Atividade bioinseticida e mecanismo de ação de vicilinas de sementes Erythrina velutina sobre moscas-das frutas Ceratitis capitata

The fruit fly Ceratitis capitata is considered the most destructive pest of the world fruitculture. Many pest management practices, mainly based on agrochemicals, have been developed to allow the world-wide commerce of fruit. Solutions to decrease the use of synthetic insecticides in agriculture are based on the development of new target-specific compounds which cause less damage to the environment, especially vegetal proteins with insecticidal effects. The aim of this work was to evaluate the deleterious effect of a purified vicilin of E. velutina (EvV) seeds to C. capitata larvae and adult insects and to investigate the mechanisms involved in these effects. EvV was purified, characterized and its deleterious effect was tested in bioassay systems. EvV mechanism of action was determined by immunodetection techniques and fluorescence localization in chitin structures that are present in C. capitata digestory system. EvV is a glycoprotein with affinity to chitin. Its molecular weight, of 216,57 kDa, was determined by gel filtration chromatography in FPLC system. Using SDS-PAGE, it was possible to observe EvV dissociation in two main subunits of 54,8 and 50,8 kDa. When it was submitted to eletrophoresis in native conditions, EvV presented only one band of acid characteristic. The WD50 and LD50 values found in the bioassays were 0,13% and 0,14% (w/w), respectively for the larvae. EvV deleterious effects were related to the binding to chitin structures presented in peritrophic membrane and gut epithelial cells, associated with its low digestibility in C. capitata digestive tract. The results described herein are the first demonstration of the larvicidal effects of plant protein on C. capitata larvae. EvV may be part of the pest management programs, in the toxic bait composition, or an alternative in plant improvement program

Atividade β-D-N-Acetilglucosaminidásica de enzimas imobilizadas extraídas da Artemia franciscana e possíveis aplicações biotecnológicas

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

Bioecology and conservation of Duck Mussel Species (Anodonta anatina L., 1758) in Northeastern Portugal

Freshwater mussel (Mollusca, Bivalvia, Unionoida) populations are one of the most endangered faunistic groups. Mussels play an important role in the functioning of aquatic ecosystems, because they are responsible for the filtration and purification of water. They have a complex life cycle, with a parasitic larvae and usually limited host fish species. The real status of these populations is still poorly understood worldwide. The objectives of the present work were the study of bioecology of duck mussel (Anodonta anatina L.) populations of Tua Basin (NE Portugal). It was made the characterization of the ecological status of Rabaçal, Tuela and Tua Rivers, selecting 15 sampling sites, equally distributed by the three rivers. Samplings were made in the winter of 2016, and several physico-chemical water parameters measured and two habitat quality indexes calculated (GQC and QBR indexes). Benthic macroinvertebrate communities were sampled based on the protocols established by the Water Framework Directive. Host fish populations for duck mussel were determined in laboratorial conditions, testing several native and exotic fish species. The results showed that several water quality variables (e.g. dissolved oxygen, conductivity, pH, total dissolved solids, and nutrients) can be used for the classification of river typology. Other responsive metrics were also determined to identify environmental degradation. For instances, hydromorphological conditions (GQC and QBR indexes) and biota related metrics (e.g. composition, distribution, abundance, diversity of invertebrate communities) contributed to the evaluation of the ecological integrity. The upper zones of Rabaçal and Tuela rivers were classified with excellent and good ecological integrity, while less quality was found in downstream zones. The host fish tests showed that only native species are effective hosts, essential for the conservation purposes of this mussel species. Threats, like pollution, sedimentation and river regularization (3 big dams are in construction or in filling phase), are the main cause of habitat loss for native mussel and fish populations in the future. Rehabilitation and mitigation measures are essential for these lotic ecosystems in order to preserve the prioritary habitats and the native species heavily threatened.

