Development of molecular approaches to assess the diversity and abundance of marine nematodes assemblages for assessment of Good Environmental Status of the estuarine and marine ecosystems

In Europe, the concerns with the status of marine ecosystems have increased, and the Marine Directive has as main goal the achievement of Good Environmental Status (GES) of EU marine waters by 2020. Molecular tools are seen as promising and emerging approaches to improve ecosystem monitoring, and have led ecology into a new era, representing perhaps the most source of innovation in marine monitoring techniques. Benthic nematodes are considered ideal organisms to be used as biological indicator of natural and anthropogenic disturbances in aquatic ecosystems underpinning monitoring programmes on the ecological quality of marine ecosystems, very useful to assess the GES of the marine environment. dT-RFLP (directed Terminal-Restriction Fragment Length Polymorphism) allows to assess the diversity of nematode communities, but also allows studying the functioning of the ecosystem, and combined with relative real-time PCR (qPCR), provides a high-throughput semi-quantitative characterization of nematode communities. These characteristics make the two molecular tools good descriptors for the good environmental status assessment. The main aim of this study is to develop and optimize the dT-RFLP and qPCR in Mira estuary (SW coast, Portugal). A molecular phylogenetic analysis of marine and estuarine nematodes is being performed combining morphological and molecular analysis to evaluate the diversity of free-living marine nematodes in Mira estuary. After morphological identification, barcoding of 18S rDNA and COI genes are being determined for each nematode species morphologically identified. So far we generated 40 new sequences belonging to 32 different genus and 17 families, and the study has shown a good degree of concordance between traditional morphology-based identification and DNA sequences. These results will improve the assessment of marine nematode diversity and contribute to a more robust nematode taxonomy. The DNA sequences are being used to develop the dT-RFLP with the ability to easily process large sample numbers (hundreds and thousands), rather than typical of classical taxonomic or low throughput molecular analyses. A preliminary study showed that the digest enzymes used in dT-RFLP for terrestrial assemblages separated poorly the marine nematodes at taxonomic level for functional group analysis. A new digest combination was designed using the software tool DRAT (Directed Terminal Restriction Analysis Tool) to distinguished marine nematode taxa. Several solutions were provided by DRAT and tested empirically to select the solution that cuts most efficiently. A combination of three enzymes and a single digest showed to be the best solution to separate the different clusters. Parallel to this, another tool is being developed to estimate the population size (qPCR). An improvement in qPCR estimation of gene copy number using an artificial reference is being performed for marine nematodes communities to quantify the abundance. Once developed, it is proposed to validate both methodologies by determining the spatial and temporal variability of benthic nematodes assemblages across different environments. The application of these high-throughput molecular approaches for benthic nematodes will improve sample throughput and their implementation more efficient and faster as indicator of ecological status of marine ecosystems.

The Theory of Social Representations in Brazilian Health Researches: a Bibliometric Profile

The study aimed to characterizing the production of national articles on health, the time frame of the past 10 years, available in the database LILACS and MEDLINE Virtual Health Library that used the Theory of Social Representations in its searches, using as descriptors the words: social representations and health. It is a descriptive study, developed in the context of ibliometrics. Of the 158 units found, 122 were considered and analyzed after removal of those that did not include the stablished inclusion criteria: articles in Portuguese,available in full and that mentioned the expression "social representations", either in the title or abstract. The journal that most published researches about the Theory of Social Representations was Science & Public Health; being the largest number of articles published in 2011. The most frequent area of knowledge covering about the Theory of Social Representations was the Public Health, with the participant group most cited health professionals. Among the data collection instruments used, the semi-structured interview was the most frequent and the kind of qualitative analysis the content analysis was the most common. Noteworthy is the growing interest for the theory and the need for greater criteria in the preparation of abstracts, considering its importance in the spread of scientific production.

Caracteres morfo-agronômicos e bioquímicos de clones elite de mandioca de mesa com raízes de polpas amarelada e rosada.

