DNA metabarcoding of the prey and microbiome of museum specimen Antarctic trematomid fishes

Registros biológicos
Versión 1.0 publicado por SCAR - Microbial Antarctic Resource System el nov 14, 2019 SCAR - Microbial Antarctic Resource System
Fecha de publicación:
14 de noviembre de 2019
Licencia:
CC-BY 4.0

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Descripción

In this dataset, stomachs and hindguts were sampled from 225 Trematomus specimens from the Natural History Museum London. Fish specimen were collected between 20 and 100 years ago and fixed in either formaldehyde or ethanol. A 313 bp fragment of the cytochrome c oxidase subunit I (COI) was amplified and sequenced for prey item identification in the stomach and a 450 bp region of the 16S rRNA gene to investigate microbiome composition in the gut system.

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Por favor, tenga en cuenta que ésta es una versión antigua del conjunto de datos.  Los usuarios deben citar este trabajo de la siguiente manera:

Sweetlove M (2019): DNA metabarcoding of the prey and microbiome of museum specimen Antarctic trematomid fishes. v1.0. SCAR - Microbial Antarctic Resource System. Dataset/Occurrence. https://ipt.biodiversity.aq/resource?r=historic_antarctic_fish_dataset_2019&v=1.0

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El publicador y propietario de los derechos de este trabajo es SCAR - Microbial Antarctic Resource System. Esta obra está bajo una licencia Creative Commons de Atribución/Reconocimiento (CC-BY 4.0).

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Este recurso ha sido registrado en GBIF con el siguiente UUID: ccba820a-bc62-4cf4-85f2-e0a991efcc6c.  SCAR - Microbial Antarctic Resource System publica este recurso y está registrado en GBIF como un publicador de datos avalado por Scientific Committee on Antarctic Research.

Palabras clave

Samplingevent

Contactos

Maxime Sweetlove
  • Originador
Data officer
Royal Belgian Institute of Natural Sciences
Rue Vautier 29
1000 Brussels
BE
Heindler Franz
  • Proveedor De Contenido
  • Proveedor De Los Metadatos
  • Punto De Contacto
University of Leuven
Leuven
BE
Hendrik Christiansen
  • Proveedor De Los Metadatos
University of Leuven
Leuven
BE
Bruno Frédérich
  • Proveedor De Los Metadatos
University of Liège
Liège
BE
Agnes Detaï
  • Proveedor De Los Metadatos
Muséum National d'Histoire Naturelle
Paris
FR
Gilles Lepont
  • Proveedor De Los Metadatos
University of Liège
Liège
BE
Gregory Maes
  • Proveedor De Los Metadatos
University of Leuven
Leuven
BE
Anton Van de Putte
  • Proveedor De Los Metadatos
Royal Belgian Institute of Natural Sciences
1000 Brussels
BE
Filip Volkaert
  • Proveedor De Los Metadatos
University of Leuven
Leuven
BE

Cobertura geográfica

Various places in the Souther Ocean and the coastal waters of the Antarctic continent. Geographic coordinates not available for all the museum specimen.

Coordenadas límite Latitud Mínima Longitud Mínima [-90, -180], Latitud Máxima Longitud Máxima [-54, 180]

Cobertura taxonómica

Fish specimen of the genus Trematomus, with DNA samples of the stomach and gut for prey composition and micro biome.

Especie Trematomus eulepidotus, Trematomus penellii, Trematomus hansoni, Trematomus eulepidotus, Trematomus brachysoma, Trematomus bernacchii, Trematomus borchgrevinki, Trematomus newnesi, Trematomus loennbergii, Trematomus scotti

Bacterial and Archaeal microbiome in the stomach and gut of the fish was profiled with the 16S ssu rRNA gene

Dominio Bacteria (Bacteria), Archaea (Archaea)

Prey (eukaryotes) of the fish were investigated with the COI marker gene

Dominio Eukaryota (Eukaryotes)

Cobertura temporal

Periodo de formación 1899-2018

Datos del proyecto

No hay descripción disponible

Título SYNTHESYS-RECTO-VERSO
Fuentes de Financiación This research received support from the SYNTHESYS Project (http://www.synthesys.info/), which is financed by European Community Research Infrastructure Action under the FP7 Capacities Program. It furthermore received funds through the Brilliant Marine Research Idea Philanthropy Award 2017 issued by the Vlaams Instituut voor de Zee (VLIZ), Belgium. Research was funded by the Refugia and Ecosystem Tolerance in the Southern Ocean project (RECTO; BR/154/A1/RECTO) as well as the Ecosystem Responses to global change—a multiscale approach in the Southern Ocean project (vERSO; BR/132/A1/vERSO) (http://rectoversoprojects.be), both funded by the Belgian Science Policy Office (BELSPO). This is contribution number 003 of the RECTO project and contribution number 029 of the vERSO project. HC was supported by a grant from the former Flemish agency for Innovation by Science and Technology (IWT), now managed through Flanders Innovation & Entrepreneurship (VLAIO, Grant No. 141328).

