Allozymic variation and differentiation of strictly parapatrically distributed European moles (Talpa europaea, n = 44) and Mediterranean moles (T. caeca, n = 34) were studied in the Val Bregaglia (Switzerland) and Val Chiavenna (Italy) to estimate interspecific separation and intraspecific gene pool structuring. Tissue samples were screened for allelic variation at 44 putative gene loci using horizontal starch gel electrophoresis. In T. europaea four and in T. caeca three regional samples separated from one another by water courses were discriminated. The proportion of polymorphic loci was 2.3 per cent in each regional sample, with polymorphisms at the Es-1 locus in T. europaea and at the Es-1 and the Es-D loci in T. caeca. Expected average heterozygosity ranged from 0.6 to 1.2 per cent in regional samples of T. europaea and from 0.9 to 1.2 per cent in T. caeca. In T. caeca one regional sample showed fixation of a unique allele (Es-D-143). This indicated paucity of migration across surrounding water courses Inbreeding coefficients (F-IS) for the Es-1 locus ranged from 0.39 to 1.0 in regional samples. Significant deviations of genotype frequencies for the Es-1 locus due to deficiency of heterozygotes were found in one T. europaea and one T. caeca regional sample, respectively. The findings are discussed with respect to the parapatric pattern of distribution and the generally low gene pool variability of Talpa species found so far.