Purificação e caracterização parcial de uma B-D glicosidade de Artemia franciscana com ação sobre celobiose e lactose

-D-glucosidase (EC 3.2.1.21) is one of the most interesting glycosidases, especially for hydrolysis cellobiose releasing glucose, is last step degradation of cellulose. This function makes the -D-glucosidase is of great interest as a versatile industrial biocatalyst, being critical to various bio-treatment / biorefinery processes, such as bioethanol production. Hen in the report, a -D-glucosidase was extracts from protein extracted of the invertebrate marine Artemia franciscana was purified and characterized with a combination of precipitation with ammonium sulfate (0 - 30%, 30 to 50%, 50 to 80%), the fraction saturated in the range of 30 to 50% (called F-II) was applied in a molecular exclusion chromatography, in Sephacryl S-200, the fractions corresponding to the first peak of activity of -D-glucosidase were gathered and applied in a chromatography of ion exchange in Mono Q; the third peak this protein obtained chromatography, which coincides with the peak of activity of -D-glucosidase was held and applied in a gel filtration chromatography Superose 12 where the first peak protein, which has activity of -D-glucosidase was rechromatography on Superose 12. This enzyme is probably multimerica, consisting of three subunit molecular mass of 52.7 kDa (determined by SDS-PAGE) with native molecular mass of 157 kDa (determined by gel filtration chromatography on Superose 12 under the system FPLC). The enzyme was purified 44.09 times with a recovery of 1.01%. Using up p-nitrophenyl-β-D-glucopiranoside as substrate obtained a Km apparent of 0.229 mM and a Vmax of 1.109 mM.60min-1.mL-1mM. The optimum pH and optimum temperature of catalysis of the synthetic substrate were 5.0 and 45 °C, respectively. The activity of the -D-glucosidase was strongly, inhibited by silver nitrate and N- etylmaleimide, this inhibition indicates the involvement of radical sulfidrila the hydrolysis of synthetic substrate. The -D-glucosidase of Artemia franciscana presented degradativa action on celobiose, lactose and on the synthetic substrate -nitrophenyl-β-D-glucopiranoside indicating potential use of this enzyme in the industry mainly for the production of bioethanol (production of alcohol from the participating cellulose), and production hydrolysate milk (devoid of milk lactose)

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

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 (AmV): purificação, caracterização, efeito deletério e mecanismo de ação para bruquídeo 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

Ecogenômica de archaea e monitoramento de comunidades de procariotos redutores de sulfato: aplicações na indústria de petróleo e gás

The human activities responsible for the ambient degradation in the modern world are diverse. The industrial activities are preponderant in the question of the impact consequences for brazilian ecosystems. Amongst the human activities, the petroliferous industry in operation in Potiguar Petroliferous Basin (PPB) displays the constant risk of ambient impacts in the integrant cities, not only for the human populations and the environment, but also it reaches the native microorganisms of Caatinga ground and in the mangrove sediment. Not hindering, the elaboration of strategies of bioremediation for impacted areas pass through the knowledge of microbiota and its relations with the environment. Moreover, in the microorganism groups associated to oil, are emphasized the sulfate-reducing prokaryotes (SRP) that, in its anaerobic metabolism, these organisms participate of the sulfate reduction, discharging H2S, causing ambient risks and causing the corrosion of surfaces, as pipelines and tanks, resulting in damages for the industry. Some ancestries of PRS integrate the Archaea domain, group of microorganisms whose sequenced genomes present predominance of extremophilic adaptations, including surrounding with oil presence. This work has two correlated objectives: i) the detection and monitoring of the gene dsrB, gift in sulfate-reducing prokaryotes, through DGGE analysis in samples of mDNA of a mangrove sediment and semiarid soil, both in the BPP; ii) to relate genomic characteristics to the ecological aspects of Archaea through in silico studies, standing out the importance to the oil and gas industry. The results of the first work suggest that the petrodegraders communities of SRP persist after the contamination with oil in mangrove sediment and in semiarid soil. Comparing the populations of both sites, it reveals that there are variations in the size and composition during one year of experiments. In the second work, functional and structural factors are the probable cause to the pressure in maintenance of the conservation of the sequences in the multiple copies of the 16S rDNA gene. Is verified also the discrepancy established between total content GC and content GC of the same gene. Such results relating ribosomal genes and the ambient factors are important for metagenomic evaluations using PCR-DGGE. The knowledge of microbiota associated to the oil can contribute for a better destination of resources by the petroliferous industry and the development of bioremediation strategies. Likewise, search to lead to the best agreement of the performance of native microbiota in biogeochemical cycles in Potiguar Petroliferous Basin ecosystem

