Hyaluronic acid improves frozen-thawed sperm quality and fertility potential in rooster

- We analyzed the effects of hyaluronic acid as a cryo-additive for rooster semen cryopreservation.
- Apoptotic status, motility, membrane functionality, lipid peroxidation and fertility rate were assessed for frozen-thawed sperm.
- 1 mM and 2 mM hyaluronic acid produced the highest quality of frozen-thawed rooster semen compared to other groups.

Beneficial effects of Hyaluronic acid (HA) has not been yet assessed for cryopreservation of rooster sperm. This study was conducted to evaluate the effects of different concentrations of HA (0, 1, 2, 4 and 8 mM) in Beltsville extender on the cryopreservation of rooster sperm. Semen samples were collected from six Ross broiler breeders (24-week) using abdominal massage, then divided into five equal aliquots and cryopreserved in Beltsville extender that contained different concentrations of HA. Motion characteristics, morphology, membrane functionality, viability, acrosome integrity, lipid peroxidation and fertility potential of sperm were assessed after thawing. HA at concentration of 2 mM (HA2) resulted in the highest (P < 0.05) total motility (55.3 ± 1.1%) and progressive motility (25.2 ± 0.8%) compared to the other groups. HA8 produced the lowest significant (P < 0.05) percentage of total (38.6 ± 1.1%) and progressive (14.7 ± 0.8%) motility. High significant percentage of membrane functionality were observed in HA1 and HA2 (43.2 ± 1.0 and 46.1 ± 1.0%, respectively) compared to HA4 (40.1 ± 1.0%) and HA8 (32.5 ± 1.0%). Moreover, HA1 and HA2 produced the higher percentage of acrosome integrity (54.8 ± 1.2 and 57.5 ± 1.2, respectively) compared to other groups. HA1 and HA2 reduced (P < 0.05) malondialdehyde formation (3.66 ± 0.08 and 3.75 ± 0.08 nmol/ml) compared to other groups. Fertility rate and hatching rate obtained from artificial insemination were significantly higher in HA1 (63.7 and 54.7%) and HA2 (67.5 and 57.7%) compared to control group (40 and 37%). Our results showed that supplementation of Beltsville extender with 1 and 2 mM HA significantly improved the quality of rooster sperm after freeze thawing.