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Association of European Marine Biological Laboratories Expanded

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Cryoprotectants synergy improve zebrafish sperm cryopreservation and offspring skeletogenesis
Publication . Diogo, Patricia; Martins, Gil; Nogueira, Rita; Marreiros, Ana; Gavaia, Paulo; Cabrita, Elsa
The synergy obtained by the combination of cryoprotectants is a successful strategy that can be beneficial on the optimization of zebrafish sperm cryopreservation. Recently, a protocol was established for this species using an electric ultrafreezer (-150 degrees C) performing cooling rate (-66 degrees C/min) and storage within one step. The ultimate objective of sperm cryopreservation is to generate healthy offspring. Therefore, the objective of this study was to select the most adequate cryoprotectant combination, for the previously established protocol, that generate high quality offspring with normal skeletogenesis. Among the permeating cryoprotectant concentrations studied 12.5% and 15% of N,N-dimethylformamide (DMF) yielded high post-thaw sperm quality and hatching rates. For these two concentrations, the presence of bovine serum albumin (10 mg/mL), egg yolk (10%), glycine (30 mM) and bicine (50 mM) was evaluated for post-thaw sperm motility, viability, in vitro fertilization success and offspring skeletal development (30 days post fertilization). Higher concentration of permeating cryoprotectant (15%) decreased the incidence of deformed arches and severe skeletal malformations, which suggests higher capacity to protect the cell against cold stress and DNA damage. Extender containing 15% DMF with Ctrl, Bicine and egg yolk were the non-permeating cryoprotectants with higher post-thaw quality. The use of these compounds results in a reduction in vertebral fusions, compressions and severity of skeletal malformations in the offspring. Therefore, these extender compositions are beneficial for the quality of zebrafish offspring sired by cryopreserved sperm with 66 degrees C/min freezing rate. To the best of our knowledge, this is the first report on skeletal development of the offspring sired by cryopreserved sperm performed with different freezing media compositions in zebrafish.
Microplastic retention by marine vegetated canopies: simulations with seagrass meadows in a hydraulic flume
Publication . de los Santos, Carmen B.; Krång, Anna-Sara; Infantes, Eduardo
Marine canopies formed by seagrass and other coastal vegetated ecosystems could act as sinks of microplastics for being efficient particle traps. Here we investigated for the first time the occurrence of microplastic retention by marine canopies in a hydraulic flume under unidirectional flow velocities from 2 to 30 cm s-1. We used as model canopy-forming species the seagrass Zostera marina with four canopy shoot density (0, 50, 100, 200 shoots m-2), and we used as microplastic particles industrial pristine pellets with specific densities from 0.90 to 1.34 g cm-3 (polypropylene PP; polystyrene PS; polyamide 6 PA; and polyethylene terephthalate PET). Overall, microplastics particles transported with the flow were retained in the seagrass canopies but not in bare sand. While seagrass canopies retained floating microplastics (PP) only at low velocities (<12 cm s-1) due to a barrier created by the canopy touching the water surface, the retention of sinking particles (PS, PA, PET) occurred across a wider range of flow velocities. Our simulations revealed that less dense sinking particles (PS) might escape from the canopy at high velocities, while denser sinking particles can be trapped in scouring areas created by erosive processes around the eelgrass shoots. Our results show that marine canopies might act as potential barriers or sinks for microplastics at certain bio-physical conditions, with the probability of retention generally increasing with the seagrass shoot density and polymer specific density and decreasing with the flow velocity. We conclude that seagrass meadows, and other aquatic canopy-forming ecosystems, should be prioritized habitats in assessment of microplastic exposure and impact on coastal areas since they may accumulate high concentration of microplastic particles that could affect associated fauna.
New vectors of TTX analogues in the North Atlantic Coast: the edible crabs Afruca tangeri and Carcinus maenas
Publication . Lage, Sandra; ten Brink, Felicitas; Canário, Adelino V. M.; Silva, José Paulo da
