The city of Salvador, capital of Bahia, presents a population with the highest prevalence of HTLV-I infection in Brazil. Until now, only one study has investigated this infection in other cities of this state, even though by using a small sample. With objective to evaluate the prevalence of HTLV-I/II antibodies in four cities of the state of Bahia. Serum samples from 1,539 individuals who lived in Catolândia, Ipupiara, Jacobina and Prado were screened by ELISA, and repeatedly reactive samples confirmed by Western Blot. Forty-seven (3.1%) samples were positive by ELISA, and 44 of them were tested by Western blot: 5 (0.3%) were positive, 8 (0.5%) were indeterminate (all of them from Jacobina) and 31 were negative. The overall prevalence of HTLV-I antibodies was 0.3%. This prevalence varied from 0.0% (Prado) to 0.7% (Jacobina), but differences were not statistically significant (p > 0.21). None of these individuals presented HTLV-II antibodies. Jacobina showed the highest prevalence of HTLV-I infection among the cities studied, although the overall prevalence was low. In conclusion, further epidemiological, clinical and virological studies will be of paramount importance to obtain a better understanding of the natural history of this infection in Jacobina.