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Background Bioassessment of rivers is a fundamental method to determine surface water quality. One of the groups most commonly employed as bioindicators of aquatic ecosystems are benthic macroinvertebrates. Their conventional assessment is based on morphological identification and entails several limitations, such as being time‐consuming and requires trained experts for taxonomic identification. The use of genetic tools to solve these limitations offers an alternative way to evaluate rivers status. The use of environmental DNA (eDNA) metabarcoding has increased in recent years for different purposes, but its use in water quality evaluation is yet to be tested. Here, morphological and eDNA based inventories of macroinvertebrates were compared from the same seven sampling sites in the Upper Nalón River Basin (Asturias, Spain). Materials & Methods High‐Throughput Sequencing (HTS) of the cytochrome oxidase subunit 1 (COI) gene was carried out on DNA from water samples using an Ion Torrent platform. Biotic water quality indices were calculated from morphological and molecular data and compared with independent physico‐chemical habitat assessment to validate eDNA based approach. Results Highly positive and significant correlation was found between eDNA metabarcoding and morphological methods (Morphological and eDNA indices, r = 0.798, 5 degrees of freedom d.f., P = 0.031;) and a highly significant negative correlation was found between molecular and habitat quality indices (Stress score & eDNA, ρ = ‐0.878 and P = 0.009; Stress score & Visual, ρ = ‐0.949 and P = 0.0002). Discussion The similarity of results from the two approaches and the correlation of eDNA metabarcoding data with the habitat quality indices, suggest that eDNA performs as well as conventional methods for calculating biotic indices in this system, positioning eDNA metabarcoding of macroinvertebrate communities to transform how river bioassessment can be achieved. Conclusion The usefulness of eDNA metabarcoding to assess rivers water quality based on macroinvertebrates assessment has been demonstrated in a dammed river basin. Rivers are impacted ecosystems that need to be monitored regularly. Here, we have tested a metabarcoding approach to compare the results on water quality monitoring with conventional assessments for benthic macroinvertebrates. Water quality monitoring in freshwaters using macroinvertebrate eDNA performs equally well than conventional morphological‐based monitoring.

Africa accounts for nearly 30% of the discovered world’s mineral reserves, with half of the world’s platinum group metals deposits, 36% of gold, and 20% of cobalt being in Southern Africa (SA). The intensification of heavy-metal production in the SA region has exacerbated negative human and environmental health impacts. In recent years, mining waste generated from industrial and artisanal mining has significantly affected the ecological integrity of SA aquatic ecosystems due to the accelerated introduction and deposition of heavy metals. However, the extent to which heavy-metal pollution associated with mining has impacted the aquatic ecosystems has not been adequately documented, particularly during bioassessments. This review explores the current aquatic ecological impacts on the heavily mined river basins of SA. It also discusses the approaches to assessing the ecological risks, inherent challenges, and potential for developing an integrated ecological risk assessment protocol for aquatic systems in the region. Progress has been made in developing rapid bioassessment schemes (RBS) for SA aquatic ecosystems. Nevertheless, method integration, which also involves heavy-metal pollution monitoring and molecular technology, is necessary to overcome the current challenges of the standardisation of RBS protocols. Citizenry science will also encourage community and stakeholder involvement in sustainable environmental management in SA.

Rapid shifts in biotic communities due to environmental variability challenge the detection of anthropogenic impacts by current biomonitoring programs. Metacommunity ecology has the potential to inform such programs, because it combines dispersal processes with niche-based approaches and recognizes variability in community composition. Using intermittent rivers—prevalent and highly dynamic ecosystems that sometimes dry—we develop a conceptual model to illustrate how dispersal limitation and flow intermittence influence the performance of biological indices. We produce a methodological framework integrating physical- and organismal-based dispersal measurements into predictive modeling, to inform development of dynamic ecological quality assessments. Such metacommunity-based approaches could be extended to other ecosystems and are required to underpin our capacity to monitor and protect ecosystems threatened under future environmental changes.

Freshwater Algae: Identification and Use as Bioindicators provides a comprehensive guide to temperate freshwater algae, with additional information on key species in relation to environmental characteristics and implications for aquatic management. The book uniquely combines practical material on techniques and water quality management with basic algal taxonomy and the role of algae as bioindicators.  Freshwater Algae: Identification and Use as Bioindicators is divided into two parts. Part I describes techniques for the sampling, measuring and observation of algae and then looks at the role of algae as bioindicators and the implications for aquatic management. Part II provides the identification of major genera and 250 important species.  Well illustrated with numerous original illustrations and photographs, this reference work is essential reading for all practitioners and researchers concerned with assessing and managing the aquatic environment.

