969 resultados para CNPQ::CIENCIAS BIOLOGICAS: BIOTECNOLOGIA
Resumo:
The growing interest and applications of biotechnology products have increased the development of new processes for recovery and purification of proteins. The expanded bed adsorption (EBA) has emerged as a promising technique for this purpose. It combines into one operation the steps of clarification, concentration and purification of the target molecule. Hence, the method reduces the time and the cost of operation. In this context, this thesis aim was to evaluate the recovery and purification of 503 antigen of Leishmania i. chagasi expressed in E. coli M15 and endotoxin removal by EBA. In the first step of this study, batch experiments were carried out using two experimental designs to define the optimal adsorption and elution conditions of 503 antigen onto Streamline chelating resin. For adsorption assays, using expanded bed, it was used a column of 2.6 cm in diameter by 30.0 cm in height coupled to a peristaltic pump. In the second step of study, the removal of endotoxin during antigen recovery process was evaluated employing the non-ionic surfactant Triton X-114 in the washing step ALE. In the third step, we sought developing a mathematical model able to predict the 503 antigen breakthrough curves in expanded mode. The experimental design results to adsorption showed the pH 8.0 and the NaCl concentration of 2.4 M as the optimum adsorption condition. In the second design, the only significant factor for elution was the concentration of imidazole, which was taken at 600 mM. The adsorption isotherm of the 503 antigen showed a good fit to the Langmuir model (R = 0.98) and values for qmax (maximum adsorption capacity) and Kd (equilibrium constant) estimated were 1.95 mg/g and 0.34 mg/mL, respectively. Purification tests directly from unclarified feedstock showed a recovery of 59.2% of the target protein and a purification factor of 6.0. The addition of the non-ionic surfactant Triton X-114 to the washing step of EBA led to high levels (> 99%) of LPS removal initially present in the samples for all conditions tested. The mathematical model obtained to describe the 503 antigen breakthrough curves in Streamline Chelanting resin in expanded mode showed a good fit for both parameter estimation and validation steps. The validated model was used to optimize the efficiencies, achieving maximum values of the process and of the column efficiencies of 89.2% and 75.9%, respectively. Therefore, EBA is an efficient alternative for the recovery of the target protein and removal of endotoxin from an E. coli unclarified feedstock in just one step.
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The Rachycentron canadum species, commonly known as beijupirá or cobia is the only representative of Rachycentridae family which has been increasingly used in marine fish farming, in intensive cultivation. As advantageous features it has easy adaptation, prolific behavior, early growth in captivity and high commercial value. Additionally, specie of Lutjanidae family (Lutjanus synagris, Lutjanus jocu, Lutjanus analis, Lutjanus alexandrei and Ocyurus chrysurus) represents an important fisheries resource in all areas of its occurrence. In Brazil, the commercial exploitation of Lutjanidae which begun in the 60's and 80's, already has showed a decline in catch volumes. This fact suggests that the snappers must have a conservative management. Despite the economic potential, little is known about the genetic and cytogenetic characteristics of these species, especially with respect to repetitive DNA analysis, which represents the major part of the eukaryotes genome, playing important evolutionary roles in the fish genome. Cytogenetic data is increasingly being used in population studies and biotechnological purposes in fishes. The cytogenetical analyzes were performed using classical methods such as Giemsa staining, C-banding and Ag-NORs, fluorochromes base-specific staining (DAPI and MM) and physical mapping of repetitive sequences among which, telomeric sequences, transposons (Tol2), retrotransposons (Rex1 and Rex3), repetitive DNA (microsatellites and Cot-1) and transcriptionally active regions of the 18S and 5S ribosomal genes and histone (H3 and H2BA) by in situ hybridization with fluorescent probes (FISH). The chromosomal patterns obtained contributed to the organization of repetitive sequences in the genome of the species, as well as karyotypical differentiation. Unusual patterns of histone sequences expansion depict the first occurrence in marine fishes. The obtained data provided subsides to the genetic knowledge of the important fisheries resource represented by the species here analyzed, seeking the marine pisciculture improvement.
