In Greece, the implementation of the Water Framework Directive for rivers is based mainly on benthic macroinvertebrates and uses a semi-quantitative method with a D-frame net, which is certified by the International Organization for Standardization. Before the official adoption of this method, a quantitative shovel sampler (\"Cretan shovel\") was used in southern Greece (Crete), which has been implemented for almost three decades due to the specific river habitats found in Crete (e.g., seasonal flow, narrow riverbeds, and coarse substrates). In this study, we compared community metrics, diversity indices, feeding groups, locomotion types, and ecological quality derived from data collected using timed semi-quantitative kick samples and quantitative shovel samples collected from the same sites simultaneously. In total, 20 samples from the north and south of Greece were collected. The majority of community metrics, diversity indices, and traits were comparable between samplers. However, there were statistically significant differences in the relative abundance of Ephemeroptera, Plecoptera, and Trichoptera, passive filterers\' and the (semi) sessil groups, and Pielou\'s index. Most differences in the ecological quality between the kick and shovel samples were observed in 50% of the sites in northern Greece because the shovel is less effective at capturing motile zoobenthos. The ecological quality assessment in Crete by the Cretan shovel is comparable with the D-frame net in 75% of the samples. Thus, the Cretan shovel could efficiently sample the Cretan streams, which are characterized by coarse, narrow, turbulent, and hydrologically fluctuating river habitats. Such comparisons could improve sampling effectiveness and make additional data available to assess ecological quality.

The paper reports the results obtained after 4 years of aquatic angiosperm transplants in areas of the Venice Lagoon (North Adriatic Sea, Mediterranean) where meadows almost disappeared due to eutrophication, pollution and overexploitation of clam resources. The project LIFE12 NAT/IT/000331-SeResto, funded by the European Union, allowed to recolonize the Habitat 1150* (coastal lagoons) in the northernmost part of the lagoon, by extensive manual transplants of small sods or single rhizomes of Zostera marina, Zostera noltei, Ruppia cirrhosa and, in some stations also of Cymodocea nodosa. Over the 4 years of the project more than 75,000 rhizomes were transplanted in 35 stations with the support of local stakeholders (fishermen, hunters and sport clubs). Plants took root in 32 stations forming extensive meadows on a surface of approx. 10 km2 even if some failures were recorded in areas affected by outflows of freshwater rich in nutrients and suspended particulate matter. The rapid recovery of the ecological status of the involved areas was the result of this meadow restoration, which was in compliance with Water Framework Directive (WFD 2000/60/EC) objectives. Moreover, the monitoring of environmental parameters in the water column and in surface sediments allowed to identify the best conditions for successful transplants. Small, widespread interventions and the participation of local stakeholders in the environmental recovery, make this action economically cheap and easily transposable in other similar environments.

This study presents an approach to the determination of nutrient sensitive areas (SA) based on National Urban Wastewater Treatment Regulation harmonized under the Water Framework Directive (WFD). A three-stage approach that covers designation of potential sensitive areas (PSA) which are under the risk of eutrophication, physicochemical and biological monitoring at PSA, and application of the suitable biotic indices is used for the determination of sensitive areas. PSA are identified as a result of either available monitoring data or if this is not available according to indexing method which developed in this study based on pressure and impact analysis. The Gediz River Basin, one of the most polluted basins in Turkiye, is chosen as a pilot area. According to the results of the study, 40% of the total drainage area of the basin was designated as sensitive area.

Chemical monitoring data sets such as those provided by the implementation of the Water Framework Directive (WFD) offer opportunities to evaluate the ecological risks of pesticides under large spatiotemporal scales and to evaluate the protectiveness of the current prospective risk-assessment framework. As a case study, we used the monitoring data set for the insecticide chlorpyrifos to perform a probabilistic risk assessment for Iberian surface-water ecosystems. The specific objectives of the study were 1) to assess the occurrence of chlorpyrifos in relation to different agricultural production land uses, 2) to assess the spatiotemporal variation in the exceedance of the European WFD short- and long-term environmental quality standards (maximum allowable concentration environmental quality standard [MAC-EQS] and annual average [AA] EQS), and 3) to perform a probabilistic risk assessment for freshwater invertebrates. A database that contains chlorpyrifos concentrations from 14 600 surface water samples taken between 2012 and 2017 in the Iberian Peninsula (Spain and Portugal) was analyzed, and chlorpyrifos was detected in 21% of these samples. The MAC-EQS was exceeded in 2% of the cases, whereas the AA-EQS was exceeded in 18% of the cases. The majority of the exceedances took place in the littoral areas of the eastern and southeastern parts of the Iberian Peninsula, particularly in areas with dominant citrus production during late spring, late summer, and autumn. The present study indicates unacceptable risks posed by chlorpyrifos to Iberian surface waters over the study period, although it was approved for use in Europe. The present study supports the need to perform further postregistration monitoring assessments with other pesticides following similar approaches, which can help to identify possible pesticide-misuse practices and improvements of the prospective risk-assessment framework. Environ Toxicol Chem 2021;40:500-512. © 2020 SETAC.