A greater understanding and effective management of biological invasions is a priority for biodiversity conservation globally. Many freshwater ecosystems are experiencing the colonization and spread of multiple co-occurrent alien species. Here the implications of both the relative abundance and richness of alien invaders on aquatic macroinvertebrate taxonomic and functional richness, ecosystem quality, and functional redundancy are assessed using long-term data from rivers in England. Based on the most common aquatic invaders, results indicated that their richness, rather than abundance, was the most important factor negatively affecting aquatic macroinvertebrate biodiversity. However, the response of functional redundancy was negatively affected by invader abundance at the river basin scale. The response of communities varied as the number of invading taxa increased, with the most marked reductions following the colonization of the first few invaders. Results indicate that different facets of multiple biological invasions influence distinct aspects of aquatic biodiversity. Preventing the establishment of new invaders and limiting invader taxa richness within a community should therefore be a conservation priority. These findings will assist river scientists in understanding mechanisms driving changes in biodiversity and facilitate the testing of ecological theories while also ensuring environmental managers and regulators can prioritize conservation / management opportunities.

DNA metabarcoding of freshwater communities typically relies on PCR amplification of a fragment of the mitochondrial cytochrome c oxidase I (COI) gene with degenerate primers. The advantage of COI is its taxonomic resolution and the availability of an extensive reference database. However, when universal primers are used on environmental DNA (eDNA) isolated from water, benthic invertebrate read and OTU numbers are typically “watered down,” that is, under represented, compared to whole specimen “bulk samples” due to greater co‐amplification of abundant nontarget taxa (e.g., fungi, algae, and bacteria). Because benthic stream invertebrate taxa are of prime importance for regulatory biomonitoring, more effective ways to capture their diversity via eDNA isolated from water are important. In this study, we aimed to improve benthic invertebrate assessment from eDNA by minimizing nontarget amplification. Therefore, we generated eDNA data using universal primers BF2/BR2 on samples collected throughout 15 months from a German Long‐Term Ecological Research site (Rhine‐Main‐Observatory, Kinzig River) to identify most abundant nontarget taxa. Based on these data, we designed a new reverse primer (EPTDr2n) with 3’‐specificity toward benthic invertebrate taxa and validated its specificity in silico together with universal forward primer fwhF2 using available data from GenBank and BOLD. We then performed in situ tests using 20 Kinzig River eDNA samples. We found that the percentage of target reads was much higher for the new primer combination compared to two universal benthic invertebrate primer pairs, BF2/BR2 and fwhF2/fwhR2n (99.6% versus 25.89% and 39.04%, respectively). Likewise, the number of detected benthic invertebrate species was substantially higher (305 versus 113 and 185) and exceeded the number of 153 species identified by expert taxonomists at nearby sites across two decades of sampling. While few taxa, such as flatworms, were not detected, we show that the optimized primer avoids the nontarget amplification bias and thus significantly improves benthic invertebrate detection from eDNA. Capturing benthic invertebrate biodiversity from environmental DNA (eDNA) isolated from water is difficult because of abundant nontarget taxa PCR amplification. A new primer combination maximizes target‐specific eDNA amplification for freshwater macroinvertebrate species. It detects especially more species than reported for two decades of extensive morphotaxonomic research at the studied Long‐Term Ecological Research (LTER) site.

Dispersal is one of the key mechanisms affecting the distribution of individuals, populations, and communities in nature. Despite advances in the study of single species, it has been notoriously difficult to account for dispersal in multispecies metacommunities, where it potentially has strong effects on community structure beyond those of local environmental conditions. Dispersal should thus be directly integrated in both basic and applied research by using proxies. Here, we review the use of proxies in the current metacommunity research, suggest new proxies, and discuss how proxies could be used in community modelling, particularly in freshwater systems. We suggest that while traditional proxies may still be useful, proxies formerly utilized in transport geography may provide useful novel insights into the structuring of biological communities in freshwater systems. We also suggest that understanding the utility of such proxies for dispersal in metacommunities is highly important for many applied fields such as freshwater bioassessment, conservation planning, and recolonization research in the context of restoration ecology. These research fields have often ignored spatial dynamics and focused mostly on local environmental conditions and changes therein. Yet, the conclusions of these applied studies may change considerably if dispersal is taken into account.

