Upland rivers across Europe still exhibit undisturbed conditions and represent a treasure that we cannot afford to lose. We hypothesize that the combination of pristine and modified conditions could demonstrate biological responses along the stressor gradients. Thus, the response of aquatic macrophyte communities to anthropogenic stressors along upland rivers in Bulgaria was studied. Six stressors were selected out of 36 parameters grouped into hydromorphological, chemical variables and combined drivers (catchment land use). The stressors strongly affected species richness on the basis of biological type (bryophytes vs. vascular plants) and ecomorphological type (hydrophytes vs. helophytes). Hydrological alteration expressed by the change of the river\'s base flow and altered riparian habitats has led to a suppression of bryophytes and a dominance of riverbank plant communities. Seventy-five percent of mountain sites were lacking bryophytes, and the vegetation at semi-mountainous sites was dominated by vascular plants. It can be concluded that hydropeaking, organic and inorganic pollution, and discontinuous urban structures caused important modifications in the aquatic macrophyte assemblages. Macrophyte abundance and the biological and ecomorphological type of aquatic macrophytes reflect multi-stressor effects in upland rivers.

Human activities put stress on our oceans and with a growing global population, the impact is increasing. Stressors rarely act in isolation, with the majority of marine areas being impacted by multiple, concurrent stressors. Marine spatial cumulative impact assessments attempt to estimate the collective impact of multiple stressors on marine environments. However, this is difficult given how stressors interact with one another, and the variable response of ecosystems. As a result, assumptions and generalisations are required when attempting to model cumulative impacts. One fundamental assumption of the most commonly applied, semi-quantitative cumulative impact assessment method is that a change in modelled cumulative impact is correlated with a change in ecosystem condition. However, this assumption has rarely been validated with empirical data. We tested this assumption using a case study of seagrass in a large, inverse estuary in South Australia (Spencer Gulf). We compared three different seagrass condition indices, based on survey data collected in the field, to scores from a spatial cumulative impact model for the study area. One condition index showed no relationship with cumulative impact, whilst the other two indices had very small, negative relationships with cumulative impact. These results suggest that one of the most commonly used methods for assessing cumulative impacts on marine systems is not robust enough to accurately reflect the effect of multiple stressors on seagrasses; possibly due to the number and generality of assumptions involved in the approach. Future methods should acknowledge the complex relationships between stressors, and the impact these relationships can have on ecosystems. This outcome highlights the need for greater evaluation of cumulative impact assessment outputs and the need for data-driven approaches. Our results are a caution for marine scientists and resource managers who may rely on spatial cumulative impact assessment outputs for informing policy and decision-making.

Benthic macroinvertebrate communities are used as indicators for anthropogenic stress in freshwater ecosystems. To better understand the relationship between anthropogenic stress and changes in macroinvertebrate community composition, it is important to understand how different stressors and species traits are associated, and how these associations influence variation in species occurrence and abundances. Here, we show the capacity of the multivariate technique of double constrained correspondence analysis (dc-CA) to analyse trait-environment relationships, and we compare it with the redundancy analysis method on community weighted mean values of traits (CWM-RDA), which is frequently used for this type of analysis. The analyses were based on available biomonitoring data for macroinvertebrate communities from the Danube River. Results from forward selection of traits and environmental variables using dc-CA analyses showed that aquatic stages, reproduction techniques, dispersal tactics, locomotion and substrate relations, altitude, longitudinal and transversal distribution, and substrate preferendum were significantly related to habitat characteristics, hydromorphological alterations and water quality measurements such as physico-chemical parameters, heavy metals, pesticides and pharmaceuticals. Environmental variables significantly associated with traits using the CWM-RDA method were generally consistent with those found in dc-CA analysis. However, the CWM-RDA does neither test nor explicitly select traits, while dc-CA tests and selects both traits and environmental variables. Moreover, the dc-CA analysis revealed that the set of environmental variables was much better in explaining the community data than the available trait set, a kind of information that can neither be obtained from CWM-RDA nor from RLQ (Environment, Link and Trait data), which is a close cousin of dc-CA but not regression-based. Our results suggest that trait-based analysis based on dc-CA may be useful to assess mechanistic links between multiple anthropogenic stressors and ecosystem health, but more data sets should be analysed in the same manner.

Contaminants of emerging concern and ocean changes are key environmental stressors for marine species with possibly synergistic, but still unexplored, deleterious effects. In the present study the influence of a simulated ocean acidification scenario (pH = 7.6) was investigated on metabolism and sub-lethal effects of carbamazepine, CBZ (1 µg/L), chosen as one of the most widely diffused pharmaceuticals in marine organisms. A multidisciplinary approach was applied on mussels, M. galloprovincialis, integrating measurement of drug bioaccumulation with changes in the whole transcriptome, responsiveness of various biochemical and cellular biomarkers including immunological parameters, lipid and oxidative metabolism, onset of genotoxic effects. Chemical analyses revealed a limited influence of hypercapnia on accumulation and excretion of CBZ, while a complex network of biological responses was observed in gene expression profile and functional changes at cellular level. The modulation of gamma-aminobutyric acid (GABA) pathway suggested similarities with the Mechanism of Action known for vertebrates: immune responses, cellular homeostasis and oxidative system represented the processes targeted by combined stressors. The overall elaboration of results through a quantitative Weight of Evidence model, revealed clearly increased cellular hazard due to interactions of CBZ with acidification compared to single stressors.

