The mutagenicity of human bile was investigated in the Ames test after blue rayon treatment. In the present study, 52 and 59 bile samples were collected from the high and the low risk population for gallbladder cancer (GBC), respectively. The bile mutagenicity was detected only when the blue rayon extracts of bile were assayed with Salmonella typhimurium TA98 in the presence of S9 mix. Thirty-two (61.5%) of 52 samples obtained from the high risk population were mutagenic. In our previous study (Mano et al., 1993), the mutagenicity was observed in 14 (58.3%) of 24 samples. After combining this data with the results of the present study, 46 (60.5%) of 76 samples revealed the mutagenicity. On the other hand, the mutagenicity was detected in only 7 (11.9%) of 59 samples collected from the low risk population. Therefore, we found a significant geographical difference in the bile mutagenicity.