High-throughput sequencing (HTS) studies on invertebrates commonly use ethanol as the main sample fixative (upon collection) and preservative (for storage and curation). However, alternative agents exists, which should not be automatically neglected when studies are newly designed. This review provides an overview of the application of propylene glycol (PG) in DNA-based studies of invertebrates, thus to stimulate an evidence-based discussion. The use of PG in DNA-based studies of invertebrates is still limited (n = 79), but a steady increase has been visible since 2011. Most studies used PG as a fixative for passive trapping (73%) and performed Sanger sequencing (66%; e.g. DNA barcoding). More recently, HTS setups joined the field (11%). Terrestrial Coleoptera (30%) and Diptera (20%) were the most studied groups. Very often, information on the grade of PG used (75%) or storage conditions (duration, temperature) were lacking. This rendered direct comparisons of study results difficult, and highlight the need for further systematic studies on these subjects. When compared to absolute ethanol, PG can be more widely and cheaply acquired (e.g. as an antifreeze, 13% of studies). It also enables longer trapping intervals, being especially relevant at remote or hard-to-reach places. Shipping of PG-conserved samples is regarded as risk-free and is authorised, pinpointing its potential for larger trapping programs or citizen science projects. Its property to retain flexibility of morphological characters as well as to lead to a reduced shrinkage effect was especially appraised by integrative study designs. Finally, the so far limited application of PG in the context of HTS showed promising results for short read amplicon sequencing and reduced representation methods. Knowledge of the influence of PG fixation and storage for long(er) read HTS setups is currently unavailable. Given our review results and taking difficulties of direct methodological comparisons into account, future DNA-based studies of invertebrates should on a case-by-case basis critically scrutinise if the application of PG in their anticipated study design can be of benefit.

The detection of species using environmental DNA (eDNA) relies on our capacity to identify DNA from the sampled environment. Once eDNA is released into the environment, the physical degradation of individual eDNA molecules over time directly affects our ability to detect species (eDNA decay). Therefore, interpreting eDNA data requires an explicit understanding of eDNA decay to accurately infer contemporary presence or absence of a given species in the study ecosystem. Most research to date on eDNA decay has focused on single‐species assays (predominantly quantitative PCR), and thus little is known on how eDNA decay affects the interpretation of metabarcoding datasets. Here, we used eDNA metabarcoding (targeting a section of the eukaryotic cytochrome c oxidase subunit I gene) of water samples from controlled tanks to examine eDNA decay in a wide variety of marine metazoan species. After the stocked organisms were removed from these tanks, we observed a sharp decrease in amplicon sequence variant (ASV) richness within the first 48 hours. Furthermore, there was a substantial change in beta diversity between 24 and 48 hours, and after 48 hours, most stocked species became undetectable. Interestingly, the estimated decay rate for each study species, calculated using a linear regression of reads over time, differed across organisms, with up to 2–3‐fold difference among species. Our results showed that, for marine temperate species, the most substantial change in eDNA metabarcoding detectability occurred within the first 48 h, and after that, eDNA from several taxa became undetectable. These findings inform the ecological interpretation of metabarcoding datasets and provide estimates of eDNA decay rate that are valuable for both simulation studies and the design of future ecological surveys. Ecological interpretations of eDNA metabarcoding data require an explicit understanding of eDNA decay across different study systems and species. Here, we used eDNA metabarcoding of water samples to examine eDNA decay in a wide variety of temperate marine metazoan species. We found the most substantial effects of eDNA decay on diversity occurred within 24–48 hours and showed that eDNA decay rate constants varied among the detected species.

Periphyton is the dominant primary producer in mountain streams and sustains the higher trophic levels. While certain periphyton groups, particularly diatoms, have received extensive study, the comprehensive characterization of the entire community has been largely neglected. This study aims to investigate the temporal pattern of biofilm in mountain streams characterized by different water thermal regimes. A one-year quantitative campaign, involving monthly samplings, was conducted in five subalpine streams in Northern Italy’s Orobic Alps to collect epilithic biofilm from a wide surface area. The total biomass was quantified and the periphyton was analyzed both for composition (diatoms, green algae, cyanobacteria, and red algae) and for pigments. Disturbance, water temperature, physico-chemical conditions, nutrients, substrate diversity, and light availability were assessed concurrently with biofilm samplings. Results show sharp biofilm variations over months. In all sites, the disturbance was the primary factor reducing biomass and pigment content. Annually, all sites experienced similar turnover in periphyton composition mainly associated with light and water temperature. Overall, the study indicates that frequent quantitative investigations of biofilm help understand intra-annual variations and identify key drivers. Such information is useful to understand the ecosystem processes and the food web dynamics.