Metagenômica: busca de novos genes envolvidos com a biodegradação de hidrocarbonetos

Industrial activities, oil spills and its derivatives, as well as the incomplete combustion of fossil fuels have caused a great accumulation of hydrocarbons in the environment. The number of microorganisms on the planet is estimated at 1030 and prokaryotes the most abundant. They colonized diverse environments for thousands of years, including those considered extreme and represent an untapped source of metabolic and genetic diversity with a large biotechnological potential. It is also known that certain microorganisms have the enzymatic capacity to degrade petroleum hydrocarbons and, in many ecosystems, there is an indigenous community capable of performing this function. The metagenomic has revolutionized the microbiology allowing access uncultured microbial communities, being a powerful tool for elucidation of their ecological functions and metabolic profiles, as well as for identification of new biomolecules. Thus, this study applied metagenomic approaches not only for functional selection of genes involved in biodegradation and emulsification processes of the petroleum-derived hydrocarbons, but also to describe the taxonomic and metabolic composition of two metagenomes from aquatic microbiome. We analyzed 123.116 (365 ± 118 bp) and 127.563 sequences (352 ± 120 bp) of marine and estuarine metagenomes, respectively. Eight clones were found, four involved in the petroleum biodegradation and four were able to emulsify kerosene indicating their abilities in biosurfactants synthesis. Therefore, the metagenomic analyses performed were efficient not only in the search of bioproducts of biotechnological interest and in the analysis of the functional and taxonomic profile of the metagenomes studied as well

Estrutura da população de Microcerotermes exiguus ( Isoptera: Termitidae) e sua partuicipação no consumo de madeira em uma área de caatinga, Nordeste, Brasil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Estudos in silico da proteína AlkB e prospecção de bactérias alcanotróficas em solos da bacia petrolífera potiguar

Some microorganisms from virgin ecosystems are able to use petroleum it as source of carbon and energy. The knowledge of microbial biodiversity can help to reveal new metabolic systems for utilization alkanes with biotechnological importance. The aim of this study is: i) Accomplish an in silico study of the AlkB protein aimed to understand the probable mechanism involved on selectivity of alkanes in Gram positive and Gram negative bactéria. ii) prospect and analyze the response of the microbial alkanotrophics communities in soil and mangrove sediments of BPP RN and soil of Atlantic forest in the Horto Dois Irmãos Reserve area/PE using the molecular biomarker, gene alkB; with the PCR and PCR-DGGE approach

Qualidade de água como tema para a socialização do conhecimento científico em região semi-árida brasileira

During the dry and rainy periods, in the Piranhas-Assu river hydrographic basin, variations in bacterioplankton and trophic state indexes were studied in the lotic and lentic environments. The means of total bacterial density wavered in the order of 107 organisms. mL-1 and they were higher in the rainy period. Cocci bacteria were numerically predominant in all sampling points, influencing the total abundance of bacterioplankton. Total bacterial biomass varied from 659 to 1997,3 µgC.L-1 due to the high values of filaments cellular volume. Total phosphorus amounted 108 µg.L-1 and it had a positive correlation with chlorophyll a (r = 0,94; p < 0,05) during dry period, when these variables presented higher concentrations. There wasn t a high spatial variation. The low transparency of water and the phosphorus and chlorophyll concentrations indicated eutrophic or hipereutrophic levels in studied points of reservoir. These variables, associated with high density and biomass values found, alert to the necessity of a constant monitoring of the semi-arid aquatic ecosystems, where the quality of water is affected by dry occurrence

Vegetação dunar: caracterização estrutural de dunas do município de Natal-RN como subsídio para implantação de técnicas de reflorestamernto, recuperação e conservação do ecossistema

The Brazilian coast has a wide variety of complex environments and ecosystems along the coast, about 80% are represented by sandbanks and dunes. The coastal ecosystems were the first to suffer the impacts man and places, as the very fragile ecosystems, are somehow altered. Are few areas of restinga well as natural features, very few protected in conservation units. Only in the last two decades the Brazilian restinga have been studies that are showing their importance for biodiversity of the country, though its economic importance remains largely unknown. In Rio Grande do Norte in the restinga vegetation and dune environments extend for almost the entire coast. The dunes are distinguished in the coastal landscape of the state due to the exuberance of its forms, heights and coating plants. The dune system is of fundamental importance for the maintenance of coastal urban settlements, especially for the city of Natal, acting on the hydrological dynamics of water table and reducing the effect of wind and movement of grains of sand to the interior and thus avoiding the burial City. However, the ecosystem of restinga and dune environments have been weakened and destroyed according to the intense urbanization and the knowledge of the vegetation of restinga installed on the dunes are still scarce. Thus, the objective of this study was to characterize the structure and floristic composition of vegetation established on a dune in the Dunes State Park Christmas and gather information to develop a model of recovery of the dune ecosystem. This dissertation is composed of 2 chapters, the first being: Structure of the vegetation of the dunes Dunes State Park in Natal, RN with the objective of describing the structure and composition of species of tree-shrub vegetation of restinga dunes of the Parque das Dunas and second: Recovery of degraded areas in a sand dune, which aimed to review the terms and concepts used in the theme of recovery and the techniques for recovery of degraded areas with emphasis on sandy environments and poor in nutrients, reporting some experiences within and external to Brazil the country, mainly in the Northeast and dunes positive and negative aspects that should be followed in building a model to be adopted for the recovery of local dunes

