Tributyltin pollution biomonitoring under the Water Framework Directive: Proposal of a multi-species tool to assess the ecological quality status of EU water bodies

The Water Framework Directive (WFD) is a key legislative action developed by the European Union in order to protect aquatic ecosystems. One of the concerning pollutants, listed in this directive as a priority hazardous substance, is tributyltin (TBT), a biocide largely used in antifouling paints and identified as a causative agent of imposex/intersex in gastropods. In order to integrate TBT pollution monitoring within this legislative framework, a practical exercise is here proposed to assess the evolution of surface water ecological status in Ria de Aveiro (NW Portugal). Three bioindicators – the caenogastropods Nucella lapillus, Nassarius reticulatus and Littorina littorea – were used under the general WFD benthic invertebrate quality element, and the vas deferens sequence index (VDSI) and the intersex index (ISI) were selected as biomarkers for the purpose of assessing the condition of this quality element regarding the impact of TBT pollution. Levels of VDSI in N. lapillus and N. reticulatus, and ISI in L. littorea, were surveyed in 2013 and compared with previous data available for the same species and study area in 1998 and 2005, providing a time lapse for a period of 15 years. VDSI and ISI values were converted into Ecological Quality Ratios (EQR) and EQR boundaries were set for each species in order to define the five ecological status classes (High, Good, Moderate, Poor and Bad). We propose N. lapillus as key bioindicator, however the combined use of further species is very useful to cover a wider study area. Based on the proposed method, it is concluded that the ecological status of the surface waters surveyed in Ria de Aveiro, concerning the impact of TBT pollution on the above benthic invertebrate taxa, improved considerably since 1998 and achieved a Good Ecological Status in 2013, thus meeting the WFD environmental objectives for this priority hazardous substance even before 2015.     

On the hoplonemertean,Kameginemertes parmiornatus (Iwata, 1957) gen. n., comb. n. from Sagami Bay,Japan

Two different tributyltin (TBT)-induced virilisation phenomena in prosobranch snails – intersex in Littorina littorea and imposex in Nucella lapillus – are compared in order to facilitate their simultaneous use in geographical large scale effect monitoring surveys. Imposex in dogwhelks is a more sensitive biomarker and should be used in areas that are only slightly or moderately contaminated with TBT (ambient TBT concentrations < 2.0 ng as Sn l-1). The assessment of intersex intensities in periwinkle populations has considerable advantages in areas with higher TBT concentrations and should be used also wherever dogwhelks are absent irrespective of the TBT exposure level. The intersex index (ISI) and vas deferens sequence (VDS) index are proposed as the most suited parameters for effect monitoring purposes. The geographical uniformity of intersex and imposex is analysed and proven for the coasts of Ireland, France, and Germany. A relative loss of TBT sensitivity in females can be found, but to a varying extent in both species. The implications of this result for biological TBT effect monitoring programmes are discussed in light of the fact that intersex and imposex have both been found to be irreversible. Because it is the objective of these programmes to assess current TBT contaminations and resulting biological effects, only relatively young specimens should be considered in the sampling strategy.     

Imposex and butyltin burden in Bolinus brandaris (Mollusca, Gastropoda) and sediment from the Tunisian coast

The present study aimed at analyzing the imposex incidence and the presence of butyltins namely tributyltin (TBT) with its di- and mono-substituted metabolites in Bolinus brandaris whole tissues and in surface sediments at seven sites from the Tunisian coast during one campaign in May 2010. Butyltin levels were evaluated using isotope dilution GC–MS. Except the population collected from Zarat site, imposex was found in snails from the remained six sites with a maximal incidence and sterility (closure of the vaginal opening) registered in Carrier bay. Both imposex indices VDSI and RPLI showed a positive correlation with tissue concentrations of TBT. Total butyltin concentrations in sediments were higher in sites located in the vicinity of shipping areas with levels of TBT high enough to cause environmental concern if there is no legislative restriction and enforcement for the sale and use of these chemicals in Tunisia. These results further confirmed that B. brandaris is a good bioindicator of butyltin pollution in the studied areas. In addition, this study provided recent and new data on sediment butyltin concentrations that could serve for long-term monitoring of TBT pollution in Tunisia and the Mediterranean Sea.     