The complex and dynamic nature of transitional ecosystems pose problems for the assessment of the Ecological Quality Status required by the European Water Framework Directive (WFD; 2000/60/EC). In six Adriatic lagoons, Ecological Quality Status was studied by comparing a biotic index based on macrophytes (MaQI), and three indices based on invertebrates (M-AMBI, M-bAMBI, and ISD). Ecological Status evaluated though MaQI and ISD resulted in quite degraded ecosystems (moderate/poor/bad), with only opportunistic algae and macrobenthic communities dominated by small size classes. Those results were supported by physico-chemical parameters, indicating high nutrients inputs, and anthropogenic pressures related with agriculture and fishery activities. Ecological Status obtained with M-AMBI and M-bAMBI was higher, with some sites reaching even the \"good\" status. The best response to anthropogenic pressures, in terms of a pressure index, was obtained by M-AMBI and M-bAMBI. Nevertheless, the response of used metrics (such as AMBI and bAMBI) to environmental variables not related to anthropogenic impact, and the high heterogeneity of physical-chemical conditions within lagoons, represent potential problems for the correct evaluation of Ecological Status of transitional waters. When different metrics give different responses it becomes a problem for managers who cannot easily make a decision on the remedial measures. The disagreement among indices arose because of the different response of biological elements to different stressors, and because the different indices based on macroinvertebrates focused on different aspects of the community, providing complementary information. So urge the need to find alternative approaches for a correct assessment of Ecological Status, with the combination of different biological elements, and considering the development of new indices (e.g. M-bAMBI) or refinement of the existing ones.

Extensive studies on the taxonomic resolution required for bioassessment purposes have determined that resolution above species level (genus, family) is sufficient for their use as indicators of relevant environmental pressures. The high-throughput sequencing (HTS) and meta-barcoding methods now used for bioassessment traditionally employ an arbitrary sequence similarity threshold (SST) around 95% or 97% to cluster sequences into operational taxonomic units, which is considered descriptive of species-level resolution. In this study, we analyzed the effect of the SST on the resulting diatom-based ecological quality index, which is based on OTU abundance distribution along a defined environmental gradient, ideally avoiding taxonomic assignments that could result in high rates of unclassified OTUs and biased final values. A total of 90 biofilm samples were collected in 2014 and 2015 from 51 stream sites on Mayotte Island in parallel with measures of relevant physical and chemical parameters. HTS sequencing was performed on the biofilms using the rbcL region as the genetic marker and diatom-specific primers. Hierarchical clustering was used to group sequences into OTUs using 20 experimental SST levels (80%-99%). An OTU-based quality index (IdxOTU) was developed based on a weighted average equation using the abundance profiles of the OTUs. The developed IdxOTU revealed significant correlations between the IdxOTU values and the reference pressure gradient, which reached maximal performance using an SST of 90% (well above species level delimitation). We observed an interesting and important trade-off with the power to discriminate between sampling sites and index stability that will greatly inform future applications of the index. Taken together, the results from this study detail a thoroughly optimized and validated approach to generating robust, reproducible, and complete indexes that will greatly facilitate effective and efficient environmental monitoring.

Chemicals in the environment occur in mixtures rather than as individual entities. Environmental quality monitoring thus faces the challenge to comprehensively assess a multitude of contaminants and potential adverse effects. Effect-based methods have been suggested as complements to chemical analytical characterisation of complex pollution patterns. The regularly observed discrepancy between chemical and biological assessments of adverse effects due to contaminants in the field may be either due to unidentified contaminants or result from interactions of compounds in mixtures. Here, we present an interlaboratory study where individual compounds and their mixtures were investigated by extensive concentration-effect analysis using 19 different bioassays. The assay panel consisted of 5 whole organism assays measuring apical effects and 14 cell- and organism-based bioassays with more specific effect observations. Twelve organic water pollutants of diverse structure and unique known modes of action were studied individually and as mixtures mirroring exposure scenarios in freshwaters. We compared the observed mixture effects against component-based mixture effect predictions derived from additivity expectations (assumption of non-interaction). Most of the assays detected the mixture response of the active components as predicted even against a background of other inactive contaminants. When none of the mixture components showed any activity by themselves then the mixture also was without effects. The mixture effects observed using apical endpoints fell in the middle of a prediction window defined by the additivity predictions for concentration addition and independent action, reflecting well the diversity of the anticipated modes of action. In one case, an unexpectedly reduced solubility of one of the mixture components led to mixture responses that fell short of the predictions of both additivity mixture models. The majority of the specific cell- and organism-based endpoints produced mixture responses in agreement with the additivity expectation of concentration addition. Exceptionally, expected (additive) mixture response did not occur due to masking effects such as general toxicity from other compounds. Generally, deviations from an additivity expectation could be explained due to experimental factors, specific limitations of the effect endpoint or masking side effects such as cytotoxicity in in vitro assays. The majority of bioassays were able to quantitatively detect the predicted non-interactive, additive combined effect of the specifically bioactive compounds against a background of complex mixture of other chemicals in the sample. This supports the use of a combination of chemical and bioanalytical monitoring tools for the identification of chemicals that drive a specific mixture effect. Furthermore, we demonstrated that a panel of bioassays can provide a diverse profile of effect responses to a complex contaminated sample. This could be extended towards representing mixture adverse outcome pathways. Our findings support the ongoing development of bioanalytical tools for (i) compiling comprehensive effect-based batteries for water quality assessment, (ii) designing tailored surveillance methods to safeguard specific water uses, and (iii) devising strategies for effect-based diagnosis of complex contamination.