Resumo:
To clarify the functional mechanisms of habitat use is necessary to analyze it in conjunction with the conduct performed by animals. The occurrence, distribution and use of space are characteristic of a species resulting from habitat selection that is in search of conditions favorable to its survival. One can relate the physical and biological factors of the environment with the ecological characteristics of the species, since these factors act by regulating the ecological success of organisms, and from there you can get important information about the habitat use and behavior of individuals. This study aimed to characterize the use of habitat and diurnal activity expressed by the Guiana dolphin, Sotalia guianensis in an estuarine area of Sergipe state, Brazil, analyzing the influence of tide and time days on the occurrence of animals and behavior s state, and group s size and composition in this cetacean species. From March 2009 to February 2010, focal groups observations of dolphins were made from fixed - point and records snapshots of data taken every 5 min. in the interval from 6 a.m to 6 p.m, in alternating shifts. The results showed that the constant presence of animals in the area of the Sergipe River estuary indicates that this is an important area of occurrence of S. guianensis, which use the region mainly in the morning, at low tide and as a feeding. As in other regions of northeastern Brazil, small groups formed 2-12 individuals were most common, with adults and immatures. The high frequency of immature animals may indicate that this area of the estuary is used as brood area and parental care of pups and young animals, since the immature animals were very associated with adults and monitoring the activities of foraging / feeding may be related to a form of learning or training of such behavior
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This work focuses the familiar cajuculture at Serra of Mel (RN) that presents a geographic and climatic structure favourable to the development of the cajuculture; the interaction between the ambient factors and agriculture in the region provides a propitious environment to the culture of the cashew; the objective of present work was associate ambient, economic, social and cultural factors of this municipy with the possibility of a sustainable agriculture associated with the vegetal biotechnology
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The mangrove is a coastal ecosystem of the big ecological importance, showing high fragility front by natural process and the human interventions in the coastal zone. This research has objective to analyses the relation between mangrove species distribution and geochemical parameters variation of the water and soil in Apodi/Mossoro estuary, located in the Rio Grande do Norte state north coastline. The results were obtained from floristic and structural analysis of the vegetation and Quick Bird satellite images interpretation (collected in 2006 year), manipulated with ENVI 4.3 and ArcGIS 9.2 software s. This estuary was characterized by to presents a gradient of the salinity around 40 kilometers extension, finding amount between 50 and 90 g/l-1. Will be identified the formation of the mix vegetation formation in the estuary mount, where the water salinity no show express wide variation on seawater (36 g/l-1), finding species: Rhizophora mangle L., Laguncularia racemosa (L.) C. F. Gaertn, Avicennia schaueriana Stap. & Leechman e Avicennia germinans L. Along of the estuary, have a streak formation of the vegetation composed by Avicennia spp. and L. racemosa. In high estuary, where the salinities value stay above 60 g/l-1, only A. germinans predominate in dwarf form. In this sense, the salinity is as a limiting factor of stress on the mangrove vegetation as it enters the estuary, this parameter should be taken into account when drawing up management plans and environmental restoration in the estuary in question
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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
Resumo:
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
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This paper aims to describe the familiar cotton culture in the districts of Tangará and Triunfo Potiguar, located in the state of Rio Grande do Norte. It relates specific problems that small cotton agriculturists face in the cotton production and sale as well as their perception in relation to public policies that have been put into practice in these districts. The research revealed soils with potentiality for the cotton culture, but that are not being cultivated for anything else but subsistence, not being able to become productive, specifically through the cotton culture. That activity should be practiced so that it preserves the environment and the familiar farmers themselves, since it was verified they are constant1y exposed to health problems due to insecticides use. In what refers to fomentation policies it was observed that bureaucracy delays the liberation of the