A mandioca (Manihot esculenta Crantz) é considerada uma espécie relevante como fonte alimentícia para a população mundial, principalmente para os países subdesenvolvidos e emergentes. A mandioca é fornecedora de energia a partir do amido acumulado em suas raízes de reserva, mas é também importante destacar a presença dos carotenóides com atividade antioxidante. Nesse contexto, o presente trabalho teve como objetivo caracterizar, por meio de descritores morfológicos, agronômicos e bioquímicos, clones elite de mandioca de mesa de polpa aparelhada e rosada do programa de melhoramento genético de mandioca da Embrapa Cerrados. Foram caracterizados durante duas safras, 13 clones de mandioca de mesa com polpa amarelada e 8 clones com polpa rosada, em comparação com a variedade testemunha IAC 576-70 (BGMC 753). Para avaliar as características morfológicas foram obtidos 40 descritores qualitativos para cada clone. Tanto nos clones de polpas amarelada quanto naqueles de raízes de polpas rosada, houve diferenças morfológicas, demostrando que nenhum clone apresentou 100% de similaridade. O fator ano/safra não influenciou a expressão fenotípica dos caracteres aferidos. Com base no coeficiente cofenético, verificou-se elevado ajuste entre a representação gráfica via dendrograma de r = 0,80 nas raízes de polpa amarelada e r = 0,92 na rosada e a matriz de dissimilaridade genética. Entre os caracteres aferidos, os que apresentaram maior entropia nas raízes amarelada foram, a coloração da epiderme externa, forma do lóbulo central da folha e cor do córtex da raiz, ao passo que na rosada foi à cor do disco, forma do lóbulo central e cor do pecíolo. Foi realizada também a caracterização com base na altura da planta, altura da primeira ramificação, peso da parte aérea sem a cepa, produtividade em raízes, índices de amido nas raízes determinados por meio do método da balança hidrostática, tempo para a cocção e teor de ácido cianídrico nas raízes. Com base nos caracteres avaliados, os clones que se destacaram com polpa amarelada e rosada respectivamente, no caractere altura da primeira ramificação (273/08 e 259/08) e (390/08, 345/08 e a testemunha IAC 576-70), altura da planta (90/08, 272/08, 273/08, 497/08, 259/08 e 450/08) e (390/08, 345/08 e 378/08), peso da parte aérea sem a cepa (94/08 e 272/08) e (390/08, 406/08, 390/08, 378/08 e 341/08), porcentagem de amido nas raízes (26/08, 272/08, 259/08 e 450/08) e (378/08, 413/08, 390/08 e a testemunha IAC 576-70), produtividade de raízes (215/08) e (testemunha IAC 576-70, 341/08, 406/08, 390/08 e 387/08). Com relação ao tempo de cocção na safra 2011/2012, todos os clones necessitaram de tempo inferior a 30 minutos. Em relação ao teor de carotenóides totais nas raízes os clones de amarelada que se destacaram foram 91/08, 94/08, 215/08, 246/08, 272/08 e 497/08, e, naqueles de raízes rosada, os clones 406/08 e 341/08. Em relação ao teor de proteínas nas raízes amarelada, os clones 26/08, 90/08 e 91/08, foram os melhores enquanto nas raízes rosada se destacaram os clones 406/08 e a testemunha IAC 576-70. Os teores de HCN nas raízes de reserva de mandioca foram inferiores a 100 mg kg-1 em todos os clones avaliados. Diferenças significativas entre clones de mandioca de polpas amarelada e rosada foram verificadas para todas as características agronômicas, morfológicas e bioquímicas avaliadas. Os clones tiveram bom desempenho nas avaliações para o cultivo comercial na região do Cerrado e, alguns destes, têm potencial para utilização no melhoramento