How, what, and where you sample environmental DNA affects diversity estimates and species detection

Environmental DNA (eDNA) is a complex mixture of DNA, varying in particle
sizes and distributed patchily in aquatic systems. Optimizing eDNA
sampling is crucial for maximizing species detection, particularly in
high-risk scenarios like invasive species management. This study compares
two eDNA capture methods (grab sample vs. tow net) at the opposite ends of
the spectrum for volume and particle size to ascertain what most
influences 1) the detection of invasive species (Dreissenid mussels and
Burmese pythons) and 2) total diversity monitoring of metazoan, fungi, and
protist with COI marker and plant communities with an ITS2 marker.
Sampling was conducted across a wide geography and diverse aquatic
environments in Minnesota and Florida, USA and Switzerland. The tow net
samples were consistently higher in eDNA yield compared to grab samples;
however, they exhibited equal or lower alpha diversity of ZOTUs
(operational taxonomic units), challenging an assumption that there is a
relationship between eDNA yield and diversity. We found biodiversity
patterns were significantly influenced by the capture method, especially
for COI communitymetazoan diversity in all three regions. The two capture
methods measured different beta diversity of COImetazoan communities
across all three regionlocations, highlighting the confounding impact of
sample volume and filtration pore size on the diversity of eDNA captured.
Interestingly, the beta diversity of plant eDNA was less impacted by the
capture method compared to themetazoan beta diversity of COI communities.
We found no clear difference in detection for the two invasive species
targets with respect to the eDNA capture method. These results underscore
the need for pilot studies before conducting biodiversity inventory and
monitoring, and a need for a greater understanding of not just how much,
but also what eDNA is captured depending on method choice, considering
both spatial and particle size heterogeneity. For biodiversity
inventories, further research is needed to understand what causes
differences in detection, and what are the optimal eDNA sampling
strategies.