The aim of this study was the identification of the genetic diversity and paternal origin of Peruvian creole cattle. A panel of 7 Y-chromosome specific markers (INRA189, UMN0103, BM861, UMN307, BYM-1, DDX3Y_1STR and ZFY10) were analyzed in 229 cattle from 6 regions of the Peruvian highlands. The creole cattle exhibited low genetic diversity (H= 0.50) mostly explained by within-population variation (98%) and absence of population structure (FST = 0.019) in the analyzed regions. These results are in concordance to other studies in Spanish cattle populations. The overall frequency and distribution of the major B. taurus haplogroups: Y1 (19%) and Y2 (81%), suggests that Peruvian creole cattle derived from the Iberian Peninsula cattle. Furthermore, our results some degree of male-mediated African cattle influence in the Peruvian creoles, supporting the findings of other studies in South American creole cattle populations. Altogether, our results revealed unique genetic characteristics of Peruvian creole cattle that may have important implications for future conservation programs.