Exploring hidden parasite diversity during a ParasiteBlitz across a coastal habitat gradient using environmental DNA metabarcoding

Abstract

Environmental DNA (eDNA) metabarcoding has lagged in parasite biodiversity assessments. We implemented this method to examine parasite diversity in sediment and water from 4 physically connected aquatic habitats in coastal South Carolina, USA, as part of a ParasiteBlitz in April 2023. Sediment was collected using a syringe corer, and water was sampled using active filtration and passive collection. Five amplicon libraries, using primers targeting portions of the mitochondrial COI of platyhelminths and 18S ribosomal RNA genes of nematodes, myxozoans, microsporidians, and protists, successfully yielded parasite sequences. Out of >5.8 million sequences, we identified >1,000 parasite amplicon sequence variants (ASVs) corresponding to ~600 parasite operational taxonomic units, from 6 parasite groups. Most diversity was observed among the microsporidians, whose assay demonstrated the highest fidelity. Actively-filtered water samples captured ASVs of all 6 groups, whereas sediment captured only 4, despite yielding 3× as many ASVs. Low DNA yields from passive water samples resulted in fewer, but some unique, ASVs representing 3 parasite groups. The most efficient sampling method varied with respect to parasite group across habitats, and the parasite communities from each habitat were distinct regardless of sampling method. We detected ASVs of 9 named species, 4 of which may represent introductions to the US. The abundance of our results demonstrates the effectiveness and efficiency of eDNA metabarcoding for assessing parasite diversity during short, intensive surveys, and highlights the critical need for more comprehensive sequence databases and the development of primers for those parasite taxa that elude detection using eDNA methods.