Monitoring the Prestige oil spill impacts on some key species of the Northern Iberian shelf

Selected key components of the continental shelf benthic and demersal communities were monitored for the two years following the Prestige oil spill (POS) in order to identify the possible ecological effects of the oil. This work includes the first results regarding changes in abundance, distribution and food habits of hake (Merluccius merluccius), four-spot megrim (Lepidorhombus boscii), Norway lobster (Nephrops norvegicus) and Pandalid shrimp (Plesionika heterocarpus) populations of Galician and Cantabrian Sea shelves following the POS. Significant reductions in the abundance of Norway lobster, Plesionika heterocarpus and four-spot megrim were detected in the POS maximum impact area, located over the Galician shelf. Noteworthy recoveries were observed in the 2004 abundance indices of four-spot megrim and Plesionika. On the other hand, no significant effects were detected in the abundance or distribution of hake juveniles even though the tar aggregates were bound by the same oceanographic drift events as the hake recruits were during the winter of 2003 (Navidad current) in different water column layers of the Cantabrian Sea. Feeding patterns of the four species analysed did not present apparent modifications that can be related to the POS.     

Occurrence of perfluorinated compounds in water, sediment and mussels from the Cantabrian Sea (North Spain)

Perfluorinated compounds (PFCs) have emerged as significant global environmental pollutants with persistent, bioaccumulative and toxic properties. The aim of this study was to determine the occurrence of PFCs in water (wastewater, submarine emissaries and port-waters), sediment and transplanted mussels in estuarine areas of high urban and industrial impact from Northern Spain. Five PFCs of industrial use were studied: perfluorooctanesulfonate, perfluorohexanesulfonate, perfluorobutanesulfonate, perfluorooctanoate acid and perfluorononanoate acid. After selective extraction, samples were analyzed by Ultra Performance Liquid Chromatography coupled to tandem mass spectrometry. ΣPFCs ranged from 0.06 to 10.9 ng/L in water, with higher levels in wastewater treatment plants effluents and port waters than in submarine emissaries. Little accumulation was observed in sediments and mussels with ΣPFCs ranging from 0.01-0.13 ng/g dw and 0.01-0.06 ng/g ww, respectively. Most ubiquitous compounds were PFOS and PFOA. Mass fluxes of PFCs to the Cantabrian Sea are estimated and the impact to the coastal ecosystem is discussed.     

Aspects of the digestive gland cells of the mussel Mytilus galloprovincialis, in relation to lysosomal enzymes, lipofuscin presence and shell size: contribution in the assessment of marine pollution biomarkers

The present study investigates the histochemical localization of N-acetyl-β-hexozaminidase (Hex), acid phosphatase (AcP) and β-glucuronidase (β-Gus) in the digestive gland of mussels Mytilus galloprovincialis, as well as the clarification of suitable enzyme for biomarkers' application dealing with lysosomes. The results show more intense and homogenous localization of Hex, in relation to AcP and β-Gus and, thus, Hex histochemistry is supported as more suitable procedure for the evaluation of "lysosomal membrane stability" and "morphometrical alterations of lysosomes". The affection of lipofuscin granules on lysosomal enzymes' activity is also discussed. Additionally, the present study examines the response of small- and large-sized mussels M. galloprovincialis by assessing the "lysosomal membrane stability", "morphometrical alterations of lysosomes", "lysosomal response index (LRI)" and "structural epithelial changes in digestive tubules". The results indicate appreciable alterations of the above parameters in large-sized mussels, supporting their greater influence by the environmental factors, in relation to small-sized ones.     

Lysosomal membrane stability and metallothioneins in digestive gland of Mussels (Mytilus galloprovincialis Lam.) as biomarkers in a field study

The lysosomal membrane destabilization and the metallothionein content in the digestive gland cells of mussels (Mytilus galloprovincialis Lam.), collected along the east coast of the North Adriatic (Istrian and Kvarnerine coast, Croatia), were examined over a period of four years (1996–1999). The lysosomal membrane stability, as a biomarker of general stress, showed that the membrane labilization period in mussels from polluted, urban- and industrial-related areas was significantly decreased (p<0.05) when compared to mussels from control, clean sea water sites. In the harbour of Rijeka, the most contaminated site, the lysosomal membrane stability was reduced by more than 70% compared to the control. This method also proved to be a useful biomarker for detection of additional stress caused by short-term hypoxia that occurred once during this study inside the polluted and periodically quite eutrophic Pula Harbour. The concentration of metallothioneins in the mussel digestive gland, as a specific biomarker of exposure to heavy metals, did not reveal significant differences (p<0.05) between sites covered by this study.     