Quantifying uncertainties in biologically-based water quality assessment: A pan-European analysis of lake phytoplankton community metrics

Lake phytoplankton are adopted world-wide as a sensitive indicator of water quality. European environmental legislation, the EU Water Framework Directive (WFD), formalises this, requiring the use of phytoplankton to assess the ecological status of lakes and coastal waters. Here we provide a rigorous assessment of a number of proposed phytoplankton metrics for assessing the ecological quality of European lakes, specifically in response to nutrient enrichment, or eutrophication, the most widespread pressure affecting lakes. To be useful indicators, metrics must have a small measurement error relative to the eutrophication signal we want them to represent among lakes of different nutrient status. An understanding of variability in metric scores among different locations around a lake, or due to sampling and analytical variability can also identify how best this measurement error is minimised.To quantify metric variability, we analyse data from a multi-scale field campaign of 32 European lakes, resolving the extent to which seven phytoplankton metrics (including chlorophyll a, the most widely used metric of lake quality) vary among lakes, among sampling locations within a lake and through sample replication and processing. We also relate these metrics to environmental variables, including total phosphorus concentration as an indicator of eutrophication.For all seven metrics, 65–96% of the variance in metric scores was among lakes, much higher than variability occurring due to sampling/sample processing. Using multi-model inference, there was strong support for relationships between among-lake variation in three metrics and differences in total phosphorus concentrations. Three of the metrics were also related to mean lake depth. Variability among locations within a lake was minimal (<4%), with sub-samples and analysts accounting for much of the within-lake metric variance. This indicates that a single sampling location is representative and suggests that sub-sample replication and standardisation of analyst procedures should result in increased precision of ecological assessments based upon these metrics.For three phytoplankton metrics being used in the WFD: chlorophyll a concentration, the Phytoplankton Trophic Index (PTI) and cyanobacterial biovolume, >85% of the variance in metric scores was among-lakes and total phosphorus concentration was well supported as a predictor of this variation. Based upon this study, we can recommend that these three proposed metrics can be considered sufficiently robust for the ecological status assessment of European lakes in WFD monitoring schemes.     

An appraisal of a biocontamination assessment method for freshwater macroinvertebrate assemblages; a practical way to measure a significant biological pressure?

Freshwater invasive or alien species (IAS) can have a major impact on benthic macroinvertebrate assemblage structure and diversity. This has implications for accurate biological monitoring, the assessment of the ecological quality status of rivers and achievement of Water Framework Directive (WFD) objectives. Although IAS constitutes a major biological pressure to WFD objectives, current approaches to ecological status assessment tend to ignore their presence. This problem is compounded as biotic indices such as the Biological Monitoring Working Party (BMWP) score do not distinguish between native and IAS, when IAS tend to be more tolerant of organic pollution than the natives they replace. Biocontamination is the presence of an IAS in a system, and we tested a new method of biocontamination assessment, designed to be used alongside current routine water quality monitoring techniques, by applying it to biological monitoring data from the river monitoring programme of a small Island, The Isle of Man. Although 54% of monitoring sites exhibited no biocontamination, 19% showed low or moderate biocontamination and 27% high or severe biocontamination. Richness contamination was low (only two contaminated families being recorded), but abundance contamination was high in some sites (87% of individuals being IAS). Sites with a greater relative abundance of IAS individuals exhibited lower BMWP water quality. Within invaded sites BMWP monitoring was not responsive to changing chemical water quality, whereas within uninvaded sites it was. In invaded sites, the relative abundance of IAS increased as ammonia and BOD₅ increased. Our study shows current monitoring approaches mask the presence of AIS within assemblages, with some highly biocontaminated sites registering high BMWP biological quality. This new index represents a simple way to integrate the IAS biological pressure into established WFD monitoring programmes, to produce more comprehensive estimates of ecological quality status than are currently being realised.     

On the Use of Mixture Toxicity Assessment in REACH and the Water Framework Directive: A Review

This review seeks to connect the scientific theory of mixture toxicity to its implementation within different regulatory frameworks. The aim is to demonstrate how mixture toxicity assessment can be more thoroughly integrated into the European chemical regulations, REACH, and the Water Framework Directive (WFD), using the experiences gained through other regulatory frameworks. The article consists of (1) an examination of the scientific underpinnings of the common mixture toxicity assessment methods; (2) a discussion of how these methods have been used in regulatory frameworks; and (3) a discussion of how the methods could be applied within REACH and the WFD. It is concluded that concentration addition should be applied as a default model for mixture toxicity assessment. Furthermore, it is concluded that REACH and the WFD only include mixture toxicity assessments in specific situations. However, it is shown that it is scientifically feasible and regulatorally practicable to integrate a more holistic mixture toxicity approach into both legislations. In this connection, the experience gained from the U.S. frameworks on mixture toxicity assessment could be useful. The construction of a database that includes data on chemicals in the European environment could be used for mixture toxicity assessment of the chemicals with individual PEC/PNECs > 0.1.     

