Abstract Portuguese Traditional Music is often overlooked in formal teaching at music schools in Portugal. Furthermore, there has been little research into its use in the classroom and the informal learning practices associated with it. Thus, an Educational Project was developed and implemented in 2021/2022, at the Regional Conservatory of Ponta Delgada (CRPD), involving 7 violin students aged between 9 and 14 years and focused on learning songs from the Traditional Music of the Azores. These songs were prepared in class using informal learning practices - auditory learning and analysis, the use of recordings, memorisation without the use of sheet music, improvisation, etc. - and presented at a concert. Additionally, a research study was conducted to analyse the effects of such intervention on the motivation, learning and attitudes of the students, as well as the perceptions developed by the students, their parents, the teachers, and the general public involved in this experience. The study also aimed to assess the feasibility of combining formal and informal learning practices in violin teaching at the CRPD and to evaluate its implications for the preservation of regional cultural identity. To this end, qualitative data were collected, through direct observation of classes with field notes, audiovisual recordings, semi-structured interviews with the students, anonymous questionnaires to the parents and the audience present at the concert, and 27 violin teachers. This data was subjected to content analysis, namely by organising it into categories and sub-categories. The results show that the inclusion of Azorean Traditional Music, using informal learning practices, had a positive effect on the students' motivation, enhancing the development of technical, musical, cognitive, and social skills. In addition, the Azorean Traditional Music concert proved to be an effective means of disseminating and valuing the region's cultural heritage, boosting the students' identity development, and preserving the region's cultural identity.

Fluorescence spectroscopy is an extremely sensitive analytical technique that has been widely used to elucidate problems that require low detection limits. Although fluorescence spectroscopy is widespread in industry and academia, there is a lack of teaching materials demonstrating its basic principles, as well as the influence of instrumental parameters involved in the technique, which limit its correct use and the full potential of it. For this reason, this work aims to explore the principles of fluorescence spectroscopy and correlate them in function of different instrumental variables to carry out a fluorescence experiment. The spectrofluorometer parameters such as the excitation wavelength, excitation and emission slits, step, integration time, detection geometry as well as concentration of the samples were varied to illustrate how the best resulting spectra can be registered free of artifacts. To do so, two organic dyes with structured vibronic bands (pyrene) and another with unstructured bands (rhodamine 6G) were used. Another study carried out involved the use of 3D spectra, excitation/emission spectra, of cadmium telluride (CdTe) quantum dots. In this case, it was possible to show the excitation spectrum of nanomaterials, collecting emissions at different wavelengths, as well as analyzing the particle size distribution. limits academia reason experiment wavelength slits step time artifacts so pyrene (pyrene rhodamine 6G G D excitationemission CdTe (CdTe dots case nanomaterials wavelengths distribution

Resumo Objetivo Neste estudo, investigamos o impacto de uma intrusão marinha nas características da coluna d'água da lagoa de Jurubatiba, um ecossistema costeiro tropical, por 12 meses após um evento de abertura da barra de areia, a fim de elucidar a duração de seus efeitos na lagoa. Métodos Amostramos mensalmente as características físicas, químicas e biológicas ao longo do perfil vertical da coluna d'água após a abertura da barra de areia e entrada de água do mar. Avaliamos a associação temporal (mensalmente) e espacial (ao longo do perfil de profundidade) entre essas variáveis e a correlação temporal na heterogeneidade vertical dessas variáveis e condições ambientais que promovem a estratificação. Resultados A intrusão marinha resultou na formação de cunhas salinas em curto prazo, levando à estratificação na concentração de oxigênio dissolvido, pH, salinidade e clorofila-a ao longo da coluna de água por, pelo menos, quatro meses. No entanto, a temperatura atmosférica e a incidência solar também emergiram como fatores cruciais associados à heterogeneidade vertical nas variáveis limnológicas, influenciando a densidade da água e o pH ao longo do período de estudo. A precipitação e a velocidade do vento tiveram apenas efeitos marginais nas variações térmicas e ectogênicas, provavelmente mediadas pela cor da água, entrada de nutrientes, profundidade da coluna d'água e orientação da lagoa na paisagem. Conclusões Apesar de rasas e expostas a ação dos ventos, as lagoas costeiras podem apresentar uma considerável heterogeneidade vertical nas condições limnológicas devido a vários fatores climáticos e hidrodinâmicos. Notavelmente, os efeitos de intrusões marinhas na salinidade e na estratificação da coluna d'água podem persistir por vários meses, afetando a qualidade da água e, provavelmente, as comunidades bióticas e o funcionamento do ecossistema. Uma vez que as atividades humanas e as mudanças climáticas podem intensificar as intrusões marinhas em lagoas costeiras, compreender seus efeitos na dinâmica limnológica é fundamental para nossa capacidade de prevê-los e conservar ecossistemas tão vulneráveis. estudo dágua d Jurubatiba tropical 1 físicas mar (mensalmente prazo dissolvido clorofilaa clorofila menos entanto ectogênicas nutrientes paisagem ventos hidrodinâmicos Notavelmente prevêlos prevê los vulneráveis