Fungos ectomicorrízicos em áreas de Mata Atlântica do Nordeste do Brasil

Ectomycorrhizal associations are poorly known from tropical lowlands of South America. Recent field trips to the reserve Parque Estadual das Dunas in Natal, in Rio Grande do Norte state, Brazil, revealed a undocumented community of ectomycorrhizal fungi. This type of Mycorrhizal association is frequently in the north hemisphere in temperate and boreal forests. The aim of this work is to analyze the occurrence of ectotrophic areas in atlantic rainforest. Collections along and around the trails in the reserve revealed six genera of putatively ECM fungi which belong to the basidiomycete, Amanitaceae, Boletaceae, Russulaceae, Entolomataceae, and Sclerodermataceae family which are poorly documented in Brazil. Plants belonging to Myrtaceae, Polygonaceae, Leguminosae/Caesalpinioideae, Erythroxylaceae, Malphigiaceae, Bromeliaceae, Loganiaceae, Sapotaceae e Celastraceae were found living next to the species of fungi analized. Our results suggest that the area studied is an ectotrophic environment which shows high diversity of putatively ECM fungi and some plants probably host ECM. The tropical lands are a potential focus to study reinforced by the new records of Scleroderma in Brazil and Northwest of Brazil

Biodegradação de hidrocarbonetos aromáticos policíclicos: prospecção metagenômica e modelagem computacional 3-D de proteínas

Knowledge of the native prokaryotes in hazardous locations favors the application of biotechnology for bioremediation. Independent strategies for cultivation and metagenomics contribute to further microbiological knowledge, enabling studies with non-cultivable about the "native microbiological status and its potential role in bioremediation, for example, of polycyclic aromatic hydrocarbons (HPA's). Considering the biome mangrove interface fragile and critical bordering the ocean, this study characterizes the native microbiota mangrove potential biodegradability of HPA's using a biomarker for molecular detection and assessment of bacterial diversity by PCR in areas under the influence of oil companies in the Basin Petroleum Geology Potiguar (BPP). We chose PcaF, a metabolic enzyme, to be the molecular biomarker in a PCR-DGGE detection of prokaryotes that degrade HPA s. The PCR-DGGE fingerprints obtained from Paracuru-CE, Fortim-CE and Areia Branca-RN samples revealed the occurrence of fluctuations of microbial communities according to the sampling periods and in response to the impact of oil. In the analysis of microbial communities interference of the oil industry, in Areia Branca-RN and Paracuru-CE was observed that oil is a determinant of microbial diversity. Fortim-CE probably has no direct influence with the oil activity. In order to obtain data for better understanding the transport and biodegradation of HPA's, there were conducted in silico studies with modeling and simulation from obtaining 3-D models of proteins involved in the degradation of phenanthrene in the transport of HPA's and also getting the 3-D model of the enzyme PcaF used as molecular marker in this study. Were realized docking studies with substrates and products to a better understanding about the transport mechanism and catalysis of HPA s

Resistência do solo à penetração e ao cisalhamento em diversos usos do solo em áreas de preservação permanente

The soil resistance to penetration and shear can be used as indicators of soil compaction and to indicate the susceptibility of a soil to erosion. The objective of this study was to quantify and compare the impact provided by different land uses in a haplic cambissol in areas of permanent preservation, from the soil resistance to penetration and shear. The experimental area was located in an area of permanent preservation, the sub-basin of the Ribeira Iguape River - SP, with different land uses: banana cultivation - CBAN, degraded pasture - PDEG, silvopastoral system - MPIS and native forest - MNAT. The test for resistance to penetration was accomplished with a digital penetrometer compaction of manual effort, to a depth of 40 cm. The soil shear strength was determined by Vane Test at a depth between 0 and 5 cm. The degraded pasture was similar to native forest, with less resistance to penetration. The banana cultivation and silvopastoral system were the land uses with the highest resistance to penetration, bringing serious risk of erosion in areas of permanent preservation. The soil under native forest had lower shear strength. The cultivation of bananas, degraded pasture and silvopastoral system were the land uses with higher shear strength of soil, indicating that the use of these soils in areas of permanent preservation is promoting the same compression.

