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ÿþThe puma is an iconic predator that ranges throughout puma shoes the Americas, occupying diverse habitats. Previous phylogeographic analyses have revealed that it exhibits moderate levels of genetic structure across its range, with few of the classically recognized subspecies being supported as distinct demographic units. Moreover, most of the species' molecular diversity was found to be in South America. To further investigate the phylogeographic structure and demographic history of pumas we analyzed mtDNA sequences from 186 individuals sampled throughout their range.

Pumas have remarkable dispersal capabilities ( Maehr 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 puma trainers 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.

Indicating that most of the species' historical subdivision occurred in that subcontinent. In puma suede addition, the genetic diversity within South America was found to be larger than in Central and North America, suggesting that pumas from the latter subcontinent actually derive from a recent re-colonization event, following extinction in North America in the Late Pleistocene (ca. 10,000 12,000 years ago).Within the mtDNA, the NADH dehydrogenase subunit 5 ( ND5 ) gene has been successfully used in phylogenetic and phylogeographic studies of felids and other carnivores ( e.g. Culver et al.

2000 ; Trinca et al. , puma sliders 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 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 .

In the present study we employ this longer ND5 segment to investigate the evolutionary history of P. concolor , with emphasis on South American populations, which were previously found to harbor high levels of diversity and inferred to have played a key role in the historical demography of this species ( Culver et al. , 2000 ). Given that the geographic sampling of South American puma uk pumas was limited in that first study, we aimed here to expand the representation of the various regions of this sub-continent.

The availability of ND5 sequences for the extinct felid Miracinonyx trumani ( Barnett et al. , 2005 ) was an additional asset of this segment, allowing the inclusion of this fossil taxon in some analyses.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 ), followed by manual verification and editing.