Kinetin improves motility, viability and antioxidative parameters of ram semen during storage at refrigerator temperature

Abstract

The present experiment was conducted to evaluate the effect of kinetin on ram semen quality during cold storage. Ejaculates were collected using an artificial vagina from five Qezel rams. Ejaculates which met the criteria (volume of 0.75-2 ml; minimum spermatozoa concentration of 2.5 × 109 spermatozoa/ml and forward progressive motility of 80%), were pooled, diluted with extender without kinetin (control) or enriched with 25 (K 25), 50 (K 50), 100 (K 100) and 200 (K 200) μM kinetin at a final concentration of 500 × 106 spermatozoa per mL. Spermatozoa motion characteristics were evaluated by computer-assisted sperm analysis. In addition, percent of viability (spermatozoa showing no color was considered to be alive) and spermatozoa with intact plasma membrane (spermatozoa with curled/swollen tail was considered healthy) were determined. Moreover, amounts of malondialdehyde (MDA), total antioxidant activity (AOA), nitric oxide (NO) and superoxide dismutase (SOD) activity were determined in the seminal plasma and spermatozoa at 0, 24, 48 and 72 h of storage. Higher percent of total and forward progressive motility was observed in K 25, K 50 and K 100 groups compared to control group at 72 h of storage (P < 0.001). Moreover, K 25 (78.61 ± 1.11%), K 50 (82.46 ± 1.08%) and K 100 (82.96 ± 1.49%) groups showed higher percentage of viability compared to control (72.38 ± 1.49%) at 72 h of storage (P < 0.05). Semen enrichment with kinetin resulted in the higher percent of intact plasma membrane of spermatozoa at 48 and 72 h (P < 0.001). Amounts of MDA were lower and amounts of AOA were higher in K 50 and K 100 groups compared to control at 48 and 72 h (P < 0.05). There were no significant differences in NO levels and SOD activities of seminal plasma and spermatozoa among groups during the experiment. The present experiment revealed that kinetin at proper concentration could enhances spermatozoa kinematics, viability, spermatozoa plasma membrane functionality and amounts of AOA and reduces the level of lipid peroxidation during chilled storage of ram semen.

Effects of different extenders and additives on liquid storage of Awassi ram semen

“…The efficiency of TL-HEPES and PBS in sperm processing (16,17) and in short-term storage (18)(19)(20) was identified in previous studies. Tris-citrate is a commonly used extender (1), and it is also usually used as the control or main extender in ram sperm-extender studies (21)(22)(23)(24). TEST, TL-HEPES, and PBS extenders were found to be effective in protecting the sperm against cold shock when freezing rat sperm and in equilibration at 4 °C (20,25).…”

Influence of idebenone on ram semen quality stored at 4°C

“…Reports have shown that ram semen enrichment with butylated hydroxytoulene, taurine, catalase, quercetin, palmitoleic acid, and kinetin enhances motility or viability compared to the control when stored as liquid at refrigerator or after freeze‐thawing process (Banday et al, ; Câmara et al, ; Eslami et al, ; Rather et al, ; Zadeh Hashem & Eslmi, ). The results of the present experiment indicated that Id at 1, 2 and 4 µM enhanced total motility, FPM, and VCL at 48 and 72 hr of storage.…”

“…Reports have shown that ram semen enrichment with butylated hydroxytoulene, taurine, catalase, quercetin, palmitoleic acid, and kinetin enhances motility or viability compared to the control when stored as liquid at refrigerator or after freeze-thawing process (Banday et al, 2017;Câmara et al, 2011;Eslami et al, 2017;Rather et al, 2016;Zadeh Hashem & Eslmi, 2018 experiment. Previous study revealed a correlation between capacities of spermatozoa in fertilization with spermatozoa motility assessed by CASA (Larsen et al, 2000).…”

Liquid Storage of Ram Semen: Associated Damages and Improvement

“…As for motility, ram semen liquid storage is associated with a gradual decrease of sperm viability [46] [47] and such decrease is negatively correlated with LPO [45] and can be attributed to the endogenous production of ROS [48].…”

“…The antioxidative properties of kinetin have been described [95]. Kinetin has been tested as an additive to ram semen extender during chilled storage up to 72 h. It was concluded that kinetin at appropriate concentrations (25 -100 µM) improves spermatozoa kinematics, viability, sperm plasma membrane functionality and antioxidative parameters [48].…”