Integrated Constructed Wetlands (ICW) working at the landscape scale: The Anne Valley project,Ireland

Availability of clean water in Europe has become a topic of great concern. The Water Framework Directive (WFD) is putting pressure on EU Member States to provide water quality of a high standard throughout the Union. An effective way of tackling this problem is to use Integrated Constructed Wetlands (ICWs) at the landscape scale. Over the last 23 years, 15 ICWs have been constructed within the Anne Valley catchment in Waterford, Ireland, to address both point-source and diffuse pollution at the catchment scale. ICWs address a diversity of pollution problems including the treatment of sewage effluent from small villages, wastewater from livestock and waste material from creameries. A series of aerial photographs shows the development of these wetlands within the catchment over time. A catchment scale GIS is currently being developed in Ireland to address the WFD concerns. The integration of ICWs into these catchment models will provide a mechanism for measuring their effectiveness at the landscape scale and for identifying strategic sites for their implementation.     

A review on the ecological quality status assessment in aquatic systems using community based indicators and ecotoxicological tools: what might be the added value of their combination?

The European Water Framework Directive (WFD) represents a transformation of the guidelines for water quality assessment and monitoring across all EU Member States. At present, it is widely accepted that the WFD requires holistic and multidisciplinary ecological approaches by integrating multiple lines of evidence. Within the scope of the WFD, the scientific community identified clear opportunities to take advantage of an ecotoxicological line of evidence. In this context, ecotoxicological tools, namely biomarkers and bioassays, were proposed to contribute to the integration of the chemical and biological indicators, and thus to provide an overall insight into the quality of a water body. More than one decade after the publication of the WFD, we reviewed the studies that have attempted to integrate ecotoxicological tools in the assessment of surface water bodies. For this purpose, we reviewed studies providing an ecological water status assessment through more conventional community based approaches, in which biomarkers and/or bioassays were also applied to complement the evaluation. Overall, from our review emerges that studies at community level appear suitable for assessing the ecological quality of water bodies, whereas the bioassays/biomarkers are especially useful as early warning systems and to investigate the causes of ecological impairment, allowing a better understanding of the cause–effect-relationships. In this sense, community level responses and biomarkers/bioassays seem to be clearly complementary, reinforcing the need of combining the approaches of different disciplines to achieve the best evaluation of ecosystem communities’ health.     

The Water Framework Directive’s “percentage of surface water bodies at good status”: unveiling the hidden side of a “hyperindicator”

The Water Framework Directive (WFD) has provided the means of standardizing the way surface water bodies are monitored throughout the European Union (EU), using a common evaluation measure, the percentage of surface water bodies at good status, based largely on the structure and functioning of aquatic ecosystems. However, the evaluation of good status is based on the way the WFD is implemented, which differs in each country. In this article, we analyze how the WFD is implemented in France, how the water agencies divide up the water bodies, the areas covered by their monitoring networks, and the modalities of obtaining data to provide the EU with the percentage of water bodies at good status. This analysis reveals that it is this hyperindicator itself that is at stake, obtained by successively aggregating values measured in time and space, from the monitoring station to the River Basin District (RBD), reducing vast amounts of information to a single measure per RBD, while long-term monitoring of the major European rivers and their sedimentary budgets, which show improvements in certain quality aspects, are largely overlooked by the WFD. When drawing up the indicator, the agencies identify certain biases but not others. This raises the question of its use and relevance for managers and politicians, at both national and European level.     

Use of marker pigments and functional groups for assessing the status of phytoplankton assemblages in lakes

Phytoplankton is a key biological quality element for the establishment of the Water Framework Directive (WFD) ecological status in reservoirs and lakes. In freshwaters, inverted microscope examination is the traditional standard method for estimating phytoplankton and assessing taxonomic composition. Based on the enumeration of algal units and measurements for biovolume calculation, this technique is cumbersome and time-consuming. In large monitoring programmes, such as the application of the WFD in lakes and reservoirs, chemotaxonomy (HPLC pigment analysis and CHEMTAX treatment) is ideally suited as an alternative method because it allows the rapid processing of large numbers of samples from numerous locations and depths, thereby providing ideal temporal and spatial resolution. The low taxonomical detail obtained by HPLC and CHEMTAX (phytoplankton classes or phyla) can easily be overcome by a rapid inverted microscope screening with identification of the dominant species. Combining HPLC and microscopy provides a useful method for monitoring phytoplankton assemblages, which can be used to implement the WFD with respect to phytoplankton. Here, we present the application of a method combining marker pigments and microscopy to phytoplankton samples from 12 Belgian reservoirs. This method substantially reduced the workload and enabled us to assess the status of the phytoplankton assemblage in these lakes. The method complies with the WFD, as it takes into account taxonomic composition, assesses abundance and biomass of the phytoplankton taxa, and easily detects blooms. Additionally, a set of templates of probability of occurrence of phytoplankton functional groups at the maximal ecological potential for reservoirs from the Central/Baltic region is presented, based on reference conditions defined for natural lakes from other regions.     

