The effects of three sweeteners, sodium saccharin, aspartame and stevioside, on urinary bladder carcinogenesis in rats initiated by N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) were evaluated. Male F344 rats were given 0.01% BBN in their drinking water for 4 weeks and then the test sweeteners in their diet for 32 weeks. All surviving rats were sacrificed after 36 weeks, and examined histologically. Treatment with sodium saccharin significantly increased the incidence and extent of preneoplastic lesions, papillary or nodular (PN) hyperplasia, in rats treated with BBN for 4 weeks. Administration of 5% aspartame or 5% stevioside in the diet did not, however, affect the incidence or extent of PN hyperplasia in BBN-treated rats. No preneoplastic or neoplastic lesions of the urinary bladder were observed in rats treated with the test sweeteners only. The results with sodium saccharin were consistent with those in our previous experiments. The data also suggest that aspartame and stevioside do not promote bladder carcinogenesis.