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Skin swabbing as a new efficient DNA sampling technique in amphibians, and 14 new microsatellite markers in the alpine newt (Ichthyosaura alpestris)

Authors

  • J. PRUNIER,

    1. Ecosphère, 3bis rue des Remises, 94100 Saint Maur des Fossés, France
    2. UMR5023 Ecologie des Hydrosystèmes Naturels et Anthropisés, Université de Lyon, Université Lyon 1, ENTPE, CNRS, Villeurbanne, F-69622, France
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  • B. KAUFMANN,

    1. UMR5023 Ecologie des Hydrosystèmes Naturels et Anthropisés, Université de Lyon, Université Lyon 1, ENTPE, CNRS, Villeurbanne, F-69622, France
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  • O. GROLET,

    1. UMR5023 Ecologie des Hydrosystèmes Naturels et Anthropisés, Université de Lyon, Université Lyon 1, ENTPE, CNRS, Villeurbanne, F-69622, France
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  • D. PICARD,

    1. Université d’Angers, Laboratoire d’Etudes Environnementales des Systèmes Anthropisés, 2 Boulevard Lavoisier, 49045 Angers Cedex 01, France
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  • F. POMPANON,

    1. Université Joseph Fourier, Laboratoire d’Ecologie Alpine, CNRS, UMR 5553, 2233 Rue de la Piscine, 38041 Grenoble Cedex 9, France
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  • P. JOLY

    1. UMR5023 Ecologie des Hydrosystèmes Naturels et Anthropisés, Université de Lyon, Université Lyon 1, ENTPE, CNRS, Villeurbanne, F-69622, France
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Jérôme Prunier, Fax: +33-145-11-24-37; E-mail: jerome.prunier@ecosphere.fr

Abstract

This study introduces a novel DNA sampling method in amphibians using skin swabs. We assessed the relevancy of skin swabs relevancy for genetic studies by amplifying a set of 17 microsatellite markers in the alpine newt Ichthyosaura alpestris, including 14 new polymorphic loci, and a set of 11 microsatellite markers in Hyla arborea, from DNA collected with buccal swabs (the standard swab method), dorsal skin swabs and ventral skin swabs. We tested for quality and quantity of collected DNA with each method by comparing electrophoresis migration patterns. The consistency between genotypes obtained from skin swabs and buccal swabs was assessed. Dorsal swabs performed better than ventral swabs in both species, possibly due to differences in skin structure. Skin swabbing proved to be a useful alternative to buccal swabbing for small or vulnerable animals: by drastically limiting handling, this method may improve the trade-off between the scientific value of collected data, individual welfare and species conservation. In addition, the 14 new polymorphic microsatellites for the alpine newt will increase the power of genetic studies in this species. In four populations from France (= 19–25), the number of alleles per locus varied from 2 to 16 and expected heterozygosities ranged from 0.04 to 0.91. Presence of null alleles was detected in two markers and two pairs displayed gametic disequilibrium. No locus appeared to be sex-linked.

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