Clonagem e expressão do gene que codifica o inibidor de quimotripsina de Erythrina velutina WILLD. - caracterização e avaliação de seu potencial farmacológico

Proteinases are enzymes distributed widely founded in several organisms and perform many different functions, from maintaining homeostasis to the worsening of some diseases such as cancer, autoimmune diseases and infections. The proteins responsible of controlling the action of these enzymes are the inhibitors, that are classified based on their target proteases and are founded since simple organisms, such as bacteria, to higher organisms, such as larger plants and mammals. Plant proteinase inhibitors act by reducing or inactivating the activity of target proteases, thus, these proteins have been studied as potential tools in the treatment of diseases related to protease activities. In this context, an inhibitor of chymotrypsin from Erythrina velutina, called EvCI was previously purified and it was observed that this protein plays in vitro anticoagulant activity and anti-inflammatory activity in in vivo model. Aiming to reduce the environmental impact caused by the purification EvCI in high amounts and to facilitate the process of obtaining this protein, the recombinant chymotrypsin inhibitor from Eryhrina velutina was produced after cloning and expression in Escherichia coli. The bacteria were grown in LB medium and after induction of the expression this material was subjected to procedures for cell lysis and the product was applied on Nickel-affinity column. The proteins adsorbed were digested by thrombin and applied on Chymotrypsin-Sepharose affinity column, obtaining the purified inhibitor, named recEvCI. After electrophoresis, the recombinant inhibitor showed an approximately molecular mass of 17 kDa, and reduced the chymotrypsin and elastase activities in vitro. The recombinant inhibitor was sequenced and was found similar amino acids residues when compared to other inhibitors deposited in the database, with some modifications. recEvCI showed high stability under pH variations and reducing conditions, maintaining its activity around 80%. This protein increased the blood coagulation time in vitro by acting on the intrinsic pathway and did not show cytotoxicity against strains of mouse 3T3 fibroblasts and RAW 264.7 macrophages. recEvCI showed microbicide activity related to release of nitric oxide and consequently the activation of macrophages, futhermore having proinflammatory effects assessed by increased release of TNF-α. These results indicate that recEvCI can be biotechnologically used as a new tool in the control of coagulation-related diseases as well as can be an activating agent of the immune system in immunosuppressed individuals

Influência do lodo de ETE na massa para fabricação de cerâmica vermelha

The WTP produce many kinds of residue on your treatment stages, but the sludge is the more problematic from the final disposition point view. The actual rate of residue production deriving from technological evolution and the crescent population needs prevents the subtle equilibrium generation between consumption and recycling/reuse, creating problems of pollution resulting from inappropriate management of residues. Thus, is necessary achieve a new equilibrium between the grow from raw materials and energy and the residue generation. This equilibrium should be achieved by technical and economic feasibility of environmental supported models through recycling and reuse. The red ceramic industry stand out in residue absorption question as raw material due their clay mass heterogeneity, constituted by clay minerals and non-clay minerals with wide mineralogical variation, allowing residue inclusion which act like plastic or non-plastic materials, contributing to retain heavy metals contained in residues in the vitreous mass formed during the burning of the ceramic bodies. This work propose the study of the influence of incorporation of 25 wt.% sludge from wastewater treatment plant, according preliminary results, in the mass to produce ceramic bodies. The raw materials was characterized through chemical composition analyses by XRF, mineralogical analyses by XRD, thermal analyses by TG and DTA, Atterberg limits and thermodilatometry. Subsequently was composed the mass with 75 wt.% of clay and 25 wt.% of dried wastewater sludge from UFRN WWTP. Samples with 6,0 x 2,0 x 0,5 cm was produced with unidirectional compacting under pressure of 20MPa and burned in temperatures between 950 and 1,200ºC. After fired, the ceramic bodies have been submitted to physical and mechanical analyses through the measure of firing shrinkage, water absorption, density, apparent porosity and flexural strength; crystallographic analyses through XRD and microstructure analyses by SEM. The technological properties obtained was satisfactory to production of roof tiles with 25 wt.% at 1,200 ºC, but the production of others products at lower temperatures was not feasible

Degradacao termica e catalitica da borra oleosa de Petroleo com materiais nanoestruturados al-mcm-41 e AL-SBA-15

