We report on the polymorphisms exhibited by three hypervariable regions within the D-loop of Columba livia (pigeon) mitochondrial DNA. A total of 131 samples were taken from 131 randomly selected birds and used in the analyses of SNPs, a variable number of tandem repeats (VNTR) and an STR locus using CE. The number of repeats for the VNTR ranged from 2 to 8 producing 21 haplotypes, with 54 individuals exhibiting heteroplasmy. The STR locus exhibited multiple and continuous repeats within each individual and these patterns were not reproducible with individuals of the same maternal lineage, where different haplotypes were noted. Combining the SNP and VNTR loci produced 38 haplotypes, with the power of discrimination being 0.93. The polymorphic regions of D-loop observed in this study are potential markers for maternal relationship identification.