This study aimed to determine the pattern of antibiotic and heavy metal resistance in Gram-negative bacteria isolated from five different sites in the Seyhan Dam Lake and Seyhan River in Adana, Turkey. The susceptibility of 268 isolates to 16 different antibiotics and five heavy metals was investigated by agar diffusion and dilution methods, respectively. The most common species isolated from the samples were Aeromonas hydrophila (17.5 %), Aeromonas caviae (8.9 %) and Citrobacter freundii (8.9 %). There was a high incidence of resistance to ampicillin (80.2 %), streptomycin (71.6 %) and cefazolin (60.4 %). Multiple antibiotic resistance indices ranged from 0.2 to 0.81, suggesting exposure to antibiotic contamination. The isolates showed tolerance to different concentrations of heavy metals. These results indicate that antibiotic and heavy metal resistance among the Seyhan Dam Lake and Seyhan River bacteria may pose a risk to the fish population and public health. At the same time, the finding in the aquatic environments of different combinations of resistance genes suggests their involvement in the spread of multidrug-resistant strains.