Beatriz Echavarri-Erasun

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.

Assessing the effects of treated and untreated urban discharges to estuarine and coastal waters applying selected biomarkers on caged mussels

To assess effects of urban discharges, biomarkers were measured in caged mussels in northern Iberian Peninsula. Lysosomal membrane stability and histopathology of gonad and digestive gland were analysed as general effect biomarkers. Exposure to specific pollutants was evaluated by autometallographical detection of metals, peroxisomal acyl-CoA oxidase activity, micronucleus test and transcription levels of vitellogenin and MT20 genes. Health status of mussels was impaired after 3 days of caging at the untreated outfall discharge and at the waste water treatment plant effluent discharge to the estuary. The most relevant finding was the significant up-regulation of vitellogenin gene transcription in male mussels exposed to the untreated outfall discharge. Metals and xenoestrogenic endocrine disruptors were bioavailable in some discharges and disturbed the health status of mussels. Biomarkers were effective in the assessment of effects of urban discharges and could be implemented in operative controls required to assess the risks associated to effluent discharges.

Medium-term assessment of the effects of the Prestige oil spill on estuarine benthic communities in Cantabria (Northern Spain, Bay of Biscay)

A specific monitoring program was implemented in the estuaries of Cantabria (northern Spain) in order to assess the medium-term effects (2003-2005) of the Prestige oil spill (POS) on benthic macroinvertebrate communities. A control-impact design was adopted, examining four unaffected and five oil-affected estuaries. High mortalities and significant differences in overall richness and diversity between the control and impacted estuaries were not detected. Some changes in the temporal evolution of species abundance were observed for some key species, but these could not be related to the spillage event. It was not possible to ensure that low magnitude effects had not occurred, due to the high range of natural variability of benthic communities, the confounding effects of other contamination sources and the absence of previous reference conditions.

The combined use of the PLHC-1 cell line and the recombinant yeast assay to assess the environmental quality of estuarine and coastal sediments.

Sediment contamination poses a potential risk for both ecosystems and human health. Risk assessment is troublesome as sediments contain complex mixtures of toxicants, and traditional chemical analyses can neither provide information about potential hazards to organisms nor identify and measure all present contaminants. This work combines the use of the PLHC-1 cell line and the recombinant yeast assay (RYA) to assess the environmental quality of estuarine and coastal sediments. The application of multiple endpoints (cytotoxicity, generation of oxidative stress, presence of CYP1A inducing agents, micronucleus formation and estrogenicity) revealed that the organic extracts of those sediments affected by industrial activities or collected near harbours and untreated urban discharges showed significant cytotoxicity, micronuclei and CYP1A induction. The study highlights the usefulness of the applied bioassays to identify those sediments that could pose risk to aquatic organisms and that require further action to improve their environmental quality.

Measuring biological responses at different levels of organisation to assess the effects of diffuse contamination derived from harbour and industrial activities in estuarine areas

To evaluate the effects of diffuse contamination, biological measurements were applied in a scrap cargo harbour, a marina and an industrial area. Metal accumulation and biomarkers (survival in air, digestive gland and gonad histopathology, lysosomal membrane stability, intralysosomal metal accumulation, transcription of vitellogenin and MT20, peroxisome proliferation and micronuclei formation) were measured in transplanted mussels, together with metrics of benthic invertebrates. Benthic species were classified into ecological groups and univariate indexes were calculated. The marina showed high richness (16) and percentage of opportunistic species (55.1%) and low metal accumulation. Mussels in the scrap cargo harbour showed high metal accumulation, up-regulation of MT20 transcription, reduced health status (LP<6 min) and increased micronuclei frequencies (up to 11.3‰). At the industrial area, low species richness (4) and badly organised assemblages were detected and chemical analyses indicated significant amounts of bioavailable metals. Overall, selected biological measurements showed potential for the assessment of diffuse contamination.

Coastal outfalls, a sustainable alternative for improving water quality in north-east Atlantic estuaries.

