Prospecção de genes de interesse biotecnológico : uma abordagem metagenômica

The total number of prokaryotic cells on Earth has been estimated at 4 to 6x1030 and only about 1% of microorganisms present in the environment can be cultivated by standard techniques of cultivation and plating. Therefore, it is a huge biological and genetic pool that can be exploited, for the identification and characterization of genes with biotechnological potential. Within this perspective, the metagenomics approach was applied in this work. Functional screening methods were performed aiming to identify new genes related to DNA repair and / or oxidative stress resistance, hydrocarbon degradation and hydrolytic activities (lipase, amylase and protease). Metagenomic libraries were built utilizing DNA extracted from soil samples collected in João Câmara RN. The libraries were analyzed functionally using specific substrate containing solid medium (hydrolytic activity), supplemented with H2O2 (DNA repair and / or resistance to oxidative stress) and liquid medium supplemented with light Arabian oil (activity, degradation of hydrocarbons). After confirmation of activity and exclusion of false-positive results, 49 clones were obtained, being 2 positive for amylase activity, 22 resistant to oxidative stress generated by H2O2 and 25 clones active for hydrocarbons degradation. Analysis of the sequences showed hypothetical proteins, dienelactona hydrolase, DNA polymerase, acetyltransferase, phosphotransferase, methyltransferase, endonucleases, among other proteins. The sequence data obtained matched with the functions tested, highlighting the success of metagenomics approaches combined with functional screening methods, leading to very promising results

Análise de grafos aplicada a relatos de sonhos: ferramenta diagnóstica objetiva e diferencial para psicose esquizofrênica e bipolar

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Comportamento alimentar do camarão marinho farfantenaeus subtilis em condições laboratoriais

Shrimp culture represents an important activity to brazilian economy. The northeastern region has presented high levels of production because of its climatic conditions. An important factor for the activity´s major development is related to the introduction of the species Litopenaeus vannamei. The use of an exotic species can disturb the ecosystem. In the last decades, L. vannamei has been the only species cultivated in Brazilian farms, there not being an alternative species for shrimp culture. So, there is an urgent need to developing new studies with the native species, which might represent an alternative concerning shrimp production, with emphasis on Farfantepenaeus subtilis. Another important aspect related to the activity is feeding management, once it is quite usual that feed offer on the pond does not take into account either the species´ physiology and behavior or the influence of environmental variables, such as light cycle and substrate. That knowledge may optimize management and so reduce the impact of effluents in the environment. This study´s objective was characterizing feeding behavior of F. subtilis in laboratory. For that, an ethogram was developed, using 20 wild animals which were observed through ad libitum and all occurrences methods. Two experiments were developed in order to register feeding behavior on different substrates, along 15 days, each. In the first experiment, 40 animals were distributed in eight aquaria, half being observed during the light phase of the 24 hour cycle and the other half in the dark phase, both in halimeda substrate. In the second experiment, 20 animals were distributed in four aquaria, under similar conditions as the previous ones, but in sand substrate. In both experiments, animals were observed respectively one, four, seven and ten hours after the beginning of the phase, for light phase, for the dark phase, in ten minute observation windows, before and immediately after feed offer. The following behaviors were registered: feed ingestion, ingestion of other items, inactivity, exploration, vertical exploration, swimming, crawling, digging, burrowing, and moving by the animals. Observation windows after feed offer also included latency to reach the tray and to ingest feed. Nineteen behaviors were described for the species. F. subtilis presented more behavioral activities in halimeda substrate even in the light phase, while burrowing was predominant in sand substrate. In both substrates, moving, crawling and exploration were more frequent after feed offer, but inactivity and burrowing were more frequent before that. Feed ingestion was more frequent in halimeda, both in light and dark phases. Weight gain was also more prominent in that substrate. In sand substrtate, ingestion was more frequent in the dark phase, which suggests that higher granulometry facilitates feed ingestion in F. subtilis juveniles. Our results demonstrate the importance of studies for the better knowledge of the species, specially its response to environmental stimuli, in order to improve animal management

Efeitos do substrato e da densidade populacional sobre as atividades comportamentais e níveis de homócitos em relação à densidade em Litopenaeus vannamei (BOONE, 1931)

