Pumas have remarkable dispersal capabilities ( Maehr puma creepers et al. , 2002 ; Beier et al. , 2003 ) and successfully occupy a diverse array of habitats, illustrating their potential to adapt to the breadth of environmental conditions occurring across the continent, from tropical forests and marshes to dry scrubland and cold Andean or Patagonian biomes ( Redford and Eisenberg, 1992 ; Nowak, 1999 ). Pumas are solitary and territorial, with large home ranges.

In parallel, molecular data ( Johnson et al. , 2006 ) have led to an estimate of its divergence from the sister-species P. yagouaroundi of 4.17 MYA (C.I.: 3.16 6.01MYA), suggesting a much longer history as a distinct evolutionary puma rihanna lineage. The speciation event that separated these lineages may have occurred in North or South America, with the molecular dating estimate supporting the former, as it tends to predate the Great American Biotic Interchange (GABI) (ca. 2.5 3.5 puma fenty MYA).

However, since the credibility interval of this estimate slightly overlaps the timing of the GABI, this issue is still not fully settled. Interestingly, Barnett et al. (2005) provided molecular evidence indicating that the extinct North American felid Miracinonyx trumani is the puma's closest relative, with a divergence time estimated at 3.19 MYA. This finding would support the hypothesis of a North American origin for the puma, with subsequent colonization of South America by this puma slides species.

In a thorough study of puma phylogeography, Culver et al. (2000) assessed the current and historical genetic diversity present in this species, based on a large sampling of individuals from across its range. That study indicated that most of the 32 classical puma subspecies did not correspond to definable genetic units and reduced the number of recognized subspecies to six. Four of these subspecies were distributed in South America.

2000 ; Trinca et al. , 2012 ). In a previous study focusing on pumas ( Culver et al. , 2000 ), three mtDNA segments were employed ( ND5 , 16S and ATP8 ). Of these, ND5 showed the highest polymorphic content in this species, based on a segment spanning 318 bp. A new primer set puma fenty slides for this gene was designed specifically for carnivores ( Trigo et al. , 2008 ), amplifying a longer fragment ( ca. 750 bp) and exhibiting successful amplification across several families .

Sequence electropherograms were visually inspected and edited using Chromas Lite 2.01 or FinchTV 1.4.0. Sequences were aligned with the CLUSTALW algorithm ( Higgings et al. , 1996 ) implemented in MEGA 4 ( Tamura et al. , 2007 )To investigate the evolutionary relationships among puma mtDNA lineages, haplotype networks were built using the median-joining approach ( Bandelt et al. , 1999 ), as implemented in the software Network 4.5.1.6.