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The Joint Danube Survey (JDS) is a regular transnational survey to monitor the quality of the Danube and its main tributaries, in accordance with the EU Water Framework Directive. During the JDS4 in 2019, conventional methods to monitor selected biological quality elements were complemented with DNA metabarcoding.All together 72 phytobenthos samples were collected along the Danube and its major tributaries within the JDS4, using light microscopy and DNA metabarcoding amplifying a fragment of the rbcL marker gene. The (i) applicability of DNA metabarcoding to identify diatom communities compared to microscopy; (ii) diversity metrics between DNA metabarcoding and microscopy analysis and (iii) the usability of DNA metabarcoding for routine monitoring and assessment of the Danube under future JDS surveys were investigated.Diatom communities resulting from light microscopy and DNA metabarcoding assessments share 26.5% of all taxa which corresponds to 64.3% when considering relative abundances. Discrepancies originate from biases both from metabarcoding, e.g. missing taxa from the reference library, and from microscopy, e.g. overlooking of hardly visible taxa. Microscopy detected more taxa in total but metabarcoding revealed a higher alpha diversity, detecting also very rare taxa in a given sample. Molecular and microscopy based Specific Pollution Sensitivity Index (IPS) values correlated significantly but differences were detected at several sites due to the differences in community composition and the overestimation of large taxa by metabarcoding. Although both methods showed a decreasing trend of IPS along the Danube, the metabarcoding based IPS covered a higher range of quality classes indicating lower values for downstream sites and the tributaries.We suggest that metabarcoding provides a standardisable and efficient tool in biomonitoring, being more distinctive among quality classes than microscopy. Due to the high sequencing depth, it is able to detect a higher diversity on genetic level in a time- and cost-efficient manner that should be implemented in future quality assessment tools. We recommend its use in future biomonitoring surveys, for now, as a complementary method to conventional ones.

Mediterranean lakes present particular challenges when developing ecological assessment techniques, due partly to long histories of human activity in their catchments but also to strong interactions with climate, intensifying in recent decades due to global warming. A survey of diatoms in lakes in Greece revealed both a strong nutrient gradient and a salinity gradient. It is necessary to disentangle these when assessing ecological status in order that appropriate measures for restoring good ecological status are applied. Two approaches were used to develop a Water Framework Directive-compatible metric for assessing ecological status using diatoms. The strongest relationship was obtained using weighted averaging, but an alternative metric, generated by rescaling a well-established and widely used diatom metric performed only slightly less well. The limited number of lakes in Greece resulted in a small dataset, creating a risk of overfitting models and the rescaled metric has the advantage of drawing on a broader autecological knowledge base. Supplementary metrics to evaluate salinity are also proposed in order to ensure that lakes subject to this pressure are clearly differentiated from those where nutrient impacts predominate. Approaches to defining reference conditions and status class boundaries draw on established criteria developed for Water Framework Assessments in Europe to ensure that status assessments in Greece reflect similar levels of ambition to those established elsewhere in the European Union.

The species composition of benthic algae changes as water phosphorus concentrations increase, and these changes can be used for ecological status assessment according to the Water Framework Directive. Natural background phosphorus concentrations in rivers and streams that are unaffected by anthropogenic impacts are usually low. Running waters draining catchments with deposits of marine clay, however, may have enhanced phosphorus concentrations, because the clay is naturally rich in apatite. Almost all clay rich areas have been cultivated for centuries, however, and fertilization has increased the soil phosphorus levels. It has, therefore, been difficult to disentangle natural from anthropogenically enhanced phosphorus in streams draining clay rich areas. We compared water phosphorus concentrations, and the Periphyton Index of Trophic Status PIT, between clay and non-clay, impacted and unimpacted rivers in Norway. We found that water phosphorus concentrations and the PIT index were higher in unimpacted clay rivers than in unimpacted non-clay rivers, indicating that natural phosphorus concentrations in clay rivers are indeed enhanced compared to rivers without deposits of marine clay. In addition, phosphate-P contributed 18–23% to total phosphorus in unimpacted clay rivers, but 33–37% in unimpacted and impacted non-clay rivers and clay rivers affected by agriculture. This indicates that the total phosphorus in unimpacted clay rivers is less bioavailable than in non-clay rivers and in impacted clay rivers. Water total phosphorus concentrations in unimpacted clay rivers significantly increased with catchment clay cover. Based on these findings, we derived new status class boundaries for the PIT index in clay rivers. Clay rivers are suggested to be assessed in only two status classes, i.e., “good or better” or “moderate or worse”, respectively. The good/moderate status class boundary for the PIT index was shown to increase with increasing catchment clay cover.