Purificação, caracterização e efeitos imunomodulatório e antiproliferativo de uma lectina do fungo Clavaria cristata (Holmsk.) Pers

A 140,0 kDa lectin was purified and characterized from the mushroom Clavaria cristata. The purification procedures from the crude extract of the mushroom comprised gel filtration chromatography on Sephacryl s200 and ion exchange on Resource Q column. The purified lectin agglutinated all types of human erythrocytes with preference for trypsinized type O erythrocytes. The haemagglutinating activity is dependent of Ca 2+ ions and was strongly inhibited by the glycoprotein bovine submaxillary mucin (BSM) up to the concentration of 0, 125 mg/mL. The C. cristata lectin (CcL) was stable in the pH range of 2,5-11,5 and termostable up to 80 °C. CcL molecular mass determined by gel filtration on a Superose 6 10 300 column was approximately 140,3 kDa. SDS polyacrilamide gel electrophoresis revealed a single band with a molecular mass of approximately 14,5 kDa, when the lectin was heated at 100 ⁰C in the presence or absence of β-mercaptoethanol. CcL induced activation of murine peritoneal macrophages in vitro resulting in the release of nitric oxide (NO), reaching the maximum production at 24 h. In experimental paw oedema model in mice, CcL showed proinflammatory activity being able to induce oedema formation. Cell viability of HepG2, MDA 435 e 3T3 cell lines was examined after 72 h of incubation with CcL in different concentrations (0,5-50 μg/mL). CcL inhibited HepG2 cells growth with an IC50 value of 50 μg/mL. In the present work, the observed immunomodulatory and antiproliferative effects indicate CcL as a possible immunomodulator compound, interfering in the macrophages immune response, taking possible anti-parasitic, anti-tumoral effects or diagnostic and/or therapeutic

Vicilinas de sementes de leguminosas selvagens:purificação, caracterização, efeito de deletério e mecanismo de ação para bruquídeos e para fungos fitopatogênicos

Globulins fractions of legume seeds of Crotalaria pallida, Erytrina veluntina and Enterolobium contortisiliquum were isolated and submitted to assays against serine, cysteine and aspartic proteinases, as also amylase present in midgut of C. maculatus and Z. subfasciatus. Hemagglutination assays indicated presence of a lectin in E. veluntina globulin fractions. This lectin had affinity to human erythrocytes type A, B and O. Vicilins were purified by chromatography on Sephacryl S-300 followed of a chromatography on Sephacryl S-200, which was calibrated using protein markers. Vicilins from C. pallida (CpV) and E. veluntina (EvV) seeds had a molecular mass of 124.6 kDa and E. contortisiliquum a molecular mass of 151kDa. Eletrophoresis in presence of SDS showed that CpV was constituted by four subunities with apparent molecular mass of 66, 63, 57 and 45 kDa, EvV with three subunities with apparent molecular mass of 45kDa and EcV four subunities, two with 37.1 kDa and two with 25.8 kDa. Non denaturantig eletrophoresis displayed single bands with high homogeneity, where CpV had lower acidic behavior. All vicilins are glycoproteins with carbohydrate contents at 1 to1.5%. Bioassays were done to detect deleterious effects of vicilins against C. maculatus and Z. subfasciatus larvae. CpV, EvV and EcV exhibited a WD50 of 0.28, 0.19 and 1.03%; LD50 0.2, 0.26, and 1.11% respectively to C. maculatus. The dose responses of CpV, EvV and EcV to Z. subfasciatus were: WD50 of 0.12, 0.14, 0.65% and LD50 of 0.09, 0.1, and 0.43% respectively. The mechanism of action of these proteins to bruchids should be based on their properties of bind to chitin present in mid gut of larvae associated with the low digestibility of vicilin. In assays against phytopatogenous fungus, only EcV was capable of inhibit F. solani growth at concentrations of 10 and 20 µg and its action mechanism should be also based in the affinity of EcV to chitin present in the fungi wall

Vicilina de sementes da leguminosa selvagem Anadenanthera macrocarpa: purificação, caracterização, efeito deletério e mecanismo de ação para Callosobruchus maculatus

