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

Effect of Urucu oil (Brazilian Amazon) on the biomass of the aquatic macrophyte Eichhornia crassipes (Mart.) Solms (Pontederiaceae)

Os rios e lagos de várzea da província petrolífera de Urucu, na Amazônia Central, são amplamente colonizados por macrófitas aquáticas, que podem ser afetadas por acidentes durante a exploração e o transporte de petróleo. Entre as macrófitas, a espécie flutuante Eichhornia crassipes (aguapé) ocorre abundantemente na região; OBJETIVO: O objetivo desse estudo foi verificar o efeito de diferentes dosagens do petróleo de Urucu (0; 0,5; 1,5 e 3,0 L.m-2) na biomassa viva e morta de E. crassipes e em algumas características físicas e químicas da água; MÉTODOS: O experimento teve oitenta e quatro dias de duração. A cada sete dias foi determinada a biomassa (viva e morta) de E. crassipes e os valores de temperatura, pH, condutividade elétrica e oxigênio dissolvido da água; RESULTADOS: A dosagem de 0,5 L.m-2 foi suficiente para causar mortalidade parcial (48%) em E. crassipes após trinta e cinco dias de exposição ao petróleo. A dosagem de 3,0 L.m-2 causou mortalidade total (100%) em E. crassipes em oitenta e quatro dias de exposição. A decomposição do petróleo e da biomassa morta de E. crassipes provocam a redução do oxigênio dissolvido e do pH, e aumento da condutividade elétrica e de fósforo total na água; CONCLUSÕES: Nós concluímos que um derramamento de petróleo pode provocar mortalidade total em uma população de uma espécie de macrófita, mas não em uma outra. Isto pode alterar a diversidade de espécies de macrófitas na região impactada. No caso de Eichhornia crassipes e Pistia stratiotes, um derramamento de petróleo de Urucu pode favorecer E. crassipes, a espécie menos sensível ao petróleo.

Rhamnolipid produced from agroindustrial wastes enhances hydrocarbon biodegradation in contaminated soil

A crude biosurfactant solution was produced by Pseudomonas aeruginosa growing on agroindustrial wastes as the substrate and used to study its effect on hydrocarbon biodegradation by the indigenous soil microflora under laboratory conditions. Two concentrations were studied at first and 1 mg of biosurfactant/g of soil showed to be the most efficient for the total petroleum hydrocarbon reduction, which reached 85% at the first 20 days in soil microcosms. Respirometric and microbial analyses showed that the biosurfactant added did not have toxic effects over the microbial population. The use of a biosurfactant for bioremediation has been limited because of its high cost production. Biosurfactants produced from cost-free by-products combines waste minimization with economic potential bioremediation process.

Biosurfactants production by yeasts using soybean oil and glycerol as low cost substrate

Biosurfactants are bioactive agents that can be produced by many different microorganisms. Among those, special attention is given to yeasts, since they can produce many types of biosurfactants in large scale, using several kinds of substrates, justifying its use for industrial production of those products. For this production to be economically viable, the use of residual carbon sources is recommended. The present study isolated yeasts from soil contaminated with petroleum oil hydrocarbons and assessed their capacity for producing biosurfactants in low cost substrates. From a microbial consortium enriched, seven yeasts were isolated, all showing potential for producing biosurfactants in soybean oil. The isolate LBPF 3, characterized as Candida antarctica, obtained the highest levels of production - with a final production of 13.86 g/L. The isolate LBPF 9, using glycerol carbon source, obtained the highest reduction in surface tension in the growth medium: approximately 43% of reduction after 24 hours of incubation. The products obtained by the isolates presented surfactant activity, which reduced water surface tension to values that varied from 34 mN/m, obtained from the product of isolates LBPF 3 and 16 LBPF 7 (respectively characterized as Candida antarctica and Candida albicans) to 43 mN/m from the isolate LPPF 9, using glycerol as substrate. The assessed isolates all showed potential for the production of biosurfactants in conventional sources of carbon as well as in agroindustrial residue, especially in glycerol.