The city of Santander ceased the discharge of sewage effluents into the bay of Santander in June, 2001 and began discharging at a site 2.4 km offshore in the nearby coastal area (Virgen del Mar, Bay of Biscay) at a water depth of about 40 m. The present study investigates the effects of the new outfall discharges on the water quality of the high-energy coastal area and the recovery of the perturbed temperate estuarine area now only affected by combined sewer overflows (CSOs). Nutrients, phytoplankton biomass and urban pollution indicators were analysed. No significant spatial or temporal change in water quality variables was found in the coastal area around the outfall. No signs of nutrification or increases in chlorophyll-a were observed throughout the study period, although a slight increase in phosphates, suspended solids and turbidity were observed two years after the relocation of the discharge. These changes were not attributed to outfall discharge but to a regional increase also observed at control stations and nearby coastal areas. Considerable reductions in indicators of urban discharges were observed in the estuary after the relocation of discharges, even at stations located around CSOs. Results from this study support the efficiency of ecological quality-driven designs of sanitation systems, which are used as management tools for sensitive and environmentally valuable coastal ecosystems in the north-east Atlantic.

Relationships between lines of evidence of pollution in estuarine areas: Linking contaminant levels with biomarker responses in mussels and with structure of macroinvertebrate benthic communities.

Data obtained in a pollution survey performed in estuarine areas were integrated using multivariate statistics. The sites selected for the study were areas affected by treated and untreated urban discharges, harbours or industrial activities as well as reference sites. Mussels were transplanted to each site and after different times of exposure, samples of water, sediments and mussels were collected. Biomarkers were analysed on mussels after 3 and 21 days of transplant whereas concentrations of contaminants were measured in water, sediments and mussels after 21 days of transplant. The structure of macroinvertebrate benthic communities was studied in sediment samples. Studied variables were organised into 5 datasets, each one constituting a line of evidence (LOE): contaminants in water, contaminants in sediments, contaminants accumulated by transplanted mussels, biomarkers in transplanted mussels and changes in the structure of macroinvertebrate benthic communities of each sampling site. Principal Component Analysis (PCA) identified the variables of each LOE best explaining variability among sites. In order to know how LOEs relate to each other, Pearsons correlations were performed. Contaminants in sediments were not correlated with the rest of LOEs. Contaminants in water were significantly correlated with contaminants and biomarkers in mussels and with structure of macroinvertebrate benthic communities. Similarly, significant correlations were found between contaminants and biomarkers in mussels and between biomarkers in mussels and structure of macroinvertebrate benthic communities. In conclusion, biomarker responses give relevant information on pollution in estuarine areas and provide a link between chemical and ecological statuses of water bodies in the context of the Water Framework Directive.

Assessment of the effects of discontinuous sources of contamination through biomarker analyses on caged mussels

The present study analysed potential adverse effects of discontinuous sources of contamination, namely the discharge of a combined sewer overflow (CSO) and of runoff in an urban area, the Bay of Santander (North Iberian Peninsula). Water samples and caged mussels were used to analyse concentrations of contaminants and biological responses. Mussels (Mytilus galloprovincialis) were transplanted to a marina receiving runoff from a petrol station and to a CSO discharge site. Samples were collected in synchrony with heavy rains along 62days. Lysosomal membrane stability (LMS) and acyl-CoA oxidase (AOX) activity were measured as core biomarkers and were analysed at all sampling times. Histopathology of digestive gland and gonads, transcription levels of vitellogenin gene, volume density of black silver deposits and micronuclei formation were measured at initial and final stages of the transplant. Chemical analyses of metals, polycyclic aromatic hydrocarbons (PAHs) and endocrine disruptors were performed in water samples and mussel flesh. Mussels accumulated low concentrations of contaminants, which is in accordance with results obtained from exposure biomarkers. AOX activity decreased in all transplanted mussels after the first heavy rain, but this change seems to be related to the seasonal pattern of the enzyme activity. Mussels located close to the CSO discharge site showed a reduction in LMS after the first rain event, when compared to mussels before the transplant and to mussels from the reference location. However, this was attributable to natural environmental changes rather than to pollution. Values of the rest of analysed biomarkers were below threshold values reported for the study area.