Assessment of the eutrophication status at Mediterranean sub-basin scale, within the European Marine Strategy Framework Directive.

The aim of this work is to define harmonized reference conditions and assessment thresholds for selected criteria elements of the Marine Strategy Framework Directive (MSFD) Descriptor 5 (Eutrophication) in the Eastern Mediterranean Sea and to test if a tool for integrated assessment of the status of marine systems can be used as a common methodological approach. In this frame, we tested two statistical approaches in order to set threshold values for four criteria of Descriptor 5: nutrients, chlorophyll a, transparency and dissolved oxygen in the bottom waters. It is noteworthy that this work revealed the need to apply common procedures in data treatment and assessment evaluation. This is the first attempt to set common methods for the assessment of eutrophication in the Eastern Mediterranean, which is essential in marine environments, especially those shared by several countries. To this end, we have applied common criteria and metrics and established thresholds "Good" and "Moderate" for nutrients, chlorophyll a, transparency and dissolved oxygen in the bottom waters for the different Water Types of the Adriatic and Aegean Seas (I, II, IIIW, IIIE), based on datasets provided by Italy, Slovenia, Croatia and Greece. The selected criteria elements were common for all countries, providing a unified approach to GES assessment of two case study areas: the Adriatic Sea and the Saronikos Gulf. Dissolved Inorganic Nitrogen (DIN) threshold values of 15.6, 6.85, 1.61 and 2.11 µmol L-1 were set for the Water Types I, II, IIIW and IIIE, respectively. We also tested if an aggregation tool for GES assessment, such as Nested Environmental status Assessment Tool (NEAT), could be used as a common methodological approach. The comparison of NEAT with TRIX showed good comparability. In this end, NEAT can be used as a useful and much needed assessment tool for assessing eutrophication status of the marine.

Metal concentrations in transitional and coastal waters measured by passive (Diffusive Gradients in Thin-films) and spot sampling: MONITOOL Project Dataset.

The MONITOOL project (2017-2023) was carried out to describe the relationships between total dissolved and labile metal concentrations measured in spot water samples and in concurrently deployed Diffusive Gradients in Thin-films (DGTs) passive samplers, respectively. The ultimate aim was to adapt existing marine metal Environmental Quality Standards (EQS marine water) for DGTs, enabling their use in the context of the European Directives (the Water Framework Directive (WFD) and the Marine Strategy Framework Directive (MSFD)). Time-integrated metal concentrations provided by DGTs, representing several days, are an advantage compared to conventional spot sampling, especially in highly dynamic systems, such as transitional waters. Hence, the MONITOOL project aimed to provide a robust database of dissolved and labile metal concentrations in transitional and coastal waters, based upon co-deployments of DGTs and collection of spot water samples at several sampling sites (England, France, Ireland, Italy, Northern Ireland, Portugal, Scotland and Spain), followed subsequently by DGT and water metal analysis. Samplings were carried out in 2018 and 2022, following agreed protocols developed in the framework of the project. The MONITOOL dataset includes metal concentrations from DGTs, measured with Inductively Coupled Plasma Mass Spectrometry (ICP-MS: Cd, Co, Cu, Fe, Mn, Ni, Pb, Zn) and in concurrently collected spot water samples by ICP-MS (Al, Cd, Co, Cu, Mn, Ni, Pb, Zn) and Anodic/Cathodic Stripping Voltammetry (ASV/CSV: Cd, Pb, Ni). Moreover, data on seawater physical-chemical parameters (salinity, temperature, dissolved oxygen, pH, turbidity, total suspended solids, dissolved organic carbon, and total organic carbon) is provided. This database presents the results obtained using, concurrently, different forms of sampling and analytical techniques, enabling the comparison of the results obtained by these strategies and allowing the adaptation of EQS in marine water (EQS marine water) to DGTs (EQS DGT), in the context of the WFD. Moreover, due to the large number of sampling sites, it could also be used for other types of research, such as those dealing with metal speciation or the determination of baseline levels.

An international intercomparison exercise on passive samplers (DGT) for monitoring metals in marine waters under a regulatory context.