Molecular analysis of the bacterial diversity in a specialized consortium for diesel oil degradation

Diesel oil is a compound derived from petroleum, consisting primarily of hydrocarbons. Poor conditions in transportation and storage of this product can contribute significantly to accidental spills causing serious ecological problems in soil and water and affecting the diversity of the microbial environment. The cloning and sequencing of the 16S rRNA gene is one of the molecular techniques that allows estimation and comparison of the microbial diversity in different environmental samples. The aim of this work was to estimate the diversity of microorganisms from the Bacteria domain in a consortium specialized in diesel oil degradation through partial sequencing of the 16S rRNA gene. After the extraction of DNA metagenomics, the material was amplified by PCR reaction using specific oligonucleotide primers for the 16S rRNA gene. The PCR products were cloned into a pGEM-T-Easy vector (Promega), and Escherichia coli was used as the host cell for recombinant DNAs. The partial clone sequencing was obtained using universal oligonucleotide primers from the vector. The genetic library obtained generated 431 clones. All the sequenced clones presented similarity to phylum Proteobacteria, with Gammaproteobacteria the most present group (49.8 % of the clones), followed by Alphaproteobacteira (44.8 %) and Betaproteobacteria (5.4 %). The Pseudomonas genus was the most abundant in the metagenomic library, followed by the Parvibaculum and the Sphingobium genus, respectively. After partial sequencing of the 16S rRNA, the diversity of the bacterial consortium was estimated using DOTUR software. When comparing these sequences to the database from the National Center for Biotechnology Information (NCBI), a strong correlation was found between the data generated by the software used and the data deposited in NCBI.

Acordos internacionais sobre a produção compartilhada de recursos naturais: a unitização interestatal na área do petróleo e gás sob o enfoque constitucional brasileiro

The apportionment of natural resources between sovereign States is a subject that relates many aspects of International law, as long as Constitutional Law, at the execution and application phases of international treaties that regulates the exploration of common goods. In this sense, because of their natural characteristics that creates an environment of constant migration and fixation in transboundary regions, terrestrial or maritime, the petroleum and the natural gas bound a complex juridical apparatus that can control the sovereign rights involved. This research is aim at accomplishing a study concerning the international agreements that enable the non-unilateral action, specifically the unitization treaties between sovereign States, as a manner to resolve situations related to the individualization of oil and/or gas reservoirs that go across their national borders. These agreements will be analyzed considering the international public law sources theory, bearing in mind yet the already existed experiences in this sense, not disregarding the way that this fact could affect Brazil. It will begin with an historical incursion over the unitization institute, covering its main characteristics and its formation and execution procedures, and finally it will address the Brazilian legal system and the comparative law threats the institute. The clauses of these relevant agreements will be analyzed in details, concerning its particularities and its contents. Because these agreements are international obligatory rules of law, it is indispensable that they are considered under the auspices of the international law system, focusing their nature and the subjects of international law and establishing them as sources of the international law, analyzing them, then, as international rules and the applicable law to these juridical relations, the conventional established, the consolidated international custom and the applicable International Law principles, appearing the State s responsibility as an important subject for the verification of the acts lawful practiced by States. The analysis of the apportionment of these natural resources ends with the individualization of possible exploitable marine oil fields located between the exclusive economic zone and the continental platform ends and the region administrated by the International Seabed Authority. At last, the Brazilian constitutional system appears as the mechanism of integration, application and execution of the international unitization agreements in Brazil, detaching the format and the proceedings that the international treaties take to acquire validity at the national legal system, passing through the treaties interpretation and the applicable constitutional principles, coming to its application in Brazil, considering the existing constitutional peculiarities and the role played by the National Agency of Petroleum, Natural Gas and bio-fuel ANP

O princípio constitucional da redução das desigualdades regionais e o transporte de gás natural

