ÿþThe puma is an iconic predator that ranges throughout the Americas, all black puma shoes 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, with emphasis on South America.

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 marshes to dry scrubland and cold all black puma sneakers Andean or Patagonian biomes ( Redford and Eisenberg, 1992 ; Nowak, 1999 ). Pumas are solitary and territorial, with large home ranges.

This finding would support the hypothesis of a North American origin for the puma, with subsequent colonization of South America by this species, all puma shoes in parallel with that of the jaguarundi.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, indicating that most of the species' historical subdivision occurred in that subcontinent.

These samples were all red puma shoes 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 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 all white puma shoes 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 ).