Females tend to occupy puma alife smaller 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.

In addition, the genetic diversity within South America was found to be larger than puma athletic shoes 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 puma basket black 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 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. puma basket bow , 2008 ), amplifying a longer fragment ( ca. 750 bp) and exhibiting successful amplification across several families ( e.g. Felidae, Mustelidae, Mephitidae, Procyonidae [unpublished data]).In the present study we employ this longer ND5 segment to investigate the evolutionary history of P. concolor , with emphasis on South American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 ).

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 the network, one with the P. yagouaroundi sequences generated here, and the other employing M.

Finally, we conducted a set of analyses to investigate the demographic history of pumas. We performed puma basket heart black neutrality tests (Tajima's D, Fu & Li' D* and F*, Fu's Fs) with DnaSP, as well as a mismatch distribution analysis ( Rogers and Harpending, 1992 ; Schneider and Excoffier, 1999 ) with Arlequin. In addition, we used the program Beast 1.6.1 ( Drummond and Rambaut, 2007 ) to perform estimates of coalescence times and past demography. We defined the best model of nucleotide substitution for our data set, which was the HKY ( Hasegawa et al. , 1985 ) G model, using the Akaike Information Criterion (AIC, Akaike, 1974 ) implemented in jModelTest 0.1 ( Posada, 2008 ).