money destined to production, seeds delivery and other actions related to production and commercialization. Some procedures such as information and training about productivity development and attention with the environrnent and infrastructure are recommended. The results indicate lack of investments, rural credit and technical help, main1y in Triunfo Potiguar. There is unanimity, from the rural farmers of that district in what refers to the desire of improving the production and remaining in that activity. In this sense, the contribution of this research for the most appropriate culture production can be materialized through the technique of tissue's culture, aimed at obtaining plants that are resistant to diseases. The data indicated that the hormonal supplementations used induced regenerative calluses making in vitro morphogenesis possible in alI tested varieties. The use of the activated coal antioxidant was efficient, reducing oxidation, however not suppressing it. The results lead to an opportunity for the familiar tàrmers from Rio Grande do Norte enlarge the cotton culture, because to obtain plants that are resistant to diseases implies in insecticides reduction, propitiating smaller impact to the atmosphere and less cost
Resumo:
The umbu tree (Spondias tuberosa Arruda) is a fruit native to the northeast of Brazil with great economic, social and ecological importance for the northeastern semi-arid region. Despite its role, the umbu tree has suffered negative pressure thanks to cluttered extractivism and to negative selection of its fruits, which as the deforestation and the dormancy of seeds contribute to the decrease of its production year after year, making necessary studies that contribute to the improvement of this specie and its conservation. Given the risks to the conservation of the specie and its usefulness to the population, the association between plant biotechnology, for being a tool that can be used to increase its production. and the perception of gathering communities, by valuing the point of view and the knowledge of the population, can facilitate its conservation. This work aimed to develop methods of propagation for umbu tree as well as contribute to its conservation by using biotechnology, with specific objectives to contribute to the conservation of this species; determine concentrations of BAP and ANA in the formation of buds; testing the efficiency of different substrates and concentrations of gibberellic acid on germination in vitro and ex vitro, as well as capture the perception of families in communities that engage in the gathering of umbu. To study the germination, the seeds were inoculated in different substrates (vermiculite, vermiculite + clay, clay, clay + manure and manure + vermiculite) and in different concentrations of gibberellic acid (0 mg, 250 g and 500 mg). For the formation of buds BAP to 0.1 mg-1 was associated with different concentrations of ANA (0.2; 0.4; 0.8mg.L-1). The study of perception was conducted by applying semi-structured questionnaire with Malhada Vermelha community. The experiments resulted in vermiculite and concentration of 500 mg gibberellic acid as the best for germination. The association of 0.1 mg.L-1 of BAP to 0.2 mg.L-1 of ANA provided better formation of buds. As to the application of questionnaires, they revealed that the population understands the decreased amount of umbu plants and umbu fruit in the region, as well as shows concern for its conservation.
Resumo:
A β-D-N-acetilglucosaminidase extracted and partially isolated from crustacean Artemia franciscana by ammonium sulfate precipitation and filtration gel chromatography Bio Gel A 1.5m. the enzyme was immobilized on ferromagnetic Dacron yielding a insoluble active derivative with 5.0 units/mg protein and 10.35% of the soluble enzyme activity. β-D-N-acetilglucosaminidase-ferromagnetic Dacron was easily removed from the reaction mixture by a magnetic field, it was reused for ten times without loss in its activity. The ferromagnetic Dacron was better activated at pH 5.0. The particles visualized at scanning electron microscope (SEM) had presented different sizes, varying between 721nm and 100µm. Infra red confirmed immobilization on support, as showed by primary amino peaks at 1640 and 1560 cm-1 . The immobilize enzyme presented Km of 2.32 ± 0.48 mM and optimum temperature of 50°C. Bought presented the same thermal stable of the soluble enzyme and larger enzymatic activity at pH 5.5. β-D-N-acetilglucosaminidase-Dacron ferromagnético showed sensible for some íons as the silver (AgNO3), with loss of activity. The β-D-N acetilglucosaminidase activity for mercury chloride (HgCl2), whom is one of the most toxic substance joined in nature, it was presented activity already diminished at 0,01mM and lost total activity at 4mM, indicating sensitivity for this type of metal. β-D-N-acetilglucosaminidase-ferromagnetic Dacron showed degradative capacity on heparan sulfate, the enzyme still demonstrated degradative capacity on heparan sulphate, suggesting a possible application to produce fractions of this glycosaminoglycan
