Article

Applicability of mitochondrial DNA for the identification of Arvicolid species from faecal samples: A case study from the threatened Cabrera's vole

Molecular Ecology Resources (Impact Factor: 3.71). 02/2011; 11(2):409 - 414. DOI: 10.1111/j.1755-0998.2010.02939.x

ABSTRACT

Arvicolid mitochondrial genomes evolve faster than in any other mammalian lineage. The genetic diversity exhibited by these rodents contrasts sharply with their phenotypic homogeneity. Furthermore, faecal droppings from Arvicolid rodents of similar body size are almost undistinguishable on the basis of pellet morphology and content. In this study, we advantaged from their high genetic diversity vs. phenotypic homogeneity to document the applicability of mtDNA extraction from vole droppings for latter identification of such via a rapid and efficient nested PCR-based technique using the threatened Microtus cabrerae as a model species. We sequenced the mitochondrial control region from 75 individuals belonging to 11 species of Arvicolinae from Spain, Portugal, Greece and Italy, and an additional 19 sequences from ten Microtus species from other countries were downloaded from Genbank. Based on these control region sequences, we successfully designed and applied a nested PCR for M. cabrerae-specific and arvicolid-generic mtDNA markers to differentiate Cabrera’s vole faecal samples among other species of the Arvicolinae subfamily. Although this study used Cabrera’s vole as a model species, similar techniques based on mtDNA sequences may find a broader applicability for noninvasive genetic conservation of vole species and their populations.

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Available from: Ramón C. Soriguer
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    • "PCR and sequencing of the mtDNA control region Polymerase chain reactions (PCRs) to amplify a section of the mitochondrial control region (d-loop) were performed using arvicolid specific primers as described by Alasaad et al. (2011). The 30 ml reaction contained 2 ml of template DNA (from bone or tissue samples), 0.25 mM of Pro þ (Haring et al. 2000) and MicoMico primers (Alasaad et al. 2011) "
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    • "There have been particular efforts to obtain DNA from carnivore scats. Considering nuclear gene amplification , generally, there has been similar DNA amplification success for fresh carnivore scats (71.7%, Murphy et al. 2007; 79.2%, Oliveira et al. 2010) and fresh rodent faeces (78%, our study; 90%, Alasaad et al. 2011; 83.3%, Moran et al. 2008). However, it is difficult to confirm such a pattern for exposed faecal samples, as, to our knowledge, there is no previous literature making use of rodent faeces collected in the field to amplify nuclear loci. "
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