DNA metabarcoding-based diet survey for the Eurasian otter (Lutra lutra): Development of a Eurasian otter-specific blocking oligonucleotide for 12S rRNA gene sequencing for vertebrates


Autoři: Priyanka Kumari aff001;  Ke Dong aff001;  Kyung Yeon Eo aff003;  Woo-Shin Lee aff004;  Junpei Kimura aff005;  Naomichi Yamamoto aff001
Působiště autorů: Department of Environmental Health Sciences, Graduate School of Public Health, Seoul National University, Seoul, South Korea aff001;  Institute of Health and Environment, Graduate School of Public Health, Seoul National University, Seoul, South Korea aff002;  Conservation and Research Center, Seoul Zoo, Gwacheon, South Korea aff003;  Department of Forest Sciences, College of Agriculture and Life Science, Seoul National University, Seoul, South Korea aff004;  College of Veterinary Medicine, Seoul National University, Seoul, South Korea aff005
Vyšlo v časopise: PLoS ONE 14(12)
Kategorie: Research Article
doi: 10.1371/journal.pone.0226253

Souhrn

The Eurasian otter (Lutra lutra) is an endangered species for which diet analyses are needed as part of its conservation efforts. Eurasian otters feed on vertebrates, such as fishes, and invertebrates, such as crustaceans, but their detailed taxonomies are not fully understood in part due to limited resolving power of traditional morphological identification methods. Here, we used high-throughput sequencing (HTS)-based DNA metabarcoding approaches to analyze diet profiles of Eurasian otters inhabiting a marshy estuary area in Korea. We investigated their diet profiles based on spraint sampling followed by DNA metabarcoding analyses targeting 12S rRNA gene region for vertebrates, 16S rRNA gene region for invertebrates, and cytochrome c oxidase 1 (COI) gene region for fishes. For the vertebrate analysis, a blocking oligonucleotide (OBS1) was designed to suppress amplification of DNA fragments derived from the otters. The 12S rRNA gene sequencing assay detected species belonging to fishes (95%) and amphibians (3.3%). Fishes detected by 12S rRNA gene sequencing included crucian carp (Carassius auratus), mullets (Mugil spp.), bluegill (Lepomis macrochirus), and northern snakehead (Channa argus), which were also detected by COI gene sequencing. Among invertebrates, mud flat crabs (Helicana spp.) and shrimps (Palaemon spp.) were abundant. The designed blocking oligonucleotide OBS1 effectively inhibited amplification of the otter’s DNA, with only up to 0.21% of vertebrate sequence reads assigned to the otter. This study demonstrated that HTS-based DNA metabarcoding methods were useful to provide in-depth information regarding diet profiles of the otters at our sampling site. By using HTS-based DNA metabarcoding approaches, future research will explore detailed taxonomies of their diets across locations and seasons.

Klíčová slova:

DNA – Gene sequencing – Invertebrates – Oligonucleotides – Otters – Ribosomal RNA – Sequence databases – Vertebrates


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2019 Číslo 12