In a country of continental dimensions as Brazil, one of the top challenges to its economic growth is the logistic related to energetical demand supply. We live now in the era of environmental protection and, in this new context of priorizations, it passes trough the search for alternative energies for the energetic matrix, due the petroleum elevated costs in the global market (and its finitude), but also due its pollution over the environment. This attempt of substitution needs solutions related to the national reality, into a national long term developing plan and based at a juridical-economic analysis of its realization. This study will look for, also based in an economical analysis, the juridical legitimity of choosing natural gas as the new protagonist of national economic growth (as a substitute of petroleum) and the necessary boost that must be done by law, based on an economic policy focused strictly for that fact, as a modifying agent of this reality. This study, therefore, will always be turned to a constitutional aspect, respecting the principles of economic order and the goal of reducing regional inequalities, which must influence the making off of a developing plan. At the end, it will try to demonstrate the juridical viability of such undertaking, tuned in jus-economical criteria. Another goal is related to the analysis of the natural gas industry, due the regulation of its transport has a major importance for national energetic integration, not only because this activity be characterized as a net industry, still under control of a natural monopoly, but also because the competitive or cooperative profile that should be priorized at the beginning of the economic planning for this activity (such as investment policies and its own rules that will submit private agents)

A tutela constitucional da exploração de petróleo em águas internacionais em face do princípio do desenvolvimento sustentável: a possibilidade de responsabilização civil do dano futuro

The demands brought by a society doomed to the constant production of global risks, which whose effects are not immediately noticed effects are not perceived immediately, claim from the Law a new Theory about the Risk, that would offer a broad environmental protection, at the same time it would still be compatible with the idea of economic efficiency, required by the Modern Industry. The expansion of the methods and technologies regarding the exploitation and production of oil causes the constant expansion of the exploitable boundaries, especially in ultra-deep waters with the Pre-salt layer, in Brazil, or the still incipient research about the polymetallic nodules and other mineral sources in international waters, like the Atlântico Sudoeste, by the Programme on Ocean Science in Relation do Non Living Resources (OSNLR), a global study performed in partnership with the Intergovernmental Oceanographic Commission, from UNESCO (IOC UNESCO) and also with the Division of Ocean Affairs and Law of the Sea (UNDOALOS). Thus, we aim to analyze the correlation, and possible collisions between the right to a balanced environment and the free exercise of economic activity and the occurrence of environmental damages from the perspective of the exploitation activities of oil and other natural resources in international waters, specifically in the Area, from the constitutional principle of sustainable development and its legitimacy by the environmental international protection. Therefore, this study also aims to evaluate the legal framework for exploration and production of oil in international waters, particularly in the Area, and appraise how the constitutional instruments and mechanisms for environmental protection can impact on the international environmental protection system in order to ensure the present and future generations an ecologically balanced environment, laid down in Article 225 of the Brazilian Constitution, even with so many risks posed by the activities of exploitation and production of oil in international waters. In the meantime, we intend to also intend to investigate the possibility of future liability for environmental damage in order to ensure that constitutional principle and, consequently, and try to define the concept of environmental damage and its implications on the constitutional principle of environmental protection. Given all that was in summary, this work aims to contribute to the evolution of the new Theory of Environmental Risk, turning the law into something more than a punitive or corrective element in this society, but into a legal risk management, that may be triggered even before the consolidation of the damage

Regulação do livre acesso no transporte de gás natural à luz da ordem econômica da Constituição de 1988

In the middle of modern social changes produced by globalization and capitalism, several markets have changed. States have left the direct coordination of these markets (chiefly public utility sector in the form of monopolies), introducing regulation in order to promote competition. These changes have affected natural gas industry by promoting competition as a key factor to the development and the increase of firms in this market. The regulatory reform of natural gas industry ocurred in EUA and Europe Union and it has produced its first results. In Brazilian context, Constitutional Amendment nbr. 09 and Federal Law nbr. 9.478/97 ( Petroleum Law ) opened the natural gas market to a broad range of private economic agents and they finished the monopoly over the industry before managed by Petrobras. The new regulatory framework of Brazilian natural gas industry has designed competition as a central element to the new form of managment of business and contractual relationships of this industry. Among the regulatory instruments, open access regulation in natural gas pipelines is directed to promote competition. The questions arised about its implementation in Brazilian context are studied in the present work, in which it is discussed the constitutional rules and principles are to be applied to the open access regulation within the theme of statal regulation of economy present in constitutional economic order

A indústria petrolífera nacional e potiguar: uma análise a partir da tecnologia em E&P

