Determination of the non-point sources of fecal pollution is essential for the assessment of potential public health risk and development of appropriate management practices for prevention of further contamination. Repetitive extragenic palindromic-PCR coupled with (GTG) 5 primer [(GTG) 5-PCR] was performed on 573 Escherichia coli isolates obtained from the feces of poultry (chicken, duck and turkey) and free-living (Canada goose, hawk, magpie, seagull and songbird) birds to evaluate the efficacy of (GTG) 5-PCR genomic fingerprinting in the prediction of the correct source of fecal pollution. A discriminant analysis with the jack-knife algorithm of (GTG) 5-PCR DNA fingerprints revealed that 95%, 94.1%, 93.2%, 84.6%, 79.7%, 76.7%, 75.3% and 70.7% of magpie, hawk, turkey, seagull, Canada goose, chicken, duck and songbird fecal E. coli isolates classified into the correct host source, respectively. The results of this study indicate that (GTG) 5-PCR can be considered to be a complementary molecular tool for the rapid determination of E. coli isolates identity and tracking the non-point sources of fecal pollution.