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Temporary rivers, forming the majority of river networks worldwide, are key biodiversity hotspots. Despite their great value for maintaining biodiversity and ecosystem functioning, they are often neglected in biomonitoring programs due to several challenges, such as their variable hydromorphology and the difficulty of establishing reference conditions given their dynamic nature, resulting in highly variable communities. Disconnected pools often form in temporary rivers when flow ceases, providing refuge for aquatic taxa. Given their importance for biodiversity conservation, revising and adapting biotic indices are needed. Here, we evaluate the performance of current biological indices designed for perennial rivers (macroinvertebrates, diatoms) and functional metrics (macroinvertebrates) in assessing biological quality of disconnected pools. We sampled 55 disconnected pools in Catalonia, NE Spain, covering local (e.g., physico-chemical variables, water chemistry) and regional (e.g., human influence, hydrological variables at the water body level) natural and anthropogenic gradients. Only a few macroinvertebrate biotic indices (e.g., family richness, EPT/EPT + OCH and OCH) showed strong responses to anthropogenic predictors and were unaffected by natural predictors at both local and regional scales, making them suitable for biomonitoring. Of the newly adopted functional metrics of macroinvertebrate communities tested, only two (i.e., functional redundancy of predators and response diversity based on the total community) responded strongly to anthropogenic predictors. The rest showed varying responses to the interactive effect of anthropogenic and natural predictors, requiring calibration efforts. Models assessing these metrics explained <40 % of the total variation, likely due to the interplay of colonization/extinction dynamics and density-dependent trophic interactions governing community assemblages in disconnected pools. Although some existing biological metrics could potentially be used to monitor the ecological status of disconnected pools, we call for further development of biomonitoring tools specifically designed for these habitats since they will become more widespread with global change., 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 © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Amphibians, the most threatened vertebrates globally, face risks due to climate change, habitat loss, and fragmentation. Their sensitivity to environmental changes highlights their importance as ecological indicators. Temporary rivers, influenced by geological, climatic, and anthropogenic factors, play a critical role in shaping biodiversity and community structure. Some species of amphibians may be adapted to these temporary waters, a fact reflected in their life cycles and various biological traits. However, to develop effective conservation strategies for amphibians, it is essential to address the knowledge gaps surrounding the complex interactions between biological dynamics and fluvial habitat conditions. In this study, we investigated how trophic interactions between amphibians and other aquatic organisms (diatoms, macroinvertebrates, and fish), coupled with environmental factors (water availability and riparian structure), can affect amphibian abundance and diversity in temporary rivers. The study was conducted in a Mediterranean river network located in Sant Llorenç del Munt i l'Obac Natural Park (Catalonia, Spain). Our expectations were that habitats suitable for egg deposition, lacking predators (e.g. tadpole-predators and fish), and abundant in food sources would likely support higher amphibian abundance and diversity. However, water availability was identified as a crucial factor shaping abundance and diversity in the studied amphibian communities, even if it correlated with fish presence, and especially impacting amphibian species usually linked to permanent water bodies. Concerning biotic interactions, while our results suggested that amphibian populations in temporary rivers are more dependent on top-down than bottom-up interactions, the presence of aquatic predators was not as conclusive as expected, suggesting that in temporary rivers the fish-avoiding amphibian species can survive using microhabitats or breeding opportunities linked to natural river dynamics. Overall, our findings highlight the importance of considering multi-trophic interactions, hydroperiod and habitat heterogeneity in temporary river ecosystems for effective amphibian conservation., 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 © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Determining biological status of freshwater ecosystems is critical for ensuring ecosystem health and maintaining associated services to such ecosystems. Freshwater macroinvertebrates respond predictably to environmental disturbances and are widely used in biomonitoring programs. However, many freshwater species are difficult to capture and sort from debris or substrate and morphological identification is challenging, especially larval stages, damaged specimens, or hyperdiverse groups such as Diptera. The advent of high throughput sequencing technologies has enhanced DNA barcoding tools to automatise species identification for whole communities, as metabarcoding is increasingly used to monitor biodiversity. However, recent comparisons have revealed little congruence between morphological and molecular-based identifications. Using broad range universal primers for DNA barcode marker cox1, we compare community composition captured between morphological and molecular-based approaches from different sources - tissue-based (bulk benthic and bulk drift samples) and environmental DNA (eDNA, filtered water) metabarcoding - for samples collected along a gradient of anthropogenic disturbances. For comparability, metabarcoding taxonomic assignments were filtered by taxa included in the standardised national biological metric IBMWP. At the family level, bulk benthic metabarcoding showed the highest congruence with morphology, and the most abundant taxa were captured by all techniques. Richness captured by morphology and bulk benthic metabarcoding decreased along the gradient, whereas richness recorded by eDNA remained constant and increased downstream when sequencing bulk drift. Estimates of biological metrics were higher using molecular than morphological identification. At species level, diversity captured by bulk benthic samples were higher than the other techniques. Importantly, bulk benthic and eDNA metabarcoding captured different and complementary portions of the community - benthic versus water column, respectively - and their combined use is recommended. While bulk benthic metabarcoding can likely replace morphology using similar benthic biological indices, water eDNA will require new metrics because this technique sequences a different portion of the community., 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 © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The textile industry is one of the largest sectors globally, representing up to 20% of industrial water pollution. However, there is limited insight into how fluvial ecosystems respond and recover from this impact. From summer 2012 to spring 2013, we examined water quality and ecological status upstream and 1.5km downstream the input of a textile industry wastewater treatment plant (WWTP) in Ripoll River, NE Spain. The ecological status was determined via diversity measures and 10 biotic indices based on diatoms, macrophytes, macroinvertebrates and fish. Our results showed that the WWTP severely deteriorated water quality and biological communities at the discharge site, but that they improved at 1.5km downstream. Severity also varied across taxa and seasons, being fish the most affected taxa and spring the season with the best ecological status. The strong correlation amongst water quality variables and many biotic indices across taxa indicated that this is a chronic pollution event affecting multiple trophic levels. Thus, this study suggests that there is an urgent need to invest in wastewater treatment in this industry to preserve the ecological integrity of Ripoll River and especially its fish fauna. Likewise, it illustrates the diagnostic power of biotic indices based on diatoms, macroinvertebrates and fish, as driven by the European Water Framework Directive., (Copyright © 2016 Elsevier Inc. All rights reserved.)