O trabalho analisa a Indústria Petrolífera em três ambientes de atuação, mundial, nacional e estadual, a partir da observação de sete variáveis, a saber: caráter do petróleo; risco; economia de escala; integração; barreiras; tecnologia e ambiente. O estudo busca perceber as interações e os transbordamentos das variáveis selecionadas em cada um dos ambientes citados, em razão os aspectos econômicos e institucionais presentes nesta atividade, cujos resultados atingem diversos setores da economia e da sociedade como um todo e balizam o próprio processo de inovação da mesma. O estudo está alicerçado no arcabouço teórico neoschumpeteriano acerca dos aspectos voltados para concorrência e inovação, além da questão institucional, e se apóia em dados da Agência Nacional do Petróleo (A.N.P.), Ministério de Minas e Energia (MME), Movimento de Combate à Corrupção Eleitoral / Tribunal Superior Eleitoral (MCCE/TSE), Organization of the Petroleum Exporting Countries (OPEC), International Energy Agency (IAE), Institut Français Pétrole (IFP), United Nations Development Programme (UNDP). Avalia-se que em termos de caracterização da indústria petrolífera nos três ambientes considerados (mundial, nacional e local), há certa similitude no que se refere ao comportamento destas variáveis, com pequenas modificações suscitadas por fatores como grau de desenvolvimento e complexidade da atividade; resultado do ambiente concorrencial e inovativo; e, pelas influências institucionais acerca de atividades e grau de inserção das mesmas no ambiente socioeconômico onde estão localizadas a atividade produtiva e o seu entorno

Biodiesel etílico filtrado de óleo residual de soja: desempenho de um trator agrícola na operação de gradagem

Filtered ethyl ester biodiesel from residual soybean oil: performance of an agricultural tractor in the disk harrow operation. This test was conducted to evaluate the performance of an agricultural tractor using a mixture of biodiesel and petroleum diesel. The experiment was conducted at FCAV-Unesp, Jaboticabal, using a 4x2 AFWD tractor with 73.6 kW (100 hp @ 2,300 rpm) of power in the motor, pulling a disk harrow. The biodiesel used was of the ethyl type, filtered, and produced from residual soybean oil. The experimental design was completed randomized in factorial scheme (4x5), with 20 treatments and 4 repetitions, in which 5 ratios of biodiesel and petroleum diesel mixtures were combined (0 and 100%, 25 and 75%, 50 and 50%, 75 and 25% and 100 and 0%), with 4 displacement runs. The analyzed variables were traction bar power, engine rotation and displacement speed. The results evidenced that the mixture ratio (biodiesel and petroleum diesel) did not compromise tractor performance until the limit of 50%.

Gestão ambiental na indústria do petróleo :um estudo sobre as práticas de gestão na perspectiva da aprendizagem organizacional

This work discusses the environmental management thematic, on the basis of ISO 14001 standard and learning organization. This study is carried through an exploratory survey in a company of fuel transport, located in Natal/RN. The objective of this research was to investigate the practices of environmental management, carried through in the context of an implemented ISO 14001 environmental management system, in the researched organization, from the perspective of the learning organization. The methodology used in this work is supported in the quantitative method, combining the exploratory and descriptive types, and uses the technique of questionnaires, having as scope of the research, the managers, employee controlling, coordinators, supervisors and - proper and contracted - of the company. To carry through the analysis of the data of this research, it was used software Excel and Statistical version 6.0. The analysis of the data is divided in two parts: descriptive analysis and analysis of groupings (clusters). The results point, on the basis of the studied theory, as well as in the results of the research, that the implemented ISO 14001 environmental system in the searched organization presents elements that promote learning organization. From the results, it can be concluded that the company uses external information in the decision taking on environmental problems; that the employees are mobilized to generate ideas and to collect n environmental information and that the company has carried through partnerships in the activities of the environmental area with other companies. All these item cited can contribute for the generation of knowledge of the organization. It can also be concluded that the company has evaluated environmental errors occurrences in the past, as well as carried through environmental benchmarking. These practical can be considered as good ways of the company to acquire knowledge. The results also show that the employees have not found difficulties in the accomplishment of the tasks when the manager of its sector is not present. This result can demonstrate that the company has a good diffusion of knowledge

