Combining stable isotope analysis with DNA metabarcoding improves inferences of trophic ecology

• Published in: PloS one 2019-07, Vol.14 (7), p.e0219070-e0219070
• Main Authors: Whitaker, Melissa R L, Baker, Christopher C M, Salzman, Shayla M, Martins, Dino J, Pierce, Naomi E
• Format: Article
• Language: English
• Subjects: Animals; Ants - physiology; Biodiversity; Biology and Life Sciences; Butterflies; Butterflies & moths; Butterflies - physiology; Carbon Isotopes; Caterpillars; Chloroplasts; Deoxyribonucleic acid; Diet; DNA; DNA barcoding; DNA Barcoding, Taxonomic; DNA, Chloroplast - genetics; DNA, Chloroplast - isolation & purification; Ecological monitoring; Ecology; Ecology and Environmental Sciences; Ecosystem; Ecosystem biology; Ecosystem management; Ecosystems; Environmental science; Evolutionary biology; Fabaceae - chemistry; Fabaceae - genetics; Food intake; Gastrointestinal Microbiome - genetics; Herbivores; Herbivory; Herbivory - genetics; Insects; Isotope analysis; Isotopes; Larva - physiology; Lycaenidae; Museums; Nitrogen Isotopes; Ontogeny; Physical Sciences; Physiological aspects; Research and Analysis Methods; Stable isotopes; Strategic management; Symbiosis; Zoology
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0219070

Knowing what animals eat is fundamental to our ability to understand and manage biodiversity and ecosystems, but researchers often must rely on indirect methods to infer trophic position and food intake. Using an approach that combines evidence from stable isotope analysis and DNA metabarcoding, we assessed the diet and trophic position of Anthene usamba butterflies, for which there are no known direct observations of larval feeding. An earlier study that analyzed adults rather than caterpillars of A. usamba inferred that this butterfly was aphytophagous, but we found that the larval guts of A. usamba and two known herbivorous lycaenid species contain chloroplast 16S sequences. Moreover, chloroplast barcoding revealed high sequence similarity between chloroplasts found in A. usamba guts and the chloroplasts of the Vachellia drepanolobium trees on which the caterpillars live. Stable isotope analysis provided further evidence that A. usamba caterpillars feed on V. drepanolobium, and the possibilities of strict herbivory versus limited omnivory in this species are discussed. These results highlight the importance of combining multiple approaches and considering ontogeny when using stable isotopes to infer trophic ecology where direct observations are difficult or impossible.