Atlantic Island freshwater ecosystems: challenges and considerations following the EU Water Framework Directive

The Water Framework Directive or WFD constitutes a major step forward in the protection of the aquatic environment and associated habitats, since it legislates for the characterization of surface water bodies across defined ecoregions and the development of ecological monitoring systems based upon elements of the aquatic biota. The Macaronesian archipelagos include the Azores, Madeira and the Canary Islands. The peripheral situation of the Macaronesian islands has set them apart from many European initiatives concerning the implementation of the WFD, which is biased towards better known continental systems. As a result, they have been included in the same ecoregion as the Iberian Peninsula and the Balearic Islands for management purposes. However, because of their oceanic situation and volcanic origin, the freshwater systems of the Macaronesian islands differ strongly from continental systems in watershed morphology and the composition of the biotic assemblages, which merits separate identification rather than inclusion within the larger Iberic-Macaronesian ecoregion and special attention concerning regional implementation of programmes of measures under the Directive.This paper has not been submitted elsewhere in identical or similar form, nor will it be during the first three months after its submission to Hydrobiologia.     

The suitability of the marine biotic index (AMBI) to new impact sources along European coasts

In recent years, several benthic biotic indices have been proposed to be used as ecological indicators in estuarine and coastal waters. One such indicator, the AZTI Marine Biotic Index (AMBI), was designed to establish the ecological quality of European coasts. The index examined the response of soft-bottom benthic communities to natural and man-induced disturbances in coastal and estuarine environments. It has been successfully applied to different geographical areas and under different impact sources, with increasing user numbers in European marine waters (Baltic, North Sea, Atlantic and Mediterranean). The AMBI has been used also for the determination of the ecological quality status (EcoQ) within the context of the European Water Framework Directive (WFD).In this contribution, 38 different applications including six new case studies (hypoxia processes, sand extraction, oil platform impacts, engineering works, dredging and fish aquaculture) are presented. The results show the response of the benthic communities to different disturbance sources in a simple way. Those communities act as ecological indicators of the ‘health’ of the system, indicating clearly the gradient associated with the disturbance.     

Epiphytic lichens as biomonitors of airborne heavy metal pollution

The research aims to assess the performance of the lichen Parmotrema reticulatum as an air pollution biomonitor of four heavy metals, namely, chromium (Cr), copper (Cu), lead (Pb) and zinc (Zn). Four contrasting land use sites within the greater Auckland region of New Zealand are used. One site is located within the relatively clean air shed of conservation land, the others within residential, commercial, and industrial areas, characterised by increasingly polluted air sheds, respectively. Three groups of lichens are monitored over a two-year period using active and passive biomonitoring methods to assess ‘on-thallus’ and ‘in-thallus’ concentrations of heavy metals. Seasonal transplants are used to quantify heavy metals accumulated by the lichen during each season. Long-term transplants are used to measure how fast lichens accumulate heavy metals and to better understand how and when heavy metals within the lichen thallus achieve equilibrium with air pollutant concentrations over time. The results show that the lichens continuously accumulates pollutants from the air until equilibrium is reached, thus transplanted lichens are useful for monitoring air pollution concentrations over time. Since pollutant concentration in the transplanted lichen at equilibrium stabilises, at this point the lichen ceases to be useful for monitoring temporal trends in air pollution, but may useful for spatial air pollution monitoring. The industrial location has the highest total accumulation for all four heavy metals, followed by the commercial and residential locations, respectively. Overall, the results show that the lichen P. reticulatum may be successfully used to monitor spatial and temporal pollution patterns caused by even very low concentrations of Cr, Cu, Pb and Zn.     