Resumo:
Sulfated Polysaccharides with unique chemical structures and important biological activities has been found in a diversity of sea invertebrates. For that, to exist a huger interest on the biotechnology field in the research theses sulfated compounds isolated from sea organisms. Despite the privileged brazilian position for these compounds attainment, there are still a few scientific informations about the isolated substances and their biological activities. A head the displayed, the present work has for objectives, to evaluate the pharmacological properties of the glycosaminoglycans isolated from the sea shrimp Litopenaeus schimitti on homeostasis, blood coagulation, leukocytes migration and platelet/leukocyte adhesion. For this, yhe glycosaminoglycans were extracted from crustacean tissues by proteolysis, fractionation with acetone and later submitted to pharmacological assays. The crustacean tissues showed compounds heparin-like, with anticoagulant activity of 45 IU/mg and 90 IU/mg, respectively. These molecules showed low residual hemorrhagic effects in the tested concentration (100 µg/mL), when compared to unfractionated commercial heparin (UFH). Another dermatan sulfate-like compound, predominately constituted for disulfated disaccharides, was isolated from crustacean abdomen. This compound showed an efficient effect on leukocytes migration inhibition, in the concentration of 15 µg/mL, reducing the cellular infiltration in 65% when compared to the controlled animals. In this same concentration, the DS reduced in 60% the protein concentration of the peritoneal exudates. In the concentration, this compound of 0.5 mg/mL, it was capable to reduce in 40% platelet/leukocytes adhesion. Our data demonstrate that these sulfated polysaccharides isolated from the shrimp L. schimitti will can be used as bioactive compounds, appearing as active principles for pharmacological development, anticoagulants and inflammatory response regulators
Resumo:
The screening for genes in metagenomic libraries from soil creates opportunities to explore the enormous genetic and metabolic diversity of microorganisms. Rivers are ecosystems with high biological diversity, but few were examined using the metagenomic approach. With this objective, a metagenomic library was constructed from DNA soil samples collected at three different points along the Jundiaí-river (Rio Grande do Norte-Brazil). The points sampled are from open area, rough terrain and with the direct incidence of sunlight. This library was analyzed functionally and based in sequence. For functional analysis Luria-Bertani solid medium (LB) with NaCl concentration varied from 0.17M to 0.85M was used for functional analysis. Positives clones resistant to hypersaline medium were obtained. The recombinant DNAs were extracted and transformed into Escherichia coli strain DH10B and survival curves were obtained for quantification of abiotic stress resistance. The sequences of clones were obtained and submitted to the BLASTX tool. Some clones were found to hypothetical proteins of microorganisms from both Archaea and Bacteria division. One of the clones showed a complete ORF with high similarity to glucose-6-phosphate isomerase which participates in the synthesis of glycerol pathway and serves as a compatible solute to balance the osmotic pressure inside and outside of cells. Subsequently, in order to identify genes encoding osmolytes or enzymes related halotolerance, environmental DNA samples from the river soil, from the water column of the estuary and ocean were collected and pyrosequenced. Sequences of osmolytes and enzymes of different microorganisms were obtained from the UniProt and used as RefSeqs for homology identification (TBLASTN) in metagenomic databases. The sequences were submitted to HMMER for the functional domains identification. Some enzymes were identified: alpha-trehalose-phosphate synthase, L-ectoina synthase (EctC), transaminase L-2 ,4-diaminobutyric acid (EctB), L-2 ,4-diaminobutyric acetyltransferase (EctA), L-threonine 3 dehydrogenase (sorbitol pathway), glycerol-3-phosphate dehydrogenase, inositol 3-phosphate dehydrogenase, chaperones, L-proline, glycine betaine binding ABC transporter, myo-inositol-1-phosphate synthase protein of proline simportadora / PutP sodium-and trehalose-6-phosphate phosphatase These proteins are commonly related to saline environments, however the identification of them in river environment is justified by the high salt concentration in the soil during prolonged dry seasons this river. Regarding the richness of the microbiota the river substrate has an abundance of halobacteria similar to the sea and more than the estuary. These data confirm the existence of a specialized response against salt stress by microorganisms in the environment of the Jundiaí river
Resumo:
The brazilian-plum (Spondias tuberosa, His) is a tropical fruit tree that has been consolidated in the market for agribusiness processing, due to its characteristic flavor of fruit. Accordingly, studies to optimize the propagation of plants are necessary for production of seedlings with agronomic and quality assurance measures. This study aimed at determining the efficient techniques for uniform seed germination, as brazilian-plum seed present mechanical dormancy, and establish optimal culture media for multiplication of shoots from the in vitro micropropagation. Firstly, in a greenhouse at the Universidade Federal do Rio Grande do Norte, was evaluated the influence of different methods of breaking dormancy in the emergence of seedlings of brazilian-plum and speed of germination (IVG) of seeds. After 60 days of cultivation, it was found that splay in the distal portion of the seed was the best treatment, with rates of 85.33% in germinability and 3.415 of IVG, compared with the treatment of seed-soaking in water for 12h + humus and the control group. Subsequently, new sources of seedling explants were obtained in studies of tissue culture. Laboratory of Plant Biotechnology that the university, was used stem apex, nodal segments and internodes in search of decontamination with various concentrations of calcium hypochlorite [Ca(OCl)2] and micropropagation, inoculating them in half WPM (1980) with various concentrations of 6-benzylaminopurine (BAP). We used 10 sample units with three replications for different concentrations of [Ca(OCl)2], BAP and explants type. After thirty days, which was observed for the control of contamination, during the establishment in vitro, concentrations of [Ca(OCl)2] between 0.5% and 2.0% were effective in combating exogenous contamination of the apex. In nodal segments and internodes, concentrations of [Ca(OCl)2] between 1.0% and 2.0% and 1.5% and 2.0% were respectively, sufficient to reduce the percentage of losses in these infestations explants. For micropropagation, the culture medium supplemented with 0.1 mg.L-1 BAP promotes better development of multiple shoots per explants from nodal segment. However, success does not get to shoot training in internodal segment
Resumo:
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
Resumo:
About 40% of the earth is occupied by tropical and subtropical forests, including 42% of dry forests, where there is Caatinga Bioma, contemplating tree forests and shrubs, with xerophytic characteristics. Study and conservations of Caatinga biologic diversity is one of the greatest challenges of Brazilian science because those are, proportionally, the less studied among natural areas, with most of the scientific effort centered in very few points around the main cities in the area and also because it is the less protected natural Brazilian area. The environmental degradation is constantly increasing and has its rhythm accelerated by the men appropriation to meet or not their own needs. Therefore, species conservation should be based in three principles: the use of natural resources by present generation, waste prevention and use of the natural resources to benefit the majority of the citizens. Among the strategies to species conservation, we can mention the ex situ conservation , in which the conservation of genetic resources may be realized outside of the natural environment in which the species occur, and in situ conservation , or, in other words, in the places where the species occur. In ex situ conservation, the germplasm collections are maintained in the field and/or in laboratories (conservation chambers), and this mainly conserves intraspecific diversity (genetic variance), the ex situ collections are continuously enriched by collection activities, introduction and germplasm interchange; the in situ conservation preserving ecosystems and habitats, maintaining and recovering native population of species of interest. So, the objective of this paper is the search for strategies to the conservation of Mimosa caesalpiniifolia B. (sabiá) using instruments of environmental perception and plant biotechnology, as mechanisms of in situ and ex situ conservation. To environmental perception, were realized open, semi-structured and qualitative interviews. The questions included socioeconomic data and knowledge of Sabiá specie. To plant biotechnology, Sabiá seed collection were realized in different location to formation of a germplasm bank. The specie micropropagation was made from nodal segment of plants from the matrizeiro. About the knowledge of rural populations and the use of Sabiá plant, some preferences occurred from speeches that the plant possesses a firm wood, not attacked by termites, legalized for exploration by the Brazilian environmental organ (IBAMA), and is a native specie. This research found the rural population has knowledge about Sabiá specie and the natural resources are exhausting. The proposal that the rural community brought was the donation of the Sabiá specie seeding initiating on the rain season, in which the seeding would be plated between the lots, in individual plantations. To the formation of a matrix bank, plant biothecnology brought answers favorable to Sabiá specie seeding, with the formation of multiple shoots