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 health and well-being of nursing students in clinical practice . what prevents it? what promotes it?

Introduction: The training of nursing students in the context of clinical practice, is characterized by educational experiences, subject to various emotional stress (stress, ambivalence, frustration, conflict), sometimes making it very vulnerable student.However not all students use the same strategies minimizing their meanings and negative effects on the level of your health and well-being Objetiv:To analyze the perception that nursing students have about the determinants of their health status and well-being in clinical practice Methods: Exploratory research Results:The results reveal the complexity of the teaching / learning process in clinical practice, identified determinants that limit and / or promote health and well-being of students, or not contributing to their motivation, self-confidence and learning. All students value the presence of the following dimensions: affective-emotional (humanization in learning experiences); relational dynamics (interactions developed with all stakeholders); methods used (professional competence of the clinical supervisor and teacher); school curriculum (adaptation of learning in theory); socialization to the profession (become nurse).Conclusions: The results indicate, that although all students evidencing the dimensions described as fundamental to learning in clinical practice, the study results are dichotomous and ambivalent. Students 2nd and 3ºanos refer a low perception in clinical practice, the indicated dimensions, and for these source of concern and uncertainty in learning, such as limiting their health condition and well-being. For students of the 4th year, these dimensions are percecionadas as gifts, and sources of motivation, learning and catalysts such as promoting their health and well-being.

A enfermagem comunitária na transição para a parentalidade - visita domiciliária de enfermagem no puerpério

Atualmente, com a alta precoce após o nascimento emerge a necessidade de cuidados domiciliários no puerpério. A pertinência deste projeto consubstancia-se na reorganização da visitação domiciliária no puerpério na USF Salus, garantindo a continuidade de cuidados após a alta hospitalar. O desenvolvimento do projeto teve por base as necessidades identificadas pelos enfermeiros da USF Salus. Teve como finalidade promover a parentalidade positiva, contribuindo para a melhoria da saúde e bem-estar das puérperas, recém-nascidos e respetivas famílias, definindo-se como objetivo geral: Reorganizar a Consulta de Enfermagem do Puerpério no Domicílio, de forma a promover a parentalidade positiva nos pais com crianças até 3 meses de idade, utentes da USF Salus, até 2016. Neste relatório é feita uma análise reflexiva sobre a visita domiciliária no puerpério e apresentada a proposta de protocolo de atuação na mesma. A avaliação do projeto na perspetiva das puérperas e dos enfermeiros legitima a relevância do mesmo na criação de soluções práticas para um problema concreto; ABSTRACT: Today, with early discharge after birth emerges the need for home care in the postpartum period. The relevance of this project is consolidated in the reorganization of home visitation puerperal at USF Salus, ensuring continuity of care after discharge. The project development was based on the needs identified by nurses USF Salus. It aimed to promote positive parenting, helping to improve the health and well-being of mothers, newborns and their respective families, defining the general objective: Rearrange the Puerpério of Nursing Consultation at Home, in order to promote positive parenting in parents with children up to 3 months of age, users of USF Salus, 2016. This report is made a reflective analysis of the home visit puerperal and presented the proposed action protocol in it. The project evaluation from the perspective of mothers and nurses legitimizes the importance of it in creating practical solutions to a specific problem.