This work evidenced, using a cytogenetics approach, that Bos indicus exerted a genetic introgression of paternal origin on Creole Colombian cattle descendent from Bos taurus. Analysis of chromosome Y morphology was carried out in heparinized blood samples of 67 bulls belonging to seven Colombian breeds. We report 4 sires belonging to the Romosinuano breed (40%) and 10 bulls of the Casanareno breed (100%) with acrocentric Y chromosome which is characteristic of Bos taurus. This finding indicates a high degree of genetic introgression in these two breeds probably caused by the continuous input of zebu stallions in the Colombian Creole breeds. In other five Creole breeds (Blanco Orejinegro -BON-, Chino Santandereano, Costeno con Cuernos, Harton del Valle and Sanmartinero), the bulls had a submetacentric Y chromosome characteristic of Bos taurus.