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Understanding the transformations of the climate system may help to predict and reduce the effects of global climate change. The geological record provides a unique archive that documents the long-term fluctuations of environmental variables, such as seasonal change. Here, we investigate how seasonal variation in seawater temperatures varied in the Mediterranean Sea during the early Pleistocene, approaching the Early-Middle Pleistocene Transition (EMPT) and the beginning of precession-driven Quaternary-style glacial–interglacial cycles. We performed whole-shell and sclerochemical stable isotope analyses (δ 18 O, δ 13 C) on bivalves, collected from the lower Pleistocene Arda River marine succession (northern Italy), after checking shell preservation. Our results indicate that seawater temperature seasonality was the main variable of climate change in the Mediterranean area during the early Pleistocene, with the Northern Hemisphere Glaciation (NHG) exerting a control on the Mediterranean climate. We show that strong seasonality (14.4–16.0 °C range) and low winter paleotemperatures (0.8–1.6 °C) were likely the triggers leading to the establishment of widespread populations of so called “northern guests” (i.e., cold water taxa) in the Mediterranean Sea around 1.80 Ma. The shells postdating the arrival of the “northern guests” record a return to lower seasonal variations and higher seawater paleotemperatures, with seasonality increasing again approaching the EMPT; the latter, however, is not associated with a corresponding cooling of mean seawater paleotemperatures, showing that the observed seasonality variation represents a clear signal of progressive climate change in the Mediterranean Sea. [ABSTRACT FROM AUTHOR]