Forested catchments are generally assumed to provide higher quality water in opposition to agricultural and urban catchments. However, this should be tested in various ecological contexts and through the study of multiple variables describing water quality. Indeed, interactions between ecological variables, multiple land use and land cover (LULC) types, and water quality variables render the relationship between forest cover and water quality highly complex. Furthermore, the question of the scale at which land use within stream catchments most influences stream water quality and ecosystem health remains only partially answered. This paper quantifies, at the regional scale and across five natural ecoregions of Wallonia (Belgium), the forest cover effect on biological water quality indices (based on diatoms and macroinvertebrates) at the riparian and catchment scales. Main results show that forest cover - considered alone - explains around one third of the biological water quality at the regional scale and from 15 to 70% depending on the ecoregion studied. Forest cover is systematically positively correlated with higher biological water quality. When removing spatial, local morphological variations, or population density effect, forest cover still accounts for over 10% of the total biological water quality variation. Partitioning variance shows that physico-chemical water quality is one of the main drivers of biological water quality and that anthropogenic pressures often explain an important part of it (shared or not with forest cover). The proportion of forest cover in each catchment at the regional scale and across all ecoregions but the Loam region is more positively correlated with high water quality than when considering the proportion of forest cover in the riparian zones only. This suggests that catchment-wide impacts and a fortiori catchment-wide protection measures are the main drivers of river ecological water quality. However, distinctive results from the agricultural and highly human impacted Loam region show that riparian forests are positively linked to water quality and should therefore be preserved.

Diatom-based ecological quality assessment methods have been implemented and used regularly in the Water Framework Directive. These indices use the species\' abundance profiles along a specific environmental gradient, which they aim to assess. However, this approach has several problematic issues including the unstable and fast-changing diatom taxonomy. The use of traits can be a solution if their responses to the environmental pressure are well-defined. Here, we developed taxonomy-based and trait-based diatom assemblage indices to assess the ecological status of riverine sites on a tropical island. The two indices are based on two sub-indices that measure the diatom assemblage response to a nutrient and organic matter/turbidity gradient. Both taxonomy- and trait-based indices correlated significantly with the selected environmental gradients of the test database, which was not used during index development. We showed that traits could be used for quality assessment of the Mayotte rivers and require much less effort than taxonomy-based indices. There were differences between the two types of indices, which are discussed in this paper. As a perspective for further studies, tests of trait-based indices among different eco-regions would be challenging.

The inventory of long-term braiding activity is a useful tool for detecting alterations in a rivers\' hydromorphological state and for a river\'s management in the context of the Water Framework Directive on integrated river basin management for Europe. Our study focuses on braided sectors of rivers in South-Eastern Subcarpathians (Romania). The inventory evaluates types of alterations based on the spatial analysis of fluvial morphology indicators (i.e., length of the river sector forming a braided pattern; width of the braided active channel), and vegetation cover (i.e., length of banks covered by forest and shrubs; area of in-stream patches of shrubs) accumulated over the last century. Furthermore, we performed a regional scale hierarchical cluster analysis to estimate the degree of alteration when compared to an historical baseline. In South-Eastern Subcarpathians, the studied rivers experienced a decrease of braiding activity revealed by the shortening and narrowing of their braided sectors, expansion of riparian forests, and the diminishment of vegetated islands\' areas. We separated three types of river clusters, corresponding to low (cluster 1), moderate (cluster 2), and high (cluster 3) degree of alteration. Moreover, the clusters demonstrate the evolutionary path of the braided pattern alterations until the functioning of another channel pattern. The inventory is relevant for differing types and levels of alterations. Additionally, this tool may serve as a first step toward the restoration of altered sectors by identifying rivers in cluster 1 as potential candidates of present-day reference sites for altered rivers with similar natural conditions as in cluster 3.