Entendendo as interações entre povos pesqueiros, manguezal e área protegida: RDS estadual Ponta do Tubarão (RN, Brasil)

The mangrove ecosystem has a great importance to maintaining marine biodiversity, and to the livelihoods of communities living around it. Despite its importance, many mangrove areas have been extensively cleared and converted to other uses. There are few places where communities can retain their traditional uses of mangrove. A good example of conservation of traditional costumes are the fishing communities of the Sustainable Development Reserve (Reserva de Desenvolvimento Sustentável RDS) Ponta do Tubarão. The creation of the reserve came from the own population, and the devastation of a mangrove area, which would be destined for activity shrimp, was one of the reasons that led the population to ask authorities the establishment of a legally protected area. Thus, this study aimed to investigate the environmental perception of the fishing communities of RDS Ponta do Tubarão with respect to the mangroves in a biological, ecological and social perspective as well as evaluate the influence of the reserve in the mangrove conservation and quality of life. To meet objectives of this study was required the appropriation of some method and approaches of ethnosciences and environmental perception. Data were collected through direct observation, and semi-structured interviews. Through content analysis found that the population has a strong dependence on the mangrove ecosystem, as well as provides a good ecological knowledge of ecosystem functions. It was also found that the reservation has a good performance in the conservation of the mangroves; however, need to outline strategies to conciliate both the biological and cultural conservation.

Sustentabilidade ambiental do cultivo intensivo de Tilápias (Oreochromis niloticus) em tanques-rede e a capacidade de suporte de quatro reservatórios em uma região semi-árida tropical

Intensive production of tilápias, in cages or net tanks, has been proposed as an alternative to increase fish production, that would generate income and offer animal protein in different tropical and subtropical countries. However, this system of production enriches the aquatic environment with nutrients, principally nitrogen and phosphor derived from the dejections of the metabolism of food and eventual food surpluses consumed by the fishes; the alimentation of the fishes in this production modality is dependent on fish food. The emission of these nutritions in levels above the limit that the system is capable metabolize can provoke a phenomenon called eutrofization, putting in risk the quality of water for public and for fish production activities. In this context the work had as a goal to evaluate the trófico state of the four reservoirs for intensive production of tilapias in net-tanks, in other words, he maximum fish production that the reservoirs are able to hold, keeping the desired quantity of nutrient concentrations in water for public use. The results of the four ecosystems in the Boqueirão de Parelhas reservoir showed that it s possible to have an intensive production of tilapias in net tanks, in this environment the annual average concentration of phosphor, was below the considered critical limits to deflagrate the process of eutrofization in semi-arid regions. The carrying capacity of the Boqueirão de Parelhas reservoir depends on the conversion of the feeding facts and phosphor content in the food but it should vary between 100 and 300 tons per year over a variation in the conversion feeding factor of 1,7 to 2,0:1 and a variation in the P in the food of 0,7 to 0,9%

Por qué hacer un trabajo de campo? Experiencia de alumnos del profesorado en ciencias biologicas

Los trabajos de campo son experiencias que docentes y alumnos pueden utilizar como estrategia didáctica para desarrollar destrezas y aumentar la motivación al abordar el tema ciencias. En el presente trabajo se describe una metodología puesta en práctica con alumnos del Profesorado en Ciencias Biológicas, que se propone a los docentes para planificar sus salidas de campo. A partir del trabajo de campo desarrollado se pudo evidenciar la importancia de éste tipo de actividades didácticas extra-áulicas para el desarrollo de actitudes y destrezas, tales como la organización de tareas, colaboración entre pares, redacción de informes científicos, además, sobre la base de hipótesis y objetivos previos, el abordaje interdisciplinario para la observación y análisis del ambiente en estudio.

