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The warmest global temperatures of the past 85 million years occurred during a prolonged greenhouse episode known as the Early Eocene Climatic Optimum (52–50 Ma). The Early Eocene Climatic Optimum terminated with a long-term cooling trend that culminated in continental-scale glaciation of Antarctica from 34 Ma onward. Whereas early studies attributed the Eocene transition from greenhouse to icehouse climates to the tectonic opening of Southern Ocean gateways, more recent investigations invoked a dominant role of declining atmospheric greenhouse gas concentrations (e.g., CO₂). However, the scarcity of field data has prevented empirical evaluation of these hypotheses. We present marine microfossil and organic geochemical records spanning the early-to-middle Eocene transition from the Wilkes Land Margin, East Antarctica. Dinoflagellate biogeography and sea surface temperature …