The effect of Zostera noltei recolonization on the sediment mercury vertical profiles of a recovering coastal lagoon.

Mercury's extreme toxicity and persistence in the environment justifies a thorough evaluation of its dynamics in ecosystems. Aveiro Lagoon (Portugal) was for decades subject to mercury effluent discharges. A Nature-based Solution (NbS) involving Zostera noltei re-colonization is being tested as an active ecosystem restoration measure. To study the effect of Zostera noltei on the sediment contaminant biogeochemistry, seasonal (summer/winter) sediment, interstitial water and labile mercury vertical profiles were made in vegetated (Transplanted and Natural seagrass meadows) and non-vegetated sites (Bare-bottom area). While no significant differences (p > 0.05) were observed in the sedimentary phase, Zostera noltei presence reduced the reactive/labile mercury concentrations in the top sediment layers by up to 40% when compared to non-vegetated sediment, regardless of season. No differences were found between vegetated meadows, highlighting the fast recovery of the contaminant regulation ecosystem function provided by the plants after re-colonization and its potential for the rehabilitation of historically contaminated ecosystems.

Zostera noltei response to transplantation into historically Hg-contaminated sediments (A mesocosm experiment): Growth, bioaccumulation and photosynthetic performance.

The present study aimed to evaluate the effectiveness of seagrass recolonization as a nature-based solution for the recovery of a coastal area historically contaminated by mercury (Laranjo Bay, Ria de Aveiro, Portugal). A mesocosm approach was employed to assess the resistance of Zostera noltei to transplantation into contaminated sediments collected in-situ (0.5-20 mg kg-1 Hg). At each sampling time (15, 30, 60, 120 and 210 days), the resistance of transplanted Z. noltei was evaluated through growth parameters (biomass and coverage area), photosynthetic performance and elemental composition. Although some significant differences (p ≤ 0.05) were observed between treatments, essentially associated with the elemental composition of plant tissues, the most relevant variations were associated with seasonality. Overall, plants were found to not be affected by sediment contamination, under the tested concentrations, suggesting that recolonization with Z. noltei can be an effective restoration strategy for historically contaminated coastal areas.

Evaluation of species-specific dissimilarities in two marine fish species: mercury accumulation as a function of metal levels in consumed prey.

The aim of this research was to compare mercury (Hg) accumulation (total and organic) and tissue distribution in two marine fish species with contrasting feeding tactics. Thus, juvenile specimens of European sea bass and Golden grey mullet were surveyed in an estuary historically affected by Hg discharges. Total Hg was preferentially accumulated in intestine, muscle, and liver, whereas gills and brain presented the lowest Hg levels observed in both species. Significant differences between species were only verified for muscle, with D. labrax's levels being greater than L. aurata's. Muscle accounted for >87% of the Hg relative tissue burden, whereas liver did not exceed 11%. Organic Hg accumulation occurred mainly in liver and muscle, with D. labrax evidencing significantly greater loads. Moreover, organic Hg in consumed prey items was also significantly greater in D. labrax. Accumulation of organic Hg in liver, intestine, and muscle seemed to vary as a function of the consumed prey items contamination, suggesting fish feeding strategies as the dominant factor determining metal accumulation. For both fish species, a stable ratio was observed between Hg increments from the reference to the contaminated site, possibly indicating that the organic Hg content of diet may regulate the internal levels of this contaminant. Thus, this ratio might prove to be a useful contamination predictor tool in early life stages of fish.

Mercury bioaccessibility in fish and seafood: Effect of method, cooking and trophic level on consumption risk assessment.

