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Mitigating ongoing losses of insects and their key functions (e.g. pollination) requires tracking large-scale and long-term community changes. However, doing so has been hindered by the high diversity of insect species that requires prohibitively high investments of time, funding and taxonomic expertise when addressed with conventional tools. Here, we show that these concerns can be addressed through a comprehensive, scalable and cost-efficient DNA metabarcoding workflow. We use 1815 samples from 75 Malaise traps across Germany from 2019 and 2020 to demonstrate how metabarcoding can be incorporated into large-scale insect monitoring networks for less than 50 € per sample, including supplies, labour and maintenance. We validated the detected species using two publicly available databases (GBOL and GBIF) and the judgement of taxonomic experts. With an average of 1.4 M sequence reads per sample we uncovered 10,803 validated insect species, of which 83.9% were represented by a single Operational Taxonomic Unit (OTU). We estimated another 21,043 plausible species, which we argue either lack a reference barcode or are undescribed. The total of 31,846 species is similar to the number of insect species known for Germany (~35,500). Because Malaise traps capture only a subset of insects, our approach identified many species likely unknown from Germany or new to science. Our reproducible workflow (~80% OTU-similarity among years) provides a blueprint for large-scale biodiversity monitoring of insects and other biodiversity components in near real time., (Molecular Ecology Resources© 2024 The Author(s). Molecular Ecology Resources published by John Wiley & Sons Ltd.)

While macroinvertebrate dispersal operates at the individual level, predictions of their dispersal capabilities often rely on indirect proxies rather than direct measurements. To gain insight into the dispersal of individual specimens, it is crucial to mark (label) and capture individuals. Isotopic enrichment with 15 N is a non-invasive method with the potential of labelling large quantities of macroinvertebrates. While the analysis of 15 N is widely utilised in food web studies, knowledge on the specific utility of isotopic enrichment with 15 N for mass labelling of macroinvertebrate individuals across different taxa and feeding types is limited. Previous studies have focused on single species and feeding types, leaving gaps in our understanding of the broader applicability of this method. Therefore, this study aimed to test and compare isotopic mass enrichment across several macroinvertebrate taxa and feeding types. We released 15 NH 4 Cl at five stream reaches in North-Rhine Westphalia, Germany, and successfully enriched 12 distinct macroinvertebrate taxa (Crustacea and Insecta). Significant enrichment was achieved in active and passive filter feeders, grazers, shredders and predators, and predominantly showed positive correlations with the enrichment of the taxa's main food sources phytobenthos and particulate organic matter. Enrichment levels rose rapidly and peaked at distances between 50 m and 300 m downstream of the isotopic inlet; significant enrichment occurred up to 2000 m downstream of the isotopic inlet in all feeding types. Macroinvertebrate density estimates on the stream bottom averaged to a total of approximately 3.4 million labelled individuals of the 12 investigated taxa, thus showing the high potential of isotopic ( 15 N) enrichment as a non-invasive method applicable for mass labelling across different macroinvertebrate feeding types. Hence, isotopic enrichment can greatly assist the analysis of macroinvertebrate dispersal through mark-and-recapture experiments, as it allows to measure the movement at the level of individual specimens., Competing Interests: The authors declare that they have no competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)