Carciniculture in Brazil occupies world-wide prominence due to shrimp culture, and the state of Rio Grande do Norte has presented the best results in the culture of the Litopenaeus vannamei in the last decade. This species has been shown to adapt easily to different environments and is between the five most cultivated penaeids of the world. The ponds are usually constructed in areas close to water courses and estuaries. Stock density and substrate ponds can pollute environment, causing losses in the growth and survival of the shrimps, being considered stress factors. Shrimps in inadequate densities and substrates can result reduced productivity of the farm; and favor diseases. So, it is important to verify how these variables influence the development of the animals in the culture farms. Our objective was to study the influence of the type of substrate and the stock density on the behavior and haemocyte count of the L. vannamei. Individually marked juvenile shrimps were kept in aquaria with 30 L of seawater and continuous aeration, in 12L-12D photoperiod. They were observed through Ad libitum and focal sampling instantaneous methods during thirty days, five times per week, six times per day (8:00 to 18:00) in windows of 15 minutes every two hours. The marking of carapace permitted quantifying molting and the feeding was supplied three times a day. Two experiments were carried out: the first one tested animals in the three different substrates (fine sand, smaller rocks-SPP and biggest rocks-SGR) with 33 shrimp/m2. In the second one, the animals were tested in three stock densities (26, 52 and 66 shrimp/m2) in fine sand substrate. At the end of experiment, biometry (first and second ones) and haemocyte count (second one) were made. The behavior of the L. vannamei seems to have been influenced by substrate and stocking density. In low granulometry of the substrate; the exploratory behavior became more frequent and inactivity of the shrimps was reduced. Burrowing was registered in sand substrate, specially in the initial period of the day. Cleaning was gradually higher along the day, presenting the biggest levels as the dark phase approached. The ingestion of feeding was more frequent in low density, and the animals were bigger and heavier at the end of the experiment. In the fine sand condition, the animals presented better growth, probably associated with the burrowing. The molting was equivalent in all types of substrate, but it was more frequent in high densities. Mortality of the shrimps was more frequent in high densities, and cannibalism and diseases were also registered in that condition. The clinical signals were similar to the ones of infectious mionecrosis (IMNV), generally associated with environment and physical stress. The haemocyte count was low for the hematologic standards of the penaeid, which we attributed for greater dilution of haemolymph in the postmolting phase. Smaller shrimps presented lower levels of haemocytes in relation to the bigger animals, count was also low in 26 shrimp/m2 density. The study demonstrates that stocking density and the granulometry of the substrate can affect the welfare, the health and the behavior of the L. vannamei. The sand substrate and low stocking density can be important tools in the management systems of shrimp production

Influência da composição química do substrato no cultivo de Pleurotus florida

Com o objetivo de avaliar a influência da composição química do substrato no cultivo do cogumelo comestível Pleurotus florida, seis resíduos agrícolas foram testados: palha de arroz, palha de feijão, palha de trigo, folha de bananeira, palha de sorgo e sabugo de milho. O substrato palha de feijão apresentou resultados semelhantes para a produção (189,8 g kg-1), eficiência biológica (89,2%) e número de cogumelos (12) à palha de arroz, substrato utilizado tradicionalmente no cultivo de Pleurotus em escala comercial. Não foi possível atribuir apenas a um fator químico as altas produções e eficiências biológicas observadas em palha de arroz e palha de feijão e muito menos para a baixa produção em palha de sorgo (77,8 g kg-1) e sabugo de milho (53,2 g g-1). No geral, substratos com relação C/N em torno de 45 (N = 1,0%), maior conteúdo de cinzas, acrescido de altos teores de P, K, Ca, Mg and Mn foram os melhores para o cultivo de P. florida.

Síntese enzimática de biosurfactante e sua aplicabilidade na indústria do petróleo

Enzymes have been widely used in biosynthesis/transformation of organic compounds in substitution the classic synthetic methods. This work is the first writing in literature of enzymatic synthesis for attainment the biossurfactants, the use glucose sucrose, ricinoleic acid e castor oil as substratum, and as biocatalyst, used immobilized lipase Thermomyces lanuginose, Rhizomucor miehei and the Candida antarctica lipase B; alkaline protease and neutral protease from Bacillus subtillis and yeast Saccharomyces cerevisiaeI. The analysis of HPLC (high performance liquid chromatography) showed that highest conversions were reached of used the alkaline protease from Bacillus subtillis. Laboratory tests, to evaluate the applicability, indicated that the produced biosurfactantes had good stability in presence of salts (NaCl) and temperature (55 e 25°C), they are effective in the reduction of the superficial tension and contac angle, but they have little foaming capacity, when compared with traditional detergents. These results suggest that the prepared surfactants have potential application as wetting agent and perforation fluid stabilizer

Matrizes poliméricas puras e modificadas para adsorção do gás sulfeto de hidrogênio

