Genotyping of Brazilian Erysipelothrix spp. strains by amplified fragment length polymorphism

One hundred and fifty-one Erysipelothrix spp. isolates from diseased and carrier swine from Brazil were identified by PCR, submitted to serotyping and analyzed by amplified fragment length polymorphism with a single enzyme (AFLP). Reference strains from Australia and the United Kingdom were also examined. The 151 strains were classified into 18 different serotypes (1a, 1b, 2a, 2b, 4, 5, 6, 7, 8, 10, 11, 12, 15, 17, 19, 21, 24 and 25), being serotype 2b the most frequent (39.7%). By associating serotyping and PCR results, it was possible to identify 146 strains as E. rhusiopathiae and five strains as E. tonsillarum. Despite the fact that for this genus AFLP did not cluster all isolates according to serotype, origin, disease or isolation data, the execution of the technique was easy and fast, demonstrating high discriminatory power. The results produced by the AFLP analysis of Erysipelothrix spp. could also support its use as a discriminatory tool for E. rhusiopathiae and E. tonsillarum species. (C) 2010 Elsevier B.V. All rights reserved.

Effect of culture media on porcine embryos produced by in vitro fertilization or parthenogenetic activation after oocyte maturation with cycloheximide

This study evaluated the effects of reversible meiotic inhibition and different culture media (PZM3 or NCSU23) on production of porcine embryos by either in vitro fertilization (IVF) or parthenogenetic activation (PA). Oocytes from abattoir-derived ovaries were allocated into two groups for maturation: CHX (5 mu g/ml cycloheximide for 10 h) or Control (no CHX). The percentage of metaphase II (MII) oocytes was determined at 36, 40 or 44 h of in vitro maturation. For IVF and PA, denuded oocytes were fertilized with purified sperm for 6 h or activated by electric stimuli. Zygotes were then subdivided into two culture groups: NCSU23 or PZM3. No effect of treatment with CHX and culture media was observed on cleavage (D3) and blastocyst (D7) rates in IVF and PA groups. There are no differences of quality or development rates between IVF-derived embryos cultured in NCSU23 or PZM3. However, we observed high quality PA embryos in PZM3 compared with NCSU23. Maturation arrest with CHX decreased the average blastocyst cell number in IVF while it was increased in PA embryos. As older oocytes are more effectively activated, CHX-blocked oocytes reached the mature stage faster than the control group. In conclusion, the CHX treatment for 10 h, followed by oocyte maturation for 40 h, is an efficient protocol to produce high quality parthenote embryos, especially when they are cultured in PZM3. However, this protocol is not satisfactory for IVF embryos production. In this case, a shorter maturation period could provide better embryo quality.

Vitamin B12 and hepatitis C: Molecular biology and human pathology

Cobalamins are stored in high concentrations in the human liver and thus are available to participate in the regulation of hepatotropic virus functions. We show that cyanocobalamin (vitamin B12) inhibited the H(IV internal ribosome entry site (IRES)-dependent translation of a reporter gene in vitro in a dose-dependent manner without significantly affecting the cap-dependent mechanism. Vitamin B12 failed to inhibit translation by IRES elements from encephalomyocarditis virus (EMCV) or classical swine fever virus (CSFV), We also demonstrate a relationship between the total cobalamin concentration in human sera and HCV viral load (a measure of viral replication in the host), The mean viral load was two orders of magnitude greater when the serum cobalamin concentration was above 200 pM (P < 0.003), suggesting that the total cobalamin concentration in an HCV-infected liver is biologically significant in HCV replication.

Effect of periparturient diseases accompanied by excessive vulval discharge and weaning to mating interval on sow reproductive performance

Objective To evaluate the effect of periparturient disease accompanied by vulval discharge, and weaning-to-mating intervals, on sow fertility and litter size. Design Reproductive data were collected and analysed from 19 Hungarian swine herds over a 4 year period. Conception rates, farrowing rates and litter sizes of sows with periparturient disease accompanied by vulval discharge were used to evaluate the relationship between duration of vulval discharge and subsequent fertility and litter size. The possibility of interactions between weaning-to-mating intervals and duration of vulval discharges was investigated to determine if there was any effect on subsequent fertility and litter size. Results and conclusions Both parity 1 and parity 2 to 8 sows having had periparturient disease accompanied by vulval discharge in excess of 6 days duration had significantly (P < 0.001) lower subsequent fertility (conception, farrowing and adjusted farrowing rates) compared with sows of similar parity where the duration of vulval discharge was < 4 or 4 to 6 days. There was no difference in fertility rates between sows, in both parity categories, with vulval discharge for < 4 days compared with 4 to 6 days. A duration of vulval discharge in excess of 6 days in parity 1 sows significantly reduced litter size (total born and live-born) in subsequent farrowings, but not in parity 2 to 8 sows. There was no interaction between the duration of vulval discharge and post-weaning to mating intervals. However sows with weaning to mating intervals between 7 and 10 days had smaller (P < 0.001) subsequent litter sizes compared with 3 to 6 or 11 to 14 day intervals. It was concluded that the duration of vulval discharge in excess of 6 days was an indication of a severe persistent endometritis adversely affecting fertility of sows.

