The South American fauna has endured challenges such as illegal hunting, cattle expansion, climate change, and landscape fragmentation. Despite ongoing conservation efforts that aim to address these issues, genetic diversity remains overlooked. We genotyped 126 Lama guanicoe individuals from an area of 2 million km(2) in Patagonia and Cuyo using the 60K SNPs Axiom-Camelids, and we assessed population viability through genetic diversity, inbreeding, and population size. We then explored population connectivities through network analyses, genetic differentiation, and Bayesian approaches. Our findings reveal six genetic clusters: Cuyo, Somuncura, North Patagonia, Central Patagonia, South Patagonia, and Tierra del Fuego Island (TDFI). Cuyo and TDFI guanacos differed from others due to isolation-by-distance and geographical barriers, respectively. Cuyo exhibited the highest degree of allele conservation, but TDFI faced heightened vulnerability due to isolation and increased homozygosity. Sharp bottlenecks in Cuyo and North Patagonia coincided with the expansion of sheep farming, while bottlenecks in the other populations occurred during the Little Ice Age. North Patagonia acted as a probable contact zone, facilitating mixing of the Cuyo, Somuncura, and Central Patagonia populations. The Central and South Patagonian populations presented the highest gene flow. Thus, genomic advances help to define distinct conservation units (genetic stocks) to guide management practices and decisions.