In order to move forward in the acceptance of a novel contaminant monitoring technique (Diffusive Gradients in Thin-films: DGT) for assessment of marine water bodies, sensu the WFD, an Inter-Laboratories Comparison (ILC) exercise (nine Europeans laboratories) was organized in the framework of the Interreg Atlantic Area MONITOOL project, which focused on the use of the DGT technique for the measurement of WFD priority metals (Cd, Ni and Pb). Reproducible results were obtained for each metal by several laboratories, supporting the assertion that DGT analysis can be performed satisfactorily by laboratories experienced in measuring metals at trace levels in marine environments, even if they have limited practice in DGT analysis. According to the Z-score analysis, among the 9 participating laboratories, 3 had 100 % of satisfactory results for Cd, Ni, and Pb, 3 had >80 % satisfactory results and 2 had about 60 % satisfactory results. This work highlights the need to clearly describe the DGT method in order to control sources of contamination during analytical steps, in particular the resin gel retrieval and the elution steps. Such international intercomparison exercise is an important step to develop the laboratory network involved in DGT analysis and contributes to the improvement of data quality.

Metals concentrations in transitional and coastal waters by ICPMS and voltammetry analysis of spot samples and passive samplers (DGT).

This study investigates the relationships among Ni, Cd and Pb's different chemical forms determined by different methodologies in coastal and transitional waters across a broad geographical scale. Concentrations were measured in spot samples and through passive sampling (DGT). High variability of metal concentrations was found among sampling sites and methodologies due to natural water fluctuations rather than to a given metal or method. Total dissolved metal concentrations in spot samples were lower than the EQS-WFD values. The labile fractions of Cd and Pb, measured in spot samples by Anodic Stripping Voltammetry and by DGT-ICPMS, were highly correlated. Similar labilities were found for Cd, while for Pb, the ASV labile fraction was ≈50% lower. These results reflect the pool of mobile and labile species available towards each technique kinetic window, and they seem not to be affected by discrete sampling flaws.

A Bayesian Network model to identify suitable areas for offshore wave energy farms, in the framework of ecosystem approach to marine spatial planning.

The production of energy from waves is gaining attention. In its expansion strategy, technical, environmental and socioeconomic aspects should be taken into account to identify suitable areas for development of wave energy projects. In this research we provide a novel approach for suitable site identification for wave energy farms. To achieve this objective, we (i) developed a conceptual framework, considering technical, environmental and conflicts for space aspects that play a role on the development of those projects, and (ii) it was operationalized in a Bayesian Network, by building a spatially explicit model adopting the Spanish and Portuguese Economic Exclusive Zones as case study. The model results indicate that 1723 km2 and 17,409 km2 are highly suitable or suitable for the development of wave energy projects (i.e. low potential conflicts with other activities and low ecological risk). Suitable areas account for a total of 2.5 TWh∙m-1 energy resource. These areas are placed between 82 and 111 m water depth, 18-30 km to the nearest port, 21-29 km to the nearest electrical substation onshore, with 143-170 MWh m-1 mean annual energy resource and having 124-150 of good weather windows per year for construction and maintenance work. The approach proposed supports scientists, managers and industry, reducing uncertainties during the consenting process, by identifying the most relevant technical, environmental and socioeconomic factors when authorising wave energy projects. The model and the suitability maps produced can be used during site identification processes, informing Strategic Environmental Assessment and ecosystem approach to marine spatial planning.

Concurrent sampling of transitional and coastal waters by Diffusive Gradient in Thin-films (DGT) and spot sampling for trace metals analysis.

This protocol was developed based on the knowledge acquired in the framework of the Interreg MONITOOL project (EAPA_565/2016) where extensive sampling campaigns were performed in transitional and coastal waters covering eight European countries. It provides detailed procedures and guidelines for the sampling of these waterbodies by concurrent collection of discrete water samples and the deployment of Diffusive Gradient in Thin-films (DGT) passive samplers for the measurement of trace metal concentrations. In order to facilitate the application of this protocol by end-users, it presents steps to follow in the laboratory prior to sampling campaigns, explains the procedures for field campaigns (including in situ measurement of supporting parameters) and subsequent sample processing in the laboratory in preparation for trace metal analyze by inductively coupled plasma-mass spectrometry (ICP-MS) and voltammetry. The protocol provides a systematic, coherent field sampling and sample preparation strategy that was developed in order to ensure comparability and reproducibility of the data obtained from each project Partner in different regions. • Standardization of the concurrent sampling of transitional and coastal waters by DGT passive samplers and spot sampling. • Robust procedures and tips based on existing international standards and comprehensive practical experience. • Links to demonstration videos produced within the MONITOOL project.

