Aspectos conceituais da regulação dos serviços de água e esgoto no Brasil

Objetiva-se, aqui, apresentar os aspectos conceituais da regulação da prestação dos serviços de água e esgoto no Brasil e analisar de forma comparativa essa regulação com a de outros setores da infraestrutura. O estudo parte do pressuposto da regulação como intervenção do Estado voltada para a eficiência e a equidade, e apresenta os fundamentos teóricos que justificam essa regulação de forma contextualizada às características do setor de água e esgoto. Mediante análise comparativa com outros setores de infraestrutura de redes, concluí-se que, em função das características físicas, econômicas e institucionais do setor de água e esgoto, será bastante complexo o estabelecimento efetivo dessa atividade conforme os princípios da lei nº 11.445/2007.

Causas do declínio acelerado da desnutrição infantil no Nordeste do Brasil (1986-1996-2006)

OBJETIVO: Descrever a variação temporal na prevalência de desnutrição infantil na região Nordeste do Brasil, em dois períodos sucessivos, identificando os principais fatores responsáveis pela evolução observada em cada período. MÉTODOS: Os dados analisados provêm de amostras probabilísticas da população de crianças menores de cinco anos estudadas por inquéritos domiciliares do programa Demographic Health Surveys realizados em 1986 (n=1.302), 1996 (n=1.108) e 2006 (n=950). A identificação dos fatores responsáveis pela variação na prevalência da desnutrição (altura para idade < -2 z) levou em conta mudanças na freqüência de cinco determinantes potenciais do estado nutricional, modelagens estatísticas da associação independente entre determinante e risco de desnutrição no início de cada período e cálculo de frações atribuíveis. RESULTADOS: A prevalência da desnutrição foi reduzida em um terço de 1986 a 1996 (de 33,9 por cento para 22,2 por cento ) e em quase três quartos de 1996 a 2006(de 22,2 por cento para 5,9 por cento ). Melhorias na escolaridade materna e na disponibilidade de serviços de saneamento foram particularmente importantes para o declínio da desnutrição no primeiro período, enquanto no segundo período foram decisivos o aumento do poder aquisitivo das famílias mais pobres e, novamente, a melhoria da escolaridade materna. CONCLUSÕES: A aceleração do declínio da desnutrição do primeiro para o segundo período foi consistente com a aceleração de melhorias em escolaridade materna, saneamento, assistência à saúde e antecedentes reprodutivos e, sobretudo, com o excepcional aumento do poder aquisitivo familiar, observado apenas no segundo período. Mantida a taxa de declínio observada entre 1996 e 2006, o problema da desnutrição infantil na região Nordeste poderia ser considerado controlado em menos de dez anos. ) Para se chegar a este resultado será preciso manter o aumento do poder aquisitivo dos mais pobres e assegurar investimentos públicos para completar a universalização do acesso a serviços essenciais de educação, saúde e saneamento

Proteção da comunidade aquática em efluente tratado por tanques de estabilização de resíduos no rio São Lourenço em São Lourenço da Serra, Vale do Ribeira, estado de São Paulo

O tratamento de esgotos de pequenas cidades por lagoas de estabilização é uma maneira simples, eficiente e de baixo custo. Os esgotos são uma fonte de contaminação das águas e solos e, conseqüentemente, contribuem para a transmissão de doenças, além de serem uma ameaça à preservação do meio ambiente. Surge a necessidade de investigar as condições dos efluentes lançados nos cursos d´água. O presente trabalho tem como objetivo realizar uma investigação da qualidade das águas residuárias tratadas por lagoas de estabilização de uma estação de tratamento de esgoto localizadas no município de São Lourenço da Serra no Vale da Ribeira no Estado de São Paulo e verificar os riscos sanitários e a comunidade aquática no Rio São Lourenço da Serra. Foram realizadas amostragens para avaliar o conjunto de lagoas anaeróbia e facultativa da estação de tratamento de esgoto do Município de São Lourenço da Serra. Os parâmetros utilizados foram pH, temperatura do ar e da água, condições climáticas, demanda bioquímica de oxigênio e nitrogênio amoniacal, bactérias termotolerantes, pigmentos fotossintéticos e comunidade zooplanctônica. Verificou-se que s sistemas de lagoa anaeróbia e facultativa foram eficientes na produção de efluente e apresentou alguns valores de acordo com a Resolução Conama nº 357, que estabelecem os valores limites para lançamento em corpos d´água. O rio São Lourenço está localizado em uma área de proteção ambiental. Os dados são comparados aos limites estabelecidos na Classe 1 e 2 e demonstram processo de eutrofização, colocando em risco à biodiversidade aquática e a saúde da população

Disponibilidade de metais pesados em Latossolo com aplicação superficial de escória, lama cal, lodos de esgoto e calcário

