Effects of fungicides on aquatic fungi and bacteria: a comparison of morphological and molecular approaches from a microcosm experiment

Background: Fungicides are frequently used in agriculture and can enter freshwater ecosystems through multiple pathways. The negative impacts of fungicides on microorganisms, fungi in particular, and their functions such as leaf decomposition have been repeatedly shown. In our previous microcosm experiment with three consecutive cycles of fungicide exposure and colonisation of leaf substrate, we found clear functional changes, but no differences in fungal community structure could be detected using morphological identification by analysing the spores of aquatic hyphomycetes. In this study, we examined the effects on fungal and bacterial community composition in detail using ITS and 16S metabarcoding and comparing the results to morphologically assessed community composition. Results: While we found fewer species with metabarcoding than with morphological identification, metabarcoding also enabled the identification of several fungal species that were otherwise unidentifiable morphologically. Moreover, by distinguishing individual amplicon sequence variants (ASVs) metabarcoding provided greater taxonomic resolution. In line with the morphological results, metabarcoding neither revealed effects of fungicides on the aquatic hyphomycetes nor on the total fungal or bacterial community composition. However, several ASVs responded significantly to fungicides, demonstrating variable tolerances within species. Conclusions: Overall, the absence of detectable effects of fungicides on the community structure despite clear functional effects, suggests a complex relationship between community structure and the ecosystem function of leaf decomposition.