The genetic groups of native donkeys in Brazil are characterized by adaptation to the local environment. However, the donkey population in the country is declining, mainly because of agricultural mechanization and transportation that has led to the abandonment and the consequent indiscriminate slaughter of these animals. There are three local genetic groups of distinct geographic and temporal formation.However, analyses of their origin, phylogenetic relationship, and population structure are scarce. Within this context, molecular markers such as the mitochondrial control region (D-loop) are useful for these analyses. This study aimed to evaluate the variation and origin of maternal lineages of groups of naturalized donkeys in Brazil (Brazilian, Nordestino and Pêga). We detected ve mitochondrial haplotypes with 18 polymorphic sites, two of them exclusively found in the Nordestino donkey. This group is more distant from the other groups. Phylogenetic analysis indicates maternal contributions of two clades (Nubian and Somali) to the formation of the genetic groups of donkeys, a fact that explains the high diversity, structure and distances of the groups. It was rstly reported here. This analysis contributes production and conservation of native donkey breeds. It also gives clues about the formation of the Iberian breeds from which Brazilian donkeys originated.Authors contribution The study conception and supervision were conducted by Gregório Miguel Ferreira de Camargo. Material preparation, data collection, analysis and rst draft were performed by Jackeline Santos Alves. Data collection was performed by Chiara Albano de Araújo Oliveira. Financial support was provided by