Spatial and temporal distribution of dissolved/dispersed aromatic hydrocarbons in seawater in the area affected by the Prestige oil spill

Seawater samples collected at three depths from 68 stations along the Northern Spanish coast were analysed for dissolved/dispersed petroleum aromatic hydrocarbons by UV-fluorescence and for 25 individual compounds by GC-MS. Sampling was performed in December 2002, just after the Prestige oil spill, and in February-March and September 2003. Higher concentrations of total aromatic hydrocarbons were found at all depths in the samples collected during December 2002 off the Galicia coast, with levels ranging between 0.19 and 28.8 microg/L eq. oil (0.1-4.8 microg/L chrysene eq.). These values decreased in the following cruises, till <0.05-2.86 microg/L oil eq. (av. 0.23 microg/L chrysene eq.) in September 2003, possibly representing the background levels for the region. However, in the Cantabrian coast they were still high at the surface in the March cruise, probably by the late arrival of the fuel-oil to this area. Some coastal hot spots were also identified, with values up to 29.2 microg/L fuel-oil eq., close to river mouths and urban areas. The individual PAH distributions in the December 2002 sampling off-Galicia were dominated by alkyl-naphthalene derivatives, consistently with the pattern distribution shown by the fuel-oil water accommodated fraction. The higher concentrations were found in the subsurface samples along the Costa da Morte, the area most heavily affected by the spill (av. 0.46 microg/L Sigma16 PAHs). The rest of the samples collected in other areas exhibited lower concentrations and a more even distribution of 2-4 ring PAHs, that ranged from 0.09 to 0.37 microg/L (av. 0.15 microg/L Sigma16 PAHs), with decreasing trends offshore and downward the water column. In September 2003, the values were rather uniform, averaging 0.09 microg/L (Sigma16 PAHs).     

Lysosomal Membrane Stability and Metallothioneins in Digestive Gland of Mussels (Mytilus galloprovincialis Lam.) as Biomarkers in a Field Study

The lysosomal membrane destabilization and the metallothionein content in the digestive gland cells of mussels (Mytilus galloprovincialis Lam.), collected along the east coast of the North Adriatic (Istrian and Kvarnerine coast, Croatia), were examined over a period of four years (1996–1999). The lysosomal membrane stability, as a biomarker of general stress, showed that the membrane labilization period in mussels from polluted, urban- and industrial-related areas was significantly decreased (p<0.05) when compared to mussels from control, clean sea water sites. In the harbour of Rijeka, the most contaminated site, the lysosomal membrane stability was reduced by more than 70% compared to the control. This method also proved to be a useful biomarker for detection of additional stress caused by short-term hypoxia that occurred once during this study inside the polluted and periodically quite eutrophic Pula Harbour. The concentration of metallothioneins in the mussel digestive gland, as a specific biomarker of exposure to heavy metals, did not reveal significant differences (p<0.05) between sites covered by this study.     

An ecotoxicological approach for the Boka Kotorska Bay (south-eastern Adriatic Sea): first evaluation of lysosomal responses and metallothionein induction in mussels

Mytilus galloprovincialis was used as a biomonitoring organism in Boka Kotorska Bay, a coastal transitional ecosystem in south-western Montenegro. Native mussels were collected in June 2008 at four sites thought to be differently impacted. Biological effects were investigated analysing both generic and specific biomarkers at cellular level (metallothionein content, lysosomal membrane stability, lipofuscin and neutral lipid accumulation, lysosomal structural changes). Trace element levels in mussels were quite low, only Cu and Zn exhibiting slight increases in the two sampling sites of Tivat Bay, the first one in front of a small working dockyard and the other in the water area of a former Naval dockyard. Mussels collected from these sites exhibited the highest values in neutral lipid and lysosomal volume density and the lowest neutral red retention times. Metallothionein content was always low, reflecting the minor body burden of the trace elements and suggesting a non-relevant environmental induction.     

