Evaluating the predictive power of field variables for species and individual molecular identification on wolf noninvasive samples

  • Mónia NakamuraEmail author
  • Raquel GodinhoEmail author
  • H. Rio-Maior
  • S. Roque
  • A. Kaliontzopoulou
  • J. Bernardo
  • D. Castro
  • S. Lopes
  • F. Petrucci-Fonseca
  • F. Álvares
Original Article


Live-trapping elusive animals is often challenging, hampering the achievement of reasonable sample sizes for molecular studies. In such cases, the use of noninvasive samples (NIS) is critical in many research fields, mostly related to ecology, management and conservation of wild species. We analysed the influence of several variables potentially associated with the quality of wolf NIS—season, weather conditions, and in situ collected site and sample characteristics—on the success rates of species and individual identification performed using mtDNA and 13 microsatellites, respectively. NIS included scats, urine and saliva collected from two areas in Portugal. Scat samples exhibited the highest success rate for both species (81%) and individual identification (59%), compared with urine (63 and 30%, respectively) or saliva samples (48 and 36%, respectively). The success rate of species identification of scats was better explained by season of collection, the presence of mucous, moisture and odour. For samples with successful species identification analysis, individual identification success was best predicted by the presence of odour. Performing a preliminary selection of scat samples with the best characteristics can increase up to 13% the success rates of molecular analysis. Urine collected on snow had a higher success rate of species identification than that collected on vegetation. To our knowledge, this was the first time that wolf urine on vegetation near ground-scratching marks is used as DNA source. Saliva samples collected with different substrate types can also be used for species identification. These results contribute to optimising noninvasive sampling procedures, maximising the success of molecular ecology studies, and ultimately minimising sampling efforts and costs.


Sample variable Noninvasive genetic sampling Saliva Faeces Urine Canis lupus 



We thank all field assistants, particularly AS Pedro, D Cadete, J Pereira, J Santos, S Pinto and V Ramiro. We thank CONGEN members (CIBIO/InBIO), JV López-Bao, S Czarnomska, N Santos and B Rodrigues for revising earlier versions of the manuscript. We also thank all anonymous reviewers for constructive comments on previous versions of the manuscript. This is contribution no. 16 from the Iberian Wolf Research Team (IWRT).

Compliance with ethical standards


R.G. and S.R. worked respectively under a Research contract (IF/564/2012) and a PhD grant (SFRH/BD/12291/2003) from Fundação para a Ciência e a Tecnologia. This work was supported by ACHLI (Associação de Conservação do Habitat do Lobo Ibérico), VentoMinho Energias Renováveis S.A and Project ‘Genomics applied to genetic resources’ cofinanced by North Portugal Regional Operational Programme 2007/2013 (ON.2—O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF).

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  1. 1.CIBIO/InBIO, Centro de Investigação em Biodiversidade e Recursos Genéticos, Universidade do PortoVairãoPortugal
  2. 2.Departamento de Biologia, Faculdade de CiênciasUniversidade do PortoPortoPortugal
  3. 3.Grupo Lobo, Faculdade de Ciências da Universidade de LisboaLisbonPortugal
  4. 4.CE3C, Centre for Ecology, Evolution and Environmental ChangeFaculdade de Ciências da Universidade de LisboaLisbonPortugal

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