Mathematical modelling of schistosomiasis japonica: comparison of control strategies in the People's Republic of China

We present the first mathematical model on the transmission dynamics of Schistosoma japonicum. The work extends Barbour's classic model of schistosome transmission. It allows for the mammalian host heterogeneity characteristic of the S. japonicum life cycle, and solves the problem of under-specification of Barbour's model by the use of Chinese data we are collecting on human-bovine transmission in the Poyang Lake area of Jiangxi Province in China. The model predicts that in the lake/marshland areas of the Yangtze River basin: (1) once-early mass chemotherapy of humans is little better than twice-yearly mass chemotherapy in reducing human prevalence. Depending on the heterogeneity of prevalence within the population, targeted treatment of high prevalence groups, with lower overall coverage, can be more effective than mass treatment with higher overall coverage. Treatment confers a short term benefit only, with prevalence rising to endemic levels once chemotherapy programs are stopped (2) depending on the relative contributions of bovines and humans, bovine treatment can benefit humans almost as much as human treatment. Like human treatment, bovine treatment confers a short-term benefit. A combination of human and bovine treatment will dramatically reduce human prevalence and maintains the reduction for a longer period of time than treatment of a single host, although human prevalence rises once treatment ceases; (3) assuming 75% coverage of bovines, a bovine vaccine which acts on worm fecundity must have about 75% efficacy to reduce the reproduction rate below one and ensure mid-term reduction and long-term elimination of the parasite. Such a vaccination program should be accompanied by an initial period of human treatment to instigate a short-term reduction in prevalence, following which the reduction is enhanced by vaccine effects; (4) if the bovine vaccine is only 45% efficacious (the level of current prototype vaccines) it will lower the endemic prevalence, but will not result in elimination. If it is accompanied by an initial period of human treatment and by a 45% improvement in human sanitation or a 30% reduction in contaminated water contact by humans, elimination is then possible. (C) 2002 Elsevier Science B.V. All rights reserved.

Amino acids 1-20 of the hepatitis C virus (HCV) core protein specifically inhibit HCV IRES- dependent translation in Hep G2 cells, and inhibit both HCV IRES- and cap-dependent translation in HuH7 and CV-1 cells.

A self-modulating mechanism by the hepatitis C virus (HCV) core protein has been suggested to influence the level of HCV replication, but current data on this subject are contradictory. We examined the effect of wild-type and mutated core protein on HCV IRES- and cap-dependent translation. The wild-type core protein was shown to inhibit both IRES- and cap-dependent translation in an in vitro system. This effect was duplicated in a dose-dependent manner with a synthetic peptide representing amino acids 1-20 of the HCV core protein. This peptide was able to bind to the HCV IRES as shown by a mobility shift assay. In contrast, a peptide derived from the hepatitis B virus (HBV) core protein that contained a similar proportion of basic residues was unable to inhibit translation or bind the HCV IRES. A recombinant vaccinia-HCV core virus was used to examine the effect of the HCV core protein on HCV IRES-dependent translation in cells and this was compared with the effects of an HBV core-recombinant vaccinia virus. In CV-1 and HuH7 cells, the HCV core protein inhibited translation directed by the IRES elements of HCV, encephalomyocarditis virus and classical swine fever virus as well as cap-dependent translation, whereas in HepG2 cells, only HCV IRES-dependent translation was affected. Thus, the ability of the HCV core protein to selectively inhibit HCV IRES-dependent translation is cell-specific. N-terminal truncated (aa 1-20) HCV core protein that was expressed from a novel recombinant vaccinia virus in cells abrogated the inhibitory phenotype of the core protein in vivo, consistent with the above in vitro data.

