A rapid PCR-based assay for identification of cryptic Myotis spp. (M. mystacinus, M. brandtii and M. alcathoe)
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The development of a quick PCR-based method to distinguish European cryptic Myotis spp., Myotis mystacinus, Myotis brandtii and Myotis alcathoe is described. Primers were designed around species-specific single nucleotide polymorphisms (SNP’s) in the ND1 mitochondrial gene, and a pair of control primers was designed in the 12S mitochondrial gene. A multiplex of seven primer combinations produces clear species-specific bands using gel electrophoresis. Robustness of the method was tested on 33 M. mystacinus, 16 M. brandtii and 15 M. alcathoe samples from across the European range of these species. The method worked well on faecal samples collected from maternity roosts of M. mystacinus. The test is intended to aid collection of data on these species through a rapid and easy identification method with the ability to use DNA obtained from a range of sources including faecal matter.
KeywordsCryptic species Chiroptera Myotis
The authors would like to thank John Altringham’s group, Conor Kelleher, Wigbert Schorcht, Julia Prüger, Laurent Arthur, Yann Le Bris, Nicolas Chenaval, Olivier Farcy, Arnaud Le Houédec and Vincent Lecoq for sending us samples. Thanks to Yannis Kazoglou, Elena Papadatou, Niko Xega, Xavier Grémillet and other volunteers as well as the Society for the Protection of Prespa (SPP) for assisting us during samples’ collection in the field. Samples collection by the authors was done under permits from the National Parks and Wildlife Service (Licence No. 74 C/2008), Ireland; the Greek Ministry of Agricultural Development and Food (Permit No. 104694/2439), Albanian Ministry of Environment, Forestry and Water Administration and French Direction Régionale de l’Environnement, de l’Aménagement et du Logement (Arrêté n° 2009-11). This research was carried out through the Centre for Irish Bat Research, funded by the National Parks and Wildlife Service.
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