Abstract Aim In this study, we investigated the impact of an episodic marine intrusion on the water column characteristics of Jurubatiba lagoon, a tropical coastal ecosystem, for 12 months following a sandbar breaching event to elucidate the duration of its effects. Methods Monthly, we sampled the physical, chemical, and biological characteristics along the water column for one year after a sandbar breaching event and seawater inflow. We assessed the temporal (monthly) and spatial (along depth profiles) associations among these variables and the temporal correlation among vertical heterogeneities in these variables and environmental drivers of stratification. Results Marine intrusion resulted in salt wedges formation in the short-term, leading to stratification in dissolved oxygen, pH, salinity, and chlorophyll-a concentration along the water column for at least four months. However, temperature and solar incidence also emerged as crucial factors associated with vertical heterogeneity in limnological variables, influencing water density and pH throughout the study period. Precipitation and wind speed had only marginal effects on thermal and ectogenic stratifications, probably mediated by water colour, nutrient inputs, water column depth and lagoon’s orientation in the landscape. Conclusions Our data unveiled that, despite their shallowness and wind exposure, coastal lagoons can exhibit considerable vertical heterogeneities in limnological variables due to various climatic and hydrodynamic aspects. Notably, the effects of marine intrusions on salinity and ectogenic column stratification can persist for several months, affecting water quality and, probably, biotic communities and ecosystem functioning. Once human activities and climate change may intensify marine intrusions on coastal lagoons, understanding spatiotemporal dynamics and their drivers is fundamental to anticipating their effects and conserving such vulnerable ecosystems. lagoon 1 Monthly physical chemical inflow monthly (monthly profiles shortterm, shortterm short term, term short-term oxygen chlorophylla chlorophyll However period stratifications colour inputs s landscape that exposure aspects Notably functioning ecosystems

Resumo A investigação sobre a aprendizagem musical tem mostrado que as atividades musicais em conjunto podem ter um impacto positivo na aprendizagem do instrumento de música. Neste contexto, um campo ainda pouco estudado são as aulas de grupo de instrumento, nomeadamente, como metodologia de apoio às aulas individuais. Por isso, neste artigo vai debater-se a pertinência da participação dos alunos em aulas de grupos de instrumento, e procurar entender se estas aulas podem ser um modo de desenvolver competências de performance, e outras, como o pensamento crítico. Foram realizados 16 questionários a alunos que frequentaram este tipo de aulas e 4 questionários a professores que as administraram. Foram observadas três aulas deste tipo numa instituição de ensino superior de música. Os dados qualitativos foram sujeitos ao que é chamado na literatura de “análise de padrão”, ou seja, analise à base da categorização dos dados recolhidos. Os resultados obtidos indicam que as aulas de grupo instrumentais são uma metodologia relevante de apoio à aprendizagem instrumental, nomeadamente, no auxílio à preparação da performance. Estas aulas ajudam os alunos a superar obstáculos - como aprender a lidar com o nervosismo - e ajuda-os a desenvolverem competências, como a concentração, competências metacognitivas e o pensamento crítico. As aulas de grupo mencionadas neste estudo permitem a simulação da performance real, o que constitui um momento de teste, correção, e avaliação do estado da preparação individual para a performance. A existência de feedback nestas aulas enriquece a aprendizagem, catalisa a aquisição de competências e ajuda os alunos a crescerem técnica, musical e humanamente.

