Tracking the phenology of riverine insect communities using environmental DNA

Aquatic insects are iconic and ecologically highly relevant inhabitants of riverine ecosystems. They are also often the target of monitoring programs to assess the ecological status of these lotic habitats. Environmental DNA (eDNA) techniques have been widely and successfully implemented to investigate freshwater insects and other macroinvertebrates. Commonly, such monitoring is conducted at one or two timepoints per year, despite the known strong seasonality and phenology of aquatic insects. Here, we assessed if and how eDNA can capture the temporal changes of the orders Ephemeroptera (mayflies), Plecoptera (stoneflies), Trichoptera (caddisflies) and Diptera (true flies). We carried out eDNA sampling at roughly monthly intervals from April to October at 25 sites across a whole river catchment in the northeastern part of Switzerland. We found pronounced, cyclic phenological trends in all orders but Trichoptera: the communities diverged from spring to summer and then in fall gradually returned closer to the spring state. The four orders exhibited different predominance in gains or losses of species detection throughout this time interval. Lastly, we found that field replicates, despite showing a relatively high local stochasticity, were able to provide a more complete assessment of aquatic communities and could thus be used as a proxy for the frequency of observation of a species through the seasons. In fact, this approach yielded comparable temporal patterns to the ones extracted from the Global Biodiversity Information Facility (GBIF). Overall, our findings demonstrate that eDNA techniques can be used to reveal intra-annual dynamics of aquatic insects. Given the current necessity to assess and monitor the biodiversity status of ecosystems, we therefore show that eDNA methods are a viable option to obtain a deeper understanding of the structuring of freshwater communities over time.