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The historical site of the Monte Mario lower Pleistocene succession (Rome, Italy) is an important marker of the Pliocene/Pleistocene boundary. Recently, the Monte Mario site was excavated and restudied. A spectacular angular unconformity characterizes the contact between the Monte Vaticano and the Monte Mario formations, which marks the Pliocene/Pleistocene boundary. Biostratigraphical analyses carried out on ostracod, foraminifer, and calcareous nannofossil assemblages indicate an Early Pliocene age (topmost Zanclean, 3.81–3.70 Ma) for the underlying Monte Vaticano Formation, whereas the Monte Mario Formation has been dated as early Pleistocene (Santernian, 1.66–1.59 Ma). Palaeomagnetic analyses point to C2Ar and C1r2r polarity chrons for the Monte Vaticano and the Monte Mario formations, respectively. The Monte Mario Formation consists of two obliquity-forced depositional sequences (MM1 and MM2) characterized by transgressive systems tracts of littoral marine environments at depths, respectively, of 40–80 m and 15–20 m. The data obtained from foraminifer and ostracod assemblages allow us to reconstruct early Pleistocene relative sea-level changes near Rome. At the Plio/Pleistocene transition, a relative sea-level drop of at least 260 m occurred, as a result of both tectonic uplift of the central Tyrrhenian margin and glacio-eustatic changes linked to early Pleistocene glaciation (Marine Isotope Stage 58).