Background: The golden eagle, Aquila chrysaetos is one of the 65 species of birds of prey found in Appendix II of the Convention on International Trade in Endangered Species of Wild Fauna and Flora. It has been estimated that the world population is 250 thousand specimens. The operant conditioning to obtain ejaculates of A. chrysaetos is described and to contribute to assisted reproduction protocols in captivity.Methods: In the reproductive season, the procedure was performed twice a week on 5 adult birds, in order to obtain ejaculates.Result: The study lasted 17 weeks, in which 160 contacts were made in total. It was possible to obtain 20 ejaculates, which were obtained from 3 birds. 4% success was achieved in obtaining ejaculates. Additionally, behaviors of clinical utility for diagnosis, care and well-being were conditioned in birds. The above should be considered in assisted reproduction programs, since, as demonstrated by the results of this study, not all the birds provide sufficient ejaculates, for which reason, the evaluation of the individuals that present the greatest advance in operant conditioning to include them in assisted reproduction programs.