This investigation aimed to evaluate the quality of cryopreserved semen from 21 Curraleiro Pé Duro bulls in a germplasm bank, categorizing the doses of the sires according to parameters related to sperm viability, and verifying the fertilizing capacity in vitro of the spermatozoids. All the samples were thawed and analyzed as to morphology, kinetics, plasma membrane integrity and acrosome integrity. Samples of only four bulls were used for evaluating the fertilization rates. The results indicated that the Curraleiro Pé Duro semen doses, stocked in the Brazilian Animal Germplasm Bank, showed variable sperm quality and high in vitro fertilizing capacity, making it possible to design appropriate strategies for assisted reproduction in order to conserve this Brazilian genetic resource, which is in danger of extinction.