The underground natural gas found associated or not with oil is characterized by a mixture of hydrocarbons and residual components such as carbon dioxide (CO2), nitrogen gas (N2) and hydrogen sulfide (H2S), called contaminants. The H2S especially promotes itself as a contaminant of natural gas to be associated with corrosion of pipelines, to human toxicity and final applications of Natural Gas (NG). The sulfur present in the GN must be fully or partially removed in order to meet the market specifications, security, transport or further processing. There are distinct and varied methods of desulfurization of natural gas processing units used in Natural Gas (UPGN). In order to solve these problems have for example the caustic washing, absorption, the use of membranes and adsorption processes is costly and great expenditure of energy. Arises on such findings, the need for research to active processes of economic feasibility and efficiency. This work promoted the study of the adsorption of sulfide gas in polymer matrices hydrogen pure and modified. The substrates of Poly(vinyl chloride) (PVC), poly(methyl methacrylate) (PMMA) and sodium alginate (NaALG) were coated with vanadyl phosphate compounds (VOPO4.2H2O), vanadium pentoxide (V2O5), rhodamine B (C28H31N2O3Cl) and ions Co2+ and Cu2+, aiming to the adsorption of hydrogen sulfide gas (H2S). The adsorption tests were through a continuous flow of H2S in a column system (fixed bed reactor) adsorption on a laboratory scale. The techniques used to characterize the adsorbents were Infrared spectroscopy (FTIR), thermogravimetry analysis (TGA), X-ray fluorescence (XRF), the X-ray diffraction (XRD) electron microscopy (SEM). Such work indicates, the results obtained, the adsorbents modified PMMA, PVC and NaALG have a significant adsorptive capacity. The matrix that stood out and had the best adsorption capacity, was to ALG modified Co2+ with a score of 12.79 mg H2S / g matrix

Contribuição de fungos micorrízicos arbusculares autóctones no crescimento de Guazuma ulmifolia em solo de cerrado degradado

Ensaios foram conduzidos, em casa de vegetação, com solos de pastagem degradada reflorestada e cerrado preservado (controle) visando avaliar a contribuição de fungos micorrízicos arbusculares (FMA) autóctones no crescimento de mutambo (Guazuma ulmifolia Lamb.). As mudas foram transplantadas para sacos de plástico (2 kg) com substratos esterilizados na proporção 4:1 (solo:areia), e o tratamento inoculado recebeu 300 esporos de FMA por saco. A inoculação não proporcionou aumento significativo na produção da matéria seca da parte aérea, matéria fresca das raízes e altura da planta, sugerindo que a G. ulmifolia não é responsiva à micorrização.

Adsorção dos gases NH3, NO e H2S em fosfato de vanadila, hidrogeno fosfato de vanadila e fosfatos de vanadila dopados por Al, Co, Mn e Cr

In this work were synthesized the materials called vanadyl phosphate, hydrogen vanadyl phosphate and vanadyl phosphate doped by transition metals with the aim in adsorption the following compounds: ammonia, hydrogen sulfide and nitrogen oxide. To characterize the starting compounds was used DRX, FTIR, FRX and TG analysis. After the characterization of substrates, proceeded de adsorption of NH3 and H2S gases in reactor, passing the gases with continuous flow for 30 min and room temperature. Gravimetric data indicate that the matrices of higher performance in adsorption of ammonia was those doped by aluminum and manganese, obtaining results of 216,77 mgNH3/g and 200,40 mgNH3/g of matrix, respectively. The matrice of higher performance in adsorption of hydrogen sulfide was that doped by manganese, obtaining results of 86,94 mgH2S/g of matrix. The synthesis of substrates VOPO4.2H2O and MnVOPO4.2H2O with nitrogen oxide was made in solution, aiming the final products VOPO4.G.nH2O and MnVOPO4.G.nH2O (G = NO and n = number of water molecules). The thermo analytical behavior and the infrared spectroscopy are indicative of formation of VOPO4.2,5NO.3H2O compound. Results of scanning electron microscopy (SEM) and Energy dispersive spectroscopy (EDS) of materials vanadyl phosphate and vanadyl phosphate modified after reaction in solid state or in solution with the gases show morphology changes in substrates, beyond the formation of orthorhombic sulfur crystals over their respective hosts when these adsorb hydrogen sulfide

Adequação da metodologia para o teste de germinação de sementes de pitaia vermelha