visando o incremento de carotenóides. ABSTRACT: Cassava (Manihot esculenta Crantz) is considered a relevant species as a food source for the world's population, particularly for developing and emerging countries. The cassava is a provider of energy from starch accumulated in their reserve roots, but it is also important to highlight the presence of carotenoids with antioxidant activity. In this context, this study aimed to characterize, using morphological, agronomic and biochemical, descriptors elite clones from sweet cassava of yellowish and pinkish pulps from the cassava breeding program at Embrapa Cerrados. They were characterized for two crops, 13 edible cassava clones with yellowish pulp and 8 clones with pinkish pulp, compared with the control variety IAC 576-70 (BGMC 753). To evaluate the morphological characteristics were obtained 40 qualitative descriptors for each clone. Both clones the yellowish pulp as those the roots the pinkish pulp, there was morphological differences among clones, showing that no clone showed 100% similarity. The year / crop factor did not influence the phenotypic expression of measured characters. Based on cofenetic coefficient, was found high fit between the graphical representation via dendrogram of r = 0.80 in the roots of yellowish pulp and r = 0.92 in the pinkish of genetic dissimilarity matrix. Among the measured characters, those with the highest entropy in the yellowish roots were, the color of the outer epidermis, the central lobe shape of the leaf and root cortex color, whereas the pinkish was the color to disc, central lobe shape and petiole color. We also performed the characterization based on plant height, the first branch point, and shoot weight without strain, productivity in roots, and index of starch in the roots determines by the method of hydrostatic balance, time for cooking and acid cyanide content in the roots. Based on the evaluated characters, clones stood out with pulps yellowish and pinkish respectively, characters height of the first branch (273/08 and 259/08) and (390/08, 345/08 and the witness IAC 576-70), plant height (90 / 08, 272/08, 273/08, 497/08, 259/08 and 450/08) and (390/08, 345/08 and 378/08), shoot weight without strain (94/08 and 272/08) and (390/08, 406/08, 390/08, 378/08 and 341/08), percentage of starch in the roots (26/08, 272/08, 259/08 and 450/08) and (378/08, 413/08, 390/08 and the witness IAC 576-70), roots of productivity (215/08) and (witnesses IAC 576-70, 341/08, 406/08, 390/08 and 387/08). Regarding the cooking time in the 2011/2012 harvest, all clones showed time less than 30 minutes. Regarding the total carotenoid content in the pulps clones of yellowish roots that stood out were 91/08, 94/08, 215/08, 246/08, 272/08 and 497/08, and, those the clones with pulp pinkish 406/08 and 341/08. Regarding the protein content in yellowish roots the clones 26/08, 90/08 and 91/08, was the best while the pinkish roots highlight clones 406/08 and witness IAC 576-70. The levels of HCN in reserve roots of cassava were less than 100 mg kg-1em all evaluated clones. Significant differences between yellowish and pinkish of pulps cassava clones were checked for all agronomic, morphological and biochemical characteristics evaluated. The clones had well in the ratings for commercial cultivation in the Cerrado region and some of these, clones has potential for use in breeding aimed at increase of carotenoids.

Composição botânica e qualidade do pasto selecionado por ovelhas em caatinga raleada e enriquecida.