O objetivo deste trabalho foi avaliar alguns atributos químicos do solo e a disponibilidade de cádmio (Cd), cromo (Cr), níquel (Ni), mercúrio (Hg), chumbo (Pb) e arsênio (As), por meio da extração pelo DTPA,em conseqüência da aplicação superficial de escória de aciaria, lama cal e lodos de esgoto centrifugados e de biodigestores, nas doses 0 (testemunha), 2, 4 e 8 Mg ha-1 e um tratamento adicional composto pela calagem superficial na dose 2 Mg ha-1. O experimento foi conduzido em delineamento de blocos ao acaso, em condições de campo, em área sob sistema plantio direto, durante 2003 e 2004. A aplicação superficial de escória de aciaria,lama cal, lodo de esgoto centrifugado e de biodigestor, até a dose 8 Mg ha-1, assim como o calcário na dose 2 Mg ha-1, não trazem problemas de disponibilidade ao ambiente, com relação aos metais pesados Cd, Cr, Hg, Pb, Ni e As, quando aplicados sobre a superfície em Latossolo Vermelho distrófico, no sistema plantio direto. A fitodisponibilidade de metais pesados às culturas da soja e aveia-preta foi nula, quando foram aplicadas doses de até 8 Mg ha-1 de lodo de esgoto, escória e lama cal sobre a superfície do solo, no sistema plantio direto

Produção de óleo essencial e balanço nutricional em Corymbia citriodora adubado com lodo de esgoto em diferentes espaçamentos

The Corymbia citriodora is one of the most cultivated species in Brazil for wood and essential oil production. The oil is obtained by the leaves distillation and the yield is around 2%. For the sustainability of leaves extraction is necessary a good nutritional replacement, because the leaves contain a large quantity of nutrients. The sewage sludge application in Corymbia plantations is a way to apply organic fertilizer and is also a good alternative for the final disposal of this residue. In this context, the objective of this study was to evaluate the sewage sludge use as fertilizer for the essential oil production in two different Spacing. The experiment was constituted by 4 blocks and the treatments were: i) Close spacing (1m x 1m) with sewage sludge + K and B; ii) Close spacing with mineral fertilization: iii) Regular spacing (3m x 1m) with sewage sludge + K and B; and iv) Regular spacing with mineral fertilization. When the C. citriodora plots were 12, 18, 24 and 30 months old, leaves and branches biomass and also essential oil production were quantified. It was observed that the system of close planting totalized a higher biomass, 3.8 times more than the conventional system, providing 3 times more essential oil production (363 l ha(-1) at the close spacing and 116 l ha(-1) at the regular Spacing during 30 months). No difference was found in the leaves or oil production with the use of sewage sludge or mineral fertilizer any of spacing studied The use of sewage sludge as fertilizer for Corymbia citriodora plantations may be an interesting alternative. because it supplies large quantities of nutrients and ensures a positive budget of nutrients in the ecosystem.

Efeito da aplicação de fertilização nitrogenada e lodo de esgoto nas características da madeira juvenil de árvores de Eucalyptus grandis

This work evaluated the wood anatomical and physical characteristics of 24 months-old Eucalyptus grandis trees, planted in 3x2m spacing and fertilized with nitrogen (6, 12, 18 month old) and sewage sludge (planting, 8 month old). For each treatment 10 eucalypts trees were cut according to the distribution of basal area. Wood samples were collected in different percentages of the total height to analyze the anatomical (vessels and fibers) and physical (wood density) characteristics. The results showed that the wood apparent density and wood basic density of the eucalypt trees in the nitrogen and sewage sludge were larger in comparison to the control. Radial profiles of wood apparent density, were similar in the three treatments, presenting the exected characteristics of juvenile wood of 24 months-old eucalypt trees. Fiber and vessel dimensions were not affected by fertilization.

A rational protocol for the successful crystallization of l-amino-acid oxidase from Bothrops atrox

Despite the valuable contributions of robotics and high-throughput approaches to protein crystallization, the role of an experienced crystallographer in the evaluation and rationalization of a crystallization process is still crucial to obtaining crystals suitable for X-ray diffraction measurements. In this work, the difficult task of crystallizing the flavoenzyme l-amino-acid oxidase purified from Bothrops atrox snake venom was overcome by the development of a protocol that first required the identification of a non-amorphous precipitate as a promising crystallization condition followed by the implementation of a methodology that combined crystallization in the presence of oil and seeding techniques. Crystals were obtained and a complete data set was collected to 2.3 A resolution. The crystals belonged to space group P2(1), with unit-cell parameters a = 73.64, b = 123.92, c = 105.08 A, beta = 96.03 degrees. There were four protein subunits in the asymmetric unit, which gave a Matthews coefficient V (M) of 2.12 A3 Da-1, corresponding to 42% solvent content. The structure has been solved by molecular-replacement techniques.