Aiming to reduce and reuse waste oil from oily sludge generated in large volumes by the oil industry, types of nanostructured materials Al-MCM-41 and Al-SBA-15, with ratios of Si / Al = 50, were synthesized , and calcined solids used as catalysts in the degradation of oily sludge thermocatalytic oil from oilfield Canto do Amaro, in the state of Rio Grande do Norte. Samples of nanostructured materials were characterized by thermogravimetric analysis (TG / DTG), X-ray diffraction (XRD), scanning electron microscopy (SEM), absorption spectroscopy in the infrared Fourier transform (FT-IR) and adsorption nitrogen (BET). The characterization showed that the synthesized materials resulted in a catalyst nanostructure, and ordered pore diameter and surface area according to existing literature. The oily sludge sample was characterized by determining the API gravity and sulfur content and SARA analysis (saturates, aromatics, resins and asphaltenes). The results showed a material equivalent to the average oil with API gravity of 26.1, a low sulfur content and considerable amount of resins and asphaltenes, presented above in the literature. The thermal and catalytic degradation of the oily sludge oil was performed from room temperature to 870 ° C in the ratios of heating of 5, 10 and 20 ° C min-1. The curves generated by TG / DTG showed a more accelerated degradation of oily sludge when it introduced the nanostructured materials. These results were confirmed by activation energy calculated by the method of Flynn-Wall, in the presence of catalysts reduced energy, in particular in the range of cracking, showing the process efficiency, mainly for extraction of lightweight materials of composition of oily sludge, such as diesel and gasoline

Pirólise de borras oleosas de petroleo utilizando nanomateriais

The oily sludge is a complex mix of hydrocarbons, organic impurities, inorganic and water. One of the major problems currently found in petroleum industry is management (packaging, storage, transport and fate) of waste. The nanomaterials (catalysts) mesoporous and microporous are considered promising for refining and adsorbents process for environment protection. The aim of this work was to study the oily sludge from primary processing (raw and treated) and vacuum residue, with application of thermal analyses technique (pyrolysis), thermal and catalytic pyrolysis with nanomaterials, aiming at production petroleum derived. The sludge and vacuum residue were analyzed using a soxhlet extraction system, elemental analysis, thin layer chromatography, thermogravimetry and pyrolysis coupled in gas chromatography/mass spectrometry (Py GC MS). The catalysts AlMCM-41, AlSBA-15.1 e AlSBA-15.2 were synthesized with molar ratio silicon aluminum of 50 (Si/Al = 50), using tetraethylorthosilicante as source of silicon and pseudobuhemita (AlOOH) as source of aluminum. The analyzes of the catalysts indicate that materials showed hexagonal structure and surface area (783,6 m2/g for AlMCM-41, 600 m2/g for AlSBA-15.1, 377 m2/g for AlSBA-15.2). The extracted oily sludge showed a range 65 to 95% for organic components (oil), 5 to 35% for inorganic components (salts and oxides) and compositions different of derivatives. The AlSBA-15 catalysts showed better performance in analyzes for production petroleum derived, 20% increase in production of kerosene and light gas oil. The energy potential of sludge was high and it can be used as fuel in other cargo processed in refinery

Biologia alimentar e morfohistologia do tubo digestório do mussum, synbranchus marmoratus, bloch (osteichthyes: synbranchidae) no açude Marechal Dutra Gargalheiras , localizado no semi-árido brasileiro

Food habits and morpho-histology of the digestive tract of marbled swamp eel, Synbranchus marmoratus (Block, 1917) were investigated. The fish samples were captured during August, 2007 to July, 2008 in the Marechal Dutra reservoir, Acari, Rio Grande do Norte. The rain fall data was obtained from EMPARN. The fish captured, were measured, weighed, dissected, eviscerated and individual stomach weights were registered. The stomach contents analyses were carried out based on volumetric method, points, frequency of occurrence and applying the Index of Relative Importance. The degrees of repletion of the stomachs were determined besides the Index of Repletion relating to feeding activity variations and frequency of ingestion during the rainy and dry seasons. The rainfall varied from 0 mm a 335 mm with a mean value of 71.62 mm. Highest rainfall of 335.5 mm was registered in March, 2008 and August to December was the dry period. During the dry period the study species presented high degrees of repletion of the stomachs, with a peak value in the month of September (mean = 4.54; ± SD = 0.56). The minimum mean value of = 3.99 ± SD = 0.25 was registered in the month of May during the rainy period. The stomach contents of S. marmoratus registered show that this fish prefers animals, 78.22% of crustaceans 2.85% of mollusks, 3.25% of fish, 1.4% of insects and 13.5% of semi-digested organic matter, thus characterizing the study species as a carnivore with a preference for crustaceans. The morpho-histological aspects of the digestive tract of S. marmoratus indicate that the mouth is terminal adapted to open widely, thin lips with taste buds, small villiform teeth forming a single series on maxillas, four pairs of branchial arches with short and widely spaced branchial rays. The oesophagus is short and cylindrical with a small diameter. The oesophagus wall is thick with mucas surface and internal parallel folds. The stomach is retilinical in form, presenting cardiac, caecal and pyloric portions. The caecal portion is long and is intermediary in position between the cardiac and pyloric portions. The cardiac portion of the stomach is short and cylindrical formed of simple epithelial cylindrical mucus cells. The caecal portion is long with narrow walls, a big cavity and smaller folds which give rise to gastric glands. The phyloric portion has no glands and primary or secondary mucas folds. The morphohistological aspects of the digestive tract of S. marmoratus indicate its adaptation to a carnivorous feeding habit