Por qué hacer un trabajo de campo? Experiencia de alumnos del profesorado en ciencias biologicas

Los trabajos de campo son experiencias que docentes y alumnos pueden utilizar como estrategia didáctica para desarrollar destrezas y aumentar la motivación al abordar el tema ciencias. En el presente trabajo se describe una metodología puesta en práctica con alumnos del Profesorado en Ciencias Biológicas, que se propone a los docentes para planificar sus salidas de campo. A partir del trabajo de campo desarrollado se pudo evidenciar la importancia de éste tipo de actividades didácticas extra-áulicas para el desarrollo de actitudes y destrezas, tales como la organización de tareas, colaboración entre pares, redacción de informes científicos, además, sobre la base de hipótesis y objetivos previos, el abordaje interdisciplinario para la observación y análisis del ambiente en estudio.

Filogenia molecular do gênero Morganella Zeller (Fungi, Basidiomycota) utilizando marcadores ITS

With the development and improvement of techniques for molecular studies and their subsequent application to the systematic, significant changes occurred in the classification of gasteroid fungi. The genus Morganella belongs to the family Lycoperdaceae, and is characterized mainly by lignicolous habit and presence of paracapilicium. Recent data demonstrate the discovery of new species for the group and the existence of a wide variety of species occurring in tropical ecosystems. However, the phylogenetic relationships of the genus, as well as the taxonomic classification, still require revisions to be better understood, the literature studies that address this issue are still very scarce. Thus, the objective of this study was to conduct studies of molecular phylogeny with species of the genus Morganella, to enhance understanding of the phylogeny of the group by including tropical species data. For this, the specimens used both for DNA extractions as for morphological review were obtained from Brazilian and foreign herbaria. For morphological analysis were observed characters relevant to the group's taxonomy. For phylogenetic analysis the Maximum Parsimony and Bayesian Analyzes were used, using the internal transcribed spacer (ITS) of nuclear ribosomal DNA. In phylogenetic analyzes, representatives from Morganella form a monophyletic clade with good support value and based on these results the genus should not be included as subgenus of Lycoperdon. The analysis indicated that M. pyriformis was not grouped with other representatives of Morganella, and therefore should not be included in the group as representative of Apioperdon subgenus because it is a Lycoperdon representative. Moreover, M. fuliginea, M. nuda, M. albostipitata, M. velutina, M. subincarnata are grouped with high support values within the genus Morganella. Morganella arenicola based on morphological and molecular studies does not aggregate in Morganella. Morganella nuda was grouped with M. fuliginea giving indications that can be treated as an intraspecific variation. The results of the analyzes favor to a better understanding of the species of Morganella. However, additional studies using a greater number of species, as well as other molecular markers are needed for a better understanding of the phylogenetic of Morganella.

Dieta do ouriço-branco, Tripneustes ventricosus (Lamarck, 1816), no Arquipélago de Fernando de Noronha, PE

Sea urchins are benthic macroinvertebrates that inhabit shallow coastal waters in tropical and temperate zones. Urchins are usually classified as generalists or omnivores as they can adjust their diet according to the food resources available in the environment. Due to the strong grazing pressure they may exert, urchins have an important role in marine ecosystems, occupying different trophic levels and stimulating the intensification of the dynamics of communities where they occur. In 2004, a monitoring program focused on the population dynamics of the white sea urchin, Tripneustes ventricosus, has been initiated in the Fernando de Noronha Archipelago. At the same time, a surprisingly lack of information on the species biology has been noted, despite their wide geographical distribution and economic importance in many parts of its range. Hence, this work was developed to provide information on the feeding habits of T. ventricosus in the archipelago. Ten specimens were collected between December 2006 and July 2007 at two sites of the archipelago, Air France and Sueste Bay for biometrics and analysis of gut contents. Test diameters ranged from 9.19 cm (± 1.1) to 10.08 cm (± 0.58). Calculated stomach repletion index (IRE) was higher (p <0.05) in the Air France site and also during January and July. The IRE was not correlated to the gonad index. Fifteen different species of algae were detected in a total of 120 stomachs examined: 4 Chlorophytas, 4 Phaeophytas and 6 Rhodophytas. Food diversity (p <0.05) was higher in December 2006 and January 2007. Although several items had a high frequency of occurrence, they were low represented in terms of weight, and consequently, had a low level of relative importance. The brown algae Dictyopteris spp and Dictyota spp, followed by the green algae Caulerpa verticillata accounted for the greatest importance in T. ventricosus diet, comprising about 90% of the consumed items

Efeitos de manipulações ascendentes e descendentes sobre a comunidade fitoplanctônica em um reservatório eutrófico no semi-árido brasileiro

