Cloacal swab sampling is a reliable and harmless source of DNA for population and forensic genetics in tortoises
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TECHNICAL NOTE
Cloacal swab sampling is a reliable and harmless source of DNA for population and forensic genetics in tortoises Nadia Mucci • Chiara Mengoni • Elisa Berti Ettore Randi
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Received: 17 January 2014 / Accepted: 23 June 2014 / Published online: 24 July 2014 Ó Springer Science+Business Media Dordrecht 2014
Abstract Testudo graeca, Testudo hermanni and Testudo marginata are endangered species listed in the IUCN Red List and Appendix II of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). Correct identification of species and distinct population units can support sound conservation projects and forensic applications. However, the collection of biological samples is not always easy or harmless. Blood sampling is usually invasive and risky. Salivary swab sampling is commonly used in other reptiles but can be unsafe for tortoises by the reason of their head retraction escape response. Cloacal swab sampling should be easier and less risky. In this study, we genotyped 37 tortoises at eight microsatellite loci and compared the reliability of individual genotypes obtained from blood, cloacal and buccal swabs. Results showed that performances of cloacal samples are comparable with those of buccal and blood samples, and proved that cloacal sampling is an alternative and reliable source of DNA for tortoise genotyping.
Electronic supplementary material The online version of this article (doi:10.1007/s12686-014-0251-3) contains supplementary material, which is available to authorized users. N. Mucci (&) C. Mengoni E. Randi Laboratorio di Genetica, Istituto Superiore per la Protezione e la Ricerca Ambientale (ISPRA), Via Ca` Fornacetta 9, 40064 Ozzano dell’Emilia, BO, Italy e-mail: [email protected] E. Berti Centro Tutela e Ricerca Fauna esotica e selvatica Monte Adone, Bologna, Italy E. Randi Department 18/Section of Environmental Engineering, Aalborg University, Sohnga˚rdsholmsvej 57, 9000 Aalborg, Denmark
Keywords CITES Blood sampling Buccal swab Cloacal swab Microsatellite loci Tortoises
Introduction During the past decades, the use of microsatellite markers (single tandem repeats—STRs) permitted to identify the fine-scale structuring in natural populations (Perez et al. 2013; De Barba et al. 2010; Mucci et al. 2010; Fabbri et al. 2007) and supported the discovery of illegal traffics and wildlife crime in forensic genetics (Alacs et al. 2010; Mucci et al. 2014; Barbanera et al. 2012; Caniglia et al. 2010). Availability of new molecular markers such as the use of animal traces and non-invasive samplings as a source of DNA, allowed researchers to accomplish conservation projects with minimal stress on animals. Tortoise species (genus Testudo) are listed in the Appendix II of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES; www.cites.org) and the IUCN Red List. In these species, the collection of blood for DNA extraction can be unsafe for animals because the access to the main blood vessels requires a man
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