Modificações na produtividade e nas características físicas e químicas da madeira de Eucalyptus grandis causadas pela adubação com lodo de esgoto tratado

O Eucalyptus grandis é uma das espécies mais cultivadas no Brasil devido à sua produtividade e qualidade da madeira. Avaliaram-se o efeito da aplicação de lodo de esgoto tratado (0 a 40 t ha-1 base seca) e uma dose de adubo mineral nos atributos físicos e químicos da madeira de Eucalyptus grandis de árvores com cinco anos de idade, no Município de Itatinga, São Paulo, Brasil. O tipo de solo foi caracterizado como Latossolo Vermelho-Amarelo Distrófico (argila = 120 g kg-1 na camada de 0-20 cm) e o clima, como mesotérmico úmido (Cwa), segundo a classificação de Köeppen. O delineamento experimental foi o de blocos ao acaso, com seis tratamentos e quatro repetições. O diâmetro à altura do peito (DAP), a altura das árvores e o volume de madeira foram obtidos em todas as parcelas de oito árvores com DAP na classe de maior freqüência. As caracterizações físicas e químicas da madeira foram realizadas de acordo com as normas da ABTCP, TAPPI e ABNT. O lodo de esgoto diminuiu a densidade básica da madeira, mas não afetou os teores de celulose, lignina, extrativos e o poder calorífico da madeira. O decréscimo de densidade da madeira pela adubação com lodo de esgoto foi compensado pela maior produtividade de madeira.

Avaliação dos teores de metais pesados no solo e na planta de cana-de-açúcar sob adubação com lodo de esgoto e vinhaça

The objective of this work was to evaluate Cd, Cr, Ni and Pb total contents in soil and sugar cane after four annual applications of sewage sludge and vinasse to supply all N and all K required by the crop. A field experiment was carried out on clayey Typic Haplustox. It was used a randomized block design with seven treatments and three replications. The treatments consisted in: (i) rate of sewage sludge to supply 100% of the N required by the crop; (ii) rate of sewage sludge to supply 200% of the N required; (iii) rate of vinasse to supply 100% of the K required; (iv) rate of vinasse to supply 200% of the K required; ( v) combination of the treatments ( i) and ( iii); ( vi) combination of the treatments ( ii) e ( iv); (vii) control ( mineral fertilization). The contents of Cd, Cr, Ni and Pb in sewage sludge reached maximum values of 1.58, 41.66, 35.17 and 45.25 mg kg(-1), respectively. These metals were not detected in vinasse. The residues were applied annually for four years. At the end of this period, sewage sludge and vinasse accumulated rates, that get up to 51 t ha(-1) and 762 m(3) ha(-1), respectively, did not cause alteration in Cd, Cr, Ni and Pb total contents in soil, up to 50 cm of depth, and in plant. Sewage sludge and vinasse application to supply N and K for sugar cane did not offer soil pollution risk by heavy metals.

Fluorescent amplified fragment length polymorphism genotyping of human and animal Staphylococcus aureus isolates from dairy farms with manual milking

Fluorescence amplified fragment length polymorphism (fAFLP) was used to assess the genetic relatedness of 40 Staphylococcus aureus strains isolated from human and animal skin samples in seven dairy farms with manual milking. S. aureus was isolated from 11 out of 30 (36%) human skin samples and from 29 out of 100 (29%) teat skin samples from apparently healthy cows. Genomic DNA from each isolate was double-digested with EcoRI and MseI and complementary oligonucleotide adaptors were ligated to the restriction fragments. Pre-selective and selective, amplification reactions were performed, the amplified fragments were separated by electrophoresis in an ABI377 sequencer and analysed using GeneScan 3.1 and Genotyper 2.5. Three single isolates (a-c), a predominant cluster with 35 isolates (d) and another cluster with two isolates (e) were identified. Both clusters d and e included human and animal isolates genetically related, because the profiles had 90-100% homology. Since no cluster was comprised uniquely of human or animal isolates and given the close genetic relatedness among human and animal samples in the farms, the present findings support the. hypothesis that dairy workers can spread S. aureus through manual milking. (C) 2005 Elsevier B.V. All rights reserved.