A regula????o no setor de saneamento: compara????o entre Fran??a, Inglaterra e Brasil

O artigo discute as caracter??sticas da regula????o dos servi??os p??blicos no setor de saneamento, apresentando comparativamente as formas de presta????o adotadas por Fran??a, Inglaterra e Brasil ??? e como esses pa??ses disciplinam a participa????o privada no setor. Enquanto a Fran??a segue um padr??o hist??rico de regula????o marcado pelo protagonismo das autoridades locais, utilizando contratos como instrumento por excel??ncia de disciplina dos servi??os, a Inglaterra introduziu, atrav??s de ambicioso processo de privatiza????o, marco regulat??rio em que ag??ncias do Governo Central s??o os atores principais. Ainda sem um modelo definido, o Brasil, atrav??s de inova????es legais, enfrenta os desafios de atrair investimentos privados para um setor sob a presen??a de conflitos federativos entre estados e munic??pios acerca da titularidade dos referidos servi??os p??blicos.

A água, o saneamento básico e os residuos sólidos urbanos no desenvolvimento de São Tomé e Príncipe

RESUMO: O trabalho que se apresenta no âmbito desta dissertação, é direcionado para a problemática da Água, do Saneamento Básico e dos Resíduos Sólidos Urbanos «RSU» em São Tomé e Príncipe. Num contexto de desenvolvimento e indo ao encontro dos anseios da Organização das Nações Unidas «ONU» e da sua perspetiva de alcançar os Objetivos do Milénio nesta área tão importante. Elegeu-se como primordial objetivo, conhecer as indicações técnico políticas instituídas em São Tomé e Príncipe, para a gestão dos problemas acima enumerados. Entender esses problemas, identificar as dificuldades sentidas pelo governo e pela generalidade dos seus habitantes no acesso à água, ao saneamento básico, à recolha e tratamento de RSU. Outra vertente será direcionada para apontar caminhos nestas áreas, onde a capacidade institucional tarda em dar resposta às necessidades básicas destes setores, inviabilizando um desenvolvimento sustentado destes ramos. Esta dissertação, assenta ainda no reconhecimento e na importância estratégica em se valorizar e consolidar redes técnico-científicas no âmbito da Linha de Investigação em Estudos Africanos e Pós-Coloniais, inserida na Unidade de Estudos e Investigação em Ciência, Tecnologia e Sociedade «UEICTS» da Universidade Lusófona de Humanidades e Tecnologias «ULHT».ABSTRACT: The work presented in this dissertation, is directed to the problem of Water Sanitation and Solid Waste «RSU» in Sao Tome and Principe. In a context of development, fulfillment of the wishes of the United Nations «UN» and its prospect of achieving the Millennium Goals in this important area. The prime objective, is, to know the indications and technical policies in place in Sao Tome and Principe for the management of the problems listed above. Understanding these problems, identifying the difficulties faced by government and by most of its residents in relation, to access to water, sanitation, collection and treatment of Solid Waste. Another aspect, is directed, to point, to ways, in these subjects where institutional capacity is slow to respond to basic needs of these sectors, preventing a sustained development of these industries. This dissertation focus on the recognition and strategic importance in considering and consolidate technical and scientific networks in the line of Research in African and Lusophone, inserted at the Unit for Studies and Research in Science, Technology and Society «UEICTS» Lusophone University of Humanities and Technology «ULHT».

Sistemas de abastecimento de água e saneamento básico

O presente relatório de estágio enquadra-se no âmbito do Trabalho Final de Mestrado (TFM) do curso de Engenharia Civil, área de especialização de Hidráulica, do Instituto Superior de Engenharia de Lisboa, e baseia-se na temática dos sistemas de abastecimento de água e drenagem de águas residuais. O estágio, intitulado de “Sistemas de Abastecimento de Agua e Saneamento Básico”, decorreu numa empresa de consultaria de engenharia denominada ENGIDRO – Estudos de Engenharia, Lda., vocacionada para a realização de estudos e projectos na área de Hidráulica, com particular incidência na Hidráulica Urbana e Saneamento Básico. O estágio iniciou-se com um adequado enquadramento profissional na empresa e incidiu principalmente no desenvolvimento de trabalhos de concepção e dimensionamento, a nível de projectos de execução, de sistemas de abastecimento de água e de saneamento básico, para 21 localidades da província de Lunda Sul, em Angola, com prestação de serviços para o Governo Provincial de Lunda Sul – Direcção Provincial da Energia e Aguas, juntamente com empresa CENGA – Consultores de Engenharia de Angola, S.A. Na prestação de serviços à entidade contratante foram efectuados trabalhos de campo, que consistiram principalmente em reconhecimentos do terreno, levantamentos topográficos e recolha de informação relevante sobre elementos condicionantes dos projectos (origens e disponibilidades de água, natureza e declive dos terrenos, tipos de infra-estruturas locais) e trabalhos de gabinete para compilação e análise da informação recolhida na elaboração dos projectos de execução, incluindo pecas desenhadas (desenhos pormenorizados) e peças escritas (memórias descritivas e justificativas, medições e orçamentos). Sendo os projectos destinados a aglomerados populacionais pouco desenvolvidos e com carências e condicionantes de diversa ordem (falta de energia eléctrica, de acessos, de telecomunicações, de meios técnicos e materiais, entre outros), prestou-se especial atenção aos aspectos da concepção, privilegiando soluções de baixa tecnologia, mais fáceis de explorar e manter com os recursos locais disponíveis.

