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Integrated 40Ar/39Ar, trace‐element, stratigraphic, palaeontological and palaethnological data provide geochronological and biochronological constraints for the sedimentary and tectonic history of a Middle Pleistocene fluvial–lacustrine basin near Rome (Cretone Basin, central Italy), which has yielded a significant record of mammal fossil remains and Palaeolithic industry. This work is a case study of the interplay between tectonics and glacio‐eustacy, which strongly influenced the evolution of the Tyrrhenian Sea margin of central Italy. Dating of tephra layers interbedded within the Cretone Basin lacustrine succession and reconstruction of relict terraced surfaces allow correlation with similar, geochronologically constrained, marine isotopic stages (MIS) 15–5 terraced deposits along the coast. Coupled extensional tectonics and regional uplift over the last 600 ka caused the progressive uplifting and westward migration of the main fluvial–lacustrine basin and the formation of a smaller satellite basin at its eastern margin. Here, stable environmental conditions during MIS 13–5 indicated continuous human and large mammal frequentation, as testified by lithic industry and fossil remains ascribable to the Acheulean and later early Middle Palaeolithic technocomplexes and Galerian–Aurelian mammal faunas, respectively. In addition to providing independent age constraints to glacial sea‐level oscillations of this region, the reconstructed chronostratigraphic setting for the Cretone Basin provides evidence for one of the oldest Acheulean lithic assemblage of central Italy, as well as new biochronological and palaeobiogeographical data for some Middle Pleistocene mammal species of Italy.

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We revise the chronostratigraphy of several sedimentary successions cropping out along a 5 km-long tract of the Aniene River Valley in Rome (Italy), which yielded six hominin remains previously attributed to proto- or archaic Neanderthal individuals, as well as a large number of lithic artefacts showing intermediate characteristics somewhere between the local Acheulean and Mousterian cultures. Through a method of correlation of aggradational successions with post-glacial sea-level rises, relying on a large set of published 40Ar/39Ar ages of interbedded volcanic deposits, we demonstrate that deposition of the sediments hosting the human remains spans the interval 295–220 ka. This is consistent with other well constrained ages for lithic industries recovered in England, displaying transitional features from Lower to Middle Paleolithic, suggesting the appearance of Mode 3 during the MIS 9-MIS 8 transition. Moreover, the six human bone fragments recovered in the Aniene Valley should be regarded as the most precisely dated and oldest hominin remains ascribable to Neanderthal-type individuals in Europe, discovered to date. The chronostratigraphic study presented here constitutes the groundwork for addressing re-analysis of these remains and of their associated lithic industries, in the light of their well-constrained chronological picture.