Petroleum can pollute pristine shorelines as a consequence of accidental spills or chronic leaks. In this study, the fate of petroleum hydrocarbons in soft pristine sediment of Caleta Valdés (Argentina) subject toex situsimulated oil pollution was assessed. Sedimentary columns were exposed to medium and high concentrations of Escalante Crude Oil (ECO) and incubated in the laboratory during 30 days. Levels of aliphatic hydrocarbons at different depths of the sedimentary column were determined by gas chromatography. Oil penetration was limited to the first three centimetres in both treatments, and under this depth, hydrocarbons were clearly biogenic (terrestrial plants) as in the whole sedimentary column of the control assay. Bioturbation by macrobenthic infauna was strongly impacted by oil pollution which resulted in reduced sediment oxygenation and low burial of petroleum hydrocarbons. This may partly explain the limited hydrocarbon biodegradation observed, as indicated by the relatively high values of the ratios nC17/pristane,nC18/phytane, and total resolved aliphatic hydrocarbons/ unresolved complex mixture. Correspondingly, at the end of the experiment the most probable number of hydrocarbon-degrading bacteria reached ~ 10^3 MPN g−1 dry weight. These values were lower than those found in chronically polluted coastal sediments, reflecting a low activity level of the oil-degrading community. The results highlight the low attenuation capacities of Caleta Valdés pristine sediments to recover its original characteristics in a short time period if an oil spill occurs. In this work, we present a novel and integrative tool to evaluate the fate of petroleum hydrocarbons and their potential damage on pristine sediments.