programs are functioning.

The main objective of this research project was to study the growth and development of the floral nectar spur of Centranthus ruber (L.) DC. Nectar spurs are tubular floral outgrowths, generally derived from the perianth organs, which typically contain secreted floral nectar. The morphological characteristics of the spur, particularly the length, determine which floral visitors will be able to access the nectar reward pooled at the spur tip. Therefore, nectar spurs are ecologically important for the development of specialised pollinator interactions and have been demonstrated to act as key innovations in the evolution of some taxa. Morphological and anatomical characteristics of the spur and floral nectary were investigated using light and scanning electron microscopy. Ultrastructural features of the nectar spur, particularly the floral nectary within, were assessed using transmission electron microscopy. Nectar in C. ruber is produced by a trichomatous nectary which runs along the entire, inner abaxial surface of the spur. The nectary is aligned with the single vascular bundle which runs along the abaxial side of the spur, through the sub-nectary parenchyma, and back up the adaxial side. The secretory trichomes are unicellular and, in late development, they develop a thick layer of secondary wall ingrowths which vastly increases the surface area of the plasma membrane for nectar secretion. Elongate, non-secretory trichomes occupy the entire remaining circumference of the spur’s inner epidermis, but their density is reduced compared to the secretory trichomes. The cellular basis for spur growth is poorly characterized in the literature. Until recently, it was assumed that all nectar spurs grow by the constant production of new cells via up to three potential meristematic regions (the meristem hypothesis, Tepfer 1953). The cellular basis for spur growth in C. ruber was investigated by cell file counts and cell length and width measurements along the lateral side of nectar spurs in each of the developmental stages. DAPI stained spurs were also examined with Confocal/Apotome microscopy to determine the timing and position of cell division activity throughout spur development. It was determined that elongation of the spur epidermal cells contributes much more to spur growth than cell division. In early development, division is the primary driver of spur growth and the cells are isotropic. However, as development progresses, cell division activity slows down and the spur cells become increasingly anisotropic until anthesis. The patterns of nectar secretion were determined by assessing the volume, solute concentration and carbohydrate composition of the nectar throughout flowering phenology in two C. ruber plants. Nectar volumes and solute amounts rose initially, followed by an eventual decline in both as phenology progressed towards senescence. Because this study was conducted on greenhouse grown plants, it can be assumed that nectar was not removed by insects, suggesting that it is likely reabsorbed following secretion. High performance liquid chromatography (HPLC) analysis determined that C. ruber's nectar is sucrose dominant and that nectar composition remains stable following anthesis throughout floral phenology.