Females tend to occupy smaller puma alife areas and disperse shorter distances, thus being more philopatric than males ( Logan and Sweanor, 2001 ; Maehr et al. , 2002 ). Overall, the species' ecological flexibility and dispersal capabilities have the potential to induce broad genetic connectivity across large geographic areas, unless historical barriers have limited gene flow among populations.Fossil evidence indicates that pumas were already present in North America 0.4 million years ago (MYA) ( Kurtén and Anderson, 1980 ). 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 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 puma athletic shoes 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 ( Woodburne, 2010 ; Eizirik, 2012 ). However, since the credibility interval of this estimate slightly overlaps the timing of the GABI, this puma basket black 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.

We obtained blood and tissue samples from 77 pumas including wild individuals captured during field-ecology projects, caught by farmers or road-killed, as well as captive animals with known geographic origin ( Table S1 ). In addition, we also collected data from 109 additional individuals whose DNA was already available in the participant laboratories, some of which had been used puma basket bow 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.To select a marker that would provide suitable information levels, we initially examined the mtDNA fragments used in previous studies, especially those involving Neotropical felids ( e.g. Eizirik et al. , 1998 , 2001 ; Johnson et al. , 1998 , 1999 ; Culver et al. , 2000 ).

The resulting novel puma sequences were deposited in GenBank (accession numbers {"type":"entrez-nucleotide-range","attrs":{"text":"KF460496-KF460523","start_term":"KF460496","end_term":"KF460523","start_term_id":"572098819","end_term_id":"572098873"}} KF460496-KF460523 ).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. We explored two outgroup options for rooting puma basket heart black the network, one with the P. yagouaroundi sequences generated here, and the other employing M.

We assessed the t MRCA for four different sample sets: (i) complete sample; (ii) South America only; (iii) North and Central Americas (NA NCA); and (iv) a subset of 24 NA NCA samples that formed a regionally endemic subclade in the haplotype network, suggestive of autochthonous diversification (see Results). The BSP analyses were performed only for sets (i) and (ii), as their larger sample size allowed for more robust demographic inferences. The MCMC parameters were the same as in the first set of Beast analyses.