Resumo: O presente estudo foi conduzido com o objetivo de determinar a composição botânica e a qualidade da dieta selecionada por ovelhas, através da técnica de micro-histologia fecal, em caatinga raleada e enriquecida com capim massai (Panicum maximum cv. Massai), recebendo diferentes quantidades de concentrado (0; 200; 350 e 500 g de concentrado por dia), e em diferentes períodos do ano (águas, transição água-seca e seca). Foram estimados também o consumo e digestibilidade dos nutrientes, bem como a degradabilidade de espécies forrageiras ingeridas pelas ovelhas. Os experimentos foram realizados na Fazenda Crioula do Meio, pertencente a Embrapa Caprinos e Ovinos em Sobral, CE no período de março a novembro de 2013. No Experimento 1, para a determinação da composição botânica e qualidade da dieta selecionada, foram utilizadas dezesseis ovelhas Somalis brasileira, gestantes, multíparas e peso médio de 30,58+2,48 kg. O acompanhamento da ingestão do pasto pelas ovelhas foi feito em três períodos (águas, transição água-seca e seca), referentes aos meses de abril, junho e agosto de 2013, respectivamente. Amostras das plantas foram coletadas para o preparo das lâminas de referência, e posterior identificação e caracterização dos descritores epidérmicos. O mesmo foi feito para as fezes coletadas nas ovelhas. Com base na proporção de cada espécie identificada nas lâminas fecais que compuseram a dieta, e na composição química das forrageiras identificadas, foi possível determinar a qualidade da dieta ingerida. De 76 espécies observadas no pasto, 33 foram identificadas na dieta das ovelhas, destacando as espécies sabiá (Mimosa caesalpiniaefolia), centrosema (Centrosema sp.), ervanço (Alternanthera brasiliana), massai (Panicum maximum cv Massai) e paco-paco (Wissadula rostrata) como as mais selecionadas pelos animais ao longo dos períodos, chegando a compor mais de 50% da dieta selecionada. Com a chegada do período seco, espécies indesejáveis como o marmeleiro (Croton sonderianus) e o mofumbo (Combretum lepreosum), também fizeram parte das plantas selecionadas. Quanto ao valor nutritivo da dieta selecionada, os animais selecionaram uma dieta com valor nutritivo superior ao amostrado no pasto. No Experimento 2, na mesma condição do experimento anterior, trinta e duas ovelhas Somalis brasileira foram utilizadas para determinação do consumo e digestibilidade dos nutrientes, realizado em três ensaios (abril - terço final de gestação; junho - lactação e agosto - desmame). Para predição do consumo, o indicador LIPE® foi utilizado. Pesagens quinzenais foram realizadas para avaliação do desempenho das ovelhas e dos cordeiros nascidos. O concentrado oferecido favoreceu a maior ingestão e digestibilidade da MS e PB, com efeito substitutivo em relação ao consumo de pasto (P<0,05). Para o período seco, menores consumos foram observados em relação aos períodos das águas e de transição água-seca (P<0,05). Maiores consumos e digestibilidades dos constituintes fibrosos foram observados para as ovelhas não suplementados (P<0,05). Na avaliação do desempenho, a suplementação oferecida determinou os maiores pesos verificados durante a lactação, ao desmame e para os pesos ao nascer e ao desmame dos cordeiros (P<0,05). No Experimento 3, dois ovinos Morada Nova foram utilizados para determinação da degradabilidade da matéria seca (MS), proteína bruta (PB) e fibra em detergente neutro (FDN) de cinco das forrageiras selecionadas pelas ovelhas no Experimento 1: M. caesalpiniaefolia, A. brasiliana, P. maximum cv. Massai, jurema-preta (Mimosa tenuiflora), C. leprosum, nos tempos 0, 6, 24, 48, 72 e 96 horas de incubação. Para cada forrageira, foram determinadas equações para o desaparecimento da MS, PB e FDN. Também foi feito o fracionamento da proteína em suas porções degradáveis e não degradáveis no rúmen. Foi observado maior desaparecimento da MS, PB e FDN, além dos melhores níveis de proteína efetivamente degradada no rúmen para A. brasiliana, seguido pelo P. maximum cv. Massai e M. caesalpiniaefolia. Com as informações obtidas, conclui-se que a micro-histologia fecal apresenta-se como uma técnica viável para avaliações da composição botânica da dieta selecionada por ovinos na caatinga. Ovelhas na caatinga possuem uma grande habilidade de selecionar a dieta, modificando-a ao longo das fases fenológicas, sempre na tentativa de estabelecer uma dieta com melhor valor nutritivo. Forrageiras como A. brasiliana, M. caesalpiniaefolia e o P. maximum cv. Massai, podem ser consideradas um interessante recurso alimentar, em virtude de seu