Abstract The research on musical learning has shown that group musical activities can have a positive impact on the learning of an instrument music. In this context, a field still little studied is the instrumental group classes, namely as a methodology to support individual classes. So, this article discusses the relevance of having students engaging in such instrument group classes and seeks to understand whether these can be a way to develop performance skills, and others, such as critical thinking. Sixteen questionnaires were conducted to students who attended this type of classes and four questionnaires to teachers who gave them. Three classes of this kind were observed in a higher music institution. Qualitative data were subject to what is called in the literature as "pattern analysis", that is, analysis based on the categorization of the collected data. The results obtained indicate that instrumental group classes are a relevant methodology to support instrumental learning, namely in helping performance preparation. These classes help students to overcome obstacles - such as learning to deal with nervousness - and help them to develop skills such as concentration, metacognitive skills, and critical thinking. The group classes mentioned in this study allow the simulation of real performance, which creates a moment of testing, correction, and evaluating the state of the individual preparation for performance. The existence of feedback in these classes enriches learning, catalyzes the acquisition of skills, and helps students to grow technically, musically, and humanly.

Abstract In the studies on musical learning, group musical practice has not been the object of in-depth study. Little is currently known about the impact of playing in an orchestra on students' motivation or learning to study the instrument. In this study, the objective was to understand the phenomenon of musical practice in orchestra to answer the research questions: Does playing in an orchestra generate intrinsic motivation in students? Does it have implications for individual learning and study of the instrument? Semi-structured interviews were conducted with 15 orchestral students to learn the attitude, feelings and perceptions of the students involved. The content analysis involved: (1) a systematic approach to the data analysis process; (2) the use of data reduction methods; and 3) coding the data into categories. This categorization resulted in what is called “pattern analysis.” The qualitative data analysis indicates that students feel motivated for orchestral practice and that playing in the orchestra can positively affect the student's learning and motivation. The orchestra practice seems to increase the practice time and help develop musical and social skills. Self-determination theory is the one that explains the obtained results. According to this theoretical perspective, to feel motivated for a given task, the individual must fulfil three basic psychological needs: autonomy, competence, and establishment of social bonds. For the students, playing in orchestra seems to be a favorable context for developing autonomy, perception of competence, and establishing social bonds with their peers. Playing in orchestra seems to be the challenge in which the balance between the perception of the difficulty of the task and the perception of being competent to perform it successfully is present. That is an excellent level of challenge.

RESUMO Objetivo: o artigo apresenta um framework para a condução de pesquisa interpretativa usando algoritmos de aprendizado profundo que dissolvem os limites entre abordagens qualitativas e quantitativas. O trabalho mostra como a pesquisa pode se beneficiar de uma abordagem integrada que usa ferramentas computacionais para superar limitações tradicionais. Proposta: a maior disponibilidade e diversidade de dados tem aumentado o valor dos algoritmos como ferramentas de pesquisa que podem ser utilizadas pelos cientistas sociais. Além disso, ajustar e usar esses artefatos computacionais pode requerer e se beneficiar de procedimentos interpretativos. Tais circunstâncias viram de cabeça para baixo o tradicional debate sobre pesquisa quantitativa e qualitativa: a estratégia de pesquisa que provavelmente produz a maior quantidade de assertividade e rigor é aquela que requer esforço interpretativo mais vigoroso. Nesses termos, a codificação de palavras por meio de redes neurais pode ajudar os pesquisadores a ler com mais proximidade os dados, antes e depois de interpretá-los. Conclusões: para aproveitar a oportunidade produzida por esses novos algoritmos, pesquisadores poderiam ampliar as concepções existentes e adotar um ponto de vista epistemológico mais participativo e agnóstico. O entrelaçamento de métodos computacionais e interpretativos tem potencial de integrar e viabilizar interpretação e mensuração com rigor analítico nos contextos de abundância de dados que devem caracterizar a pesquisa em ciências sociais nas próximas décadas. Objetivo quantitativas tradicionais Proposta disso qualitativa vigoroso termos interpretálos. interpretálos interpretá los. los interpretá-los Conclusões agnóstico décadas

ABSTRACT Objective: the paper introduces a framework for conducting interpretive research using deep learning algorithms that blur the boundaries between qualitative and quantitative approaches. The work evidences how research might benefit from an integrated approach that uses computational tools to overcome traditional limitations. Proposal: the increased availability and diversity of data raises the utility of algorithms as research tools for social scientists. Furthermore, tuning and using such computational artifacts may benefit from interpretive procedures. Such circumstances turn the traditional debate between quantitative and qualitative research on its head: the research strategy that likely yields the most assertiveness and rigor is the one that may require vigorous hermeneutic effort. Along these lines, neural word embeddings can be instrumental in allowing researchers to read the data closely before and after interpretation. Conclusions: to take advantage of the opportunity generated by these new algorithms, researchers may broaden their previous conceptions and adopt a participative point of view. In the coming decades, the interweaving of computational and interpretive methods has the potential to integrate rigorous social science research. Objective approaches limitations Proposal scientists Furthermore procedures head effort lines interpretation Conclusions view decades

