This study was carried out with the aim to investigate the presence of carriers of diphtheria bacillus in a sample of 1970 healthy children, 6-14 years aged, residing in eight Italian towns. Three non-toxigenic strains of Corynebacterium diphtheriae biotype mitis were isolated from as many healthy children. Molecular characterization by ribotyping showed close genetic relation of two of the wild strains with the C7(b) reference strain whereas one of the wild strains showed close genetic relation with two collection strains isolated in the same geographic area (Emilia-Romagna) from diphtheria patients in the seventy years. This supports the hypothesis of the persistence of some non toxigenic C. diphtheriae strains derived from ancient endemic strains under the selective pressure of mass immunization against diphtheria. The persistence of carriers of diphtheria bacilli, although non toxigenic, suggests that high levels of immunity must be maintained, not only in children, but also in adults by booster vaccination.