European Journal of Wildlife Research

, Volume 54, Issue 2, pp 253–261 | Cite as

A non-invasive genetic method to identify the sympatric mustelids pine marten (Martes martes) and stone marten (Martes foina): preliminary distribution survey on the northern Iberian Peninsula

  • Aritz Ruiz-González
  • Jonathan Rubines
  • Oskar Berdión
  • Benjamín J. Gómez-Moliner
Original Paper


The closely related mustelids European pine marten (Martes martes) and stone marten (Martes foina) sympatrically inhabit a large area of Europe. However, given our limited knowledge of their bioecological relationships, their extremely elusive behaviour and the fact that their faeces cannot be distinguished on the basis of morphology alone, it is very difficult to monitor their populations. In this study, we describe a reliable non-invasive polymerase chain reaction (PCR)–restriction fragment length polymorphism (PCR-RFLP) method for distinguishing between M. martes and M. foina based on the analysis of deoxyribonucleic acid extracted from faeces samples. The method was specifically designed to avoid possible interference from potential prey mammals and other sympatric carnivores. The procedure consists of PCR amplifying a mitochondrial D-loop region followed by digesting the resulting 276-bp-long amplicons with the restriction enzymes HaeIII and RsaI. To assess the efficiency of this technique, we conducted a preliminary field study across the potential sympatric distribution areas of both marten species in the northern Iberian Peninsula. Out of 359 faeces samples collected, we identified 80 as specimens from the stone marten and 235 from the pine marten. Unequivocal species identification was thus possible in 88% of the faeces samples collected. These findings reveal the combined use of non-invasive genetic sampling and GIS technology to be a reliable and cost-effective procedure for improving our knowledge of the spatial distributions of sympatric marten species. This protocol could also be used to identify and improve information gaps, to develop effective research and management programmes and in population and landscape genetics studies of marten species.


Non-invasive genetic sampling Genetic species identification Faecal DNA 



This study was funded by the Biodiversity Section, Dept. of Territorial Planning and Environment of the Basque Government. A. Ruiz-González holds a Ph.D. fellowship awarded by the Dept. of Education Universities and Research (Basque Government). The authors wish to thank the following persons and institutions for supplying tissue and faecal samples: the technical staff of the National Parks of Ordesa and Monte Perdido (E. Villagrasa), Picos de Europa (A. Mora) and Aigüestortes i Estany de Saint Mauricy (J. Canut); J.Herr (Department of Biology and Environmental Science, University of Sussex.), Dr. A. Abramov (Zoological Institute, Russian Academy of Sciences), A. Fernandez (Doñana Biological Station-University of Oviedo), Dr. I. Barja and his research group (UAM), F. López-Giraldez (DNA and Tissue Collection, Pompeu Fabra University), P. Aymerich (PN Alt Pirineu), J.M. Fernandez (IAN), G. Belamendia (CEA–MCN Álava), P. Lizarraga and L. Lorza (CRF Martioda-DFA), J. Pinedo (DFA), I. Amigo (DFV), H. Aguirre, P. Pérez and G. Dominguez. We are also indebted to Mikel Gurrutxaga (Dept. of Natural Environment and Geographical Information System, IKT) for his help with the GIS treatment of data and preparing maps.

Supplementary material


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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Aritz Ruiz-González
    • 1
  • Jonathan Rubines
    • 1
  • Oskar Berdión
    • 1
  • Benjamín J. Gómez-Moliner
    • 1
  1. 1.Department of Zoology and Animal Cell Biology, Zoology Laboratory, Facultad de FarmaciaUniversidad del País Vasco (UPV-EHU)VitoriaSpain

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