How much is enough? Effects of technical and biological replication on metabarcoding dietary analysis.
bat ecology
metabarcoding
molecular diet analyses
replication
sampling design
trophic ecology
Journal
Molecular ecology
ISSN: 1365-294X
Titre abrégé: Mol Ecol
Pays: England
ID NLM: 9214478
Informations de publication
Date de publication:
01 2019
01 2019
Historique:
received:
29
01
2018
revised:
16
05
2018
accepted:
23
05
2018
pubmed:
26
6
2018
medline:
13
11
2019
entrez:
26
6
2018
Statut:
ppublish
Résumé
DNA metabarcoding is increasingly used in dietary studies to estimate diversity, composition and frequency of occurrence of prey items. However, few studies have assessed how technical and biological replication affect the accuracy of diet estimates. This study addresses these issues using the European free-tailed bat Tadarida teniotis, involving high-throughput sequencing of a small fragment of the COI gene in 15 separate faecal pellets and a 15-pellet pool per each of 20 bats. We investigated how diet descriptors were affected by variability among (a) individuals, (b) pellets of each individual and (c) PCRs of each pellet. In addition, we investigated the impact of (d) analysing separate pellets vs. pellet pools. We found that diet diversity estimates increased steadily with the number of pellets analysed per individual, with seven pellets required to detect ~80% of prey species. Most variation in diet composition was associated with differences among individual bats, followed by pellets per individual and PCRs per pellet. The accuracy of frequency of occurrence estimates increased with the number of pellets analysed per bat, with the highest error rates recorded for prey consumed infrequently by many individuals. Pools provided poor estimates of diet diversity and frequency of occurrence, which were comparable to analysing a single pellet per individual, and consistently missed the less common prey items. Overall, our results stress that maximizing biological replication is critical in dietary metabarcoding studies and emphasize that analysing several samples per individual rather than pooled samples produce more accurate results.
Identifiants
pubmed: 29940083
doi: 10.1111/mec.14779
pmc: PMC7379978
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
165-175Subventions
Organisme : EDP Energias de Portugal
Pays : International
Organisme : Fundação para Ciência e Tecnologia
ID : IF/00497/2013
Pays : International
Organisme : Fundação para Ciência e Tecnologia
ID : LTER/BIA-BEC/0004/2009
Pays : International
Organisme : Fundação para Ciência e Tecnologia
ID : PD/BD/113462/2015
Pays : International
Organisme : Fundação para Ciência e Tecnologia
ID : PD/BD/52606/2014
Pays : International
Organisme : EU Horizon 2020
ID : 668981
Pays : International
Informations de copyright
© 2018 The Authors. Molecular Ecology Published by John Wiley & Sons Ltd.
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