The aim of this work was to evaluate the bioremediation potential of microorganisms from intertidal sediments of a sandy beach affected by a major oil spill 7 years before and subject to chronic petroleum contamination since then. For that, the response of microorganisms to a new oil contamination was assessed in terms of community structure, abundance, and capacity to degrade hydrocarbons. Experiments were carried out under laboratory-controlled conditions by mixing sediment with crude oil with three different nitrogen supplementations in 50 ml serum bottles under constant shake for 15 days. Autochthonous microorganisms were able to respond to the new oil contamination by increasing their abundance (quantified by DAPI) and changing the community structure (evaluated by DGGE). This response was particularly clear for some specific bacterial groups such as Pseudomonas, Actinomycetales, and Betaproteobacteria. These communities presented an important potential for hydrocarbon degradation (up to 85 % for TPHs and 70 % for total PAHs), being the biodegradation stimulated by addition of an appropriate amount of nitrogen.