Parasite communities in Boops boops (L.) (Sparidae) after the Prestige oil-spill: detectable alterations

Environmental pollution affects parasite populations and communities, both directly and through effects on intermediate and final hosts. In this work, we present a comparative study on the structure and composition of metazoan parasite communities in the bogue, Boops boops, from two localities (Galician coast, Spain) affected by the Prestige oil-spill (POS). We focus on the distribution of both individual parasite species and larger functional groupings by using both univariate and multivariate analyses. Our results indicate directional trends in community composition that might be related to the Prestige oil-spill disturbance of the natural coastal communities off Galicia. Endoparasite communities in B. boops reflected a notable change in the composition and abundance of the benthic fauna in the localities studied post-spill probably due to organic enrichment after the POS.     

Influences of ammonia-nitrogen and dissolved oxygen on lysosomal integrity in green-lipped mussel Perna viridis: laboratory evaluation and field validation in Victoria Harbour, Hong Kong

Lysosomal integrity in mussels has been applied as a biomarker to detect the pollution of trace organics and metals in the natural environments. However, few studies have examined the effects of water quality on the response of lysosomal integrity, in particular total ammonia-nitrogen (TAN) and dissolved oxygen (DO). This study demonstrated that high level of TAN (2.0 mg/l) and low DO (2.5 mg O₂/l) could significantly reduce the lysosomal integrity in green-lipped mussel Perna viridis, respectively by 33% and 38%, whereas the mussel lysosomal integrity decreased by 70% in the combined treatment of TAN and low DO under laboratory conditions after one week. The mussel lysosomal integrity of all treatment groups could return to the control level after a three week recovery period. In the field validation in Victoria Harbour, Hong Kong during an one-year study period, lysosomal integrity in P. viridis identified the cleanest site east to the harbour, where the lowest TAN and highest DO concentrations were found. While lysosomal integrity in mussels seemed not affected by seasonal changes, approximately 40% of the variation of this biomarker could be attributable to the changes in TAN and DO in seawater. In conclusion, the response of the mussel lysosomal integrity can be confounded by both TAN and DO prevailing in the natural environments and thus caution must be exercised in relating the observed changes in lysosomal integrity to any specific pollutant in coastal water quality monitoring studies.     

The Elbe flood 2002--toxic effects of transported contaminants in flatfish and mussels of the Wadden Sea

Cellular changes in livers from flounder (Platichthys flesus L.) and digestive glands of blue mussels (Mytilus edulis) were studied to analyze whether the Elbe flood catastrophe in August 2002 had any deleterious effects on animal health. Flatfish and mussels were sampled at different locations in the Elbe estuary and the Wadden Sea after the flood wave had reached the German Bight. Internationally used core biomarkers (lysosomal membrane stability, biotransformation enzymes) were applied to assess the toxic effects of putatively increased pollution levels. In comparison to earlier data from long-term studies at the same stations, we found a significant impairment in the function of cell organelles (lysosomes) involved in the detoxification and elimination of pollutants in fish liver. Concentrations of relevant contaminants (organochlorines, polychlorinated biphenyls) were analyzed in parallel with cellular biomarkers, and conspicuously raised concentrations of insecticides metabolites were detected. Cell recovery and a clear reduction in contaminant concentrations were observed in fish livers five months after the flood at all sampling sites except the Helgolander Tiefe Rinne.     

An ecotoxicological approach for the Boka Kotorska Bay (south-eastern Adriatic Sea): First evaluation of lysosomal responses and metallothionein induction in mussels

Mytilus galloprovincialis was used as a biomonitoring organism in Boka Kotorska Bay, a coastal transitional ecosystem in south-western Montenegro. Native mussels were collected in June 2008 at four sites thought to be differently impacted. Biological effects were investigated analysing both generic and specific biomarkers at cellular level (metallothionein content, lysosomal membrane stability, lipofuscin and neutral lipid accumulation, lysosomal structural changes). Trace element levels in mussels were quite low, only Cu and Zn exhibiting slight increases in the two sampling sites of Tivat Bay, the first one in front of a small working dockyard and the other in the water area of a former Naval dockyard. Mussels collected from these sites exhibited the highest values in neutral lipid and lysosomal volume density and the lowest neutral red retention times. Metallothionein content was always low, reflecting the minor body burden of the trace elements and suggesting a non-relevant environmental induction.     