Assessing variability in the ratio of metal concentrations measured by DGT-type passive samplers and spot sampling in European seawaters.

The current study evaluates the effect of seawater physico-chemical characteristics on the relationship between the concentration of metals measured by Diffusive Gradients in Thin films (DGT) passive samplers (i.e., DGT-labile concentration) and the concentrations measured in discrete water samples. Accordingly, Inductively Coupled Plasma Mass Spectrometry (ICP-MS) was used to measure the total dissolved metal concentrations in the discrete water samples and the labile metal concentrations obtained by DGT samplers; additionally, lead and cadmium conditional labile fractions were determined by Anodic Stripping Voltammetry (ASV) and total dissolved nickel was measured by Cathodic Stripping Voltammetry (CSV). It can be concluded that, in general, the median ratios of DGT/ICP and DGT/ASV(CSV) were lower than 1, except for Ni (median ratio close to 1) and Zn (higher than 1). This indicates the importance of speciation and time-integrated concentrations measured using passive sampling techniques, which is in line with the WFD suggestions for improving the chemical assessment of waterbodies. It is the variability in metal content in waters rather than environmental conditions to which the variability of the ratios can be attributed. The ratios were not significantly affected by the temperature, salinity, pH, oxygen, DOC or SPM, giving a great confidence for all the techniques used. Within a regulatory context such as the EU Water Framework Directive this is a great advantage, since the simplicity of not needing to use corrections to minimize the effects of environmental variables could help in implementing DGTs within monitoring networks.

Contaminants of emerging concern in the Basque coast (N Spain): Occurrence and risk assessment for a better monitoring and management decisions.

The study of the presence in the aquatic environment of certain substances considered as contaminants of emerging concern (CEC) is a preliminary step to the analysis of the possible harmful effects on aquatic ecosystems and the establishment of the corresponding environmental quality standards. In order to monitor the occurrence of CECs in the aquatic environment, the European Commission established in 2015 and 2018 two watch-list of substances for Union-wide monitoring in the field of water policy (Decision (EU) 2015/495 and Decision (EU) 2018/840). In the coast of the Basque Country, southeast of the Bay of Biscay, 19 of these watch list substances were monitored quarterly from May 2017 to March 2019. Water samples were collected at the effluent of three wastewater treatment plants and five control points associated with receiving waters (transitional and coastal water bodies). The most frequently quantified substances were azithromycin (91%), imidacloprid (82%), clarithromycin (80%), diclofenac (78%) and erythromycin (73%), with frequencies of quantification higher in wastewaters (83-100%) than in receiving waters (70-85%). In general, concentrations in wastewater were also higher than in receiving waters, indicating a dilution effect in the environment. In receiving waters, six out of the nineteen substances monitored exceeded their respective Predicted No-Effect Concentrations: azithromycin (34%), imidacloprid (9%), 17ß-estradiol (E2) (9%), clarithromycin (7%), ciprofloxacin (7%), and diclofenac (5%); and therefore, their levels could pose an environmental risk.

'The past is the future of the present': Learning from long-time series of marine monitoring.

Using a long-term (1995-2014) monitoring network, from 51 sampling stations in estuaries and coasts of the Basque Country (Bay of Biscay), the objective of this investigation was to assess the responsiveness of 83 variables in water (18), sediments (27), biota (26), phytoplankton (2), macroinvertebrates (5) and fishes (5) to different human pressures and management actions. We used a total of 3247 series of data to analyse trends of improvement and worsening in quality. In a high percentage of the cases, the management actions taken have resulted in positive effects in the environment, as shown by the trend analysis in this investigation. Overall, much more trends of improvement than of worsening have been observed; this is true for almost all the media and biological components studied, with the exception of phytoplankton; and it applies as well to almost all the stations and water bodies, with the exception of those corresponding to areas with water treatment pending of accomplishment. In estuaries with decreasing human pressures during the period, the percentage of series showing quality improvement was higher (approx. 30%) than those showing worsening of quality (12%). Moreover, in those water bodies showing an increase of pressure, variables which can be considered indicators of anthropogenic effects showed negative trends (quality worsening). On the other hand, some of the variables analysed were more affected by natural variability than by changes in pressures. That was the case of silicate, nitrate and suspended solids, which followed trends correlated to salinity, which, in turn, was related to the rainfall regime during the study period.