Gestão ambiental na indústria do petróleo : fatores associados ao comportamento pró-ambiental de empregados de uma empresa brasileira de óleo e gás

This survey develops a study about the pro-environment behavior of a brazilian company of oil and gas sector, emphazing the Petróleo Brasileiro S/A PETROBRAS in its Unit of Business of Exploration and Production in the State of Rio Grande do Norte and Ceará − UN-RNCE. The work s development adopt the bibliographic research methodology with the descriptive exploratory analisys, with the purpouse of identifying related points with the knowledge and the analysed enterprise s staff conduct, by the aplication of the interview with a instrument of data colecting with closed variable questions from answers that varies from o to 10.The answers allowed us to know the employees knowledge about the related aspects of environmental responsability, knowing also the involvement of the employees with the politics of environmental managment that has been implemented in the operating area of the company. The research led us to an important verification of the pro-environmental behavior of the people interviewed. The results depends of a wise acting, not only inside the company, but also outside, where the results of this acting will be more perceived for the society. Finally, we conclude that the company has a politics of management with a great importance, emphasinzing that the employees are been in constant training about their behavior resulting in satisfaction

Técnicas de inteligência artificial para a geração dinâmica de set points para uma coluna de destilação

Artificial Intelligence techniques are applied to improve performance of a simulated oil distillation system. The chosen system was a debutanizer column. At this process, the feed, which comes to the column, is segmented by heating. The lightest components become steams, by forming the LPG (Liquefied Petroleum Gas). The others components, C5+, continue liquid. In the composition of the LPG, ideally, we have only propane and butanes, but, in practice, there are contaminants, for example, pentanes. The objective of this work is to control pentane amount in LPG, by means of intelligent set points (SP s) determination for PID controllers that are present in original instrumentation (regulatory control) of the column. A fuzzy system will be responsible for adjusting the SP's, driven by the comparison between the molar fraction of the pentane present in the output of the plant (LPG) and the desired amount. However, the molar fraction of pentane is difficult to measure on-line, due to constraints such as: long intervals of measurement, high reliability and low cost. Therefore, an inference system was used, based on a multilayer neural network, to infer the pentane molar fraction through secondary variables of the column. Finally, the results shown that the proposed control system were able to control the value of pentane molar fraction under different operational situations

Implantação de um sistema de detecção de vazamentos em oleodutos com tratamento de alarmes falsos

The pumping of fluids in pipelines is the most economic and safe form of transporting fluids. That explains why in Europe there was in 1999 about 30.000 Km [7] of pipelines of several diameters, transporting millíons of cubic meters of crude oil end refined products, belonging to COCAWE (assaciation of companies of petroleum of Europe for health, environment and safety, that joint several petroleum companies). In Brazil they are about 18.000 Km of pipelines transporting millions of cubic meters of liquids and gases. In 1999, nine accidents were registered to COCAWE. Among those accidents one brought a fatal victim. The oil loss was of 171 m3, equivalent to O,2 parts per million of the total of the transported volume. Same considering the facts mentioned the costs involved in ao accident can be high. An accident of great proportions can bríng loss of human lives, severe environmental darnages, loss of drained product, loss . for dismissed profit and damages to the image of the company high recovery cost. In consonance with that and in some cases for legal demands, the companies are, more and more, investing in systems of Leak detection in pipelines based on computer algorithm that operate in real time, seeking wíth that to minimize still more the drained volumes. This decreases the impacts at the environment and the costs. In general way, all the systems based on softWare present some type of false alarm. In general a commitment exists betWeen the sensibílity of the system and the number of false alarms. This work has as objective make a review of thé existent methods and to concentrate in the analysis of a specific system, that is, the system based on hydraulic noise, Pressure Point Analyzis (PPA). We will show which are the most important aspects that must be considered in the implementation of a Leak Detection System (LDS), from the initial phase of the analysis of risks passing by the project bases, design, choice of the necessary field instrumentation to several LDS, implementation and tests. We Will make na analysis of events (noises) originating from the flow system that can be generator of false alarms and we will present a computer algorithm that restricts those noises automatically