We studied three commonly used bioindicator groups: phytoplankton, phytobenthos and benthic invertebrate communities’ structure patterns in transitions between lotic and lentic habitats as consequences of two consecutive large dams constructed in the early twentieth century on the intermediate reach of a lowland river in Normandy (NW, France), the Selune. According to sampling station position along the river, including two in the two reservoir lakes, we defined three sorts of transitions, from upstream to downstream: lentic to lotic transition (hereafter len-lot), lentic to lentic transition (hereafter len-len), within the two reservoir lakes, and lentic to lotic transition (hereafter len-lot).Lot-len transition, imposed by damming, and the recovery of lotic character of the river afterward dams, promoted different diversity patterns for the studied organisms’ groups. Phytobenthos’ α diversity significantly incresed in the len-lot transition, and the highest β diversity score was observed in the lot-len. Both, habitat conditions and spatial controls drove phytobenthos community structure. Benthic invertebrate community was mainly shaped by habitat conditions. Invertebrate’s α diversity significantly changed in the lot-len and len-lot transitions, with the β diversity score being the highest in the lot-len transition. Community’s traits composition of invertebrates best fitted predictions on lotic- and lentic- type communities. Phytoplankton’s α diversity remained unchanged in the transitions and subsequently β diversity scores were low.According to the results, we proposed three classes of organisms based on their response to river damming: i) organisms indifferent or tolerant to dams effect, like phytoplankton communities in this study, that were neither hampered by changes in water flow velocity, and did not display dispersal limitation neither; ii) organisms sensitive to habitat conditions shift but not limited in their dispersal capacity by the dam, like benthic invertebrates and phytobenthos in this study. Benthic invertebrate community and phytobenthos communities differed in their response likely due to their differential dispersal capabilities and niche breadth; iii) organisms limited in their dispersal by the dam, not represented in this study.Riverine organisms best tracked transitions, while β diversity assessment revealed as the most useful metric to disentangle the relative contribution of dispersal limitation or habitat modification provoked by multiple dams.