The bioaccessible Hg fraction in fish and seafood commonly present in the Mediterranean diet was assessed through three distinct in vitro extraction schemes. Each extraction method provided different results, highlighting the lack of a universal methodology to estimate mercury (Hg) bioaccessibility in those matrices. Bioaccessible Hg fractions ranged from 10 to nearly 90% of total mercury (T-Hg) and increased in predator species (Swordfish - Xiphias gladius, Blue Shark - Prionace glauca and Tuna - Thunnus sp.). Among the three extractions tested, the Unified Bioaccessibility Method (UBM) provided the highest estimation of Hg bioaccessibility for consumers. The tested cooking procedures (frying, grilling and steaming) considerably reduced the bioaccessible fraction. Results indicate that bioaccessible Hg found in ingested fish and seafood is far below the levels set by the current safety risk assessment legislation. These findings highlight the importance of integrating bioaccessibility measurements in food safety legislation.

Metal(oid)s accumulation (Hg and As) and their biochemical effects in Halimione portulacoides (Ria de Aveiro, Portugal).

For decades, mercury (Hg) and arsenic (As) were discharged into the Aveiro Lagoon. This study was designed to assess the natural attenuation process evolution by: (1) evaluating Hg and As concentration in sediments and in Halimione portulacoides, and assess the long-term evolution of a historically contaminated salt-marsh system; (2) evaluating the stress levels imposed by Hg and As, and the mechanisms triggered to squash oxidative damage. Concentrations of Hg and As in sediments varied considerably between sampling locations. H. portulacoides did not bioconcentrate Hg and As, restricted translocation to aerial biomass and immobilized contaminants in cell walls. The ions that reached the cytosol induced oxidative stress, restrained by antioxidant mechanisms, especially SOD and CAT activity. Results show that after 25 years of natural attenuation, contamination decreased in the sediment, but remains above non-contaminated systems and continues to induce toxicity in the saltmarsh halophyte community, evidencing the persistent effect of Hg and As contamination.

Contribution of primary producers to mercury trophic transfer in estuarine ecosystems: possible effects of eutrophication.

There is an ongoing eutrophication process in the Ria de Aveiro coastal lagoon (Portugal), with progressive replacement of rooted primary producers for macroalgae. Taking advantage of a well-defined environmental contamination gradient, we studied mercury accumulation and distribution in the aboveground and the belowground biomass of several salt marsh plants, including the seagrass species Zostera noltii and the dominant green macroalgal species Enteromorpha sp. The results of these experiments were then placed into the context of the estuarine mercury cycle and transport from the contaminated area. All salt marsh plants accumulated mercury in the root system, with Halimione portulacoides showing the highest levels, with up to 1.3 mg kg(-1) observed in the most contaminated area. Belowground/aboveground ratios were generally below 0.4, suggesting that salt marsh plants are efficient immobilizers and retainers of mercury agents. Moreover, due to their sediment accretion capacities, salt marsh plants seem to play an important role in the sequestration of mercury in estuarine sediments. Seagrasses, on the other hand, accumulated considerable amounts of mercury in the aboveground biomass with belowground/aboveground ratios reaching as high as 1.4. These results may be due to their different routes of uptake (roots and foliar uptake) which suggests that seagrass meadows can be an important agent in the export of mercury from contaminated areas, considering the high aboveground biomass replacement rates. Rooted macrophytes accumulate less mercury in their aboveground biomass than macroalgae. The change of primary producer dominance due to eutrophication can originate a 4- to 5-fold increase in primary producer associated mercury. This mercury would be available for export, making it bioavailable to estuarine food webs, which stresses the need to reverse the current eutrophic status of estuarine systems.

Mercury pollution in Ria de Aveiro (Portugal): a review of the system assessment.