Grains and legume seeds are foods that form the basis of the diets of many cultures around the world, winch contritbute to the daily nutrient requirements of humans. Vicilins (7S globulin) are storage proteins found in legume seeds, and may have an additional function constitutive defense of the embryo against pests and pathogens. In this work the vicilin from Anadenanthera macrocarpa - AmV (red-angico), was purified and partially characterized, its effect on development and larval survival and adult emergence of Callosobruchus maculatus was evaluated by determination of LD50, WD50 and ED50 in system bioassay. Purification of vicilin was initiated by the chitin affinity chromatography and then gel filtration (Superdex 75 Tricorn 10x300 mm) FPLC system followed by reverse phase chromatography (C8 phenomenex) on HPLC system. Bioassays WD50 and LD50 for larvae were 0.32% and 0.33% (w:w) respectively, since the ED50 for adults was 0.096%. The probable mechanism of action was evaluated by testing digestibility of AmV in vitro, and observed for the involvement of two fragments vicilins immunoreactive against polyclonal Anti-vicilin from Erythrina velutina (Anti-EvV) about of 22 and 13 kDa chitin binding. The AmV in its native form has been recognized by the anti-EvV, indicating that there is a conserved region in the vicilin and is probably corresponding to the chitin binding domains. These results point to a new vicilin chitin binding that can subsequently be used as a possible biopesticide protein source, in order to control insect pest C. maculatus and confirm literature findings that demonstrate vicilin in the presence of different kinds of ligands to conserved regions chitin not yet characterized

Adição de superplastificante e anti-segregante à base de CMC em pastas de cimento para Poços de Petróleo

Oil well cementing materials consist of slurries of Special class Portland cement dispersed in water. Admixtures can be used to provide the necessary fluidity, so the material can be efficiently pumped down as well as penetrate porous rocks with controlled filter loss. Construction admixtures can be used to modify the properties of oil well cements provided they can withstand and hold their properties at the higher than ambient temperatures usually encountered in oil fields. In civil construction, superplasticizer play the role of dispersants that reduce the facto r of water cement improve mechanical properties and fluidity of the cement, whereas anti-segregation agents improve the workability of the slurry. In the present study, oil well cement slurries were produced adding both a dispersant and an anti-segregation agent conventionally used in Portland CPII-Z-32 RS cement aiming at materials for primary cementing and squeeze operations. Three basic aspects were evaluated: fluidity, filter loss and the synergetic effect of the admixtures at two temperatures, i.e., 27°C and 56°C, following API RP 10B practical recommendations. The slurries were prepared using admixture concentrations varying from 2.60 Kgf/m3 (0.02 gallft3) to 5.82 Kgf/m3 (0.045 galJft3) BWOC. The density of the slurries was set to 1.89 g/cm3 (15.8 Ib/gal). 0.30 to 0.60% BWOC of a CMC-based anti-segregation agent was added to the cement to control the filter loss. The results showed that the addition of anti-segregation at concentrations above 0.55% by weight of cement resulted in the increased viscosity of the folders in temperatures evaluated. The increasing the temperature of the tests led to a reduction in the performance of anti-segregation. At concentrations of 5.20 kgf/m3 (0,040 gallft3) and 5.82 Kgf/m3 (0,045 gal/ft 3) observed a better performance of the properties evaluated in the proposed system. At low temperature was observed instability in the readings of rheology for all concentrations of anti-segregation. Contents that increasing the concentration of anti¬-segregation is limited concentrations greater than 0.55 % BWOC of the CMC in temperature analyzed. The use of the system with CMC promoted a good performance against the properties evaluated. The principal function of anti¬-segregation was optimized with increasing concentration of superplasticizer, at temperatures above the 2rC. The study of the behaviour of systemic additives, resulting in slurries of cement, which can be optimized face studies of other intrinsic properties in oil fields

Existência de subgrupos livres em grupos finitamente apresentados com mais geradores do que relações

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Exatas Departamento de Matemática, 2016.

Obtenção de filtro cerâmico a partir da diatomita e casca de arroz carbonizada visando tratamento de efluente têxtil

