This study analyzes ancient DNA from El Mirón Cave, Spain, to explore human and animal life during the Last Glacial Maximum, 26.5 to 19 thousand years ago. Focusing on the Late Mousterian to Initial Magdalenian periods, the research identified 29 species, including 14 not previously detected archaeologically. Findings include the presence of hyenas during the Magdalenian and evidence of mitochondrial DNA continuity among humans throughout the Solutrean, underscoring the Iberian Peninsula’s importance in understanding Pleistocene survival.. A sedimentary ancient DNA perspective on human and carnivore persistence through the Last Glacial Maximum in El Mirón Cave, Spain.

Caves are primary sites for investigating human and carnivore occupation and subsistence patterns throughout the Palaeolithic. Iberia served as a critical human and animal refugium in Europe during the Last Glacial Maximum (LGM), 26.5 to 19 thousand years before the present (cal. kya), marked by significant climatic cooling and environmental changes. As such, the Iberian Peninsula is a key location for understanding human population dynamics before, during and after these climatic events. We recovered and analysed sedimentary ancient DNA (sedaDNA) data from the lower archaeological stratigraphic sequence of El Mirón cave (Cantabria, Spain), encompassing the Late Mousterian (associated with Neanderthals), Gravettian (c. 31.5 cal kya), Solutrean (c. 24.5-22cal kya), and Initial Magdalenian (d. 21-20.5 cal kya) periods, associated with anatomically modern humans. We identified 29 species of animals including humans from layers spanning the Mousterian to the Initial Magdalenian. 14 of these species had not been identified from the archaeozoological (i.e., osteological) record, including the presence of hyenas in the Magdalenian. Additionally, the phylogenetic analysis of the animal DNA retrieved from El Mirón sediments, including 72 mammalian mitochondrial sedaDNA genomes, shows the presence of simultaneous different faunal mtDNA lineages. Finally, we recovered three human mtDNA sequences from Solutrean levels that, together with published data, point to mtDNA haplogroup continuity in Iberia throughout the Solutrean period.