Use of Cassava Peel as Carbon Source for Production of Amylolytic Enzymes by Aspergillus niveus

Aspergillus niveus produced high levels of alpha-amylase and glucoamylase in submerged fermentation using the agricultural residue cassava peel as a carbon source. In static conditions, the amylase production was substantially greater than in the agitated condition. The optimized culture conditions were initially at pH 5.0, 35 degrees C during 48 hours. Amylolytic activity was still improved (50%) with a mixture of cassava peel and soluble starch in the proportion 1:1 (w/w). The crude extract exhibited temperature and pH optima approximately 70 degrees C and 4.5, respectively. Amylase activity was stable for 1 h at 60 degrees C, and at pH values between 3.0 and 7.0. The enzyme hydrolysed preferentially maltose, starch, penetrose, amylose, isomaltose, maltotriose, glycogen and amylopectin, and not hydrolysed cyclodextrin (alpha and beta), trehalose and sucrose. In the first hour of reaction on soluble starch, the hydrolysis products were glucose and maltose, but after two hours of hydrolysis, glucose was the unique product formed, confirming the presence in the crude extract of an alpha-amylase and a glucoamylase.

Characterization of Novel OmpA-Like Protein of Leptospira interrogans That Binds Extracellular Matrix Molecules and Plasminogen

Leptospira interrogans is the etiological agent of leptospirosis, a zoonotic disease of human and veterinary concern. The identification of novel proteins that mediate host-pathogen interactions is important for understanding the bacterial pathogenesis as well as to identify protective antigens that would help fight the disease. We describe in this work the cloning, expression, purification and characterization of three predicted leptospiral membrane proteins, LIC10258, LIC12880 (Lp30) and LIC12238. We have employed Escherichia coli BL21 (SI) strain as a host expression system. Recently, we have identified LIC12238 as a plasminogen (PLG)-binding receptor. We show now that Lp30 and rLIC10258 are also PLG-receptors of Leptospira, both exhibiting dose-dependent and saturating binding (K(D), 68.8 +/- 25.2 nM and 167.39 +/- 60.1 nM, for rLIC10258 and rLIC12880, respectively). In addition, LIC10258, which is a novel OmpA-like protein, binds laminin and plasma fibronectin ECM molecules and hence, it was named Lsa66 (Leptospiral surface adhesin of 66 kDa). Binding of Lsa66 to ECM components was determined to be specific, dose-dependent and saturable, with a KD of 55.4 +/- 15.9 nM to laminin and of 290.8 +/- 11.8 nM to plasma fibronectin. Binding of the recombinant proteins to PLG or ECM components was assessed by using antibodies against each of the recombinant proteins obtained in mice and confirmed by monoclonal anti-polyhistidine antibodies. Lsa66 caused partial inhibition on leptospiral adherence to immobilized ECM and PLG. Moreover, this adhesin and rLIC12238 are recognized by antibodies in serum samples of confirmed leptospirosis cases. Thus, Lsa66 is a novel OmpA-like protein with dual activity that may promote the attachment of Leptospira to host tissues and may contribute to the leptospiral invasion. To our knowledge, this is the first leptospiral protein with ECM and PLG binding properties reported to date.

Expression, crystallization and preliminary crystallographic analysis of PilZ(XAC1133) from Xanthomonas axonopodis pv. citri

Proteins containing PilZ domains are widespread in Gram-negative bacteria and have recently been shown to be involved in the control of biofilm formation, adherence, aggregation, virulence-factor production and motility. Furthermore, some PilZ domains have recently been shown to bind the second messenger bis(3'-> 5') cyclic diGMP. Here, the cloning, expression, purification and crystallization of PilZ(XAC1133), a protein consisting of a single PilZ domain from Xanthomonas axonopodis pv. citri, is reported. The closest PilZ(XAC1133) homologues in Pseudomonas aeruginosa and Neisseria meningitidis control type IV pilus function. Recombinant PilZ(XAC1133) containing selenomethionine was crystallized in space group P6(1). The unit-cell parameters were a = 62.125, b = 62.125, c = 83.543 angstrom. These crystals diffracted to 1.85 angstrom resolution and a MAD data set was collected at a synchrotron source. The calculated Matthews coefficient suggested the presence of two PilZ(XAC1133) molecules in the asymmetric unit.

Production of the refolded oligopeptide-binding protein (OppA) encoded by the citrus pathogen Xanthomonas axonopodis pv. citri

The oligopeptide-binding protein, OppA, binds and ushers oligopeptide substrates to the membrane-associated oligopeptide permease (Opp), a multi-component ABC-type transporter involved in the uptake of oligopeptides expressed by several bacterial species. In the present study, we report the cloning, purification, refolding and conformational analysis of a recombinant OppA protein derived from Xanthomonas axonopodis pv. citri (X. citri), the etiological agent of citrus canker. The oppA gene was expressed in Escherichia coli BL21 (DE3) strain under optimized inducing conditions and the recombinant protein remained largely insoluble. Solubilization was achieved following refolding of the denatured protein. Circular dichroism analysis indicated that the recombinant OppA protein preserved conformational features of orthologs expressed by other bacterial species. The refolded recombinant OppA represents a useful tool for structural and functional analyses of the X. citri protein.