Source characterization and spatio-temporal evolution of the metal pollution in the sediments of the Basque estuaries (Bay of Biscay).

According to Water Framework Directive requirements, Member States must identify and analyze effects derived from human pressures in aquatic systems. As different kind of pressures can impact water bodies at different scales, analyses of spatio-temporal evolution of water bodies becomes essential in order to understand ecosystem responses. In this investigation, an analysis of spatio-temporal evolution of sedimentary metal pollution (Cd, Cr, Cu, Hg, Ni, Pb, Zn) in 12 Basque estuaries (Bay of Biscay) is presented. Data collected in extensive sampling surveys is the basis for the GIS-based statistical approach used. The implementation of pollution abatement measures is reflected in a long-term decontamination process, mostly evident in estuaries with highest historical sediment pollution levels. Spatial evolution is determined by either naturally occurring or human driven processes. Such spatial processes are more obviously being reflected in estuaries with lower historical sediment pollution levels.

Implementation of the European Marine Strategy Framework Directive: a methodological approach for the assessment of environmental status, from the Basque Country (Bay of Biscay).

The implementation of the Marine Strategy Framework Directive (MSFD) is directing European marine research towards the coordinated and integrated assessment of sea environmental status, following the ecosystem-based approach. The MSFD uses a set of 11 descriptors which, together, summarise the way in which the whole system functions. As such, the European Commission has proposed an extensive set of indicators, to assess environmental status. Hence, taking account of the large amount of data available for the Basque coast (southern Bay of Biscay), together with a recent proposal for assessment within the MSFD, an integrated environmental status assessment approach is developed (for the first time) in this contribution. The strengths and weaknesses of the method, combined with proposals from the MSFD, are discussed.

Sensitivity comparison of laboratory-cultured and field-collected amphipod Corophium multisetosum in toxicity tests.

The feasibility of using lab-cultured amphipods Corophium multisetosum (Stock 1952) to evaluate the toxicity of contaminants present within marine sediments was studied. This was done by comparing the sensitivity of lab-cultured amphipods in a cadmium toxicity test and to toxic sediment samples, during a 10-days bioassay, with field collected individuals. Different responses were observed between field and cultured individuals. Cadmium test indicated high temporal variability in the LC(50) values of field amphipods (2.40-6.55 mg L(-1)). Sensitivity of cultured amphipods was within the seasonal range of the field individuals (5.81 mg L(-1), LC(50)). However, culture amphipods showed much lower sensitivity in toxic sediment samples. Our results indicate that sensitivity should be determined using a sediment matrix, if the assessment of toxicity is based upon bioassays performed with cultured burrower-amphipods.

Seasonal variability in gonad development in the sea urchin (Paracentrotus lividus) on the Basque coast (southeastern Bay of Biscay).

The main limiting factor to carrying out toxicological bioassays with sea urchin larvae is the restricted period for the availability of viable gametes. Although studies have been undertaken of the reproductive cycle of the sea urchin for several areas of the Bay of Biscay, only limited information exists for the southeasternmost area (the Basque coast). Furthermore, this geographical zone presents some particular environmental conditions, e.g., relatively warm waters. In this study, the gonad state of a population of Paracentrotus lividus, settled on the rocky shores of the city of Donostia-San Sebastian have been monitored. These sea urchins are observed to behave like a typical Atlantic population, with generally high gonad indices and a single yearly main spawning period, between April and May. Further, females show a shorter spawning period than males, becoming the limiting organisms for the availability of larvae.

Induction to maturation of the sea urchin Paracentrotus lividus (Lamarck, 1816) under laboratory conditions.

Laboratory experiments were carried out over two different periods, with the aim of investigating and utilizing the induction to gonadal maturation of the sea urchin Paracentrotus lividus. The final objective was to obtain viable gametes outside the period of natural spawning in the environment; which would allow the utilization of ecotoxicological bioassays with sea urchin larvae at any time of the year. The experiment consisted of maintaining some sea urchins in tanks and providing them with a natural photoperiod, unlimited food and a constant temperature of 20 degrees C. During days 0, 30, 60 and 90, gonads from 15 of these sea urchins were compared with those collected simultaneously in the natural environment. The gametes obtained were used to carry out fecundations, in order to check their viability. The final results obtained were clearly influenced by the gonadal state of the sea urchins at the initial stage of the experiment. The best results were obtained within a time period of 60 days and when the initial gonad index was low.