The effluents released by the textile industry have high concentrations of alkali, carbohydrates, proteins, in addition to colors containing heavy metals. Therefore, a filter was prepared aiming primarily to the removal of color. In order to prepare this filter, rice hulls and diatomite were used, which have in their structure, basically amorphous hydrated silica. The silica exists in three crystalline forms: quartz, tridymite and cristobalite. In accordance with the above considerations, this study was divided into two stages; the first corresponds to the preparation of the filter and the second to carry out the tests in the effluent/filter in order to verify the efficiency of the color removal. First, the raw material was subjected to a chemical analysis and XRD, and then the diatomite was mixed, via humid, with a planetarium windmill with 20 %, 40 %, 60 % and 80 % of rice husk ash. To the mixture, 5 % carboxymethylcellulose (CMC) was added as a binder at room temperature. The samples were uniaxially compacted into metallic matrix of 0.3 x 0.1 cm² of area at a pressure of 167 MPa by means of hydraulic press and then sintered at temperatures of 1,000 °C, 1,200 °C and 1,400 °C for 1 h and submitted to granulometry test using laser, linear retraction, water absorption, apparent porosity and resistance to bending, DTA, TMA and XRD. To examine the pore structure of the samples scanning electron microscope (SEM) was used. Also tests were carried out in a mercury porosimeter to verify the average size of the pores and real density of the samples. In the second stage, samples of the effluent were collected from a local industry, whose name will be preserved, located in Igapó, in the State of Rio Grande do Norte - RN. The effluent was first pretreated before filtration and then subjected to a treatment of flotation. The effluent was then characterized before and after filtration, with parameters of color, turbidity, suspended solids, pH, chemical and biochemical oxygen demand (COD and BOD). Thus, through the XRD analysis the formation of cristobalite α in all samples was observed. The best average size of pore was found to be 1.75 μm with 61.04 % apparent porosity, thus obtaining an average 97.9 % color removal and 99.8 % removal of suspended solid

Membrana de alumina anódica: comportamento da microestrutura e estudo das propriedades ópticas após tratamento térmico

Thin commercial aluminum electrolytic and passed through reactions was obtained with anodic alumina membranes nanopores. These materials have applications in areas recognized electronic, biomedical, chemical and biological weapons, especially in obtaining nanostructures using these membranes as a substrate or template for processing nanowires, nanodots and nanofibers for applications noble. Previous studies showed that the membranes that have undergone heat treatment temperature to 1300° C underwent changes in morphology, crystal structure and optical properties. This aim, this thesis, a study of the heat treatment of porous anodic alumina membranes, in order to obtain and to characterize the behavior changes structures during the crystallization process of the membranes, at temperatures ranging between 300 and 1700° C. It was therefore necessary to mount a system formed by a tubular furnace resistive alumina tube and controlled environment, applying flux with special blend of Ag-87% and 13% N2, in which argon had the role of carrying out the oxygen nitrogen system and induce the closing of the pores during the densification of the membrane. The duration of heat treatment ranged from 60 to 15 minutes, at temperatures from 300 to 1700° C respectively. With the heat treatment occurred: a drastic reduction of porosity, grain growth and increased translucency of the membrane. For the characterization of the membranes were analyzed properties: Physical - thermogravimetric, X-ray diffraction, BET surface area; morphological - SEM, EDS through compositional and, optical absorbance, and transmittance in the UV-VIS, and FTIR. The results using the SEM showed that crystallization has occurred, densification and significant changes in membrane structure, as well as obtaining microtube, the BET analysis showed a decrease in specific surface area of the membranes has to 44.381 m2.g-1 to less than 1.8 m2.g-1 and in the analysis of transmittance and absorbance was found a value of 16.5% in the range of 800 nm, characteristic of the near infrared and FTIR have confirmed the molecular groups of the material. Thus, one can say that the membranes were mixed characteristics and properties which qualify for use in gas filtration system, as well as applications in the range of optical wavelength of the infra-red, and as a substrate of nanomaterials. This requires the continuation and deepening of additional study

Comportamento reológico de materiais cimentícios para poços de petróleo contendo naftaleno condensado ou policarboxilato

