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The bioassessment of aquatic ecosystems is currently based on various biotic indices that use the occurrence and/or abundance of selected taxonomic groups to define ecological status. These conventional indices have some limitations, often related to difficulties in morphological identification of bioindicator taxa. Recent development of DNA barcoding and metabarcoding could potentially alleviate some of these limitations, by using DNA sequences instead of morphology to identify organisms and to characterize a given ecosystem. In this paper, we review the structure of conventional biotic indices, and we present the results of pilot metabarcoding studies using environmental DNA to infer biotic indices. We discuss the main advantages and pitfalls of metabarcoding approaches to assess parameters such as richness, abundance, taxonomic composition and species ecological values, to be used for calculation of biotic indices. We present some future developments to fully exploit the potential of metabarcoding data and improve the accuracy and precision of their analysis. We also propose some recommendations for the future integration of DNA metabarcoding to routine biomonitoring programs. [ABSTRACT FROM AUTHOR]

• A standardized protocol yields reproducible bulkDNA metabarcoding results. • Highly abundant and large species are shared most often between laboratories. • Need for standardization when comparing alpha diversity between different studies. • Betadiversity patterns are robust to changes in the metabarcoding wetlab protocol. DNA metabarcoding can be used in marine environmental monitoring if results are reproducible between labs and robust against modifications to the lab protocol. In this interlaboratory study, we conducted a ring test where subsamples of blended macrobenthos samples were distributed to four laboratories located in Belgium, the Netherlands, Germany and Denmark. Samples were processed by a standardized lab protocol and by an adapted protocol, and the resulting datasets were analyzed with the same bioinformatics pipeline. Different biodiversity indicators were calculated. Our results show that bulkDNA metabarcoding of marine macrobenthos offers a highly reproducible assessment of alpha diversity patterns when using a standardized protocol, since comparable species numbers, Shannon indices and Inverse Simpson indices were found between laboratories. Especially high abundant species and species with large body sizes were shared between the laboratories. The need for using a standardized protocol to enhance comparability in alpha diversity between different studies was shown. Beta diversity patterns are less subjected to changes in the metabarcoding protocol and were almost identical between different laboratories, as the main clustering was always based on the macrobenthic community, independent of the used protocol or the laboratory that conducted the work. We conclude that DNA metabarcoding for marine environmental monitoring is an appropriate method when the aim is to study changes in community patterns and advocate its implementation in routine monitoring programs of national and European authorities, providing that a standardized protocol is implemented and/or a detailed description of the protocol is available. [ABSTRACT FROM AUTHOR]

We calculated a Living Planet Index (LPI) for the Netherlands, based on 361 animal species from seven taxonomic groups occurring in terrestrial and freshwater habitats. Our assessment is basically similar to the global LPI, but the latter includes vertebrate species and trends in population abundance only. To achieve inferences on trends in biodiversity more generally, we added two insect groups (butterflies and dragonflies) and added occupancy trends for species for which we had no abundance trends available. According to the LPI, the state of biodiversity has slightly increased from 1990 to 2014. However, large differences exist between habitat types. We found a considerable increase in freshwater animal populations, probably because of improvement of chemical water quality and rehabilitation of marshland habitats. We found no trend in the LPI for woodland populations. In contrast, populations in farmland and open semi-natural habitats (coastal dunes, heathland and semi-natural grassland) declined, which we attribute to intensive agricultural practices and nitrogen deposition, respectively. The LPI shows that, even in a densely populated western European country, ongoing loss of animal biodiversity is not inevitable and may even be reversed if adequate measures are taken. Our approach enabled us to produce summary statistics beyond the level of species groups to monitor the state of biodiversity in a clear and consistent way. [ABSTRACT FROM AUTHOR]