The Ria de Aveiro (Portugal) is a coast al lagoon adjacent to the Atlantic Ocean and it has an inner bay (Laranjo bay) that received a highly contaminated effluent discharged by a mercury cell chlor-alkali plant from the 1950s until 1994. The aim of this study is to review in a holistic way several research studies that have been carried out in the Ria de Aveiro, in order to evaluate the remobilization of the mercury accumulated within the system and the recovery of the lagoon. The spatial distribution of the total mercury in the surrounding terrestrial environment has also been considered. Results indicate that the main mercury contamination problems in the Ria de Aveiro are confined to the Laranjo bay. Mercury export to the coastal waters and its impact on the nearshore compartments (water column, sediment and biota) are low. No direct effects of the mercury from nearby industrial activities were detected in Aveiro's urban soils, although historical mercury contamination is still affecting soil quality in the immediate vicinity of the chlor-alkali plant, located in Estarreja. Moreover, macrophyte harvesting for human direct or indirect use and the consumption of mussels, crabs and the sea bass from the Laranjo bay may constitute a health risk. Further studies focusing on developing skills for the restoration of the ecosystem are presently underway.

Multiple regression analysis to assess the spatial distribution and speciation of mercury in surface sediments of a contaminated lagoon.

The concentrations of inorganic mercury (IHg) and methylmercury (MeHg) in surface sediments from the contaminated Aveiro Lagoon (Portugal) were determined by species-specific isotope dilution analysis. Different behaviour of IHg and MeHg was observed based upon multiple regression analysis, including geochemical characteristics of the surface sediments (fine fraction, concentrations of organic matter and metals) and non-Euclidean distances between sampling points. This data treatment method was valid over the entire concentration range of IHg and MeHg, allowing robust quantitative evaluation with respect to extrapolation. For IHg, there was statistical separation of the dispersion away from the contamination source and of Al concentration in the sediments. The MeHg concentrations followed those of IHg at high concentrations. The geochemical variables, such as concentrations of Ca (marine influence proxy), Mn and organic matter, were necessary to describe the behaviour of MeHg across the whole concentration range. The models for MeHg demonstrated that, close to the mouth of the lagoon, net production of MeHg was higher. In future, multiple regression analysis could be applied to separate and to evaluate quantitatively the effects of geochemistry and dispersion away from the contamination source in sediments contaminated with other substances.

Advances on assessing nanotoxicity in marine fish - the pros and cons of combining an ex vivo approach and histopathological analysis in gills.

The need to overcome logistic and ethical limitations of in vivo nanotoxicity evaluation in marine organisms is essential, mostly when dealing with fish. It is well established that medium/solvent conditions affect dispersion and agglomeration of nanoparticles (NPs), which represents a constraint towards a solid and realistic toxicity appraisal. In this way the pros and cons of an ex vivo approach, using a simplified exposure medium (seawater) and addressing gills histopathology, were explored. The nanotoxic potential of environmentally realistic concentrations of titanium dioxide NPs (TiO2 NPs) was also assessed, disclosing the morpho-functional effects on the gills and the possible uptake/elimination processes. Excised gills of the Senegalese sole (Solea senegalensis) were directly exposed in artificial seawater to 20 and 200 µg L-1 TiO2 NPs, for 2 h and 4 h. Semi-quantitative and quantitative histological analyses were applied. The normal morphology of the gill's epithelia was only slightly altered in the control, reflecting protective mechanisms against the artificiality of the experimental conditions, which, together with the absence of differences in the global histopathological index (Ih), corroborated that the gill's morpho-functional features were not compromised, thereby validating the proposed ex vivo approach. TiO2 NPs induced moderate severity and dissemination of histopathological lesions. After 2 h, a series of compensatory mechanisms occurred in NP treatments, implying an efficient response of the innate defense system (increasing number of goblet cells) and effective osmoregulatory ability (chloride cells proliferation). After 4 h, gills revealed signs of recovery (normalization of the number of chloride and goblet cells; similar Ih), highlighting the tissue viability and effective elimination and/or neutralization of NPs. The uptake of the TiO2 NPs seemed to be favored by the higher particle sizes. Overall, the proposed approach emerged as a high-throughput, reliable, accurate and ethically commendable methodology for nanotoxicity assessment in marine fish.

The macrobenthic community along a mercury contamination in a temperate estuarine system (Ria de Aveiro, Portugal).

