Freeze-storage of epididymal sperm is an important technique for the preservation of gametes in animals, including those becoming extinct. We froze canine sperm recovered from the cauda epididymis and investigated the fertility. The qualities of sperm from the cauda epididymis before freezing were: mean sperm motility, 89.4 ± 1.6 (SE) %; sperm viability, 89.1 ± 1.1%; and these were significantly higher than those of sperm from the caput-corpus epididymis (P<0.01, P<0.05). The number of sperm recovered from both cauda epididymides varied among animals: 6.3-122.3 × 10⁷, mean 61.5 ± 10.0 × 10⁷. Freezing was used only for sperm recovered from the cauda epididymis. The sperm motility and viability after thawing were 19.5 ± 2.5% and 53.1 ± 3.3%, respectively. These were slightly lower than those of frozen-thawed ejaculated sperm, but the differences were not significant. When 2 × 10⁸, 3 × 10⁸, or 4 × 10 ⁸ sperm were inseminated in the unilateral uterus, only one animal inseminated with 3 × 10⁸ sperm was fertilized (1/16, 6.3%). When 1 × 10⁸ sperm were inseminated in the bilateral uterine tubes, one of six animals (16.7%) was fertilized. Therefore, although the qualities of epididymal sperm after thawing were similar to those of ejaculated sperm, the conception rate obtained with frozen-thawed epididymal sperm was low in beagle dogs. It is necessary to investigate the differences in damage between epididymal sperm after thawing and ejaculated sperm and to develop a method for improving the conception rate.