A total of 330 white storks (Ciconia ciconia) and 138 black kites (Milvus migrans) were blood sampled during four consecutive years in an area heavily contaminated as a consequence of a massive spillage of toxic acid mining waste rich in heavy metals that impacted on the Doñana National Park (south western Spain), in April 1998. The alkaline single cell gel electrophoresis (Comet) assay was performed as a genotoxicity test, in order to assess whether the high level of DNA damage first detected by us 1 year after the disaster was still present in birds in each of the successive 3 years. Our results clearly show that, when compared with control individuals from non-polluted areas, white storks and black kites born in the contaminated area for a period of up to 4 years after the toxic accident have suffered an increase of at least 2- to 10-fold in the level of their genetic damage through the study period. Taken as a whole, these observations seem to indicate that the toxic spill still appears to be affecting the wildlife 4 years after the mining disaster and that attempts at cleaning up the waste have proved ineffective based on DNA damage detection.