Mercury is a widely distributed environmental pollutant and a toxic element to all living organisms. This study represents an attempt to evaluate its correlation with the macrobenthic community structure in a temperate estuarine system, the Ria de Aveiro (Portugal). The macrobenthic community structure showed significant differences along the mercury gradient. Overall, the increase of mercury contamination was associated with reduced total abundance, lower species richness, and dominance of tolerant taxa. The polychaetes Hediste diversicolor and Alkmaria romijni, and the isopod Cyathura carinata were associated with high levels of mercury in sediment, while the bivalves Scrobicularia plana, Cerastoderma edule and Abra alba showed higher density in lesser contaminated sites. Furthermore, highly contaminated sites showed substantially lower abundances of surface-deposit feeders and herbivores, and higher abundances of subsurface-deposit feeders and omnivores. Salinity was also a significant factor affecting the community, being responsible for seasonal variations in the macrobenthic assemblages.

Pattern and pathways for mercury lifespan bioaccumulation in Carcinus maenas.

Carcinus maenas is an important and exploited natural resource in temperate estuaries, being consumed by local populations and also used as fish bait. Through bioaccumulation and bioamplification processes occurring in polluted estuarine ecosystems, this species may directly or indirectly represent a major pathway for Human contamination. No information is available on the overall lifespan bioaccumulation pattern of mercury for this species, which would be an invaluable tool for assessing the risks associated with C. maenas harvest in contaminated areas. Taking that in mind, the main objectives of this work were to clarify the lifespan bioaccumulation pattern of mercury for this species. A consistent lifespan accumulation pattern was found for this species, both for total and organic mercury, and coherent differences were found between genders, suggesting different physiological responses to contamination. From our results two scenarios emerge: (a) for low mercury contamination areas where the diet is the major pathway for metal uptake, with higher levels in muscle and hepatopancreas (twice as high as in gills) and higher organic mercury fractions (as high as 80-90% in muscle), crabs bioaccumulate with age and (b) on high contaminated areas, environmental exposure is predominant, with higher levels found in gills (reaching 0.8 mg kg(-1)), lower organic fractions and no mercury increment with age. Detoxification strategies, dietary differences and species mobility may account for these two accumulation patterns.

Mobility versus retention of mercury in bare and salt marsh sediments of a recovering coastal lagoon (Ria de Aveiro, Portugal).

During decades, mercury (Hg) was discharged into the Aveiro Lagoon. Twenty-five years after the cessation of discharges, sediment cores were collected at two areas to assess the evolution of the mercury contamination status. Vertical Hg profiles differed considerably between bare and vegetated sediments. Bare sediments contained significantly less Hg (<8 mg kg-1) than historical data of 1995 (up to 40 mg kg-1), probably resulting from erosion. Salt marsh sediments were marked by a Hg sub-surface enrichment, reaching 44 mg kg-1 in the site closer to the industrial discharge point. High Hg concentrations in 2007, 2011 and 2016 were found at similar sediment layers. These results emphasise the role of halophyte plants in the cycling and retention of Hg in sediments. The persistency of high Hg in the upper marsh layers highlights the complexity in the recovery of historically contaminated marshes and the vulnerability to modifications in hydrology associated with climate changes.

Addressing the impact of mercury estuarine contamination in the European eel (Anguilla anguilla L., 1758) - An early diagnosis in glass eel stage based on erythrocytic nuclear morphology.

The decline of the European eel (Anguilla anguilla L., 1758) population throughout Europe has been partially attributed to pollution. As glass eel estuarine migration may represent a considerable threat, the impact of mercury (Hg) contamination at this stage was evaluated through an in situ experiment (7days). Total Hg (tHg) bioaccumulation was evaluated concomitantly with erythrocytic nuclear morphology alterations: erythrocytic nuclear abnormalities assay (ENA), frequency of immature erythrocytes (IE) and the erythrocytic maturity index (EMI). The ENA results suggested a genotoxic pressure at the most contaminated sites, in line with the tHg increase. The EMI data, together with IE frequency, showed that fish exposed to high levels of Hg exhibited alterations of haematological dynamics, translated into an erythropoiesis increment. Despite the presence of these compensatory mechanisms, the present findings suggest a harmful impact of Hg on genome integrity at this early development stage, potentially affecting eels' condition and ultimately the population sustainability.