Remoção de matéria orgânica, de nutrientes e de coliformes no processo anaeróbio em dois estágios (reator compartimentado seguido de reator UASB) para o tratamento de águas residuárias de suinocultura

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

First isolation and molecular characterization of Toxoplasma gondii from finishing pigs from São Paulo State, Brazil

Toxoplasma gondii infection is widely prevalent in humans in Brazil. Among the food animals, pigs are considered the most important meat source of T. gondii for infection in humans. In the present study, we report the first isolation of viable T. gondii from finishing pigs in Brazil. Antibodies to T. gondii were found in 49 (17%) of 286 pigs prior slaughter using the modified agglutination test (MAT) at a serum dilution of 1:25. Attempts were made to isolate T. gondii from 28 seropositive pigs. Samples of heart, brain, and tongue from each pig were pooled, digested in acid pepsin, and bioassayed in five mice per pig. Viable T. gondii was isolated from seven pigs; all isolates were lethal for mice. Restriction fragment length polymorphism on products of SAG2 locus amplified by PCR revealed that two isolates were Type I and five were Type III. The results indicate that phenotypically and genetically T. gondii isolates from pigs from Brazil are distinct from isolates of T gondii from pigs in the USA. (c) 2005 Elsevier B.V. All rights reserved.

Chemical attributes of a degraded soil after application of water treatment sludges

Mining is an activity that tends to degrade the environment. The restoration of mining areas, aims to accelerate and improve the succession process according to its future use. The objective of this work was to rehabilitate a soil degraded by tin mining activities in the Amazon Basin (Jamari National Forest, State of Rondonia) with the application of water treatment sludge (WTS), and verify the effect of Sludge on Values of pH (CaCl2 0.01 mol L-1), organic matter, P, Ca, Mg, K, H+Al, and soil micronutrient contents when Cultivated with native plants, legumes, and grass species. A factorial (3 x 5) experimental design was used to optimize the rehabilitation of these areas including three N rates (100, 200, and 300 mg N ka(-1) soil supplied by WTS), five plant species (grasses, legumes, and native plants), and two controls (degraded soil with no fertilizer and degraded soil fertilized with mineral fertilizers), with four replications. WTS increased pH values. The chemical products used to treat the water contributed, in greater extension, to increase soil Ca and Fe contents. The use of WTS as fertilizer proved viable, since it contains nutrients for plants; however, nitrogen Should not be used as a criterion to define the rate of Sludge application, because it is present at small amounts in the WTS.

Efeito residual de quatro aplicações anuais de lodo de esgoto e vinhaça na qualidade tecnológica da cana-de-açúcar

O principal objetivo deste trabalho foi avaliar os efeitos residuais, por quatro anos seguidos, de diferentes tipos de resíduos, aplicados de formas e em doses distintas, sobre a qualidade tecnológica da cana-de-açúcar de quinto corte (cultivar SP 81-3250). Os tratamentos testados são combinações de dois tipos de resíduos (lodo de esgoto; vinhaça; lodo de esgoto + vinhaça) com dois modos de aplicação (ao lado da linha de cultivo e em área total) e com duas doses (100 e 200%), e um tratamento-testemunha. O delineamento experimental foi em blocos casualizados, com três repetições, totalizando 39 parcelas. As variáveis analisadas foram: percentagem de sólidos solúveis no caldo extraído (Brix % C.E.), percentagem aparente de sacarose no caldo extraído (Pol % C.E.), percentagem da fibra na cana (Fibra % cana), pureza (%), percentagem aparente de sacarose na cana (Pol % cana), percentagem de açúcares redutores na cana (AR % cana), percentagem de açúcares redutores totais na cana (ART % cana) e açúcares totais recuperáveis (ATR). Os fatores de variação testados (tipo de resíduo, modo de aplicação e doses) não promoveram alterações na qualidade e não comprometeram a valorização da cana-de-açúcar.

