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Adrados, B.; Zanin, M.; Silveira, L.; Villalva, P.; Ch vez, C.; Keller, C.; Gonz lez-Borrajo, N.; Harmsen, B.J.; Rubio, Y.; Palomares, F.
Non-invasive genetic identification of two sympatric sister-species: ocelot (_Leopardus pardalis_) and margay (_L. wiedii_) in different biomes
2017  Conservation Genetics Resources (11): 203-217

The method presented in here allows for the unequivocal differentiation between non-invasive samples of ocelots and margays, when used in combination with the protocol by Roques et al. (Mol Ecol Resour 11:171-175, 2011). It reduced costs and laboratory processing, decreasing the chances of contamination, and therefore facilitating the identification of large numbers of samples by simple PCR amplification, without the need for sequencing. Our protocol was tested on 371 faeces collected in the field in 25 different sites across the distribution areas of both species, and compared to the mtDNA sequencing of 23 of the samples. The concatenation of a commonly used segment of Cytochrome b with other mitochondrial markers such as ATP-8 allows the identification of these two sister-species, but there was not enough resolution when used alone. With this new method, we could identify 98% of the field faeces, obtaining a global rate of 22.3 faeces of ocelot per margay sample in roads and trails located in potential areas of coexistence, suggesting generally lower densities of margay, although margays could be underestimated due to their arboreal habits. In Serra das Almas, located in the Brazilian Caatinga, the number of faeces of ocelot detected was especially elevated. Low densities of ocelots could be favoring higher densities of margays in areas like the Sinaloan dry forests and the Chimalapas montane forests, both in Mexico. Ocelots were found from open to forested habitats, including mosaic cropland and grassland, however margays were only found in forests where average canopy height exceeded 5 m.

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