Hylocereus undatus (Haw.), popularmente conhecida como pitaia vermelha é uma cactácea para a qual se tem registrado um aumento de consumo nos últimos anos e, por ser ainda pouco explorada, vários aspectos referentes à sua propagação ainda são desconhecidos. Até o momento, não existem critérios para a execução de testes de germinação para sementes dessa espécie, publicados nas Regras para Análise de Sementes. Nesse sentido, objetivou-se com este trabalho adequar a metodologia quanto ao substrato, temperatura e tempo de contagem inicial e final para o teste de germinação. Foram testados quatro substratos (rolo de papel, areia, vermiculita e solo) e quatro temperaturas (20, 25, 30 e 20-30°C). O efeito dos substratos no desempenho germinativo das sementes foi avaliado pelo teste de germinação e de primeira contagem instalado com quatro subamostras de 50 sementes. Foram feitas contagens diárias do número de plântulas emergidas até atingirem a estabilidade e, no final do experimento, foram avaliados a porcentagem de germinação das sementes, considerando as plântulas normais, anormais e sementes mortas. Concluiu-se que o teste de germinação de sementes de pitaia vermelha deve ser realizado à temperatura constante de 25°C em rolo de papel, com contagens inicial e final aos cinco e dez dias após a semeadura, respectivamente.

A biotecnologia e a educação ambiental no resguarde a duas espécies em vias de extinção na flora da Mata Atlântica Brasileira

The anthropical action caused destruction of great part of the Atlantic forest remaining today around 7% to 8% of the original portion. A classical example of the degradation is the situation of the Pau-Brasil (Caesalpinia echinata Lam.) which appellant exploration summarized abruptly decreased the wild occurrence of the species. Besides the economic exploration and the threat of extinction, the plant shows some characteristics that its own physiology makes difficult the survival in natural conditions. For this reason, any strategy developed to conserve it should also guarantee ways for its multiplication. In these conditions, the only reasonable technology is micropropagation in vitro based on immature segments of Catullus (explants or internodes). However, in virtue of the threat of extinction, extractivism and fenology, the disponibility of the sources of explants are reduced. Then, to turn this around, the present research showed the hypothesis that arrange these explants would have as a solution to elaborate a bank of matrixes and the utilization of produced seedlings as source of explant. Then, assuming that the soil is a fundamental element for a good formation of the seedlings, there were tests in a greenhouse on the Laboratory of plant biotechnology at UFRN, from May to June of 2008 that showed the influence of the four different soils on the production of Pau-Brasil. The objective of this work was to make possible the seedlings production of the specie and from them to establish a bank of matrixes as source of explant to guarantee the continuity of the process of micropropagation, contributing with the preservation of the specie

Conservação de espécies nativas em um assentamento rural do município de Apodi-RN: impactos socioambientais e econômicos e o uso da biotecnologia como alternativa promissora

The Caatinga biome has a high diversity of potential and their conservation constitutes one of the greatest challenges of Brazilian science. The sustainable management of the Caatinga emerges as an alternative that through the formation of systems agrossilvipastoris, enables the use of forest resources sustainably, ensuring their conservation, regeneration and recovery. In RN this technique has been developed mainly in settlements of Agrarian Reform, such as P. A. Moaci Lucena, and their impacts go beyond the environmental aspect and reverberate socially and economically on the quality of life of family farmers. Despite the efficiency of the Sustainable Management of the Caatinga in the conservation of native species, many forests species of this biome faces serious problems of propagation and for this reason have become vulnerable to extinction, as is the case of Mimosa caesalpiniifolia Benth . Thus , it is evident the need to use sustainable alternatives to overcome the difficulties of propagation of this species and enable their replacement in areas where their existence is threatened. The Plant Biotechnology is considered as a promising alternative in this sense, considering that by micropropagation enables the large-scale production of seedlings with high health genetics status. This work has the following objectives: evaluate the perception of family farmers of P. A. Moaci Lucena in relation to social, environmental and economic impacts of the Sustainable Management of the Caatinga and check the conditions of germination and in vitro propagation of Mimosa caesalpiniifolia Benth that enabling the production of seedlings of this specie on a large scale. To achieve the first objective, semi-structured interviews showed that in the perception of farmers PA Moaci Lucena, the Sustainable Management of the Caatinga was responsible for generating many social, environmental and economic impacts that affected directly in the improvement in the quality of life of the families of the Settlement Project Moaci Lucena. Have to achieve the second objective, were investigated the influence of different substrates and concentrations of growth regulator BAP in the germination and shoot induction in vitro of Mimosa caesalpiniifolia Benth. The vermiculite was presented as the most suitable substrate for germination of this species, because it provided a more rapid germination, higher growth rates and higher dry matter accumulation. Regarding micropropagation, the concentration of 17.76 μmol/L of BAP presented a more responsive in relation to multiplication rate and the number of shoots in M. caesalpiniifolia, thus constituting the most suitable concentration for the in vitro propagation of this specie

