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Using multi-marker DNA metabarcoding to reveal the diet of a scarce woodland bird
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  • Ewan Stenhouse,
  • Paul Bellamy,
  • Will Kirby,
  • Ian Vaughan,
  • Lorna Drake,
  • Angela Marchbank,
  • Trudy Workman,
  • William Symondson,
  • Pablo Orozco-terWengel
Ewan Stenhouse
Cardiff University

Corresponding Author:stenhouseeh@cardiff.ac.uk

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Paul Bellamy
Royal Society for the Protection of Birds
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Will Kirby
Royal Society for the Protection of Birds
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Ian Vaughan
Cardiff University
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Lorna Drake
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Angela Marchbank
Cardiff University College of Biomedical and Life Sciences
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Trudy Workman
Cardiff University Cardiff School of Biosciences
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William Symondson
Cardiff University
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Pablo Orozco-terWengel
Cardiff University
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Abstract

Understanding the role diet plays in the structure of food webs is vital, and dietary knowledge is key for conservation management success. There is limited knowledge of the diets of woodland bird species, due largely to difficulties in accurately identifying plant and invertebrate taxa being consumed. Here, we show the effectiveness of multi-marker faecal metabarcoding to provide the most in-depth dietary analysis of a generalist passerine, the Hawfinch (Coccothraustes coccothraustes, Linnaeus), to date. Faecal samples were obtained from 2016-2019 from Hawfinch populations prior to and during the breeding season throughout the UK. DNA was extracted from 263 samples and amplified using Internal Transcribed Spacer 2 (ITS2) and cytochrome C oxidase subunit I (COI) barcodes. Using high-throughput sequencing (HTS), we identified 49 and 97 ITS2 and COI zero radius operational taxonomic units (zOTUs) respectively which equated to reputed dietary items. The herbivorous element of Hawfinch diet was dominated by naturally occurring taxa such as beech (Fagus sylvatica, Linnaeus), hornbeam (Carpinus betulus, Linnaeus) and oak (Quercus sp., Linnaeus). The most taxon rich and commonly recorded invertebrate taxon identified was Lepidoptera. We found Hawfinch diet varied spatially, as well as between sexes. Hawfinch showed broad dietary plasticity and utilised multiple resources within their foraging environments. Our study shows the potential of multi-marker DNA metabarcoding to reveal subtle dietary differences, but also highlights the challenges of studying omnivorous species using metabarcoding methods.