valor nutricional e aproveitamento por ovelhas criadas na caatinga. Abstract: This study was conducted in order to determine the botanical composition and diet quality selected by sheep through fecal micro-histological technique, in thinned and enriched caatinga with Massai grass (Panicum maximum cv Massai.), receiving different amounts of concentrate (0; 200; 350 and 500 g of concentrate per day) at different periods (wet, transition wet-dry and dry). Were also estimated the intake, digestibility, as well as the degradability of forage species eaten by sheep. The experiments were performed in the "Fazenda Crioula do Meio", owned by Embrapa Goats and Sheep, in Sobral, Ceará State, Brazil, from march to november 2013. In Trial 1, sixteen female, pregnant, multiparous, with average body weight of 30,58+2,48 kg Somalis brasileira breed sheep were used to determine the botanical composition and the quality of the selected diet. The monitoring of pasture intake of sheep were conducted in three phenological periods of the caatinga's pasture (wet season, transition wet-dry and dry season). Plant samples were collected for the preparation of the reference slides, with subsequent identification and characterization of epidermal descriptors. The same was done for the feces collected in sheep. Considering the proportion of each species identified in fecal slides which composed the diet, and the chemical composition of forage identified, it was possible to determine the quality of the selected diet. From 76 species observed in the pasture, 33 species was identified in the sheep selected diet, emphasizing the Sabiá (Mimosa caesalpiniaefolia), centrosema (Centrosema sp.), ervanço (Alternanthera brasiliana), massai (Panicum maximum cv Massai) e paco-paco (Wissadula rostrata) as the most selected species by sheep during the study, composing more than 50% of the selected diet. In the Dry Season, undesirable species like marmeleiro (Croton sonderianus) and mofumbo (Combretum leprosum), were also constituent of the diet. Regarding to the nutritional value of selected diet, the sheep selected a diet with more protein than the sampled in the pasture. In the final late gestation, the sheep without supplementation ate a diet above 16% of CP, higher than the selected diet by treatments 350 and 500 g of concentrate per day (P<0.05). Were also observed to the non supplemented sheep, less fiber content intake (P<0.05). In the Trial 2, in the same condition of the first Trial, thirty two Somalis brasileira female sheep were used to determine the intake and digestibility, conducted in three assays (April - third late pregnancy; June - lactation and August - weaning). To predict the intake, the marker LIPE was used. Sheep and lambs were weighted every two weeks to performance evaluation. The concentrate offered to sheep favored to higher intake and digestibility of DM and CP than non supplemented sheep, with inverse relationship to the pasture intake (P<0.05). For the Dry Season, lower intake were observed than for Wet Season and Transition Wet-Dry (P<0.05). Higher intakes and digestibility of the fiber constituents were verified to non supplemented sheep (P<0.05). To performance evaluation, the offered supplementation determined the higher weights observed during lactation and weaning of sheep, and to birth and weaning weights of lambs (P<0.05). In Trial 3, two male sheep were used to determine the degradability of dry matter (DM), crude protein (CP) and neutral detergent fiber (NDF) of five forages selected by sheep in Trial 1: M. caesalpiniaefolia, A. brasiliana, P. maximum cv. Massai, jurema-preta (Mimosa tenuiflora) and C. lepreosum, at zero, 6, 24, 48, 72 and 96 hours of incubation. For each forage, were determined equations for the disappearance of DM, CP and NDF. It was also realized the protein fractions in their degradable and non-degradable in the rumen parts. Was detected a higher disappearance of DM, CP and NDF, as also better proportion of the rumen degradable protein to A. brasiliana, followed by P. maximum cv. Massai and M. caesalpiniaefolia. With the information obtained, it is concluded that the fecal micro histological technique presents as a viable technique to evaluate the selected diet by sheep in caatinga's pasture. On this pasture, the sheep are skilled to select the diet, changing during the phenological phases, trying to form a diet of better nutritive value. Forages as A. brasiliana, P. maximum cv. Massai and M. caesalpiniaefolia, can be considered an interesting food source to ewes kept in the caatinga.