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.

Mechanical properties of adobe blocks used in building construction in Huambo Province - Angola

Despite significant advances in building technologies with the use of conventional construction materials (as concrete and steel), which significantly have driven the construction industry, earth construction have demonstrated its importance and relevance, as well as it has matched in an efficient and eco-friendly manner the social housing concerns. The diversity of earth construction techniques allowed this material to adapt to different climatic, cultural and social contexts until the present time. However, in Angola, the construction with earth is still associated with population fringes of weak economic resources, for which, given the impossibility of being able to acquire modern construction materials (steel, cement, brick, among others), they resort to the use of available natural materials. Furthermore, the lack of scientific and technical knowledge justifies the negative appreciation of traditional building techniques, and the derogatory way how are considered the earth constructions in Angolan territory. Given the country's current development status, and taking into account the environmental requirements and the real socio-economic sustainability of Angola, it is considered that one of the viable and adequate options, could be the recovering and upgrading of the ancestral techniques of earth construction. The purpose of this research is to develop the technical and scientific knowledge in order to improve and optimize these construction solutions, responding to the real problems of housing quality as well as to the current social, economic and environmental sustainability requirements. In this paper, a description of the physical and mechanical characteristics of the adobes typically used in the construction of traditional houses in some localities of Huambo, province in Angola, is carried out. The methodology was based on mechanical in-situ testing in adobe blocks manufactured with traditional procedures: i) tensile strength evaluated with the bending test and compressive strength test on earth blocks specimens; and, ii) durability and erodibility test by Geelong method adopting the New Zealand standard (NZS) procedures (4297: 1998; 4297: 1998 and 4297: 1999). The results allow the characterization of the materials used in the construction of raw earth in the Huambo region, contributing to the development of knowledge of these sustainable and traditional housing constructive solutions with a strong presence in Angola [1, 2]. This study is part of a larger project in the area of Earth Construction [3], which aims to produce knowledge which can stimulate the use of environmental friendly construction materials and contribute to develop constructive solutions with improved performance, durability, comfort, safety and sustainability.