Relationship between Quantitative Indices of Individual and Digestive Cell Conditions in the Common Mussel,Mytilus edulis L., from the Biscay Coast

Some data based on quantitative measurements at the individual and histological levels have been compared in four different populations of Mytilus edulis from the Biscay coast. Each population has been divided into two groups, corresponding to animals living at high and low tide levels. Biometric measurements show evident differences between samples from these two groups at all the studied stations. This should be the result of a distinct adaptation to hydric stress. On the other hand, a positive correlation between the flesh condition (FC) index: and the mean epithelial thickness (MET) of digestive tubules has been found. So, authors conclude that both FC and MET are good populational stress indices, and the use of both parameters simultaneously can be helpful in the assessment of environmental stress.     

AOX‐Bildung bei der sonolytischen Oxidation von Salicylsäure in Gegenwart von Chlorid

AOX‐formation by the Sonochemical Treatment of Salicylic Acid in Presence of Chloride Ultrasound shows great potential for improving water, wastewater and sludge treatment processes. However, a number of questions exist: for example the influence of suspended solids or salts. In this paper the influence of chloride in view of AOX‐formation should be investigated. As organic model compound salicylic acid (0.05…1 mmol/L) was used. Formation of AOX (adsorbable organically bound halogens) was measured in dependence upon chloride concentrations (1.4…141 mmol/L) and pH 2 and 9. Ultrasound irradiation was performed in glass reactor (500 mL) at 206 kHz and 353 kHz by continuous bubbling argon:oxgen (4:1) at a flow rate of 1 L/min into the solutions. The elimination of salicylic acid and the formation of hydrogen peroxide were not influenced by the presence of chloride (up to 56 mmol/L). The rate of salicylic acid elimination and the formation rate of hydrogen peroxide are at concentrations of mmol/(L min) levels but those of AOX formation of μmol/(L min) levels. The reactions leading to chlorinated by‐products can be seen as minor reaction paths. The yield of AOX depending on frequency, pH and chloride concentration lies between 10 μg/L and 900 μg/L. In acidic medium they were twice as high as in basic medium. Below the ratio chloride : salicylic acid of 2 mol/mol, AOX could not be identified.     

Lysosomal membrane stability and metallothionein content in Mytilus galloprovincialis (L.), as biomarkers. Combination with trace metal concentrations

The simultaneous study of two biomarkers, the lysosomal membrane stability (LMS) of the digestive gland and the metallothionein (MT) content of the digestive gland, the gills and the mantle/gonad complex of the mussel Mytilus galloprovincialis was examined in an enclosed estuarine system in June and October 2001. Mussels were collected along the Gulf of Thermaikos (northern Greece) from stations displaying a pollution gradient, while Olympiada in the Gulf of Strymonikos was used as a reference station. In addition, the heavy metal (Cd, Pb, Cu and Zn) content, using atomic absorption spectrophotometry (AAS), were evaluated in the digestive gland, the gills and the mantle/gonad complex of mussels collected from the same sites and seasons. LMS values were significantly greater, and the MT content of the studied tissues were significantly less in mussels collected from the reference station compared to mussels from more polluted stations located in the Gulf of Thermaikos. Significant correlation was observed between the MT content of the gills and the mantle/gonad complex with the LMS values.     

