Gap-analysis and annotated reference library for supporting macroinvertebrate metabarcoding in Atlantic Iberia

DNA metabarcoding provides a rapid and effective identification tool of macroinvertebrate species. The accuracy of species-level assignment, and consequent taxonomic coverage, relies on comprehensive DNA barcode reference libraries, which, due to incompleteness, are currently a recognized limitation for metabarcoding applications. In this study, we assembled a comprehensive reference library of DNA barcodes for Atlantic Iberia marine macroinvertebrate species, assessed gaps in species coverage and examined data ambiguities. Initially, an Iberian species checklist for the three dominant groups of marine macroinvertebrates was compiled, comprising 2827 species (926 Annelida, 638 Crustacea and 1263 Mollusca). A total of 18162 DNA sequences of the cytochrome c oxidase subunit I barcode region (COI-5P) matching the species checklist were compiled in a BOLD dataset, where taxonomic discordances were evaluated and cases of deep intraspecific divergence flagged. Gap-analysis showed that 63% of the Iberian macroinvertebrate species still lack a DNA barcode. Coverage gaps varied considerably across taxonomic groups with Mollusca displaying the highest sequence representation in the dataset (427 species, 49% of the total number of sequences), and Crustacea the highest species coverage with 338 species barcoded (53% of the checklist). In contrast, Polychaeta displayed the lower levels of completion (288 species, 16% of the total number of sequences). In total, 1545 Barcode Index Numbers (BINs) were assigned to 1053 barcoded species, of which 66% were taxonomically concordant, 26% displayed multiple BINs and 8% were discordant. Overall, results show that there is still a large portion of marine invertebrate taxa in this region of Europe pending barcode coverage, even considering only the dominant groups. However, the most notable finding was the relevant proportion of species flagged for significant intraspecific divergence and possible hidden diversity. The annotated reference library and gap-analysis here provided can therefore contribute to prioritize marine macroinvertebrate taxa for future research efforts and barcode coverage.Fundação para a Ciência e a Tecnologia | Ref. UIDB/04050/2020Fundação para a Ciência e a Tecnologia | Ref. PD/BD/127994/2016Fundação para a Ciência e a Tecnologia | Ref. SFRH/BD/131527/201

Gap-analysis and annotated reference library for supporting macroinvertebrate metabarcoding in Atlantic Iberia

DNA metabarcoding provides a rapid and effective identification tool of macroinvertebrate species. The accuracy of species-level assignment, and consequent taxonomic coverage, relies on comprehensive DNA barcode reference libraries, which, due to incompleteness, are currently a recognized limitation for metabarcoding applications. In this study, we assembled a comprehensive reference library of DNA barcodes for Atlantic Iberia marine macroinvertebrate species, assessed gaps in species coverage and examined data ambiguities. Initially, an Iberian species checklist for the three dominant groups of marine macroinvertebrates was compiled, comprising 2827 species (926 Annelida, 638 Crustacea and 1263 Mollusca). A total of 18162 DNA sequences of the cytochrome c oxidase subunit I barcode region (COI-5P) matching the species checklist were compiled in a BOLD dataset, where taxonomic discordances were evaluated and cases of deep intraspecific divergence flagged. Gap-analysis showed that 63% of the Iberian macroinvertebrate species still lack a DNA barcode. Coverage gaps varied considerably across taxonomic groups with Mollusca displaying the highest sequence representation in the dataset (427 species, 49% of the total number of sequences), and Crustacea the highest species coverage with 338 species barcoded (53% of the checklist). In contrast, Polychaeta displayed the lower levels of completion (288 species, 16% of the total number of sequences). In total, 1545 Barcode Index Numbers (BINs) were assigned to 1053 barcoded species, of which 66% were taxonomically concordant, 26% displayed multiple BINs and 8% were discordant. Overall, results show that there is still a large portion of marine invertebrate taxa in this region of Europe pending barcode coverage, even considering only the dominant groups. However, the most notable finding was the relevant proportion of species flagged for significant intraspecific divergence and possible hidden diversity. The annotated reference library and gap-analysis here provided can thereThis study was supported by the project The NextSea [NORTE-01-0145-FEDER-000032], under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). This work was supported by the "Contrato-Programa'' UIDB/04050/2020 funded by national funds through the FCT I.P. SD and PEV work was supported by national funds through the Portuguese Foundation for Science and Technology (FCT, I.P.) in the scope of the project NIS-DNA [PTDC/BIA-BMA/29754/2017]. BRL benefitted from an FCT fellowship PD/BD/127994/2016. MALT benefitted from an FCT fellowship co-financed by European Social Fund (ESF) SFRH/BD/131527/2017

Antioxidant and anti-inflammatory extracts from sea cucumbers and tunicates induce a pro-osteogenic effect in Zebrafish Larvae