Dynamics of some limnological characteristics in pacu (Piaractus mesopotamicus) culture tanks as function of handling

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

Análise da presença de agentes mutagênicos nas águas do Rio Pitimbú/RN

The Pitimbu River is located at the oriental portion of the State of Rio Grande do Norte, including three importants cities named Macaíba, Parnamirim e Natal. Although its high importance as a water source, which supplys great part of the South Zone of the Natal city, this river receives a large quantity of domestic and industrial waste water without treatment. The Pitimbu River headhas its river-head located in the city of Macaiba, goes through Parnamirim, then it flowing into at the Jiqui Lake in Natal. The aim of this study was to evaluate, qualitatively and quantitatively, the environmental quality of the Pitimbu River by genotoxicity bio-assays, which are important tools for genetics toxicological evaluation. In this work, five samples sites, distributed along the river, were used to collect water samples. Another point site, located near Jiqui lake, was used to collect drinkable water, which was treated by CAERN, the water treatment entreprise of Rio Grande do Norte. The following assays where used to evaluate the quality of these samples: Allium cepa assay; Comet assay; Micronuclei (MN) assay; and Ames test. For the Allium cepa assay, sixteen specimes where used for each water sample from the sample sites. In this assay both microscopic, like cytogenetic damage, and macroscopic aspects, as morphological variation were evaluated. Red blood cells from periferical blood of the Crenicichla menezesi native specie were used not only for the MN assay, but also for the Comet assay. These fishes were collected at different points on the Pitimbu River and the negative control was developed using fishes of the same species that were bring to the laboratory and maintained for 100 days in the optimal experimental conditions. For the Ames test, TA100, YG1042, TA98 and YG1041 strains were used in the directed method without metabolic activation. The results found by the Allium cepa assay showed that two water sample sites induced increase of mitotic index (IM). Additionally, compared to the control, all the water samples increased the chromossomal aberrations frequency and/or micronucleus. Among the sample sites, two also showed an abnormal growth rate in its root and two samples induced morphological alteration. With the MN test in red blood cells, a high frequence of MN was observed in tree sample. By comparing all the results obtained on the water sample points and with the negative control, a significant variation on the MN frequency was observed. Positive results were also observed for the same sample to water test by the Comet assay. These results allow concluding that the proposed specie Crenicichla menezesi has a good profile as a bio-indicator for the evaluation of environmental water quality and the MN and comet test can be usuful for in situ evaluation. By the Ames test, it was possible to detect the mutagenic activity on the waters from the Pitimbu River in different levels of mutagenicity. This result suggests that this river has several substances that induced changes directly to the DNA. The mechanisms involved to this phenomenon could be by both processes, by changing of the reading frame and by nucleotide substitution. These data set indicate the presence of mutagenic agents, which can represent in risk to biot and human beens

Composição da fauna de formigas (Hymenoptera: Formicidae) de serapilheira em florestas semidecídua e de Eucalyptus spp., na região sudeste do Brasil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Uso sustentável da Copernicia prunifera (Miller) H. E Moore no semiárido potiguar: valorização de saberes e conservação dos recursos genéticos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Histórias e relatos sobre Pipa: a praia internacional do Rio Grande do Norte