Top-down (grazing) and bottom-up (nutrient, light) controls are important in freshwater ecosystems regulation. Relative importance of these factors could change in space and time, but in tropical lakes bottom-up regulation has to been appointed as more influent. Present study aimed to test the hypothesis that phytoplankton growths rate in Armando Ribeiro reservoir, a huge eutrophic reservoir in semi-arid region of Rio Grande do Norte state, is more limited by nutrient available then zooplankton grazing pressure. Bioassay was conduced monthly from September (2008) to August (2009) manipulating two levels of nutrients (with/without addition) and two level of grazers (with/without removal). Experimental design was factorial 2X2 with four treatments (X5), (i) control with water and zooplankton from natural spot ( C ), (ii) with nutrient addition ( +NP ), (iii) with zooplankton remove ( -Z ) and (iv) with zooplankton remove and nutrient addition ( -Z+NP ). For bioassay confection transparent plastic bottles (500ml) was incubate for 4 or 5 days in two different depths, Secchi`s depth (high luminosity) and 3 times Secchi`s depth (low luminosity). Water samples were collected from each bottle in begins and after incubates period for chlorophyll a concentration analysis and zoopalnktonic organisms density. Phytoplankton growths rates were calculated. Bifactorial ANOVA was performance to test if had a significant effect (p<0,005) of nutrient addition and grazers remove as well a significant interaction between factors on phytoplankton growths rates. Effect magnitude was calculated the relative importance of each process. Results show that phytoplankton growth was in generally stimulated by nutrient addition, as while zooplankton remove rarely stimulated phytoplankton growth. Some significant interactions happening between nutrient additions and grazers remove on phytoplankton growth. In conclusion this study suggests that in studied reservoir phytoplankton growth is more controlled by ascendent factors than descendent

Variação temporal da Comunidade de macroinvertebrados bentônicos em um riacho intermitente do semiárido brasileiro

In Brazilian semiarid region, the majority of water bodies are temporary and the biodiversity in these ecosystems is poorly known. The goal of this study was to describe the temporal variation of benthic macroinvertebrates in an intermittent stream in the Brazilian semiarid region. From March to July of 2009, surveys of benthic macroinvertebrates and water physiochemical parameters were done in a first order stream located at Piranhas-Açu River basin. 25 macroinvertebrates families were found, 21 belonging to Insecta class. The chironomids were the most abundant group during all study period and were represented by 19 genus. The largest densities and taxonomic richness were seen in the drying phase of the stream while the smallest values were found in the period with the heaviest rainfalls ( wet phase ). Only the families Chironomidae and Ceratopogonidae were found during all study period, suggesting that these groups have better adaptations to support floods and droughts. Furthermore, these two groups seemed to be pioneering in this stream. Among the chironomids, Paratendipes dominated the period of floods while Tanytarsus were more abundant in the drying phase . This study showed that hydrological fluctuation is the main force influencing the macroinvertebrate community in this stream, therefore, efforts seeking the conservation and management of Brazilian semiarid water bodies should consider this high natural variability in flow regime

Metagenômica comparativa de solo de regiões de mata atlântica e caatinga do estado do Rio Grande do Norte - Brasil

The microorganisms play very important roles in maintaining ecosystems, which explains the enormous interest in understanding the relationship between these organisms as well as between them and the environment. It is estimated that the total number of prokaryotic cells on Earth is between 4 and 6 x 1030, constituting an enormous biological and genetic pool to be explored. Although currently only 1% of all this wealth can be cultivated by standard laboratory techniques, metagenomic tools allow access to the genomic potential of environmental samples in a independent culture manner, and in combination with third generation sequencing technologies, the samples coverage become even greater. Soils, in particular, are the major reservoirs of this diversity, and many important environments around us, as the Brazilian biomes Caatinga and Atlantic Forest, are poorly studied. Thus, the genetic material from environmental soil samples of Caatinga and Atlantic Forest biomes were extracted by direct techniques, pyrosequenced, and the sequences generated were analyzed by bioinformatics programs (MEGAN MG-RAST and WEBCarma). Taxonomic comparative profiles of the samples showed that the phyla Proteobacteria, Actinobacteria, Acidobacteria and Planctomycetes were the most representative. In addition, fungi of the phylum Ascomycota were identified predominantly in the soil sample from the Atlantic Forest. Metabolic profiles showed that despite the existence of environmental differences, sequences from both samples were similarly placed in the various functional subsystems, indicating no specific habitat functions. This work, a pioneer in taxonomic and metabolic comparative analysis of soil samples from Brazilian biomes, contributes to the knowledge of these complex environmental systems, so far little explored

Análise metagenômica da microbiota de ambientes aquáticos do estado do Rio Grande do Norte - Brasil

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