Bone metabolic disorders such as osteoporosis are characterized by the loss of mineral from the bone tissue leading to its structural weakening and increased susceptibility to fractures. A growing body of evidence suggests that inflammation and oxidative stress play an important role in the pathophysiological processes involved in the rise of these conditions. As the currently available therapeutic strategies are often characterized by toxic effects associated with their long-term use, natural antioxidants and anti-inflammatory compounds such as polyphenols promise to be a valuable alternative for the prevention and treatment of these disorders. In this scope, the marine environment is becoming an important source of bioactive compounds with potential pharmacological applications. Here, we explored the bioactive potential of three species of holothurians (Echinodermata) and four species of tunicates (Chordata) as sources of antioxidant and anti-inflammatory compounds with a particular focus on polyphenolic substances. Hydroethanolic and aqueous extracts were obtained from animals' biomass and screened for their content of polyphenols and their antioxidant and anti-inflammatory properties. Hydroethanolic fractions of three species of tunicates displayed high polyphenolic content associated with strong antioxidant potential and anti-inflammatory activity. Extracts were thereafter tested for their capacity to promote bone formation and mineralization by applying an assay that uses the developing operculum of zebrafish (Danio rerio) to assess the osteogenic activity of compounds. The same three hydroethanolic fractions from tunicates were characterized by a strong in vivo osteogenic activity, which positively correlated with their anti-inflammatory potential as measured by COX-2 inhibition. This study highlights the therapeutic potential of polyphenol-rich hydroethanolic extracts obtained from three species of tunicates as a substrate for the development of novel drugs for the treatment of bone disorders correlated to oxidative stress and inflammatory processes.info:eu-repo/semantics/publishedVersio

Artificial reefs built by 3D printing: Systematisation in the design, material selection and fabrication

The recovery of degraded marine coasts and the improvement of natural habitats are current issues of vital importance for the development of life, both marine and terrestrial. In this sense, the immersion of artificial reefs (ARs) in the marine environment is a way to stimulate the recovery of these damaged ecosystems. But it is necessary to have a multidisciplinary approach that analyses the materials, designs and construction process of artificial reefs in order to understand their true impact on the environment. For this reason, this paper presents the manufacture of artificial reefs by 3D printing, proposing designs with a combination of prismatic and random shapes, with different external overhangs as well as inner holes. For the definition of the artificial reef designs, criteria provided by marine biologists and the results obtained from a numerical simulation with ANSYS were taken into account, with which the stability of the artificial reefs on the seabed was analysed. Three dosages of cement mortars and three dosages of geopolymer mortars were studied as impression materials. The studies included determination of the rheological properties of the mortars, to define the printability, determination of the cost of the materials used, and determination of the mechanical strength and biological receptivity in prismatic specimens that were immersed in the sea for 3 months. To evaluate the environmental impact of the materials used in the production of the mortars, a Life Cycle Assessment (LCA) was carried out. In order to choose the mortars that encompassed the best properties studied, Multi-Criteria Decision Making (MCDM) was applied and the two best mortars were used for the manufacture of the artificial reefs. Finally, the advantages and disadvantages of the 3D printing process used were analysed. The results of the studies carried out in this research show that cement mortars have better characteristics for artificial reef applications using 3D printing, and that the technique applied for the manufacture of the artificial reefs allowed the digital models to be faithfully reproduced

On the Properties Evolution of Eco-Material Dedicated to Manufacturing Artificial Reef via 3D Printing: Long-Term Interactions of Cementitious Materials in the Marine Environment

This paper deals with the evolution monitoring of biomass colonization and mechanical properties of 3D printed eco-materials/mortars immersed in the sea. Measurements of tensile strength, compressive strength, and Young’s modulus were determined on samples deployed along the Atlantic coast of Europe, in France, United Kingdom, Spain, and Portugal. The samples were manufactured using 3D printing, where six mix designs with a low environmental impact binder were used. These mortars were based on geopolymer and cementitious binders (Cement CEM III), in which sand is replaced by three types of recycled sand, including glass, seashell, and limestone by 30%, 50%, and 100% respectively. The colonization of concrete samples by micro/macro-organisms and their durability were also evaluated after 1, 3, 6, 12, and 24 months of immersion. The results showed that both biomass colonization and mechanical properties were better with CEM III compared to geopolymer-based compositions. Therefore, the mixed design optimized according to mechanical properties show that the use of CEM III should be preferred over these geopolymer binders in 3D printed concrete for artificial reef applications

Artificial reefs in the North –East Atlantic area: Present situation, knowledge gaps and future perspectives

Artificial reefs have been deployed in multiple regions of the world for different purposes including habitat restoration and protection, biodiversity and fish stock enhancement, fisheries management and recreation. Artificial reefs can be a valuable tool for ecosystem protection and rehabilitation, helping mitigate the effects of anthropogenic impacts that we face today. However, knowledge on artificial reefs is unevenly distributed worldwide, with some regions having much more quality information available and published (e.g. European Mediterranean Sea area), while others, for instance the North-East Atlantic area, do not. Here, we provide a characterization of purposely built artificial reefs in North-East Atlantic area based on all available literature (i.e. research papers and reports), highlighting the needs and gaps that are vital for establishing future perspectives for artificial reef deployment and research. In the North-East Atlantic area, sixty-one purposely built artificial reefs have been deployed since 1970, mostly between the years 1990–2009, with Spain being the country with the highest number of artificial reefs. The most reported purpose for their deployment is fisheries productivity and habitat/species protection, although, most artificial reefs are multipurpose in order to maximise the benefits of a given financial investment. The majority of artificial reefs were submerged at < 50 m, mainly between 10 and 20 m of depth. The most used designs were cubic blocks and complex designs made by an array of combined shapes, which mostly consist of concrete (79%). From all the analysed data on artificial reefs, 67% of the cases reported surveys to assess biodiversity after the deployment. However, in 26% of those cases, data was not available. When data was available, only 31% of cases reported long-term biomonitoring surveys (3 years or more). Based upon these findings, we noticed a general lack of scientifically robust data, including records of species and abundance of both fish and invertebrates, as well as macroalgae, preventing an adequate determination of the best balance between shape, construction material and bio-colonization. Critiques and suggestions are discussed in the light of currently available data in order to perform more efficient research, evaluation and functioning of future artificial reefs