Efeito do sombreamento e do substrato sobre a germinação e o crescimento de plântulas de Acacia mangium e Acacia mearnsii

The speed and uniform germination of seeds, followed by the prompt seedling emergence, are highly desirable characteristics in seedling production. Studies aiming to enable the use of organic waste such as sewage sludge and pine bark in the supply chain represent a current demand. The aim of this study was to identify conditions of shading and substrate which are more favorable to the process of seed germination and seedling growth of Acacia mangium and Acacia mearnsii. After overcoming dormancy in hot water, the seeds were sown in three substrates: substrate plantmax commercial forest (control), pine bark and pine bark mixed with biosolids (1:1). The tubes were placed in a tree nursery under three shading conditions: 50%, 30% and 0% (full sun). The evaluated parameters were: speed and germination percentage (daily until 30 days) and shoot length of seedlings (weekly from 75 to 150 days). The experiment was a completely randomized design with treatments in a factorial 3 x 3 and the results were submitted to regression analysis. The highest germination was obtained in the sowing of Acacia mangium in pine bark at 30% shading and commercial substrate at 30 and 50% shading for Acacia mearnsii in pine bark under 50% shading or commercial substrate in full sun. For both species, seedlings grew faster and reached larger size in the final substrate of pine bark with biosolids and in any condition of shading (30 e 50 %) and full sun, anticipating the production of seedlings.

Atributos físicos de um latossolo cultivado com cana-de-açúcar após aplicações de lodo de esgoto e vinhaça

O objetivo deste trabalho foi avaliar se a aplicação anual de lodo de esgoto e vinhaça, resíduos empregados com a finalidade exclusiva de fornecer a quantidade necessária de N e K para cana-de-açúcar, causaria alteração em alguns atributos físicos de um Latossolo Vermelho distroférrico argiloso, após as colheitas do 3º e 4º cortes da cultura. O experimento foi conduzido em blocos ao acaso, com três repetições. Os tratamentos foram: (i) dose de lodo de esgoto para suprir 100% do N exigido pela cultura; (ii) dose de lodo de esgoto para suprir 200% do N exigido; (iii) dose de vinhaça para suprir 100% do K exigido; (iv) dose de vinhaça para suprir 200% do K exigido; (v) combinação dos tratamentos (i) e (iii); (vi) combinação dos tratamentos (ii) e (iv); (vii) testemunha (adubação mineral recomendada). Os somatórios das doses dos resíduos foram de 39 e 51 t ha-1 de lodo de esgoto e de 870 e 1.174 m³ ha-1 de vinhaça até o 3º e 4º cortes, respectivamente. Nas camadas de 0-10; 10-20; 20-30; 30-40 e 40-50 cm de profundidade, após as colheitas do 3º e 4º cortes da cultura, foram determinados: teor de matéria orgânica, porosidade total, macroporosidade, microporosidade, densidade do solo, densidade de partícula, resistência do solo à penetração e teor de água gravimétrico. Aplicações de lodo de esgoto e vinhaça não causaram modificação nos atributos físicos, resultado associado à falta de efeito dos resíduos na matéria orgânica do solo.

Acúmulo de metais pesados em cana-de-açúcar mediante a aplicação de lodo de esgoto e vinhaça

O uso de resíduos agroindustriais pode acumular metais pesados no solo e na planta. O objetivo deste trabalho foi avaliar os efeitos do lodo de esgoto e/ou vinhaça, aplicados no solo, disponibilizando Cd, Cr, Ni e Pb ao solo e quantificar as respectivas concentrações em plantas de cana-de-açúcar, após três aplicações anuais sucessivas. O experimento foi conduzido em condições de campo, com parcelas experimentais de cinco linhas espaçadas de 1,5 m e 10 m de comprimento. O delineamento experimental adotado foi o de blocos casualizados, com três repetições. Os tratamentos avaliados foram: 1. Lodo de esgoto com 100% de N; 2. Lodo de esgoto com 200% de N; 3. Vinhaça com 100% de K; 4. Vinhaça com 200% de K; 5. Lodo de esgoto+vinhaça com 100% de N e K; 6. Lodo de esgoto+vinhaça com 200% de N e K; 7. Testemunha (fertilização mineral). Os baixos teores de metais pesados (Cd, Cr, Ni e Pb) disponíveis no solo e nas partes das plantas de cana-de-açúcar indicam que o lodo de esgoto e a vinhaça, empregados nas doses mencionadas, não apresentaram, após três aplicações anuais sucessivas, potencial de contaminação do sistema solo-planta.