Seasonal fluctuations of tissue mercury contents in the European shore crab Carcinus maenas from low and high contamination areas (Ria de Aveiro, Portugal).

The main objective was to study the seasonal variation of mercury concentrations in different tissues (muscle, hepatopancreas and gills) of Carcinus maenas from low and high Hg contaminated areas, a valuable resource in temperate estuaries and a possible pathway for human uptake. Individuals of two size classes (around 35 and 55 mm cephalothorax wide) were captured monthly between March 1999 and May 2000 in two areas of Ria de Aveiro: in the main navigation channel that connects the lagoon to the sea, and in the inner lagoon area heavily contaminated by mercury (maximum Hg in sediments of 5.4 microg g(-1)). Pronounced decreases in salinity and temperature and reduced food availability in winter seemed to be the responsible for the decline of the crab condition index (0.75-0.45) in larger individuals. Muscle and hepatopancreas exhibited higher mercury concentrations than gills, with concentrations in the contaminated site ranging from 0.03 to 0.63 microg g(-1) and 0.02 to 0.34 microg g(-1), respectively. Linear regressions between muscle and hepatopancreas (r=0.94, p<0.001) and muscle and gills (r=0.97, p<0.001) suggested a rapid redistribution of mercury inside the organism. During winter, a rapid elimination of mercury was found in the three analysed tissues followed by uptake. Larger crabs presented elimination rates from 18 to 34 ng g(-1) per week, while the smaller crabs showed lower elimination rates (10-24 ng g(-1) per week). The uptake was similar in both size classes (11-15 ng g(-1) and 8.1-15 ng g(-1) per week, respectively for large and small crabs). Our results suggest that C. maenas harvested in the contaminated areas must be considered with caution, since Hg concentrations were found to exceed the threshold concentration allowed for human consumption (0.5 microg g(-1)).

Can Nassarius reticulatus be used as a bioindicator for Hg contamination? Results from a longitudinal study of the Portuguese coastline.

The focus of this work is to explore the use of the netted whelk, Nassarius reticulatus (L.), as an indicator of mercury (Hg) contamination, by assessing the concentration of Hg in the sediments and in the whelk along the entire Portuguese coast. Total Hg concentrations ranged from below the detection limit (0.01 ng absolute mercury) up to 0.87 mg kg(-1) dry weight (dwt) in sediments and between 0.06 and 1.02 mg kg(-1) (dwt) for organisms, with no significant differences between males and females. Although organic mercury was not detected in the sediments, it represented, on average, 52% of the total Hg in the whelk tissues, and as high as 88% in some cases, suggesting mercury accumulation from dietary intake. Significant negative correlations were found between the total Hg concentrations in the sediments and the log(10) of Hg concentrations in whelk tissues males (r=-0.64; P<0.01) and females (r=-0.52; P<0.01) indicating that the species is a poor indicator of Hg contamination. Nevertheless, since the highest concentrations of organic mercury in the whelk tissues were found in the least contaminated areas, this species must be highly relevant in the trophic web, namely on the possible biomagnification of mercury. The high dietary mercury accumulation from feeding on carrion and the low bioavailability of mercury to whelks in estuarine sediments may be the basis of the mercury accumulation pattern in N. reticulatus.

Estimation of mercury background values in sediment and biota of the Bijagós archipelago, Guinea-Bissau.