Histerese térmica de sistemas magnéticos nanoestruturados

We report a theoretical investigation of thermal hysteresis in magnetic nanoelements. Thermal hysteresis originates in the existence of meta-stable states in temperature intervals which may be tuned by small values of the external magnetic field, and are controlled by the systems geometric dimensions as well as the composition. Two systems have been investigated. The first system is a trilayer consisting of one antiferromagnetic MnF2 film, exchange coupled with two Fe lms. At low temperatures the ferromagnetic layers are oriented in opposite directions. By heating in the presence of an external magnetic field, the Zeeman energy induces a gradual orientation of the ferromagnets with the external field and the nucleation of spin- op-like states in the antiferromagnetic layer, leading eventually, in temperatures close to the Neel temperature, to full alignment of the ferromagnetic films and the formation of frustrated exchange bonds in the center of the antiferromagnetic layer. By cooling down to low temperatures, the system follows a different sequence of states, due to the anisotropy barriers of both materials. The width of the thermal hysteresis loop depends on the thicknesses of the FM and AFM layers as well as on the strength of the external field. The second system consists in Fe and Permalloy ferromagnetic nanoelements exchange coupled to a NiO uncompensated substrate. In this case the thermal hysteresis originates in the modifications of the intrinsic magnetic

Evolução geomorfológica, (des)caracterização e formas de uso das Lagoas da cidade do Natal-RN

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Caracterização macro, meso e microscópica das estruturas frágeis do corpo arenítico conglomerático da região de Santana do Acaraú (CE) e seu embasamento circundante

The structural framework of the sedimentary basins usually plays an important role in oil prospects and reservoirs. Geometry, interconectivity and density of the brittle features developed during basin evolution could change the permo-porous character of the rocks involved in generation, migration and entrapment of fluid flow. Once the structural characterization of the reservois using only sub-surface data is not an easy task, many studies are focused in analogous outcrops trying to understand the main processes by which brittle tectonic is archieved. In the Santana do Acaraú region (Ceará state, NE Brazil) a pack of conglomeratic sandstone (here named CAC) has its geometry controlled mainly by NE trending faults, interpreted as related to reactivation of a precambrian Sobral Pedro II Lineament (LSP-II). Geological mapping of the CAC showed a major NE-SW trending synform developed before its complete lithification during a dextral transpression. This region was then selected to be studied in details in order of constrain the cretaceous deformation and so help the understanding the deformation of the basins along the brazilian equatorial margin. In order to characterize the brittle deformation in different scales, I study some attributes of the fractures and faults such as orientation, density, kinematic, opening, etc., through scanlines in satellite images, outcrops and thin sections. The study of the satellite images showed three main directions of the macrostructures, N-S, NE-SW and E-W. Two of theses features (N-S and E-W) are in aggreement with previous geophysical data. A bimodal pattern of the lineaments in the CAC´s basement rocks has been evidenciated by the NE and NW sets of structures obtained in the meso and microscale data. Besides the main dextral transpression two others later events, developed when the sediments were complety lithified, were recognized in the area. The interplay among theses events is responsible for the compartimentation of the CAC in several blocks along within some structural elements display diferents orientations. Based on the variation in the S0 orientation, the CAC can be subdivided in several domains. Dispite of the variations in orientations of the fractures/faults in the diferents domains, theses features, in the meso and microscopic scale, are concentrated in two sets (based on their trend) in all domains which show similar orientation of the S0 surface. Thus the S0 orientation was used to group the domains in three major sets: i) The first one is that where S0 is E-W oriented: the fractures are oriented mainly NE with the development of a secondary NW trending; ii) S0 trending NE: the fractures are concentrated mainly along the trend NW with a secondary concentration along the NE trend; iii) The third set, where S0 is NS the main fractures are NE and the secondary concentration is NW. Another analized parameter was the fault/fracture length. This attribute was studied in diferent scales trying to detect the upscale relationship. A terrain digital model (TDM) was built with the brittlel elements supperposed. This model enhanced a 3D visualization of the area as well as the spatial distribution of the fault/fractures. Finally, I believe that a better undertanding of the brittle tectonic affecting both CAC and its nearby basement will help the future interpretations of the tectonic envolved in the development of the sedimentary basins of the brazilian equatorial margin and their oil reservoirs and prospects, as for instance the Xaréu field in the Ceará basin, which subsurface data could be correlated with the surface ones