Recently, a new class of materials, the Perovskite Nanocrystals, also known as perovskite quantum dots (PQDs) has emerged. Perovskite nanocrystals are promising in optoelectronics due to their quantum confinement properties, narrow photoluminescence bands with short decay times, optically tunable bandgaps, easy surface passivation and high photoluminescent quantum yields. The intense photoluminescence covering the entire visible region of the spectrum can be easily adjusted through the composition and/or the nanocrystals sizes. This paper presents the state-of-the-art of these materials, since their discovery to current research, including their synthesis methods, properties and applications.

A estimativa da produtividade das pastagens ao longo do seu ciclo vegetativo é um indicador de grande interesse para os agricultores para efeitos de planeamento e apoio à tomada de decisão. A gestão do pastoreio animal, em termos de encabeçamento em cada parcela, da rotação dos animais entre estas, do cálculo das necessidades de aprovisionamento de forragem e de suplementação alimentar, baseia-se na produtividade. O método tradicional para medir a produtividade da pastagem envolve o corte da matéria verde numa determinada área, o que constitui um processo demorado e caro para recolher um número de amostras representativo. Neste trabalho apresentam-se os resultados de ensaios realizados entre 2007 e 2018 para calibrar uma sonda (“Grassmaster II”) em três parcelas agrícolas de montado no Alentejo. Foi utilizada a sonda em 1279 pontos de amostragem, nos quais se recolheram amostras de pastagem para determinação da produtividade (biomassa total e produção de matéria seca). Foi realizada uma análise de regressão mensal entre as leituras corrigidas da sonda (“Corrected Meter Reading”, CMR) e os valores de matéria verde e de matéria seca da pastagem (respectivamente, MV e MS, em kg ha-1). Os resultados obtidos mostraram fortes correlações entre a produtividade da pastagem e as leituras da sonda, especialmente nos meses de Fevereiro (r=0,936; p<0,01; RMSE= 297 kg ha-1 para a MS; r=0,919; p<0,01; RMSE= 2404 kg ha-1 para a MV) e de Março (r=0,807; p<0,01; RMSE= 377 kg ha-1 para a MS; r=0,886; p<0,01; RMSE= 1971 kg ha-1 para a MV). Na Primavera, à medida que o ciclo de desenvolvimento da pastagem evoluiu, verificou-se uma quebra nos coeficientes de correlação entre os parâmetros, diminuindo a fiabilidade da estimativa na altura do pico de produção (Abril-Maio), embora com coeficientes de correlação significativos. Estes resultados mostram que a sonda “Grassmaster II” pode apoiar decisões na gestão de pastagens de sequeiro e respectivo pastoreio.

Estimating pasture productivity throughout its growth cycle is an indicator of great interest to farmers in planning and supporting decision making. The management of animal grazing, in regards to animal stocking in each parcel, rotation of the animals by the different parcels, calculation of the forage supply requirements and feed supplementation of the animals are based on pasture productivity. The standard method of assessing pasture biomass is based on cutting the forage within a specific area and requires great effort and expense to collect enough samples to accurately represent a pasture. This work presents the results of tests carried out to calibrate a probe (“Grassmaster II”) in three parcels at Alentejo between 2007 and 2018. The probe was used at 1279 sampling points, in which grass samples were collected for the purpose of determining productivity (total biomass and dry matter production). A monthly regression analysis was performed between corrected meter reading (CMR) measured by the probe and the values of pasture green matter and dry matter (respectively, GM and DM, in kg ha-1). The results showed strong correlations between pasture productivity and CMR, especially in February (r = 0,936; p <0,01; RMSE = 297 kg ha-1 for DM; r = 0,919; p <0,01; RMSE = 377 kg ha-1 for GM) and March (r= 0,807; p<0,01; RMSE= 377 kg ha-1 for DM; r= 0,886; p<0,01; RMSE= 1971 kg ha-1 for GM). During the vegetative cycle of the pasture, the correlation coefficients between the parameters had the tendency to diminish, decreasing the reliability of the estimate at the peak of production (April-May), although still with significant correlation coefficients. These results show that the “Grassmaster II” probe might support decisions making in dryland pastures and grazing management.

