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31 December 2020 Multilocus Metabarcoding of Terrestrial Leech Bloodmeal iDNA Increases Species Richness Uncovered in Surveys of Vertebrate Host Biodiversity
Mai Fahmy, Kalani M. Williams, Michael Tessler, Sarah R. Weiskopf, Evon Hekkala, Mark E. Siddall
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Abstract

Leech-derived invertebrate DNA (iDNA) has been successfully leveraged to conduct surveys of vertebrate host biodiversity across the Indo Pacific. However, this technique has been limited methodologically, typically only targeting mammalian 16S rDNA, or both 16S and vertebrate 12S rDNA for leech host determination. To improve the taxonomic richness of vertebrate host species in iDNA surveys, we re-analyze datasets from Bangladesh, Cambodia, China, and Madagascar through metabarcoding via next generation sequencing (NGS) of 12S, 16S (2 types, one designed to target mammals and the other, residual eDNA), nicotinamide adenine dinucleotide hydride dehydrogenase 2 (ND2), and cytochrome c oxidase subunit 1 (COI). With our 5 primer sets, we identify 41 unique vertebrate hosts to the species level, among 1,200 leeches analyzed, along with an additional 13 taxa to the family rank. Within our 41 taxa, we note that adding ND2 and COI loci increased species richness detection by 25%. NGS has emerged as more efficient than Sanger sequencing for large scale metabarcoding applications and, with the decline in cost of NGS, our pooled sample multilocus protocol is an attractive option for iDNA biodiversity surveys.

© American Society of Parasitologists 2020
Mai Fahmy, Kalani M. Williams, Michael Tessler, Sarah R. Weiskopf, Evon Hekkala, and Mark E. Siddall "Multilocus Metabarcoding of Terrestrial Leech Bloodmeal iDNA Increases Species Richness Uncovered in Surveys of Vertebrate Host Biodiversity," Journal of Parasitology 106(6), 843-853, (31 December 2020). https://doi.org/10.1645/19-189
Published: 31 December 2020
JOURNAL ARTICLE
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KEYWORDS
biodiversity
conservation
iDNA
leeches
metabarcoding
vertebrates
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