The main goal of this paper is to study the effect of the spatio-temporal changes of Land Use/Land Cover (LULC) within the hydrologic regime of the Cervaro basin in Southern Italy. LANDSAT Thematic Mapper (TM) imagery acquisition dates from 1984, 2003, 2009, and 2011 were selected to produce LULC maps covering a time trend of 28 years. Nine synthetic bands were processed as input data identified as the most effective for the Artificial Neural Network (ANN) classification procedure implemented in this case study. To assess the possible hydrological effects of the detected changes during rainfall events, a physically-based lumped approach for infiltration contribution was adopted within each sub-basin. The results showed an increase in flood peak and a decrease of the rangelands, forests, and bare lands between 1984 and 2011, indicating a good correlation between flooding areas and land use changes, even if it can be considered negligible in basins of large dimensions. These results showed that the impact of land use on the hydrological response is closely related to watershed scale.