Resumo A partir de meados do século XVIII, no contexto das chamadas Reformas Pombalinas, as “Leis de Liberdade dos Índios” incorporaram uma vasta região no norte da América portuguesa à administração civil, transformando mais de 60 aldeamentos missionários em vilas e lugares portugueses de população majoritariamente indígena. Esta pesquisa propõe-se a investigar a evolução demográfica dos índios aldeados e da população livre não aldeada sob esse novo regime. Para uma parte da historiografia, a população das vilas e lugares indígenas, sob o Diretório (1757-1798), era mantida artificialmente pelo constante aporte de novos contingentes de povos nativos, por meio dos descimentos. No entanto, a análise dos mapas de população elaborados a partir de 1773 mostra um crescimento lento, mas consistente de todos os grupos da população das capitanias do Estado do Grão-Pará, mesmo sofrendo os danos das frequentes epidemias e do trabalho compulsório ao qual estava sujeita a maior parte da população indígena.

Abstract As of the mid-eighteenth century, in the context of the so-called Pombaline Reforms, Indian Freedom Laws incorporated a large region in northern Portuguese America into civilian administration, transforming more than sixty missionary settlements into Portuguese villages and sites with predominantly indigenous population. This paper seeks to research into the demographic evolution of “aldeado” (subject to compulsory labour) indians and the free non- “aldeado” population under this new regime. For some historians, the population of Indian villages under the Directorio (1757-1798) was artificially maintained by the constant influx of new contingents of native peoples through descimentos. However, the analysis of population tables produced since 1773 shows a slow but consistent growth of all population groups in the captaincies of the State of Grão-Pará, even suffering frequent epidemics and harm from compulsory work to which by the majority of the indigenous population was subject.

Resumen Desde mediados del siglo XVIII, en el contexto de las denominadas reformas pombalinas, las Leyes de Libertad de los Indios incorporaran una inmensa región en el norte de la América portuguesa a la administración civil y transformaron más de sesenta reducciones misioneras en municípios portugueses de población predominantemente indígena. Esta investigación se propone investigar la evolución demográfica de los indios «aldeados» y de la población libre no «aldeada» bajo el nuevo régimen. Para una parte de la historiografía, la población de las villas y lugares indígenas se mantuvo artificialmente, en el marco del Directorio (1757-1798), por el suministro constante de nuevos contingentes de pueblos nativos por medio de los descensos. Sin embargo, el análisis de los mapas de población compilados desde 1773 muestra un crecimiento lento pero consistente de todos los grupos de población de las capitanías del estado de Grão-Pará, a pesar de las epidemias frecuentes y de los daños provocados por el trabajo obligatorio que sujetaba a la mayoría de la población indígena.

ABSTRACT In wetlands, the knowledge accumulated on the role of aquatic plants in the methane cycle focused on emergent macrophytes, to the detriment of other typologies. Herein, we evaluated whether the free-floating macrophyte Salvinia auriculata Aubl. and the floating-leaved macrophyte Eichhornia azurea (Sw.) Kunth. decrease the water column methane concentrations compared to a plant-free surface. We prepared microcosms by inserting an individual of S. auriculata or of E. azurea into chambers filled with lagoon water previously bubbled with CH4. Another set of chambers was incubated only with the prepared water, representing the plant-free surface. Half of the chambers were kept in the dark and half in sunlight to simulate a diel cycle. We observed greater loss of CH4, higher O2 uptake and lower CO2 outflow in the plants treatments. The decrease in methane concentrations in the E. azurea treatments was 93.5% in the light and 77.2% in the dark. In the S. auriculata treatments, the decreases were 74.2% and 67.4% in the light and in the dark, respectively. In plant-free surface the decrease was 58.7% in the light and 36.3% in the dark. These results indicate a role of floating aquatic macrophytes in the methane cycle in the water column. Moreover, our results suggest a diel variation of methane oxidation and methane emission, according to the differences observed in O2 uptake and CO2 outflow between dark and light conditions. Thus, future predictions of global methane budget should include the role played by floating aquatic macrophytes.

Abstract This text studies the imperative nature of the prohibition of committing crimes against humanity in international law, and the obligation to prosecute, judge, and punish these crimes. In this way, it develops the complementarity of transitional justice with the abovementioned obligations. These results in the conclusion that transitional justice processes have to be built on the paradigm of "no peace without justice" and not the other way around.

