Isolamento, seleção e cultivo de leveduras com potencia biotecnológico para a produção de endo-xilaneses

Xilanases são enzimas que catalisam a hidrólise das xilanas e têm sido em grande parte, obtidas a partir de bolores e bactérias. No entanto poucos estudos têm sido relatados sobre a produção destas enzimas por leveduras. O presente trabalho teve como objetivo isolar leveduras de diferentes fontes vegetais visando à produção de xilanases, além de maximizar sua produção, estudar o uso de diferentes fontes de nitrogênio e cultivar as leveduras em meios contendo coprodutos agroindustriais. As amostras de alimentos e resíduos foram enriquecidas em caldo extrato de malte e levedura e isoladas em Ágar Nutriente Wallerstein, as leveduras isoladas foram, a seguir, avaliadas quanto à capacidade de degradar xilana presente no meio e produzir halos de hidrólise, os quais foram visualizados através do uso do corante vermelho congo. Os micro-organismos selecionados como potenciais produtores de xilanase foram crescidos em meio complexo líquido e as atividades enzimáticas de endoxilanase, β-xilosidase, carboximetilcelulase, celulase total, pH e concentração de biomassa foram avaliadas ao longo de 96 h de cultivo. Dentre as leveduras isoladas, sete foram selecionadas, e a 18Y foi a que apresentou a maior atividade de endo- xilanase (2,7 U.mL-1 ), sendo esta isolada de chicória e identificada como Cryptococcus laurentii. Esta estirpe apresentou capacidade de produzir xilanase com baixos níveis de celulase, sendo assim selecionada neste trabalho. A maximização de endo-xilanase foi avaliada fazendo uso de planejamento experimental onde primeiramente foi realizado um planejamento fracionário 2 6-2 para verificar os efeitos do pH inicial e as concentrações de xilana, peptona, (NH4)2SO4, extrato de levedura e KH2PO4 sobre a atividade enzimática. Após selecionar as variáveis xilana, peptona, pH e extrato de levedura foi realizado um delineamento composto central rotacional (24 ) onde todos os cultivos foram mantidos a 30°C, 150 rpm durante 96 h sendo retiradas alíquotas para determinação das atividades, pH e biomassa. A produção máxima foi de 6,9 U.mL-1 usando 10,0 g.L-1 de extrato de levedura, 10,0 g.L-1 de peptona, 10,0 g.L-1 de xilana, 1,0 g.L-1 de (NH4)2SO4 em pH 6,5 o que permitiu um incremento de mais de 250% sobre a atividade. Posteriormente foram realizados ensaios avaliando diferentes fontes e concentrações de nitrogênio orgânico e inorgânico. A presença de NH4NO3 e (NH₄)₂SO₄ usados na concentração de 3% proporcionaram as maiores atividades de endo-xilanase (6,2 e 6,0 U.mL-1 respectivamente). O sulfato de amônio foi selecionado e fixado em 1 g.L-1 e logo após um planejamento completo 22 foi realizado onde as variáveis xilana e extrato de levedura foram estudadas e as demais fixadas. As condições ótimas estabelecidas para a produção da enzima foram: concentração de xilana de 18,6 g.L-1 , concentração de extrato de levedura de 10 g.L-1 atingindo 14 U.mL-1 . Após a maximização enzimática estudou-se o crescimento de Pichia pastoris NRRL Y-1603 e Cryptococcus laurentti em cinco substratos agroindustriais visando a possibilidade estes substratos substituírem a xilana em cultivos para a produção de endo-xilanase. Os ensaios foram realizados utilizando os subtratos pré-tratados com NaOH 4% e não tratados. Para inserção dos mesmos aos meios de cultivo, estes foram moídos e adicionados na concentração de 2%. O pré-tratamento para todos as fontes de hemicelulose foi eficiente e promoveu aumento nas atividades produzidas. Cryptococcus laurentti apresentou maior atividade enzimática (8,7 U.mL-1 ) em farelo de arroz desengordurado e pré- tratado enquanto que a levedura Pichia pastoris NRRL Y-1603 apresentou sua melhor condição para produção de endo-xilanase quando cultivada em meio contendo casca de aveia e o farelo de arroz pré-tratados, alcançando atividades máximas de 7,6 e 7,5 U.mL-1 .