Personas asociadas al proyecto:

Heindler Franz
  • Autor

Métodos de muestreo

Fish were carefully dissected: stomachs were opened to remove stomach content and a small portion of the hindgut (1 cm) was removed. Stomach content and hindgut were stored separately in 70% ethanol.

Área de Estudio Stomach and hindgut samples of 225 specimens of the genus Trematomus were obtained from the Natural History Museum, London. Sampling dates ranged from 1901 to 1988, sampling locations were in the Southern Ocean around the Antarctic continent. Contemporary samples were caught with hook and line in the vicinity of the Gerlache Strait, Antarctic Peninsula in the season of 2017–2018.
Control de Calidad During molecular laboratory work special care was taken to prevent (cross-) contamination of samples. Workbench wipes (workbench contamination), human saliva wipes (human contamination) and no-template extractions (blanks) were included as contamination controls for amplification and sequencing.

Descripción de la metodología paso a paso:

  1. A large piece of stomach content (0.5 × 0.5 cm) or the entire piece of hindgut (1 cm) was placed into screwcap microtubes with 500 μl of Phosphate Buffered Saline (PBS) at pH 9. Tissue was fragmented thoroughly in each tube to ameliorate efficiency. Samples were heated to 100°C for 10 min, left to cool on ice for 5 min and then spun down with 20,000 × g for 5 min. PBS was carefully removed without taking any tissue and replaced by 500 μl of PBS at pH 7.2 and again heated to 100°C for 10 min. PBS was again carefully removed and further purification steps were conducted using the commercial Nucleospin® Tissue (Macherey-Nagel, Accession number: 740952) DNA extraction kit following the manufacturer's protocol.
  2. For prey identification a 313 bp region of the COI gene was amplified from the stomach content using the tailed primers NGSmlCOIint and NGSjgHCO2198. The V3 and V4 region (460 bp) of the 16S rRNA gene was amplified using the tailed primers 16s-IllumTS-F and 16s-IllumTS-R to assess the microbiome composition. The reaction mix for the amplicon PCR for COI contained 12.5 μl of MytaqTM 2x Mix (Bioline, Accession number: BIO-25041), 0.5 μl of each primer (20 μM), 10.5 μl of molecular grade water and 1 μl of DNA template with a PCR profile of 10 s of denaturation at 95°C, 30 s of annealing at 62°C and 60 s elongation at 72°C for 16 cycles with the annealing temperature dropping every cycle by 1°C, followed by 25 cycles with an annealing temperature at 46°C. The reaction mix for the amplicon PCR for 16S contained 12.5 μl of MytaqTM 2x Mix, 2.5 μl of each primer (1 μM), 2.5 μl of DNA template (5 ng ul−1) and 5 μl of molecular grade water with a PCR profile of 60 s of initial denaturation at 95°C followed by 25 cycles of 15 s denaturation at 95°C, 15 s of annealing at 55°C and 10 s elongation at 72°C, finishing with a final extension of 72°C for 300 s. PCR products were cleaned up using Agencourt AMPure XP beads (Beckman Coulter, Accession number: A63882) following the manufacturer's instructions with a bead to template ratio of 0.8 to 1. Thereafter followed an indexing PCR, which binds a unique primer barcode to each respective sample following Lange et al. (2014) with a PCR mix of 10 μl of MytaqTM 2x Mix, 0.5 μl of each forward and reverse indexing-primer (to form a unique identifiable primer combination for each sample; 20 μM) and 9 μl of DNA template with a PCR profile of an initial denaturation of 1 min at 95°C followed by 15 cycles of denaturation for 15 s at 95°C, 15 s of annealing at 51°C and 10 s of extension at 72°C finishing with a final extension of 5 min at 72°C. The PCR product was cleaned up again, then quantified using the commercial Quant-iTTM Picogreen® kit (Thermo Fisher) and pooled, if sufficient template (20 ng) was available. Sequencing took place on an Illumina MiSeq PE 3000 (Genomics Core, KU Leuven, Belgium).

Datos de la colección

Nombre de la Colección Antarctic Fish, London Natural History Museum

Referencias bibliográficas

  1. Heindler F.M., Christiansen H., Frédérich B., et al. (2018) ‘Historical DNA Metabarcoding of the Prey and Microbiome of Trematomid Fishes Using Museum Samples’. Frontiers in Ecology and Evolution 6: 151, https://doi.org/10.3389/fevo.2018.00151. https://doi.org/10.3389/fevo.2018.00151.

Metadatos adicionales