Net overboard: comparing marine eDNA sampling methodologies at sea to
unravel marine biodiversity
Abstract
Environmental DNA (eDNA) analyses are powerful for describing marine
biodiversity but must be optimized for their effective use in routine
monitoring. To maximize eDNA detection probabilities of sparsely
distributed populations, water samples are usually concentrated from
larger volumes and filtered using fine-pore membranes, often a
significant cost-time bottleneck in the workflow. This study aimed to
streamline eDNA sampling by investigating plankton net versus bucket
sampling, direct versus sequential filtration including self-preserving
filters. Biodiversity was assessed using metabarcoding of the small
ribosomal subunit (18S rRNA) and mitochondrial cytochrome c oxidase I
(COI) genes. Multi-species detection probabilities were estimated for
each workflow using a probabilistic occupancy modelling approach.
Significant workflow-related differences in biodiversity metrics were
reported. Highest amplicon sequence variant (ASV) richness was attained
by the bucket sampling combined with self-preserving filters, comprising
a large portion of micro-plankton. Less diversity but more metazoan taxa
were captured in the net samples combined with 5 µm pore size filters.
Pre-filtered 1.2 µm samples yielded few or no unique ASVs. The highest
average (~32%) metazoan detection probabilities in the
5 µm pore size net samples confirmed the effectiveness of
pre-concentrating plankton for biodiversity screening. These results
contribute to streamlining eDNA sampling protocols for uptake and
implementation in marine biodiversity research and surveillance.