The present study evaluated the mutagenic potential of surface water from the Lucrecia dam. The Tradescantia-micronucleus (Trad-MCN) test and CBMN assay in human peripheral blood lymphocytes were applied, corresponding to an in vivo and in vitro system, respectively. Heavy metals and some physicochemical properties were also measured. Water samples were collected in November 2009 (dry season) and May 2010 (rainy season) at three different points. Results of both assays for raw water showed positive responses for the points analyzed when compared to the negative control. The CBMN assay showed that diluted water was still able to induce a significant increase in micronucleus frequency. For both assays, the highest mean MN was observed in the dry season. Chemical analyses detected an increase in heavy metal levels at the sampling points and in the different seasons. These findings indicate the presence of genotoxins, such as heavy metals, in the water, which may be affecting the entire ecosystem, as well as human health. More prolonged monitoring is recommended in order to better characterize this public water supply.