In the past decade, there has been a drastic decline in the number of Eastern Black rhinoceros (black rhinoceros) (Diceros bicornis michaeli), primarily because of poaching across their natural habitats, leaving few individuals in small, isolated populations that are vulnerable to demographic extinction, disease epidemics, genetic drift and inbreeding. However, genetic consequences of the demographic decline on the remaining populations have not been investigated. Using the mitochondrial control region, this study investigated how current levels of genetic diversity relate to historical patterns, quantified genetic differentiation between extant populations and assessed the impacts of previous translocations on genetic diversity across populations. A total of 74 individual eastern black rhinoceroses were sampled from five extant populations in Tanzania and one neighbouring cross-border population in the Maasai Mara in Kenya. Six maternal haplotypes were identified, with an overall haplotype diversity of h=0.7 but low overall nucleotide diversity within populations (pi = 0.017) compared to historical populations from Tanzania (pi = 0.021). There was extensive variation in haplotype distribution between populations, with more variation exists within (65.5 %) than among the populations (35.5%), which may indicate lack of migration between populations. Specifically, some geographically close populations with different histories of introductions didnt share any haplotypes, suggesting that gene flow is currently restricted. The haplotypes were distributed among three east African haplogroups (CV, CE and EA) that have been described in previous studies, suggesting that multiple lineages have been preserved despite loss of haplotypes. One of the haplotypes was highly divergent and matched sequences previously classified as a subspecies that has not been recognised in recent years (D. b. ladoensis). We recommend that current levels of diversity be maintained by allowing natural movements of rhinoceroses between the populations, with the possibility of introducing additional variations by translocation of individuals between sites.