This work evaluates the mercury (Hg) contamination status (sediments and biota) of the Bijagós archipelago, off the coast of Guinea-Bissau. Sediments exhibited very low concentrations (<1-12ngg(-1)), pointing to negligible sources of anthropogenic Hg in the region. Nevertheless, Hg is well correlated to the fine fraction, aluminium, and loss on ignition, indicating the effect of grain size and organic matter content on the presence of Hg in sediments. Mercury in the bivalves Tagelus adansoni and Senilia senilis did not vary considerably among sites, ranging within narrow intervals (0.09-0.12 and 0.12-0.14µgg(-1) (dry weight), respectively). Divergent substrate preferences/feeding tactics may justify slight differences between species. The value 11ngg(-1) is proposed as the sediment background concentration for this West-African coastal region, and concentrations within the interval 8-10ngg(-1) (wet weight) may be considered as reference range for S. senilis and T. adansoni in future monitoring studies.

Field transplantation of the bivalve Scrobicularia plana along a mercury gradient in Ria de Aveiro (Portugal): uptake and depuration kinetics.

The bioaccumulation and depuration capabilities of mercury by the edible bivalve Scrobicularia plana was studied in a coastal lagoon (Ria de Aveiro, Portugal) through a transplantation experiment. Little information on this topic is available in the literature, especially concerning different tissues' responses to contaminant exposure, but the present study is one of the few works that can surpass this knowledge gap. Organisms from a reference area were transplanted to two different contaminated areas in the Ria de Aveiro. In both areas, the bivalves (i.e., entire organism, digestive gland and the rest of the organism) presented a similar saturation model of mercury accumulation, the digestive gland being the tissue that reached the highest concentrations after 25 days of exposure to the contaminant. During this short uptake period, the transplanted organisms reached 20-30% of the concentrations observed in resident contaminated organisms. After the exposure period, the organisms were transplanted to a clean area for more than 25 days of depuration. At the end of the transplantation period, organisms lost approximately 50% of their mercury body burden (60%: the entire organism and digestive gland; 35%: gills and 40%: the rest of the organism) and the ones from the least contaminated site almost reached the concentrations recorded in the reference area. So, the results suggest that S. plana is a promising biomonitoring species, since it accumulates the contaminant in a considerable extent quite rapidly and at the same time it has a low metal retention capacity (low biological half-life) when exposed to clean sediments.

Mercury biomagnification in a contaminated estuary food web: effects of age and trophic position using stable isotope analyses.

The main aim of this study was to ascertain the biomagnification processes in a mercury-contaminated estuary, by clarifying the trophic web structure through stable isotope ratios. For this purpose, primary producers (seagrasses and macroalgae), invertebrates (detritivores and benthic predators) and fish were analysed for total and organic mercury and for stable carbon and nitrogen isotopic signatures. Trophic structure was accurately described by δ(15)N, while δ(13)C reflected the carbon source for each species. An increase of mercury levels was observed with trophic level, particularly for organic mercury. Results confirm mercury biomagnification to occur in this estuarine food web, especially in the organic form, both in absolute concentrations and fraction of total mercury load. Age can be considered an important variable in mercury biomagnification studies, and data adjustments to account for the different exposure periods may be necessary for a correct assessment of trophic magnification rates and ecological risk.

Kinetics of the PO4-P adsorption onto soils and sediments from the Mondego estuary (Portugal).

In order to assess the status of a North Atlantic Ocean Ecoregion (Mondego system, Portugal) after the implementation of a management programme, a study on PO4-P sorption was carried out. Considering that sorption velocity markedly determines the systems promptness to phosphorus external loading, the kinetics of the sorption of PO4-P onto estuarine sediments and the upstream agricultural soils was studied. The pseudo-second order kinetic equation gave an appropriate description of PO4-P sorption onto these soils and sediments, allowing for a quantitative comparison. For soils, the kinetic constants (k2*, µg(-1) g h(-1)) ranged between 0.007 and 0.017 µg(-1) g h(-1) while, in the case of sediments these ranged between 0.008 and 0.012 µg(-1) g h(-1). Results showed that the reduction of water residence time after the management action was relevant for the system status, affecting its reaction capacity to phosphorus loading.