Conservação, manutenção e construção de património estratégico das águas de Santarém

A água é um bem essencial ao ser humano. Nos dias de hoje é de extrema importância fazer chegar este bem às populações, um papel que no nosso país é desempenhado pelos municípios, por empresas municipais e por empresas concessionadas. Este relatório, enquadrado no âmbito do trabalho final de mestrado em Engenharia Civil, especialização em Hidráulica, no Instituto Superior de Engenharia de Lisboa, cujo título é “Conservação, Manutenção e Construção de Património Estratégico das Águas de Santarém”, relata a experiência de estágio do autor do presente relatório na A.S. – Empresa das Águas de Santarém – EM, S.A. O relatório aborda as temáticas do abastecimento de água, do tratamento de águas residuais, da construção de sistemas de saneamento e estações de tratamento de águas residuais, da reparação de equipamentos da empresa e do consumo de energia dos equipamentos da empresa. São descritas as actividades desenvolvidas pelo autor, que incluem o acompanhamento activo em visitas a obras de construção e manutenção de património, a monitorização e controlo de perdas de água, bem como a actualização em suporte informático do cadastro dos equipamentos pertencentes à A.S., com a criação de fichas de equipamento individuais e de um suporte informático para registo e consulta comparativa de consumos energéticos. Esta experiência de estágio permitiu ao autor um contacto com a realidade de uma empresa em toda a diversidade das actividades anteriormente nomeadas, que será descrita objectivamente nos capítulos do presente documento. O estágio foi claramente enriquecedor para o autor, fornecendo-lhe formação objectiva e a preparação necessária para a sua integração no mercado de trabalho nesta área específica da Engenharia Civil.

Higienização de utensílios alimentares em estabelecimentos de ensino pré-escolar: um estudo em duas instituições de ensino

A transmissão de microrganismos patogénicos pode ocorrer através da sua presença em resíduos de alimentos ou água depositados sobre os utensílios que se encontram em deficientes condições de higienização. Para que seja assegurada a higiene alimentar torna-se essencial a conservação e higienização dos utensílios, desempenhando os manipuladores de alimentos um papel determinante no que diz respeito à execução de um eficaz protocolo de higienização, uma correcta manipulação e a adopção de normas higiénicas que evitem a contaminação dos utensílios. Da totalidade das amostras realizadas aos utensílios alimentares provenientes das duas instituições de ensino pré-escolar analisados neste estudo, 27% encontravam-se contaminadas e num insuficiente estado de higienização devido à presença de microrganismos mesófilos aeróbios a 30ºC em número superior a 100 UFC/peça. Foi verificado a presença de fungos e a pesquisa positiva de enterobactérias e bactérias presumíveis de E. coli. Em ambos os estabelecimentos de ensino, os utensílios constituídos por material plástico apresentaram níveis de contaminação na generalidade superiores em relação aos constituídos por aço inoxidável. O tipo de material e o estado dos utensílios são também aspectos relevantes que podem influenciar o processo de adesão microbiana e consequente formação de biofilme, pelo que na aquisição dos utensílios devem ser consideradas as características inerentes aos diferentes materiais, devendo os mesmos ser substituídos periodicamente devido ao desgaste causado pelo seu frequente uso. Para além disso, os utensílios devem passar constantemente por uma avaliação microbiológica para controlo da eficiência do processo de higienização, contribuindo para a garantia da higiene alimentar e segurança do alimento. A garantia da higiene alimentar constitui uma base importante em idades de maior vulnerabilidade.