Evaluation of the genotoxicity of cellulose nanofibers

Background: Agricultural products and by products provide the primary materials for a variety of technological applications in diverse industrial sectors. Agro-industrial wastes, such as cotton and curaua fibers, are used to prepare nanofibers for use in thermoplastic films, where they are combined with polymeric matrices, and in biomedical applications such as tissue engineering, amongst other applications. The development of products containing nanofibers offers a promising alternative for the use of agricultural products, adding value to the chains of production. However, the emergence of new nanotechnological products demands that their risks to human health and the environment be evaluated. This has resulted in the creation of the new area of nanotoxicology, which addresses the toxicological aspects of these materials.Purpose and methods: Contributing to these developments, the present work involved a genotoxicological study of different nanofibers, employing chromosomal aberration and comet assays, as well as cytogenetic and molecular analyses, to obtain preliminary information concerning nanofiber safety. The methodology consisted of exposure of Allium cepa roots, and animal cell cultures (lymphocytes and fibroblasts), to different types of nanofibers. Negative controls, without nanofibers present in the medium, were used for comparison.Results: The nanofibers induced different responses according to the cell type used. In plant cells, the most genotoxic nanofibers were those derived from green, white, and brown cotton, and curaua, while genotoxicity in animal cells was observed using nanofibers from brown cotton and curaua. An important finding was that ruby cotton nanofibers did not cause any significant DNA breaks in the cell types employed.Conclusion: This work demonstrates the feasibility of determining the genotoxic potential of nanofibers derived from plant cellulose to obtain information vital both for the future usage of these materials in agribusiness and for an understanding of their environmental impacts.

Grazing behavior of lactating Murrah buffalo cows supplemented with cupuassu byproduct.

The goal of this work was to evaluate grazing behavior of lactating Murrah buffalo cows supplemented with cupuassu byproduct containing 16.8, 51.4, 8.3, 83.5 and 3.4% of ether extract (EE), neutral detergent fiber (NDF), crude protein (CP), total digestible nutrients (TDN) and nonfibrous carbohydrate (NFC), respectively. Experimental rations were balanced for reaching levels of 22.0 % of crude protein (CP) and 80.1 % of total digestible nutrient (TDN).

Milk composition of Murrah buffalo cows supplemented with cupuassu byproduct in Western Amazon.

The aim of this study was to evaluate milk composition of Murrah buffalo cows supplemented with rations containing cupuassu byproduct in substitution of ground corn. Isoprotein and isoenergy rations were balanced with 22.0% of crude protein (CP) and 80.1% of total digestible nutrient (TDN).

Cupuassu byproduct in supplements for buffalo cows: milk yield and calf performance.

The aim of this study was to evaluate milk yield and calf performances of Murrah buffalo cows supplemented with rations containing cupuassu byproduct in substitution of ground corn.

O processo de urbanização de Mossorò: dos processos históricos à estrutura urbana atual

The general objective of this academic work is to analyze the relationship between the territorial division and the urban expansion process of Mossoró city, understood here as the production and occupation of space. The urban expansion in Mossoró, since its formation as settlement in 1772 until current days, identifies with the Brazilian urbanization process whose growth is determined by the capitalist development. Thus, the expansion was determined by several economical specializations imposed by the territorial division of work which occurs at an interregional level, and, sometimes at an international level. Then, each specialization determined a moment of the urban expansion of the city, as follows: a) The cattle farmer specialization, between 1772 and 1857, when the urban expansion was shy, is summarized to a commercial square that received goods from Aracati aiming to cover a wide rural area; b) The commercial emporium specialization, between 1857 and 1930, when the urban expansion took an important impulse with the concentration of public and private capitals; c) The salt industry and the agricultural-industrial exporter specialization inside a state of development policy, between 1930 and 1970, when the urban expansion, joined to the settlement of the working class in the urban soil, developed along an important axis -the railway; d) The render of services specialization inside a state of intervention policy, between 1970 and 1990, when the urban expansion was characterized by the accelerated growth rhythm, by the reuse of some spaces, and by space segregation of demanding people; e) The render of services policy inside a neoliberal state policy, since 1990 until current days, when the urban expansion reduced its rhythm abruptly, when only small alterations occurred in the existing spaces. It focused on social policies and on several slums eradication programs. Finally, the territorial structure is deeply articulated with others, no territorial, but economical, social and political, which happens at a national, regional and local rate. Only within a historical and conceptual panorama, it was possible to explain the urban expansion in Mossoró from its formation in 1772 until current days. Therefore, this work is a several discipline analysis of the urbanization process existing in Mossoró