The overwhelming gear that boosted tourism as the main economic activity of Pipa a district formerly agricultural and fishery, that belongs to the municipality of Tibau do Sul/RN produced significant changes in the social, economic and cultural rights of the native population of the place where social relations were narrow and justified in some cases, the line of kinship in an environment where all residents knew each other. However, we can not observe the emergence of this activity just as voracious disarticulator of old local forms of sociability that in a linear process, destroys the old, replacing it with the new. New compositions are generated by merging elements of past and present. Overcoming opposition to simplistic positive or negative impacts of tourism sought especially in this dissertation, review the history of Pipa (told in narrative form), showing how slowly this district has become a major international tourist destination and how its residents were being swallowed by the "whirlwind" that led to this reality. The research used qualitative methodology and was based on photographic, literature and survey, observation, experience, oral and written reports acquired in the field. We conducted in-depth interviews (oral history) with subjects that are the living memory of the place local residents (mostly natives). We found that before the process of "modernization" resulting from the pressure of globalization itself and the capital investment in tourism resulting from the native population is not passive. On the contrary, natives resist, creating mechanisms of material and symbolic reformulations. The present moment is always dynamic. Because of that, the identity of a place is not the crystallization of its past. Many landscapes still reveal materialities of yesteryear, such as registration of social practices in the construction of the place

Viabilidade in vitro de grãos de pólen de bacurizeiro - Clusiaceae

O bacurizeiro (Platonia insignis Mart.) é uma frutífera nativa da Amazônia, que apresenta alogamia acentuada e autoincompatibilidade esporofítica. A viabilidade in vitro tem sido utilizada para representar a capacidade potencial do pólen em completar o processo de fertilização O objetivo do presente trabalho foi estudar a viabilidade de polens de bacurizeiro por meio da germinação in vitro , por ser esta uma técnica em que o comportamento germinativo é semelhante in vivo . Os ensaios foram conduzidos em laboratório, em um delineamento inteiramente casualizado, analisados em esquema fatorial 2 x 3 x 4, em que: 2=formas de propagação (pé-franco e enxertada); 3=estágios da antese (pré-antese, antese e pós-antese), e 4=concentrações de sacarose (0; 7,5; 10 e 20%), com 10 repetições. Houve diferença significativa na germinação do pólen, sendo que, via polinização manual, o pólen deve ser coletado na antese, enquanto o melhor meio para a germinação in vitro é com 7,5 g de sacarose.

Proteinograma sérico de bovinos da raça Curraleir

Estudou-se o perfil eletroforético das proteínas séricas de bovinos sadios da raça Curraleiro por meio da técnica de eletroforese em gel de acrilamida contendo dodecil sulfato de sódio. Utilizaram-se amostras de soro sanguíneo de 228 bovinos da raça Curraleiro, 51 machos e 177 fêmeas, com idades entre sete meses e 12 anos, pertencentes a dois rebanhos localizados nos Estados de Goiás e Tocantins. Foram quantificadas proteína total e concentração plasmática de fibrinogênio. Verificaram-se variações nas concentrações das diferentes frações proteicas. Foram detectadas 26 proteínas e identificadas 10 delas. A ceruloplasmina esteve ausente em 78,1% dos indivíduos, e a α-antitripsina não foi detectada em nenhum animal. Proteína total, globulina, IgA, IgG e fibrinogênio aumentaram com a idade e houve correlação positiva entre os níveis séricos de haptoglobina e α1-glicoproteína ácida.

Padrões distintos de congruência climática em duas espécies invasoras de prosopis em zonas semi-áridas da América do Sul

Based on climate data and occurrence records, ecological niche models (ENM) are an important opportunity to identify areas at risk or vulnerable to biological invasion. These models are based on the assumption that there is a match between the climatic characteristic of native and invaded regions predicting the potential distribution of exotic species. Using new methods to measure niche overlap, we chose two exotic species fairly common in semi-arid regions of South America, Prosopis juliflora (Sw.) D.C. and Prosopis pallida (H. ; B. ex. Willd) HBK, to test the climate matching hypothesis. Our results indicate that both species occur with little niche overlap in the native region while the inverse pattern is observed in the invaded region on South America, where both species occur with high climatic overlap. Maybe some non-climate factor act limiting the spread of P. pallida on the native range. We believe that a founder effect can explain these similarities between species niche in the invaded region once the seeds planted in Brazil came from a small region on the Native range (Piura in Peru), where both species occur sympatric. Our hypothesis of a founder effect may be evident when we look at the differences between the predictions of the models built in the native and invaded ranges. Furthermore, our results indicate that P. juliflora shows high levels of climate matching between native and invaded ranges. However, conclusions about climate matching of P. pallida should be taken with caution. Our models based on climatic variables provide multiple locations suitable for occurrence of both species in regions where they still don t have occurrence records, including places of high interest for conservation.