1,153 unrelated individuals from three different areas of Italy were tested for the red cell esterase D polymorphism. The gene frequencies found in the three groups do not differ significantly from each other. The EsD2 allele in the total sample has a frequency of 14.6% but it is difficult at the present stage to know if this figure is valid for the whole Italian population. If so, the Italian EsD2 allele frequency lies at the upper limit of the range of the European values. No variant phenotypes were observed.