Exploring the potential of novel biomixtures and Lentinula edodes fungus for the degradation of selected pesticides. Evaluation for use in biobed systems

An approach to reduce the contamination of water sourceswith pesticides is the use of biopurificaction systems. The active core of these systems is the biomixture. The composition of biomixtures depends on the availability of local agro-industrial wastes and design should be adapted to every region. In Portugal, cork processing is generally regarded as environmentally friendly and would be interesting to find applications for its industry residues. In this work the potential use of different substrates in biomixtures, as cork (CBX); cork and straw, coat pine and LECA (Light Expanded Clay Aggregates), was tested on the degradation of terbuthylazine, difenoconazole, diflufenican and pendimethalin pesticides. Bioaugmentation strategies using the white-rot fungus Lentinula edodes inoculated into the CBX, was also assessed. The results obtained from this study clearly demonstrated the relevance of using natural biosorbents as cork residues to increase the capacity of pesticide dissipation in biomixtures for establishing biobeds. Furthermore, higher degradation of all the pesticides was achieved by use of bioaugmented biomixtures. Indeed, the biomixtures inoculated with L. edodes EL1were able to mineralize the selected xenobiotics, revelling that these white-rot fungi might be a suitable fungus for being used as inoculum sources in on-farm sustainable biopurification system, in order to increase its degradation efficiency. After 120 days, maximum degradation of terbuthylazine, difenoconazole, diflufenican and pendimethalin, of bioaugmented CBX, was 89.9%, 75.0%, 65.0% and 99.4%, respectively. The dominant metabolic route of terbuthylazine in biomixtures inoculated with L. edodes EL1 proceeded mainly via hydroxylation, towards production of terbuthylazine-hydroxy-2 metabolite. Finally, sorption process to cork by pesticides proved to be a reversible process,working cork as a mitigating factor reducing the toxicity to microorganisms in the biomixture, especially in the early stages.

Arquiteturas do trigo: espaços de silagem no Alentejo, do século XIX à atualidade

O presente trabalho de investigação apresenta um estudo sistemático da expressão arquitetónica que teve na região do Alentejo a implantação agro-industrial da cultura do trigo, em especial, durante a "Campanha do Trigo", iniciada em 1929 e prosseguida durante o Estado Novo, até 1969. Para tanto, procede ao mapeamento dos elementos edificados, em ligação com os processos de produção, silagem e moagem industrial do trigo, bem como ao seu transporte, estabelecendo uma análise comparativa com outras regiões e países, com o intuito de revelar o impacto destes equipamentos nos diversos territórios e paisagens. Assim, procura-se compreender as relações que se estabeleceram entre a forma arquitetónica do silo (cujo desenho resulta fundamentalmente de uma resposta programática intrinsecamente ligada à sua função primordial - armazenamento do cereal) e os lugares, onde se implantaram e com os quais se procuraram articular, e as paisagens, com as quais perspetivaram dialogar. Para tal, procede-se a descrição do estado atual de cada exemplar das denominadas "arquiteturas do trigo", com o objetivo de gerar, num primeiro nível, o entendimento de cada caso em particular e, posteriormente, estabelecer, por efeito de comparação, um confronto, quer entre os diferentes contextos em que se inserem, quer entre as suas caraterísticas formais, de modo a tirar ilações que possibilitem problematizar o seu uso futuro, equacionando diferentes perspetivas de atuação sobre o edificado. Finalmente procura-se introduzir uma discussão segundo diferentes orientações e posicionamentos, de modo a invocar perspetivas de atuação sobre os silos, que se adequem a cada situação específica, a cada contexto e a cada lugar. É no confronto entre as abordagens enunciadas, que se evidencia, mais do que a pertinência, a emergência social e cultural de uma ação diligente, quer ao nível particular de cada peça silar, quer do sistema, do qual era parte integrante a linha de caminho de ferro, que através do seu traçado permitia interligar todas as peças silares e garantir o seu funcionamento a escala industrial; ABSTRACT: This research presents a systematic study of architectural expression of wheat crop agro-industrial development, in particular the "Wheat Campaign", started in 1929 and continue during the "New State", until 1969. We first proceed to map the built elements related to production processes, silage and wheat industrial grinding, as well as the ways of transportation, establishing a comparative analysis with other countries and regions, in order to reveal the impact of these objects in various territories and landscapes. We seek to understand the relationships between the architectural form of the silo (whose design is mainly a result of an intrinsically programmatic response linked to its primary function - cereal storage) and the places where they are implemented and the landscapes which they sought to communicate and articulate. Secondly, we describe the current status of each copy of the "wheat architectures" in Alentejo, with the main objective of attain, at first, the understanding of each particular case and, then, to establish a connection between the different contexts in which they operate and their formal characteristics, in order to conceive and discuss their future use, equating different perspectives of action in the buildings. Finally seeks to introduce a discussion according to different orientations and positions in order to invoke prospects for action on the grain elevators that are appropriate to each specific situation, every context and every place. It is the clash between the stated approaches, which shows more than the relevance, it shows social and cultural emergence of a diligent action on the particular level of each grain elevator on the system, which was an integral part of the railway, which through its tracing allowed to interconnect all grain elevators and assure its operation on an industrial scale