Crystallization and preliminary X-ray diffraction analysis of a glutathione S-transferase from Xylella fastidiosa

Glutathione S-transferases (GSTs) form a group of multifunctional isoenzymes that catalyze the glutathione-dependent conjugation and reduction reactions involved in the cellular detoxification of xenobiotic and endobiotic compounds. GST from Xylella fastidiosa (xfGST) was overexpressed in Escherichia coli and purified by conventional affinity chromatography. In this study, the crystallization and preliminary X-ray analysis of xfGST is described. The purified protein was crystallized by the vapour-diffusion method, producing crystals that belonged to the triclinic space group P1. The unit-cell parameters were a = 47.73, b = 87.73, c = 90.74 angstrom, alpha = 63.45, beta = 80.66, gamma = 94.55 degrees. xfGST crystals diffracted to 2.23 angstrom resolution on a rotating-anode X-ray source.

Molecular determinants of improved cathepsin B inhibition by new cystatins obtained by DNA shuffling

Background: Cystatins are inhibitors of cysteine proteases. The majority are only weak inhibitors of human cathepsin B, which has been associated with cancer, Alzheimer's disease and arthritis. Results: Starting from the sequences of oryzacystatin-1 and canecystatin-1, a shuffling library was designed and a hybrid clone obtained, which presented higher inhibitory activity towards cathepsin B. This clone presented two unanticipated point mutations as well as an N-terminal deletion. Reversing each point mutation independently or both simultaneously abolishes the inhibitory activity towards cathepsin B. Homology modeling together with experimental studies of the reverse mutants revealed the likely molecular determinants of the improved inhibitory activity to be related to decreased protein stability. Conclusion: A combination of experimental approaches including gene shuffling, enzyme assays and reverse mutation allied to molecular modeling has shed light upon the unexpected inhibitory properties of certain cystatin mutants against Cathepsin B. We conclude that mutations disrupting the hydrophobic core of phytocystatins increase the flexibility of the N-terminus, leading to an increase in inhibitory activity. Such mutations need not affect the inhibitory site directly but may be observed distant from it and manifest their effects via an uncoupling of its three components as a result of increased protein flexibility.

Protein preparation, crystallization and preliminary X-ray analysis of Trypanosoma cruzi nucleoside diphosphate kinase 1

The flagellated protozoan parasite Trypanosoma cruzi is the aetiological agent of Chagas disease. Nucleoside diphosphate kinases (NDPKs) are enzymes that are involved in energy management and nucleoside balance in the cell. T. cruzi TcNDPK1, a canonical isoform, was overexpressed in Escherichia coli as an N-terminally poly-His-tagged fusion protein and crystallized. Crystals grew after 72 h in 0.2 M MgCl(2), 20% PEG 3350. Data were collected to 3.5 angstrom resolution using synchrotron X-ray radiation at the National Synchrotron Light Laboratory (Campinas, Brazil). The crystals belonged to the trigonal space group P3, with unit-cell parameters a = b = 127.84, c = 275.49 angstrom. Structure determination is under way and will provide relevant information that may lead to the first step in rational drug design for the treatment of Chagas disease.

Crystallization and preliminary X-ray diffraction analysis of recombinant chlorocatechol 1,2-dioxygenase from Pseudomonas putida

Chlorocatechol 1,2-dioxygenase from the Gram-negative bacterium Pseudomonas putida (Pp 1,2-CCD) is considered to be an important biotechnological tool owing to its ability to process a broad spectrum of organic pollutants. In the current work, the crystallization, crystallographic characterization and phasing of the recombinant Pp 1,2-CCD enzyme are described. Reddish-brown crystals were obtained in the presence of polyethylene glycol and magnesium acetate by utilizing the vapour-diffusion technique in sitting drops. Crystal dehydration was the key step in obtaining data sets, which were collected on the D03B-MX2 beamline at the CNPEM/MCT - LNLS using a MAR CCD detector. Pp 1,2-CCD crystals belonged to space group P6(1)22 and the crystallographic structure of Pp 1,2-CCD has been solved by the MR-SAD technique using Fe atoms as scattering centres and the coordinates of 3-chlorocatechol 1,2-dioxygenase from Rhodococcus opacus (PDB entry 2boy) as the search model. The initial model, which contains three molecules in the asymmetric unit, has been refined to 3.4 A resolution.