The complete mitochondrial DNA D-loop structure of pigeon (Columba livia) was established in this study. A strategy of amplifying three partial fragments of the D-loop and then combing the three fragments to cover the full length of the D-loop was adopted. Ten samples from pigeons were collected and were successfully amplified and sequenced. Repetitive sequences of a VNTR and an STR were both observed at the 3'-end of D-loop region. DNA sequence data revealed polymorphic sequences including indels, SNP, VNTR and STR within the D-loop. The size of the D-loop ranged from 1310 to 1327 bp from the initiation site of D-loop to the site immediately upstream of the repeat sequences depending upon the number of insertions or deletions. Each sample could be distinguished based on four genotyping procedures; being indels, SNPs, VNTRs and STRs. The polymorphic nature of the D-loop can be a valuable method for maternal identification and genetic linkage of pigeon in particular forensic science investigations.