Evaluating social and environmental impacts at Embrapa.

A strategic planning process has been implemented at the Brazilian Agricultural Research Agency (Embrapa) to introduce sustainable development objectives in all steps of agricultural Research and Development. An essential component of the institutional mission statement hence devised has called for the systematic assessment of social and environmental impacts (in addition to the traditionally studied economic ones) of all technology innovations resulting from R&D. The proposed approach emphasizes the interest of promoting close interaction between R&D teams and technology-adopting producers, under actual field contexts, in order to improve both the technology development and the demand probing processes. Given the multiplicity of technological applications ensuing from Embrapa?s very broad research encompassment, and the variety of environmental and productive contexts involved, a customized impact assessment system has been proposed. Directed at the appraisal of agricultural technology development research projects (ex-ante) as well as their ensuing innovations (ex-post), the Ambitec-Agro System comprises a set of integrated socio-environmental indicators, constructed in modules suited to Agricultural, Animal husbandry, and Agro-industrial activities, besides a specific module for Social Impact Assessment. The system has been routinely applied in technology appraisal in all of Embrapa?s Units, as a basis for their institutional performance evaluations, and toward the formulation of the annual Social Balance Report. Following the inception of this institutional technology appraisal initiative, several methodological innovations have been proposed within Embrapa, including technical improvements and applicability adaptations of the Ambitec-Agro system, and approaches to further-reaching objectives, such as the sustainable development of rural communities, and the environmental management of agricultural activities.

Carbon in humic fractions of organic matter in soil treated with organic composts under mango cultivation.

Soil organic matter (SOM) plays a key role in maintaining the productivity of tropical soils, providing energy and substrate for the biological activity and modifying the physical and chemical characteristics that ensure the maintenance of soil quality and the sustainability of ecosystems. This study assessed the medium-term effect (six years) of the application of five organic composts, produced by combining different agro-industrial residues, on accumulation and chemical characteristics of soil organic matter. Treatments were applied in a long-term experiment of organic management of mango (OMM) initiated in 2005 with a randomized block design with four replications. Two external areas, one with conventional mango cultivation (CMM) and the other a fragment of regenerating Caatinga vegetation (RCF), were used as reference areas. Soil samples were collected in the three management systems from the 0.00-0.05, 0.05-0.10, and 0.10-0.20 m layers, and the total organic carbon content and chemical fractions of organic matter were evaluated by determining the C contents of humin and humic and fulvic acids. Organic compost application significantly increased the contents of total C and C in humic substances in the experimental plots, mainly in the surface layer. However, compost 3 (50 % coconut bagasse, 40 % goat manure, 10 % castor bean residues) significantly increased the level of the non-humic fraction, probably due to the higher contents of recalcitrant material in the initial composition. The highest increases from application of the composts were in the humin, followed by the fulvic fraction. Compost application increased the proportion of higher molecular weight components, indicating higher stability of the organic matter.

Integration of by-products and waste from agri-food supply chains into the «water-energy-food» nexus (RECOW-ER): the role of fungi