Tetrodotoxin (TTX) and its analogues are naturally occurring toxins historically responsible for human poisoning fatalities in Eastern Asia. It is typically linked to the consumption of pufferfish and, to a lesser extent, marine gastropods and crabs. In the scope of a comprehensive project to understand the prevalence of emergent toxins in edible marine organisms, we report, for the first time, the detection of TTX analogues in the soft tissues of edible crabs, the European fiddler crab (Afruca tangeri) and green crab (Carcinus maenas), harvested in southern Portugal. No TTX was detected in the analyzed samples. However, three TTX analogues were detected—an unknown TTX epimer, deoxyTTX, and trideoxyTTX. These three analogues were found in the European fiddler crab while only trideoxyTTX was found in the green crab, suggesting that the accumulation of TTX analogues might be influenced by the crabs’ different feeding ecology. These results highlight the need to widely monitor TTX and its analogues in edible marine species in order to provide adequate information to the European Food Safety Authority and to protect consumers.
Rhodolith physiology across the atlantic: towards a better mechanistic understanding of Intra- and interspecific differences
Publication . Schubert, Nadine; Peña, Viviana; Salazar, Vinícius W.; Horta, Paulo A.; Neves, Pedro; Ribeiro, Cláudia; Otero-Ferrer, Francisco; Tuya, Fernando; Espino, Fernando; Schoenrock, Kathryn; Hofmann, Laurie C.; Le Gall, Line; Santos, Rui; Silva, João
Coralline algae are important components in a large variety of ecosystems. Among them, rhodoliths are a group of free-living coralline red algae that cover extensive coastal areas, from tropical to polar regions. In contrast to other ecosystem engineers, limited research efforts preclude our understanding of their physiology, underlying mechanisms, drivers and potential differences related to species under varying environments. In this study, we investigated the photosynthetic and calcification mechanisms of six Atlantic rhodolith species from different latitudes, as well as intra-specific differences in one species from four locations. Laboratory incubations under varying light levels provided simultaneous photosynthesis- and calcification-irradiance curves, allowing the assessment of inter- and intra-specific differences on the coupling between these two processes. Stable isotope analysis and specific inhibitor experiments were performed to characterize and compare carbon-concentrating mechanisms (CCMs), as well as the involvement of specific ion-transporters for calcification. Our findings showed significant differences in rhodolith physiological mechanisms that were partially driven by local environmental conditions (light, temperature). High variability was found in the coupling between photosynthesis and calcification, in CCM-strategies, and in the importance of specific ion transporters and enzymes involved in calcification. While calcification was strongly correlated with photosynthesis in all species, the strength of this link was species-specific. Calcification was also found to be reliant on hotosynthesis- and light-independent processes. The latter showed a high plasticity in their expression among species, also influenced by the local environment. Overall, our findings demonstrate that (1) rhodolith calcification is a biologically-controlled process and (2) the mechanisms associated with photosynthesis and calcification display a large variability among species, suggesting potential differences not only in their individual, but also community responses to environmental changes, such as climate change.
Genomics-metabolomics profiling disclosed marine Vvbrio spartinae 3.6 as a producer of a new branched side chain prodigiosin
Publication . Vitale, Giovanni Andrea; Sciarretta, Martina; Esposito, Fortunato Palma; January, Grant Garren; Giaccio, Marianna; Bunk, Boyke; Sproeer, Cathrin; Bajerski, Felizitas; Power, Deborah; Festa, Carmen; Monti, Maria Chiara; D'Auria, Maria Valeria; de Pascale, Donatella
A wide range of prescreening tests for antimicrobial activity of 59 bacterial isolates from sediments of Ria Formosa Lagoon (Algarve, Portugal) disclosed Vibrio spartinae 3.6 as the most active antibacterial producing strain. This bacterial strain, which has not previously been submitted for chemical profiling, was subjected to de novo whole genome sequencing, which aided in the discovery and elucidation of a prodigiosin biosynthetic gene cluster that was predicted by the bioinformatic tool KEGG BlastKoala. Comparative genomics led to the identification of a new membrane di-iron oxygenase-like enzyme, annotated as Vspart_02107, which is likely to be involved in the biosynthesis of cycloprodigiosin and analogues. The combined genomicsmetabolomics profiling of the strain led to the isolation and identification of one new branched-chain prodigiosin (5) and to the detection of two new cyclic forms. Furthermore, the evaluation of the minimum inhibitory concentrations disclosed the major prodigiosin as very effective against multi-drug-resistant pathogens including Stenotrophomonas maltophilia, a clinical isolate of Listeria monocytogenes, as well as some human pathogens reported by the World Health Organization as prioritized targets.

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European Commission

Funding programme

H2020

Funding Award Number

730984

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