The Sürgü Stream, located in the Euphrates River basin of Turkey, is used for drinking water source, agricultural irrigation and rainbow trout production. Therefore, water quality of the stream is of great importance. In this study, multivariate statistical techniques (MSTs) and water quality index (WQI) were applied to assess water quality of the stream affected by multiple stressors such as untreated domestic sewage, effluents from fish farms, agricultural runoff and streambank erosion. For this, 16 water quality parameters at five sites along the stream were monitored monthly during one year. Most of parameters showed significant spatial variations, indicating the influence of anthropogenic activities. All parameters except TN (total nitrogen) showed significant seasonal differences due to high seasonality in WT (water temperature) and water flow. The spatial variations in the WQI were significant (p < 0.05) and the mean WQI values ranged from 87.6 to 95.3, indicating \"good\" to \"excellent\" water quality in the stream. Cluster analysis classified five sites into three groups, that is, clean region, low polluted region and very clean region. Stepwise temporal discriminant analysis (DA) identified that pH, WT, Cl-, SO42-, COD (chemical oxygen demand), TSS (total suspended solids) and Ca2+ are the parameters responsible for variations between seasons, and stepwise spatial DA identified that DO (dissolved oxygen), EC (electrical conductivity), NH4-N, TN (total nitrogen) and TSS are the parameters responsible for variations between the regions. Principal component analysis/factor analysis revealed that the parameters responsible for water quality variations were mainly associated with suspended solids (both natural and anthropogenic), soluble salts (natural) and nutrients and organic matter (anthropogenic).

Contaminants found in watercourses are not only the result of anthropogenic activities but also depend on river\'s seasonal hydrodynamics. This is particularly true in Mediterranean climate regions where long dry periods are interrupted by strong rainfalls. Storm events remobilize particles from soils and sediments and, as a consequence, the load of particulate matter in rivers can be quite considerable, severely affecting water quality. Nevertheless, an absence of fieldwork studies exists concerning the simultaneous dynamics of mixtures of pollutants in river waters, particularly during strong rainfalls and floods. Our study assessed the concentrations of six families of pollutants, including pesticides, at these events, and compared them to those observed at drought sampling periods. We have used as model a typical Mediterranean coastal river from Southeast France, the Têt River, whose hydrodynamics and major elements fluxes have been fairly investigated. As expected, our results show that chemical mixtures due to human activities occur and that they are particularly relevant during storm events. But the results of our study argue that exceptional multicontamination phenomena actually happen more often than expected because they are linked to recurrent sudden intense rainfall events in the Mediterranean. In particular, combined sewer overflows are responsible for this major issue in urbanized areas, whereas runoff and leaching will be the most important sources of pollutant mixtures occurring at flood flow peak. After an overview of the sources responsible for chronic multiple stressors events in regions under a Mediterranean climate regime worldwide, we revisit best management measures to reduce risks from the presence of chemical mixtures in the environment.

A key challenge for the ecological risk assessment of chemicals has been to evaluate the relative contribution of chemical pollution to the variability observed in biological communities, as well as to identify multiple stressor groups. In this study we evaluated the toxic pressure exerted by >200 contaminants to benthic macroinvertebrates in the Danube River using the Toxic Unit approach. Furthermore, we evaluated correlations between several stressors (chemical and non-chemical) and biological indices commonly used for the ecological status assessment of aquatic ecosystems. We also performed several variation partitioning analyses to evaluate the relative contribution of contaminants and other abiotic parameters (i.e. habitat characteristics, hydromorphological alterations, water quality parameters) to the structural and biological trait variation of the invertebrate community. The results of this study show that most biological indices significantly correlate to parameters related to habitat and physico-chemical conditions, but showed limited correlation with the calculated toxic pressure. The calculated toxic pressure, however, showed little variation between sampling sites, which complicates the identification of pollution-induced effects. The results of this study show that the variation in the structure and trait composition of the invertebrate community are mainly explained by habitat and water quality parameters, whereas hydromorphological alterations play a less important role. Among the water quality parameters, physico-chemical parameters such as suspended solids, nutrients or dissolved oxygen explained a larger part of the variation in the invertebrate community as compared to metals or organic contaminants. Significant correlations exist between some physico-chemical measurements (e.g. nutrients) and some chemical classes (i.e. pharmaceuticals, chemicals related to human presence) which constitute important multiple stressor groups. This study demonstrates that, in large rivers like the Danube, the variation in the invertebrate community seems to be more related to varying habitat and physico-chemical conditions than to chemical pollution.