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Tacuarembó is a department located in the northeastern of Uruguay, whose population is the result of several migration waves from Europe and Near East as well as Africans and Afro-descents mostly from Brazil, these waves settled on the territory of various Native ethnic groups (Charrúa, Minuán, and Guaraní). In the past, this population has been the focus of genetic studies showing this tri-hybrid origin, with greater contributions of Natives and Africans than other Uruguayan regions. In this paper we aim to analyzeAluinsertions (A25, ACE, APOA1, B65, D1, F13B, PV92, TPA25) in order to provide valuable information for ancestrality and genetic differentiation (GD), and to compare with previous studies on Tacuarembó population as well as toAlufrequencies in other Uruguayan populations.The European contribution withAluand classical markers (CM) was almost equal to a previous study using 22 classical markers (63% vs 65%) while African contribution was higher(30% vs15% respectively), and the Native Americans contribution shows an important difference 7% inAluvs20%. There are no significant differences between Tacuarembó and Montevideo in GD, but there are significant differences between Tacuarembó and Basques descendants from Trinidad. Our results support the previous findings obtained with classical markers that demonstrate the tri-hybrid composition of the Tacuarembó population, correlated with the historic records. Finally,Aluinsertions provides interesting information in light of the admixture process in the Uruguayan population.