High-frequency monitoring of phytoplankton dynamics within the European water framework directive: application to metalimnetic cyanobacteria

Large, sub-alpine, stratified lakes are directly within the scope of the European Water Framework Directive (WFD) and need adapted monitoring systems. Moreover, anthropogenic eutrophication was frequently the main cause of their water quality degradation in the 20th century. This paper is primarily aimed at demonstrating how in situ sensor-equipped buoys could be the base of monitoring designs to support the WFD objectives. The core of this paper, mainly methodological, focuses on single-depth, high frequency (4 per hour) fluorescence measurements. It shows that the internal wave pattern provides additional information to the single-depth time series to assess phytoplankton dynamics in a stratified water column displaying strong, vertical biomass heterogeneity. The paper deals with the following three aspects: (1) definition of an indicator to determine whether or not the sensor actually detects the metalimnetic fluorescence peak, (2) vertical representation of chlorophyll distribution from single-depth series; (3) time evolution of the fluorescence peak (maximum value, depth and associated temperature, peak width).     

Testing the European stream typology of the Water Framework Directive for macroinvertebrates

In the EU Water Framework Directive (WFD) a typological framework is defined for assessing the ecological quality of water bodies in the future. The conditions in the Directive impose a strong demand for `new' assessment systems. During the AQEM project an assessment system was developed for European streams using macroinvertebrates. The aim of this study was to test if the typology suggested in the WFD is useful for developing an assessment system for macroinvertebrates in streams. In total 889 streams of 29 stream types were sampled in eight countries all over the major geographical gradients in Europe. These stream types fit the WFD typological demands and fit to the major European geographic regions (ecoregions). The sites included gradients from reference conditions (for the definition see Nijboer et al., 2004) to sites with bad ecological quality. Despite standardisation there were large differences between the participating countries concerning the number of taxa, the number of specimens and the taxonomic resolution. The data, including macroinvertebrates and environmental variables were analysed by using Canonical Correspondence Analysis (CCA). The observed macroinvertebrate distribution largely supported the WFD typological criteria. This means that the major macroinvertebrate distribution patterns in European streams follow climatological and geomorphological conditions and are well distinguished in terms of stream types. Furthermore, it was shown that large scale factors affected the macroinvertebrate distribution even on a very fine scale. Most explanatory variables seemed to be scale independent. Even at a fine scale major factors concerning geology, geomorphology and hydrology added to the species occurrences. Within stream types morphology together with physico-chemistry best explained the macroinvertebrates distribution. In conclusion, the WFD typology is useful for an assessment system for streams using macroinvertebrates. The large scale factors were indeed the variables that explained most of the variation in species composition. But as these factors even strongly act at the scale of stream types, a further refinement is most probably necessary to disentangle typological actors from water quality ones.     

A synthesis of the biological quality elements for the implementation of the European Water Framework Directive in the Mediterranean ecoregion: The case of Saronikos Gulf

Benthic macroinvertebrates, macroalgae and phytoplankton constitute the biological quality elements proposed in the Water Framework Directive (WFD, 2000/60/EC) to be used for the classification of the ecological status of a water body. In the context of the preparation for the implementation of WFD, classification schemes for all three elements have been developed and tested for all the European ecoregions. In the present work, the classification schemes, with the corresponding metric tools and the interpretations of the class boundaries according to the normative definitions of WFD, are presented for each biological element in Saronikos Gulf, as case study in the Mediterranean ecoregion. The combination of the three biological elements into an integrated classification for coastal water bodies has been one of the major issues addressed in the context of WFD guidelines. Results are interpreted and validated, through an ecological viewpoint, on the basis of relevant environmental data. Moreover, this work presents a way to combine the EUNIS system to the typology of water bodies in the Mediterranean ecoregion.     

Efficient sampling methodologies for lake littoral invertebrates in compliance with the European Water Framework Directive

Lake shores are characterised by a high natural variability, which is increasingly threatened by a multitude of anthropogenic disturbances including morphological alterations to the littoral zone. The European Water Framework Directive (EU WFD) calls for the assessment of lake ecological status by monitoring biological quality elements including benthic macroinvertebrates. To identify cost- and time-efficient sampling strategies for routine lake monitoring, we sampled littoral invertebrates in 32 lakes located in different geographical regions in Europe. We compared the efficiency of two sampling methodologies, defined as habitat-specific and pooled composite sampling protocols. Benthic samples were collected from unmodified and morphologically altered shorelines. Variability within macroinvertebrate communities did not differ significantly between sampling protocols across alteration types, lake types and geographical regions. Community composition showed no significant differences between field composite samples and artificially generated composite samples, and correlation coefficients between macroinvertebrate metrics calculated with both methods and a predefined morphological stressor index were similar. We conclude that proportional composite sampling represents a time- and cost-efficient method for routine lake monitoring as requested under the EU WFD, and may be applied across various European geographical regions.