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ÿþThe puma is an iconic predator that ranges new balance 501 womens throughout 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.

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 MYA) and the implied colonization of South America by any felid ( new balance 574 red Woodburne, 2010 ; Eizirik, 2012 ). 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 new balance 993 mens indicating that the extinct North American felid Miracinonyx trumani is the puma's closest relative, with a divergence time estimated at 3.19 MYA.

Four of these subspecies were distributed in South America, indicating that most of the species' historical subdivision occurred in that subcontinent. In 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 new balance 1080 mens 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. , 2012 ).

In addition to expanding the geographic coverage of South American regions to refine inferences on patterns of matrilineal subdivision, we have performed novel analyses on puma demographic history, which revealed consistent evidence of a recent population expansion in South America, prior to re-colonization of North America.

In addition, we also collected data from 109 additional individuals new balance cruz v2 whose DNA was already available in the participant laboratories, some of which had been used in earlier genetic studies employing different markers ( Culver et al. , 2000 ; Castilho et al. , 2011 ; Miotto et al. , 2011 , 2012 ). Therefore, as a whole we collected novel data from a total of 186 individuals. These samples were originated from a diverse array of geographic regions throughout most of the puma range, with greater emphasis on South America (see Table S1 ). Two samples of Puma yagouaroundi were also included to be used as outgroups in some analyses.