The mitochondrial DNA of Apis mellifera, sampled in 105 beehives and 7 geographical zones associate to 14 localities in Colombia have been studied. A fragment of the intergenic region between the RNAt leu and the subunit II of the cytochrome oxidase gene was amplified by PCR and digested with the endonuclease DraI. Thirteen haplotypes were identified: eight belonging to the African lineage A (90,5% for A1, A2, A4, A5, A6, A8, A9 and A13), 4 to European lineage M (7,6% M2, M4, M5 and M7) and 1 to C (C1, 1,9%), of the east European lineage. The presence of eight different A haplotipes, suggests the occurrence of more than one episode of hybridization, introgression and expansion of the africanized phenoma which suggests the presence of other haplotypes in other populations not included in the study. The gradient of distribution of African lineage is represented by the A1 haplotype, to the center of the country from the Caribbean coast with A4 and A6 haplotypes. The european lineages, are not represented significantly on the population studied. Colombia beekeeping activities depend on the africanized honeybee.