Resumen El texto aborda el carácter imperativo de la prohibición en el derecho internacional de la comisión de crímenes de lesa humanidad, así como la obligación que dé juzgamiento, investigación y sanción de las conductas que constituyen estos crímenes. Asimismo, analiza cómo el concepto de justicia transicional tiene una naturaleza complementaria frente a las mencionadas obligaciones, lo que hace que todo proceso de justicia de transición tenga que construirse en torno al paradigma "no hay paz sin justicia", y no en sustitución del mismo.

Tendo presente as potencialidades dos jogos para aprender e o interesse crescente dos estudantes por jogar, desenvolvemos um jogo mobile para estudantes de licenciatura, começando por conhecer as suas preferências de jogo. Neste artigo apresentam-se as etapas conducentes à criação do jogo, nomeadamente: os jogos que os estudantes universitários mais gostam de jogar, tendo-se realizado um survey no ensino superior (n=626). De seguida, procedeu-se à análise desses jogos, tendo por base os 36 princípios de aprendizagem propostos por Paul Gee, bem como as mecânicas de jogo identificadas por Ernest Adams e Joris Dormans. Por fim, descreve-se o jogo Konnecting, explicitando-se as mecânicas de jogo, e os princípios de aprendizagem subjacentes.

Keeping in mind the advantages of using games for learning and the increasing interest of students to play games, we developed a mobile game for undergraduate students. The starting point was to first know their preferences. In this paper, the steps that lead to the creation of the game will be presented, namely: the games that university students most enjoy playing, collected through a survey carried out in higher education (n=626); further, these games were analyzed according to the learning principles suggested by Paul Gee, as well as the game mechanics identified by Ernest Adams e Joris Dormans. Finally, the game Konnecting is described, explaining the mechanics involved and the underlying learning principles.

A Assembleia de Deus enfrentou uma série de pressões sociais relacionadas às mudanças que se processaram no Brasil na década de 1980: novos governos, nova Constituição, novos planos econômicos, novos movimentos políticos e manifestações culturais eram alguns dos elementos que direta ou indiretamente impunham novos desafios a essa Igreja na penúltima década do século XX. A Igreja, que até então mantivera uma postura de negação aos assuntos políticos, por julgar que esse tipo de tema pertencia estritamente ao plano terreno, recorreu a uma série de medidas para defender seus interesses e pontos de vista em uma nova conjuntura. O jornal Mensageiro da Paz teve, então, papel central como instrumento de propagação das suas mensagens. O artigo busca analisar as mensagens veiculadas nesse mensário por meio das quais a liderança assembleiana expressava seu descontentamento com os rumos políticos da nação brasileira e articulava a inserção de representantes da Igreja nas principais esferas políticas do país.

The Assembleia de Deus (Assembly of God) experienced a lot of social pressure because of the changes that occurred in Brazil in the decade of 1980: new government, new constitution, new economic plans, new political movements and cultural events were some of the elements that directly or indirectly imposed new challenges to its church in the penultimate decade of the twentieth century. The Church, that until the 80's kept a posture of denial to the political issues, for thinking that kind of theme belonging strictly to the earthly plane, used a series of actions to defend its interests and points of view in a new situation. In that context, the journal Mensageiro da Paz had a central role as an instrument of propagation of the Church's messages. This paper seeks to examine the messages of this monthly publication through which the assemblies leadership expressed their dissatisfaction with the political direction of the Brazilian nation and articulated the inclusion of representatives of the Church in the main political spheres of the country.

Introduction Molecular biology procedures to detect, genotype and quantify hepatitis C virus (HCV) RNA in clinical samples have been extensively described. Routine commercial methods for each specific purpose (detection, quantification and genotyping) are also available, all of which are typically based on polymerase chain reaction (PCR) targeting the HCV 5′ untranslated region (5′UTR). This study was performed to develop and validate a complete serial laboratory assay that combines real-time nested reverse transcription-polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP) techniques for the complete molecular analysis of HCV (detection, genotyping and viral load) in clinical samples. Methods Published HCV sequences were compared to select specific primers, probe and restriction enzyme sites. An original real-time nested RT-PCR-RFLP assay was then developed and validated to detect, genotype and quantify HCV in plasma samples. Results The real-time nested RT-PCR data were linear and reproducible for HCV analysis in clinical samples. High correlations (> 0.97) were observed between samples with different viral loads and the corresponding read cycle (Ct - Cycle threshold), and this part of the assay had a wide dynamic range of analysis. Additionally, HCV genotypes 1, 2 and 3 were successfully distinguished using the RFLP method. Conclusions A complete serial molecular assay was developed and validated for HCV detection, quantification and genotyping.