Temporary rivers are widespread in the Mediterranean region and impose a challenge for the implementation of the Water Framework Directive (WFD) and other environmental regulations. Surprisingly, an overarching analysis of their ecological status and the stressors affecting them is yet missing. We compiled data on the ecological status of 1504 temporary rivers in seven European Mediterranean region countries and related their ecological status (1) to publicly available data on pressures from the European WISE-WFD dataset, and (2) to seven more specific stressors modelled on a sub-catchment scale. More than 50 % of the temporary water bodies in the Mediterranean countries reached good or even high ecological status. In general, status classes derived from phytobenthos and macrophyte assessment were higher than those derived from the assessment of benthic invertebrates or fish. Of the more generally defined pressures reported to the WISE-WFD database, the most relevant for temporary rivers were 'diffuse agricultural' and 'point urban waste water'. Of the modelled more specific stressors, agricultural land use best explained overall ecological status, followed by total nitrogen load, and urban land use, while toxic substances, total phosphorus load and hydrological stressors were less relevant. However, stressors differed in relevance, with total nitrogen being most important for macrophytes, and agricultural land use for phytobenthos, benthic invertebrates and fish. For macrophytes, ecological quality increased with stressor intensity. The results underline the overarching effect of land use intensity for the ecological status of temporary water bodies. However, assessment results do not sufficiently reflect hydrological stress, most likely as the biological indicators used to evaluate these systems were designed for perennial water bodies and thus mainly target land use and nutrient impacts. We conclude that biomonitoring systems need to be updated or newly developed to better account for the specific situation of temporary water bodies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Climate change threatens surface waters worldwide, especially shallow lakes where one of the expected consequences is a sharp increase in their water temperatures. Phytobenthos is an essential, but still less studied component of aquatic ecosystems, and it would be important to learn more about how global warming will affect this community in shallow lakes. In this research, the effects of different climate change scenarios (SSP2-4.5 and SSP5-8.5, as intermediate and high emission scenarios) on the structure and function of the entire phytobenthos community using species- and trait-based approaches were experimentally investigated in an outdoor mesocosm system. Our results show that the forecasted 3 °C increase in temperature will already exert significant impacts on the benthic algal community by (1) altering its species and (2) trait composition (smaller cell size, lower abundance of colonial and higher of filamentous forms); (3) decreasing Shannon diversity; and (4) enhancing the variability of the community. Higher increase in the temperature (+5 °C) will imply more drastic alterations in freshwater phytobenthos by (1) inducing very high variability in species composition and compositional changes even in phylum level (towards higher abundance of Cyanobacteria and Chlorophyta at the expense of Bacillariophyta); (2) continuing shift in trait composition (benefits for smaller cell volume, filamentous life-forms, non-motile and weakly attached taxa); (3) further reducing the functional diversity; (4) increasing biofilm thickness (1.4 μm/°C) and (5) decreasing maximum quantum yield of photosystem II. In conclusion, already the intermediate emission scenario will predictably induce high risk in biodiversity issues, the high emission scenario will imply drastic impacts on the benthic algae endangering even the function of the ecosystem., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Recent results of field studies on the exposed coast of Lithuania were used to model the area occupied by the red alga Furcellaria lumbricalis using the Natural Neighbor interpolation technique, while linear regression was applied to estimate the species' standing stock. The area covered by F. lumbricalis extended for 26 km along the coast between depths of 1 and 15 m. The maximum species cover in the study area ranged between 4 and 10 m depth, which is one of the widest in the Baltic Sea. The modelled area of F. lumbricalis covered 35 ± 11 km2 with a total biomass of 7554 ± 3813 t.

Although the Water Framework Directive specifies that macrophytes and phytobenthos should be used for the ecological assessment of lakes and rivers, practice varies widely throughout the EU. Most countries have separate methods for macrophytes and phytobenthos in rivers; however, the situation is very different for lakes. Here, 16 countries do not have dedicated phytobenthos methods, some include filamentous algae within macrophyte survey methods whilst others use diatoms as proxies for phytobenthos. The most widely-cited justification for not having a dedicated phytobenthos method is redundancy, i.e. that macrophyte and phytoplankton assessments alone are sufficient to detect nutrient impacts. Evidence from those European Union Member States that have dedicated phytobenthos methods supports this for high level overviews of lake condition and classification; however, there are a number of situations where phytobenthos may contribute valuable information for the management of lakes., (Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.)

Alpine and Nordic rivers are often considered as being among the most pristine in Europe. Nevertheless, acidification and eutrophication impact surface waters in these regions. Soft-bodied, i.e. non-diatom, benthic algae are used as indicators for eutrophication and acidification in both Norway and Austria, but consistency of indicator values has never been tested. We compared species optima with respect to pH, conductivity, total phosphorus (TP), and NO₃(-)-N concentration for 21 species, derived from geographically and temporally extensive datasets from Norway and Austria, respectively. The ranges of all four measured parameters were different between Norway and Austria, with Austria having generally higher values for all measured parameters. Optima for all 21 species with respect to pH, conductivity and NO₃(-)-N were significantly different between Norway and Austria, while 5 of the 21 taxa showed no significant differences for TP. Nevertheless, species optima for Norway and Austria were significantly correlated with each other for TP, pH and conductivity. This indicates that positions of species optima relative to each other may be stable across ecoregions, in spite of the absolute values of species optima being different. In contrast, optima with respect to NO₃(-)-N were not correlated, possibly suggesting a lesser importance of NO₃(-) in shaping benthic algal assemblages than TP and pH. We conclude that the use of eutrophication and acidification models across different ecoregions may give meaningful results, but requires regional testing of species optima., (© 2013.)