Monitoring biomarkers in fish (Lepidorhombus boscii and Callionymus lyra) from the northern Iberian shelf after the Prestige oil spill

Hepatic biomarker responses were measured in two demersal fish species (Lepidorhombus boscii and Callionymus lyra) from the northern Iberian shelf associated with the massive Prestige oil spill (POS), five months after the accident. The biomarkers selected were 7-ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), glutathione reductase (GR), catalase (CAT), and DNA integrity. Interspecies differences and spatial variations in biomarker responses were observed along the shelf. GST, GR and CAT activities were significantly elevated in L. boscii in the most oil impacted area (Finisterre) and positively correlated (p<0.05) with POS tar aggregate densities. The lack of previous data from the area together with the existence of chronic background pollution of the shelf implies that the observed biomarker responses cannot be solely attributed to the petroleum hydrocarbon components of the spilled oil. This first biological effect assessment showed that L. boscii is a potentially suitable target species to be used in future biomonitoring programmes along the northern Iberian shelf.     

Recent trends of extreme precipitation indices in the Iberian Peninsula using observations and WRF model results

Spatial and temporal distributions of the trends of extreme precipitation indices were analysed between 1986 and 2005, over the Iberian Peninsula (IP). The knowledge of the patterns of extreme precipitation is important for impacts assessment, development of adaptation and mitigation strategies. As such, there is a growing need for a more detailed knowledge of precipitation climate change.This analysis was performed for Portuguese and Spanish observational datasets and results performed by the Weather Research and Forecast (WRF) model forced by the ERA-Interim reanalysis. Extreme precipitation indices recommended by the Expert Team for Climate Change Detection Monitoring and Indices were computed, by year and season. Then, annual and seasonal trends of the indices were estimated by Theil-Sen method and their significance was tested by the Mann-Kendal test. Additionally, a second simulation forced by the Max Planck Institute Earth System Model (MPI-ESM), was considered. This second modelling configuration was created in order to assess its performance when simulating extremes of precipitation.The annual trends estimated for the 1986–2005, from the observational datasets and from the ERA-driven simulation reveal: 1) negative statistically significant trends of the CWD index in the Galicia and in the centre of the IP; 2) positive statistically significant trends of the CDD index over the south of the IP and negative statistically significant trends in Galicia, north and centre of Portugal; 3) positive statistically significant trends of the R75p index in some regions of the north of the IP; 4) positive statistically significant trends in the R95pTOT index in the Central Mountains Chain, Leon Mountains and in the north of Portugal.Seasonally, negative statistically significant trends of the CWD index were found in Galicia, in winter and in the south of the IP, in summer. Positive statistically significant trends of the CWD index were identified in the Leon Mountains, in spring, and in Galicia, in autumn. For the CDD index, negative statistically significant trends were seen in Valencia, in the spring, and, in Galicia and Portugal (north and centre), in summer. Positive statistically significant trends of the CDD index were found: in the east of the IP, in the winter; in the Cantabrian Mountain, in the spring; and, in the south of the IP, in summer. Regarding to the R75p index, negative statistically significant trends were found in Galicia, in winter and positive statistically significant trends in the north of Portugal, in spring and in the Central Mountains Chain and north of Portugal, in autumn. For the R95pTOT index, negative statistically significant trends were found over the Sierra Cuenca and Sierra Cazorla, in winter and positive statistically significant trends were found over the Sierra Cebollera, in winter and in Castile-la Mancha region, in spring.The results of the annual and seasonal trends of the extreme precipitation indices performed for observational datasets and the simulation forced by ERA-Interim, are similar. The results obtained for the simulation forced by MPI-ESM are not satisfactory, and can be a source of criticism for the use of simulation forced by MPI-ESM in this type of climate change studies. Even for the relatively short period used, the WRF model, when properly forced is a useful tool due to the similar results of Portuguese and Spanish observational datasets and the simulation forced by ERA-Interim.     

Composition and daytime vertical distribution of the ichthyoplankton assemblage in the Central Cantabrian Sea shelf, during summer: An Eulerian study