Environmental impact on the bacteriological quality of domestic water supplies in Lagos, Nigeria

OBJECTIVE: To assess the impact of town planning, infrastructure, sanitation and rainfall on the bacteriological quality of domestic water supplies. METHODS: Water samples obtained from deep and shallow wells, boreholes and public taps were cultured to determine the most probable number of Escherichia coli and total coliform using the multiple tube technique. Presence of enteric pathogens was detected using selective and differential media. Samples were collected during both periods of heavy and low rainfall and from municipalities that are unique with respect to infrastructure planning, town planning and sanitation. RESULTS: Contamination of treated and pipe distributed water was related with distance of the collection point from a utility station. Faults in pipelines increased the rate of contamination (p<0.5) and this occurred mostly in densely populated areas with dilapidated infrastructure. Wastewater from drains was the main source of contamination of pipe-borne water. Shallow wells were more contaminated than deep wells and boreholes and contamination was higher during period of heavy rainfall (p<0.05). E. coli and enteric pathogens were isolated from contaminated supplies. CONCLUSIONS: Poor town planning, dilapidated infrastructure and indiscriminate siting of wells and boreholes contributed to the low bacteriological quality of domestic water supplies. Rainfall accentuated the impact.

A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations

The conjugation of antigens with ligands of pattern recognition receptors (PRR) is emerging as a promising strategy for the modulation of specific immunity. Here, we describe a new Escherichia coli system for the cloning and expression of heterologous antigens in fusion with the OprI lipoprotein, a TLR ligand from the Pseudomonas aeruginosa outer membrane (OM). Analysis of the OprI expressed by this system reveals a triacylated lipid moiety mainly composed by palmitic acid residues. By offering a tight regulation of expression and allowing for antigen purification by metal affinity chromatography, the new system circumvents the major drawbacks of former versions. In addition, the anchoring of OprI to the OM of the host cell is further explored for the production of novel recombinant bacterial cell wall-derived formulations (OM fragments and OM vesicles) with distinct potential for PRR activation. As an example, the African swine fever virus ORF A104R was cloned and the recombinant antigen was obtained in the three formulations. Overall, our results validate a new system suitable for the production of immunogenic formulations that can be used for the development of experimental vaccines and for studies on the modulation of acquired immunity.

Occupational exposure to toxigenic Aspergillus versicolor in Portuguese swines

Biological factors associated with airbome dust are the most important hazards in pig buildings and include allergenic and/or toxic compounds, as well as infectious agents such as fungi and their metabolites, like mycotoxins. Inhalation of such agents can be a potential occupationai treat. Exposure of workers from swine confinement buildings to respiratory hazards has been reported elsewhere in Europe, Asia and America. Analogous data has not been reported for Portugal and this omission has hindered the development of policies in the area of occupational health and farm safety. Aspergilius versicolor is known as being the major producer of the hepatotoxic and carcinogenic mycotoxin sterigmatocystin. The toxicity of this mycotoxin is manifested primarily in liver and kidney. This study aimed to determine occupational exposure treat due to fungal contamination caused by A. versicolor in seven Portuguese swine.

Comparison of Aspergillus species-complexes detected in different environmental settings

Purpose: Samples from different environmental sources were screened for the presence of Aspergillus, and the distribution of the different species-complexes was determined in order to understand differences among that distribution in the several environmental sources and which of these species complexes are present in specific environmental settings. Methods: Four distinct environments (beaches, poultries, swineries and hospital) were studied and analyzed for which Aspergillus complexes were present in each setting. After plate incubation and colony isolation, morphological identification was done using macro- and microscopic characteristics. The universal fungal primers ITS1 and ITS4 were used to amplify DNA from all Aspergillus isolates, which was sequenced for identification to species complex level. SPSS v15.0 for Windows was used to perform the statistical analysis. Results: Thirty-nine isolates of Aspergillus were recovered from both the sand beach and poultries, 31 isolates from swineries, and 80 isolates from hospital environments, for a total 189 isolates. Eleven species complexes were found total. Isolates belonging to the Aspergillus Versicolores species-complex were the most frequently found (23.8%), followed by Flavi (18.0%), Fumigati (15.3%) and Nigri (13.2%) complexes. A significant association was found between the different environmental sources and the distribution of the several species-complexes (p<0.001); the hospital environment had a greater variability of species-complexes than other environmental locations (10 in hospital environment, against nine in swine, eight in poultries and seven in sand beach). Isolates belonging to Nidulantes complex were detected only in the hospital environment, whereas the other complexes were identified in more than one setting. Conclusion: Because different Aspergillus complexes have different susceptibilities to antifungal drugs, and different abilities in producing mycotoxins, knowledge of the species-complex epidemiology for each setting may allow preventive or corrective measures to be taken toward decreasing professional workers or patient exposure to those agents.