Effects from aging on semen quality of fresh and cryopreserved semen in Labrador Retrievers

Characteristics of frozen sperm associated with fertility and aging have not been fully determined in dogs. The aim of this study was to determine the relationship between fresh and post-thaw sperm quality, age, among the factors of motility, viability, morphology and oxidative stress in a group of fertile stud dogs with extensive breeding records and in dogs with reported subfertility problems. Sperm parameters from 39 fertile dogs were measured in fresh semen and frozen thawed semen. Additionally, frozen semen from 9 subfertile dogs was thawed and analyzed. Subfertile dogs were defined by referring veterinarians solely on the basis of owner history; breeding records were not available to this study. Evaluation included total motility (TM) and progressive motility (PM), average path velocity (VAP), viability, morphology and presence of sperm reactive oxygen species (ROS). Fertile males' ages ranged from 1 to 10 years, subfertile males' ages ranged from 4 to 14 years. All dogs were assigned to age groups according to age as young (1-3 years) middle (4-6 years) and senior (>7 years). The effect of sperm state (fresh vs. frozen-thawed), age, and fertility history (fertile vs. subfertile) on the measured endpoints were determined using a mixed effects model. TM showed a negative correlation with age in the frozen-thawed semen. Additionally, TM in fresh semen was higher in all age groups compared to post thaw semen (P < 0.05). PM was higher in all age dogs of fresh semen compared to frozen-thawed semen (P < 0.05). As such, TM and PM in post thaw semen statistically decreased compared to fresh semen regardless of age of stud male (P < 0.05). Differences in VAP were only observed between young and senior age groups in fresh semen (P < 0.05) while young and middle age dogs from fresh semen were different in all age groups for frozen-thawed semen (P < 0.05). ROS was higher in the young frozen-thawed semen compared to the young fresh semen (P < 0.05). When comparing TM, PM and VAP of fertile vs. subfertile dogs, middle aged fertile dogs are different from middle aged and senior subfertile dogs (P < 0.05). No differences were found in measures of ROS between fertile and subfertile. For sperm viability and morphology, differences were observed between all age groups of fertile dogs in comparison to all ages of subfertile dogs (P < 0.05). Sperm motility appeared to be the most affected parameter by freezing damage than any other parameter measured in this study, while we were not able to determine a significant association between ROS production and fertility status.