ABSTRACT The aim of this study was to develop a preliminary physical map of single nucleotide polymorphisms (SNPs) in alpaca using an alpaca/hamster radiation hybrid panel and a bovine highdensity SNP genotyping microarray (BovineHD BeadChip-Illumina). The methodology included genotyping 92 alpaca/hamster hybrid cell clones, and four control samples (male alpaca, female alpaca, hamster and 1:10 DNAmixture) with the microarray. After genotyping the alpaca and hamster DNA control samples, only bovine SNPs with a call frequency of 1 were retained. The SNPs identified in the alpaca DNA samples were then filtered to remove those also found in the hamster. From the remaining alpaca SNPs, to decrease the probability of false positives, only those with a call frequency from 0.2 to 0.8 in the 92 hybrid clone samples were retained for the final analysis. The remaining alpaca specific SNPs were tabulated in MapMaker format and were analysed with the Carthagene software to identify linkage groups. The linkage groups were mapped to the reference genome Vicugna_pacos-2.0.2, using the BLAST software and the SHORTBLAST command on the Galaxy platform. The two alpaca control samples had 294 165 SNPs with positive signals in the bovine microarray. After eliminating the most common hamster SNPs and the most-likely false positives, 2924 SNPs remained. These were assigned to 33 linkage groups encompassing 216 SNPs. The estimated distance between these SNPs ranged from 65.3 and 671.9 cR, with a logarithm of the odds (LOD) >6.0. Finally, a total of 31 SNPs was found in the reference genome (E-value <0.05).

RESUMEN El objetivo del estudio fue desarrollar un mapa físico preliminar de polimorfismos de nucleótido simple (PNSs) en alpaca usando un panel celular híbrido irradiado alpaca/ hámster y una micromatriz de genotipado de alta densidad de bovino (BovineHD BeadChip-Illumina). Se realizó el genotipado de 92 clones celulares híbridos irradiados y cuatro muestras control (alpaca macho, alpaca hembra, hámster y 1:10 alpaca/hámster mezcla de ADN) con la micromatriz. Luego del genotipaje de las muestras control del ADN de alpaca y hámster solo se retuvieron PNSs de bovinos que mostraron una frecuencia de señal positiva de 1. Los PNSs identificados en el ADN de alpaca fueron filtrados para sustraer los presentes en el hámster. De estos últimos, solo se retuvieron para el análisis final aquellos que tuvieron una frecuencia de señal positiva de 0.2 a 0.8 para los 92 clones y se eliminaron los PNSs que presentaron falsos positivos. Los restantes PNSs específicos de alpaca fueron tabulados en el formato MapMaker y fueron analizados con el programa Carthagene para identificar grupos de ligamiento. Los grupos ligados fueron ubicados en el genoma referencial Vicugna_pacos-2.0.2, aplicando el programa BLAST y el comando SHORTBLAST, en la plataforma Galaxy. Las dos muestras control de alpaca registraron 294 165 PNSs, con señal positiva en la micromatriz de bovino. La cantidad de PNSs de alpaca, después de eliminar los PNSs positivos comunes con hámster, fue 50 686 y luego de eliminar los posibles falsos positivos quedaron 2924 PNSs. Esta cantidad se analizó con el programa Carthagene y resultó en 33 grupos de ligamiento, con un total de 216 PNSs. Las distancias calculadas entre estos PNSs oscilan entre 65.3 y 671.9 cR, con un logaritmo de probabilidades (LOD) >6.0. Finalmente, se encontraron un total de 31 PNSs en el genoma referencial (E-value<0.05).

Abstract To investigate the evolution and biogeography of an endemic group of South American foxes, we examined mitochondrial DNA control region sequences for 118 individuals belonging to all six extant species of the genus Lycalopex. Phylogenetic and molecular dating analyses supported the inference that this genus has undergone a very recent and rapid radiation, stemming from a common ancestor that lived ca. 1 million years ago. The Brazilian endemic L. vetulus was supported as the most basal species in this genus, whereas the most internal group is comprised by the recently diverged (ca. 350,000 years ago) Andean/Patagonian species L. griseus and L. culpaeus. We discuss the inferred phylogenetic relationships and divergence times in the context of the current geographic distributions of these species, and the likely effects of Pleistocene climatic changes on the biogeography of this group. Furthermore, a remarkable finding was the identification of multiple individuals classified as L. gymnocercus bearing mtDNA haplotypes clearly belonging to L. griseus, sampled in regions where the latter is not known to occur. At a minimum, this result implies the need to clarify the present-day geographic distribution of each of these fox species, while it may also indicate an ongoing hybridization process between them. Future testing of this hypothesis with in-depth analyses of these populations is thus a priority for understanding the history, evolutionary dynamics and present-day composition of this endemic Neotropical genus.

The puma is an iconic predator that ranges 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. 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. Our objectives were to refine the phylogeographic assessment within South America and to investigate the demographic history of pumas using a coalescent approach. Our results extend previous phylogeographic findings, reassessing the delimitation of historical population units in South America and demonstrating that this species experienced a considerable demographic expansion in the Holocene, ca. 8,000 years ago. Our analyses indicate that this expansion occurred in South America, prior to the hypothesized re-colonization of North America, which was therefore inferred to be even more recent. The estimated demographic history supports the interpretation that pumas suffered a severe demographic decline in the Late Pleistocene throughout their distribution, followed by population expansion and re-colonization of the range, initiating from South America.