Pastas a base de cimento Portland são utilizadas na cimentação de poços de petróleo. Elas consistem de uma mistura de partículas sólidas de cimento dispersas em água e aditivos. Atualmente, diversos materiais alternativos são utilizados como aditivos, objetivando a modificação e a melhoria das propriedades das pastas de cimento, especialmente no aumento da fluidez. Novos aditivos plastificantes são capazes de suportar as diversas condições de poços, promovendo propriedades no estado fluido compatíveis às condições exigidas para cimentação.Dispersantes são os componentes da pasta que garantem fluidez, além de proporcionar controle na água perdida por filtração na formação porosa, garantindo o sucesso da operação de bombeio. Em deter minados campos, além do efeito da profundidade, as condições geológicas das formações promovemvariações do gradiente de pressão e temperatura ao longo da profundidade vertical do poço. Recentemente, diversos aditivos químicos da indústria da construção civil tem sido estudados em condições de cimentação de poços de petróleo. Vários produtos testados tem apresentado desempenho superior aos produtos normalmente empregados pela indústria de petróleo com boa relação custo/benefício em função do volume de mercado da construção civil. Resultados promissores na seleção de aditivos com função dispersante da construção civil para operações de cimentação de poços de petróleo onshore foram obtidos para temperaturas até 80°C. O potencial de uso desses aditivos permite estabelecer novas soluções para problemas encontrados na cimentação de poços de petróleo HPHT, poços sujeitos à injeção de vapor, poços depletados e poços produtores de gás. Na construção civil, os superplastificantes permitem reduzir o fator água/cimento das argamassas proporcionando melhoria de propriedades como resistência mecânica e fluidez. Assim, o objetivo deste trabalho foi o estudo e a caracterização reológica de pastas constituídas de cimento Portland, água e aditivos do tipo plastificante, com função dispersante a base de naftaleno condensado e policarboxilato, na faixa de temperaturas de 58°C e 70ºC. As condições utilizadas para a avaliação dos aditivos alternativos foram baseadas em uma cimentação primária para um poço hipotético de 2200 m de profundidade e gradientes geotérmicos de 1,7°F/100 pés e 2,1°F/100 pés. Os resultados demonstraram a grande eficiência e o poder dispersivo do policarboxilato para as temperaturas estudadas. O aditivo promoveu alta fluidez, sem efeitos de sedimentação da pasta. O dispersante à base de naftaleno reduziutant o a viscosidade plástica como o limite de escoamento acimada concentração a partir de 0,13%. O modelo de Bingham descreveu bem o comportamento reológico das pastas com policarboxilato para todas as concentrações

Isolamento e caracterização biológica e bioquímica de um activador do factor x da coagulação presente no veneno da serpente Bothrops erythromelas

Dissertação de Mestrado Integrado em Medicina Veterinária

Estabilidade térmica dos compostos biologicamente ativos do colostro

Dissertação de Mestrado, Engenharia Biológica, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Resposta metabólica da ostra Crassostrea gigas na presença do parasita Amyloodinium ocellatum

Dissertação de Mestrado, Aquacultura e Pescas, Especialização em Aquacultura, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Desenvolvimento de membranas porosas de alumina para fins de tratamento de efluente industrial têxtil

Textile production has been considered as an activity of high environmental impact due to the generation of large volumes of waste water with high load of organic compounds and strongly colored effluents, toxic and difficult biodegradability. This thesis deals with obtaining porous alumina ceramic membranes for filtration of textile effluent in the removal of contaminants, mainly color and turbidity. Two types of alumina with different particle sizes as a basis for the preparation of formulation for mass production of ceramic samples and membranes. The technological properties of the samples were evaluated after using sintering conditions: 1,350ºC-2H, 1,450ºC-30M, 1,450ºC-2H, 1,475ºC-30M and 1,475ºC-2H. The sintered samples were characterized by XRD, XRF, AG, TG, DSC, DL, AA, MEA, RL, MRF-3P, SEM and Intrusion Porosimetry by Mercury. After the characterization, a standard membrane was selected with their respective sintering condition for the filterability tests. The effluent was provided by a local Textile Industry and characterized at the entry and exit of the treatment plant. A statistical analysis was used to study the effluent using the following parameters: pH, temperature, EC, SS, SD, oil and grease, turbidity, COD, DO, total phosphorus, chlorides, phenols, metals and fecal coliform. The filtered effluent was evaluated by using the same parameters. These results demonstrate that the feasibility of the use of porous alumina ceramic membranes for removing contaminants from textile effluent with improved average pore size of 0.4 micrometre (distribution range varying from 0,025 to 2.0 micrometre), with total porosity of 29.66%, and average percentages of color removal efficiency of 89.02%, 92.49% of SS, turbidity of 94.55%, metals 2.70% (manganese) to 71.52% (iron) according to each metal and COD removal of 72.80%