OBJETIVOS: Este estudo teve como objetivo avaliar, através de um experimento com intervalos amostrais curtos, (1) os efeitos da qualidade dos detritos no consumo de oxigênio dissolvido (OD), sobre a formação de carbono inorgânico dissolvido (CID) e na relação estequiométrica entre o OD consumido e o carbono mineralizado (O/C ratio) durante a lixiviação de detritos de plantas no início da decomposição; (2) a variação temporal das variáveis mencionadas acima de acordo com as alterações na qualidade de cada detrito ao longo do tempo. MÉTODOS: Os detritos de folhas, galhos e serapilheira (3 g) previamente secos foram incubados em câmaras de mineralização contendo 1 L de inóculo. As câmaras foram mantidas a 24 ± 2 °C, em condição aeróbia. As concentrações de OD, carbono orgânico dissolvido (COD) e CID foram avaliadas. O consumo de OD foi ajustado a um modelo cinético de primeira ordem. As taxas horárias de consumo de OD e produção de CID e as razões O/C foram determinadas. RESULTADOS: O consumo de OD (2,62 mg.g-1) e a produção de CID (1,20 mg.g-1) foram maiores na decomposição das folhas, e menores na serapilheira (1,50 mg.g-1 e 0,42 mg.g-1, respectivamente). Inversamente, a razão O/C foi maior na decomposição da serapilheira (3,56). As taxas de consumo de OD (0,50 mg.g-1.h-1) e de produção de CID (0,41 mg.g-1.h-1) foram maiores no início da decomposição do detrito de folhas. CONCLUSÕES: Intervalos amostrais curtos são fundamentais para a compreensão da dinâmica de decomposição na fase de lixiviação. O maior consumo de OD na decomposição das folhas indicou maior teor de compostos lábeis nesse detrito. As maiores razões O/C na decomposição da serapilheira podem ser atribuídas ao seu maior teor de substâncias refratárias. As variações nas taxas de consumo de OD e nas razões O/C sugerem que o lixiviado é heterogêneo, compreendendo substâncias lábeis e refratárias, assim como o detrito como um todo. Em termos de alterações da qualidade da água, as folhas constituíram o recurso mais crítico e a serapilheira apresentou os menores efeitos.

AIMS: This study aimed to evaluate, through an experiment with short sampling intervals, (1) the effects of detritus quality on dissolved oxygen (DO) consumption, on dissolved inorganic carbon (DIC) formation and on the stoichiometric ratio between the DO consumed and mineralized carbon (O/C ratio) during leaching of plant detritus in the early decomposition; (2) the temporal variation of the variables mentioned above according to changes in the quality of each detritus over time. METHODS: The detritus of leaves, branches and litter (3 g) previously dried were incubated in decomposition chambers containing 1 L of inoculum. The chambers were maintained at 24 ± 2 °C in aerobic condition. The DO concentrations dissolved organic carbon (DOC) and DIC were evaluated. The consumption of DO was adjusted to a first-order kinetic model. The hourly rates of DO consumption, DIC production and O/C ratios were determined. RESULTS: The DO consumption (2.62 mg.g-1) and DIC production (1.20 mg.g-1) were higher in leaves decomposition, and smaller in litter (1.50 and 0.42 mg.g-1, respectively). Inversely, the O/C ratio was higher in the decomposition of litter (3.56). The rates of DO consumption (0.50 mg.g-1.h-1) and DIC production (0.41 mg.g-1.h-1) were greatest in the early decomposition of leaves. CONCLUSIONS: Short sampling intervals are key to understanding the dynamics of decomposition in the leaching phase. Higher consumption of DO in leaves decomposition indicated a higher content of labile compounds in this detritus. The highest O/C ratios in the litter decomposition can be attributed to its higher content of refractory substances. Variations in DO consumption rates and in O/C ratios over time suggest that the leachate is heterogeneous, comprising labile and refractory fractions, analogous to the detritus as a whole. In terms of water quality alterations, leaves constituted the most critical resource and the litter presented less pronounced effects.