The skin microbiome (based on 16S) of Antarctic Humpback whales (Megaptera novaeangliae)

Последняя версия опубликовано SCAR - Microbial Antarctic Resource System мар 19, 2019 SCAR - Microbial Antarctic Resource System
Дата публикации:
19 марта 2019 г.
Опубликовано:
SCAR - Microbial Antarctic Resource System
Лицензия:
CC-BY 4.0

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Описание

Amplicon sequencing dataset (Illumina MiSeq) of the microbiome (16S) on skin samples of Humpback whales (Megaptera novaeangliae)

Версии

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Как оформить ссылку

Исследователи должны дать ссылку на эту работу следующим образом:

Bierlich K, Miller C, DeForce E, Friedlaender A, Johnston D, Apprill A (2019): The skin microbiome (based on 16S) of Antarctic Humpback whales (Megaptera novaeangliae). v1.1. SCAR - Microbial Antarctic Resource System. Dataset/Metadata. https://ipt.biodiversity.aq/resource?r=antactice_humpback_whale_skin_microbiome&v=1.1

Права

Исследователи должны соблюдать следующие права:

Публикующей организацией и владельцем прав на данную работу является SCAR - Microbial Antarctic Resource System. Эта работа находится под лицензией Creative Commons Attribution (CC-BY 4.0).

Регистрация в GBIF

Этот ресурс был зарегистрирован в GBIF, ему был присвоен следующий UUID: 0144ad97-f933-4557-a599-0dd4c736066e.  SCAR - Microbial Antarctic Resource System отвечает за публикацию этого ресурса, и зарегистрирован в GBIF как издатель данных при оподдержке Scientific Committee on Antarctic Research.

Ключевые слова

Metadata

Контакты

Kevin Bierlich
  • Originator
  • Point Of Contact
Woods Hole Oceanographic Institution
Woods Hole
US
Carolyn Miller
  • Originator
Woods Hole Oceanographic Institution
Woods Hole
US
Emelia DeForce
  • Originator
Woods Hole Oceanographic Institution
Woods Hole
US
Ari Friedlaender
  • Originator
Oregon State University
Newport
US
David Johnston
  • Originator
Duke University Marine Laboratory
Beaufort
US
Amy Apprill
  • Originator
Woods Hole Oceanographic Institution
Woods Hole
US
Maxime Sweetlove
  • Metadata Provider
Research assistent
Royal Belgian Institute for Natural Sciences
Rue Vautier 29
1000 Brussels
BE

Географический охват

Healthy humpback whales in te Gerlache Strait, Palmer Deep Canyon and Bransfield Strait (Antarctica)

Ограничивающие координаты Юг Запад [-68,077, -69,898], Север Восток [-64,046, -61,605]

Таксономический охват

Bacteria and Archaea were targeted using universal primers for the 16S ssu rRNA gene (V4 region)

Domain Bacteria (Bacteria), Archaea (Archaea)

Временной охват

Период формирования 2010-2013

Данные проекта

Описание отсутсвует

Название The skin microbiome (based on 16S) of Antarctic Humpback whales (Megaptera novaeangliae)
Финансирование Humpback whale skin biopsy samples were collected under NOAA permit 808-735 and ACA permit 2009-14 as part of NSF OPP awards ANT-07-39483 and 1250208. This project was supported by 67 donors to the “Whale Bacterial Buddies” crowd- funded project supported by WHOI, the Edna Bailey Sussman Fund, and the Michael K. Orbach Enrichment Fund awarded.

Исполнители проекта:

Kevin Bierlich

Методы сбора

Skin samples were collected from the upper flank near the dorsal fin of humpback whales via standard biopsy techniques. Specifically, a 68-kg pull crossbow and floating Ceta-Dart darts with 40-mm surgical stainless steel tips collected a small skin and blubber sample roughly 5 mm wide and 20 to 30 mm deep, depending on the angle of the sampling dart on impact. Tips were sterilized using 8.25% concentrated bleach, allowed to dry, cleaned, and then doused in 100% ethyl alcohol before each use. The darts were retrieved by hand and the tips immediately placed into a sterile bag (such that the sample was not contaminated) and on ice for no more than 10 h before transfer to a cryovial via sterile tools for freezing at 80°C. The global positioning system (GPS) location and the regional names of the bay, strait, etc. were recorded for each skin sample collected. One skin sample, w132a, was collected from an acoustic tag placed on the animal.

Охват исследования Ninety-four skin samples were collected from 89 humpback whales in 12 different locations along the Western Antarctic Peninsula (WAP) during May and June 2010 (fall/late foraging season) and January and February 2013 (early summer/early foraging season). Five individuals were sampled twice during the study; three individuals were resampled on the same day, one a week later, and one was sampled in 2010 and 2013.

Описание этапа методики:

  1. DNA was extracted from 1 to 30 mg of the top layer of the epidermis, with an average of 13 mg, from each sample using a DNeasy tissue kit (Qiagen, Valencia, CA, USA) and quantified using the Invitrogen Qubit 2.0 assay fluorometer (Life Technologies, Beverly, MA, USA). The V4 region of the SSU rRNA gene was amplified using barcoded primers (515F-Y, 5=-GTGYCAGCMGCCGCGGTAA-3=; and 806RB, 5=-GGACTACNVGGGTWTCTAAT-3=), broadly targeting bacteria and archaea. Samples and sterile water (negative control) were amplified in triplicate using PCR on a S1000 thermal cycler (Bio-Rad Laboratories, Hercules, CA, USA), as follows: 2 min at 95°C, followed by 30 to 35 cycles of 20 s at 95°C, 15 s at 55°C, and 5 min at 72°C, followed by 10 min at 72°C. Triplicate PCR mixtures contained 1 ul of DNA, 5 ul of GoTaq 5 Flexi buffer, 14.75 l of H2O, 2.5 l of a 25 nM MgCl2 solution, 200 nM each dinucleoside triphosphate (dNTP), and 0.25 l of a 5 units/ l GoTaq DNA polymerase solution per sample, along with 200 nM each primer. After PCR, amplification was assessed by mixing 5u l of each sample with 1 ul of 10,000 Sybr Safe dye run on a 1% agarose–Tris- borate-EDTA (agarose-TBE) gel. The replicate reaction mixtures were purified using Agencourt AMPure XP beads (Beckman Coulter, Inc., Pasadena, CA, USA) and quantified using the Qubit assay. Barcoded amplicons were sequenced using a 2x250-bp MiSeq Illumina format at the University of Illinois W. M. Keck Center for Comparative and Functional Genomics.

Библиографические ссылки

  1. Bierlich, K. C., Miller, C., DeForce, E., Friedlaender, A. S., Johnston, D. W., & Apprill, A. (2018). Temporal and regional variability in the skin microbiome of humpback whales along the Western Antarctic Peninsula. Appl. Environ. Microbiol., 84(5), e02574-17. https://doi.org/10 .1128/AEM.02574-17

Дополнительные метаданные

Альтернативные идентификаторы 0144ad97-f933-4557-a599-0dd4c736066e
https://ipt.biodiversity.aq/resource?r=antactice_humpback_whale_skin_microbiome