Liquid storage of avian spermatozoa is currently being employed in programs utilizing the artificial insemination to optimize the management of genetically superior males. It is mandatory to use efficient semen storage techniques in order to prevent the reduction of the fertilizing ability of stored semen. The present study was designated to evaluate the effect of oleic acid on rooster semen quality stored at 4°C for 48 h. Semen was collected from 10 roosters twice a week. Good quality ejaculates were pooled and after dilution, the semen was enriched with 0 (control), 0.125 (O 0.125), 0.25 (O 0.25), 0.5 (O 0.5), and 1 (O1) millimolar oleate. Forward progressive motility and viability of spermatozoa were evaluated at 0, 24, and 48 h. Moreover, malondialdehyde (MDA) and total antioxidant activity (AOA) levels were measured in seminal plasma and spermatozoa at the mentioned time points. Motility was 80.33 ± 1.45, 80.00 ± 2.08, and 66.00 ± 2.30% at 24 h and 56.33 ± 1.45, 57.33 ± 2.18, and 41.33 ± 2.02% at 48 h in O 0.125, O 0.25, and control, respectively (P < 0.001). Total AOA concentrations of seminal plasma were significantly higher in oleate treated groups than the control at 24 and 48 h (P < 0.03). Moreover, concentrations of AOA in spermatozoa revealed that oleate treated group showed higher AOA values compared to the control group at 24 and 48 h (P < 0.001). MDA concentrations of seminal plasma and spermatozoa were lower in oleate treated groups in comparison with control group at 24 and 48 h (P < 0.05). In conclusion, rooster semen enrichment with low doses of oleate would exert beneficial effects on the quality of semen during cooled storage.

The purpose of the present experiment was to investigate the protective effects of palmitoleate on the quality of ram semen during low temperature liquid storage. Ejaculates were collected using the artificial vagina from four Qezel rams twice a week. Ejaculates were pooled, diluted with Tris-egg yolk extender without palmitoleate (control) or supplemented with 0.125 (P 0.125), 0.25 (P 0.25), 0.5 (P 0.5) and 1 (P 1) mM palmitoleate at a final concentration of 500 × 10 spermatozoa/ml. Total motility and forward progressive motility (FPM) as well as other spermatozoa kinematics were evaluated by computer-assisted sperm analysis. Moreover, viability and membrane functionality were determined in the spermatozoa. Additionally, amounts of malondialdehyde (MDA), total antioxidant activity (AOA), nitric oxide (NO) and superoxide dismutase (SOD) activities were evaluated in the medium and spermatozoa at 0, 24, 48 and 72 hr of storage. The palmitoleate supplementation resulted in a significant (p < .05) increase in total motility and FPM with the highest increase at 0.5 mM concentration for 72 hr. P 0.5 group also resulted in the highest percentage of membrane-intact spermatozoa (76.60 ± 1.95%) and viability (75.81 ± 1.34%) at 72 hr (p < .05). The amounts of MDA and NO were lower in P 0.125, P 0.25 and P 0.5 groups compared to control at 48 hr and 72 hr (p < .05). Higher amounts of AOA were obtained in palmitoleate-treated groups in medium and spermatozoa during storage time (p < .05). Furthermore, palmitoleate supplementation increased the SOD activities in spermatozoa compared to the control (p < .05). The results of the present experiment reveal that supplementation with 0.5 mM palmitoleate improves ram spermatozoa motion characteristics, AOA levels and SOD activities during liquid storage. Then, palmitoleate could be used as an antioxidant source during liquid storage of ram semen.

Current study was designed to evaluate the effects of β-cyfluthrin, as a toxicant substance, and trans -ferulic acid ( trans - FA ), as a protective agent, on different parameters of rooster semen upon liquid storage. For this purpose, semen samples of roosters (Ross 308, n = 10, 32-wk-old) were collected twice a week. Good quality samples (≥70% progressive motion) were diluted, pooled and then divided for the purposes of the study. In the first experiment, motility of spermatozoa was evaluated following exposure to different concentrations of β-cyfluthrin (1, 2.5, 5, 10, 20, 40, and 80 µM) at 0, 24, and 48 h of storage. In the second experiment, constant doses of β-cyfluthrin (10 µM) alone or in combination with trans -FA (10, 25 mM) were assessed on motility and viability of spermatozoa at 0, 24, and 48 h time points. Moreover, amounts of malondialdehyde ( MDA ), total antioxidant capacity ( TAC ), total nitrate-nitrite, total hydroperoxide ( HPO ), and activity of superoxide dismutase ( SOD ) were evaluated in the homogenate of spermatozoa-diluent at studied time points. Results of the first experiment showed that amounts of β-cyfluthrin greater than 5 µM, significantly reduced the motility of spermatozoa at 24 and 48 h of storage ( P < 0.05). The second experiment demonstrated that, trans -FA especially at 10, 25 mM doses restored the motility and viability of spermatozoa compared to β-cyfluthrin treated group ( P < 0.05). Amounts of MDA (10, 25 mM), hydroperoxide (10, 25, and 50 mM), and nitrate-nitrite (10, 25, and 50 mM) were lower and TAC (10 and 25 mM) were greater in trans -FA + β-cyfluthrin treated groups compared to β-cyfluthrin alone treated samples ( P < 0.05). However, activity of SOD did not show significant changes by the treatment ( P > 0.05). It seems that trans -FA could ameliorate toxic effect of β-cyfluthrin via reduction of peroxidative (as evident by measurement of MDA) and nitrosative (as evident by measurement of nitrate-nitrite) reactions over cold preservation of rooster semen.

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.

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