During summer, wind driven coastal upwelling dominates in the Central Cantabrian Sea (southern Bay of Biscay). Nevertheless, atmospheric forcing is highly variable and wind pulses may cause noticeable and fast hydrographic responses in the shelf region. In this paper, the composition and vertical distribution of the summer ichthyoplankton assemblage during the daytime at a fixed station, located on the Central Cantabrian Sea shelf, are documented. Also, the impact of a short-time scale hydrographic event on the abundance and structure of the larval fish assemblage is examined. Significant small-scale temporal hydrographic variability was observed. Currents showed changes in speed and direction and significant changes in thermocline depth were also observed. A total of 34 taxa of fish larvae were identified. Engraulis encrasicolus eggs and larvae of the shelf-dwelling species Trachurus trachurus, Capros aper and E. encrasicolus dominated the ichthyoplankton assemblage. The distribution of E. encrasicolus eggs and fish larvae was vertically structured. E. encrasicolus egg concentration increased exponentially towards the surface. Fish larvae showed a subsurface peak of concentration and their vertical distribution was not conditioned by thermocline depths. The short term hydrographic event did not affect the vertical distribution of fish larvae but it accounted for significant temporal changes in larval fish assemblage structure and abundance. Results suggest that temperature and light intensity are important factors in the vertical distribution of fish larvae. They also indicate that the temporal monitoring of the larval fish assemblage in this region requires multiple sampling sites.     

Induction, adaptation and recovery of lysosomal integrity in green-lipped mussel Perna viridis

Biomarkers are generally applied to detect pollution in environmental monitoring. Such biological responses should accurately reflect the stress over time in a quantitative manner. As such, the initial and maximum responses induced by stress, as well as adaptation and recovery of these biomarkers, need to be fully understood or else erroneous false-negative or false-positive may be arrived. However, most of the biomarker studies only provided information on initially induced responses under different concentrations of toxicants, while biological adaptation and recovery were poorly known. In this study, the time required for induction, adaptation and recovery of lysosomal integrity in green-lipped mussel Perna viridis upon exposure to benzo[a]pyrene was investigated over a period of 62 days. Maximum induction occurred on day 6 when lysosomal integrity was significantly reduced by 51%, and no further change or adaptation was detected thereafter. When mussels were depurated in clean seawater after 18 days of exposure to benzo[a]pyrene, a gradual recovery was observed, with lysosomal integrity returning to its background level and showing a complete recovery after 20 days of depuration. Lysosomal integrity was significantly correlated with the body burden concentrations of benzo[a]pyrene and condition index of the mussels. The relatively fast induction (6 days) and recovery (20 days) without apparent adaptation suggested that lysosomal integrity in P. viridis can serve as a good biomarker in biomonitoring, as its response is not likely to generate both false-negative and false-positive results.     

Bioaccumulation depuration kinetics and effects of benzo(a)anthracene on Mytilus galloprovincialis

Polycyclic aromatic hydrocarbons (PAHs) are important environmental pollutants due to their persistence and bioaccumulation potential both in organisms and in sediments. In this study, bioaccumulation and depuration experiments were performed employing local Mediterranean mussel species, Mytilus galloprovincialis, with two biomarkers: filtration rate and lysosomal stability (neutral red retention) assay. Benzo(a)anthracene (BaA) was chosen as the model PAH compound due to its common presence in several matrices in the marine environment. Bioconcentration Factors (BCFs) for the mussels exposed to different BaA concentrations were calculated from both kinetic rate constants and from the experimental data and found between 27-3184 and 16-2745, respectively. Experimental QSAR values were found comparable to the previous QSAR models created for BaA. The effect of BaA exposure on the mussels showed a more clear dose-response relationship according to the results of lysosomal stability compared to the filtration rate.     

Bioaccumulation depuration kinetics and effects of benzo(a)anthracene on Mytilus galloprovincialis

Polycyclic aromatic hydrocarbons (PAHs) are important environmental pollutants due to their persistence and bioaccumulation potential both in organisms and in sediments. In this study, bioaccumulation and depuration experiments were performed employing local Mediterranean mussel species, Mytilus galloprovincialis, with two biomarkers: filtration rate and lysosomal stability (neutral red retention) assay. Benzo(a)anthracene (BaA) was chosen as the model PAH compound due to its common presence in several matrices in the marine environment. Bioconcentration Factors (BCFs) for the mussels exposed to different BaA concentrations were calculated from both kinetic rate constants and from the experimental data and found between 27-3184 and 16-2745, respectively. Experimental QSAR values were found comparable to the previous QSAR models created for BaA. The effect of BaA exposure on the mussels showed a more clear dose–response relationship according to the results of lysosomal stability compared to the filtration rate.