The agricultural sector is undoubtedly one of the sectors that has the greatest impact on the use of water and energy to produce food. The circular economy allows to reduce waste, obtaining maximum value from products and materials, through the extraction of all possible by-products from resources. Circular economy principles for agriculture include recycling, processing, and reusing agricultural waste in order to produce bioenergy, nutrients, and biofertilizers. Since agro-industrial wastes are principally composed of lignin, cellulose, and hemicellulose they can represent a suitable substrate for mushroom growth and cultivation. Mushrooms are also considered healthy foods with several medicinal properties. The thesis is structured in seven chapters. In the first chapter an introduction on the water, energy, food nexus, on agro-industrial wastes and on how they can be used for mushroom cultivation is given. Chapter 2 details the aims of this dissertation thesis. In chapters three and four, corn digestate and hazelnut shells were successfully used for mushroom cultivation and their lignocellulosic degradation capacity were assessed by using ATR-FTIR spectroscopy. In chapter five, through the use of the Surface-enhanced Raman Scattering (SERS) spectroscopy was possible to set-up a new method for studying mushroom composition and for identifying different mushroom species based on their spectrum. In chapter six, the isolation of different strains of fungi from plastic residues collected in the fields and the ability of these strains to growth and colonizing the Low-density Polyethylene (LDPE) were explored. The structural modifications of the LDPE, by the most efficient fungal strain, Cladosporium cladosporioides Clc/1 strain were monitored by using the Scanning Electron Microscope (SEM) and ATR-FTIR spectroscopy. Finally, chapter seven outlines the conclusions and some hints for future works and applications are provided.

Agro-Based Biocomposites for Industrial Applications

Leaf fibers are fibers that run lengthwise through the leaves of most monocotyledonous plants such as pineapple, banana, etc. Pineapple (Ananas comosus) and Banana (Musa indica) are emerging fiber having a very large potential to be used for composite materials. Over 150,000 ha of pineapple and over 100,000 ha of banana plantations are available in Brazil for the fruit production and enormous amount of agricultural waste is produced. This residual waste represents one of the single largest sources of cellulose fibers available at almost no cost. The potential consumers for this fiber are pulp and paper, chemical feedstock, textiles and composites for the automotive, furniture and civil construction industry.

Secagem do resíduo industrial da acerola em leito de jorro: estudos fluidodinâmicos e análise do desempenho do secador

In this research, the drying process of acerola waste was investigated by using a spouted bed drier. The process was conducted using high density polyethylene inert particles with the objective of producing an ascorbic acid-rich final product. The fruit waste was ground and used to prepare different water-maltodextrin suspensions. Initially, fluidynamical experiments were conducted in order to evaluate the feeding effect on the spouted bed drier fluidynamics behavior. The experimental planning 23 + 3 was used to investigate the effect of the following variables: solids concentration, drying air temperature, intermittence time, production efficiency, solids retention and product losses by elutriation of fine particles on drier walls. The effect of selected independent variables on the drier stability was also evaluated based on a parameter defined as the ratio between the feed suspension volume and the total inert particles volume. Finally, the powder quality was verified in experiments with fixed feed flow and varying air drying temperature, drying air velocity and intermittence time. It was observed that the suspension interferes in the spouted bed drier fluidynamics behavior, and higher air flow is necessary to stabilize the drier. The suspension also promotes the expansion of the spouted bed diameter, decreases the solid circulation and favors the air distribution at the flush area. All variables interfere in the spouted bed performance, and the solids concentration has a major effect on the material retention and losses. The intermittence time also has great effect on the stability and material retention. When it comes to production efficiency, the main effect observed was the drying air temperature. First order models were well adjusted to retention and losses data. The acerola powder presented ascorbic acid levels around 600 to 700 mg/100g. Similar moisture and ascorbic acid levels were obtained for powders obtained by spouted bed and spray drier. However, the powder production efficiency of the spray drier was lower when compared to spouted bed drier. When it comes to energetic analysis, the spray drier process was superior. The results obtained for spouted bed drier are promising and highly dependent on the operational parameters chosen, but in general, it is inferred that this drying process is adequate for paste and suspension drying

Acerola's clones of industrial interest

In order to know which clone of acerola is better for acerola industrialization, we studied the pectin methylesterase (PME) specific activity, pectin content and vitamin C content in five different clones of acerola. The pectin yield varied from 1.37 to 2.99% and the highest content of pectin occurred in clones 3 and 5. Ascorbic acid varied significantly from 1157.5 to 1735.5 mg/100 g of pulp in the five clones. The highest content of vitamin C occurred in clone 4. The PME specific activity varied from 0.79 to 2.92 units g(-1)/g of pulp and the highest values occurred in clone 2. We also studied the optimum temperature and the optimum pH of this enzyme. Clones 1, 2, 4 and 5 showed optimum temperature at 90C. Clone 3 showed practically the same specific activity at all temperatures studied. Clones 1 and 4 showed an optimum pH of 9.0 and clone numbers 2, 3 and 5 showed a pH optimum at 8.5.

Agro-Based Biocomposites for Industrial Applications

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)