The mitochondria-targeted antioxidant Mito-TEMPO conserves rooster's cooled semen quality and fertility potential.

The PUFAs content of rooster sperm cells makes them vulnerable to the thermal shocks during chilling storage, which reduces the fertility performance of cooled sperm. Extender supplementation with antioxidants is a reasonable method to conserve sperm fertility potential during cooling storage process. The aim of this study was to determine the effect of Mito-TEMPO addition to the Lake medium on rooster sperm quality and fertility potential during cooling process. Semen samples were diluted in the Lake medium and assigned into five equal aliquots and supplemented with 0, 0.5, 5, 50 and 500 μM Mito-TEMPO. Then, the samples were cooled at 5 °C and conserved up to 50 h. Total motility, progressive motility, morphology, viability, membrane integrity, lipid peroxidation and mitochondrial activity of samples were analyzed during 0, 25 and 50 h of cooling period. Artificial insemination was also conducted using 25 h-cooled semen. No significant difference was observed among different treatments during quality evaluations at 0 h storage. Extender supplementation with 5 and 50 μM Mito-TEMPO presented greater (P ≤ 0.05) total motility, progressive motility, viability, membrane integrity and lower lipid peroxidation compared to other groups during 25 and 50 h cooling storage. Mitochondrial activity was higher (P ≤ 0.05) in groups received 5, 50 and 500 μM Mito-TEMPO than others. Fertility rate of 25 h-cooled-stored samples was higher (P ≤ 0.05) in groups containing 5 and 50 μM Mito-TEMPO compared to control group. In conclusion, addition of 5 and 50 μM Mito-TEMPO as a mitochondria-targeted antioxidant to the storage medium could be a suitable method to conserve rooster semen quality against stressful conditions of cooling storage process.

Masoudi R ，Asadzadeh N ，Sharafi M 《-》

Effects of freezing extender supplementation with mitochondria-targeted antioxidant Mito-TEMPO on frozen-thawed rooster semen quality and reproductive performance.

Rooster semen cryopreservation is a useful method to utilize semen samples for artificial insemination in commercial flocks, but with use of the freezing-thawing process there is a reduction in the quality and fertilization capacity of rooster spermatozoa post-thawing. The aim of the current study was to investigate the efficacy of the mitochondria-targeted antioxidant Mito-TEMPO on rooster sperm quality and fertilization capacity after conducting the freezing-thawing processes. Semen samples were diluted and there were five equal aliquots supplemented with 0, 0.5, 5, 50 and 500 μM Mito-TEMPO. Semen samples were subsequently cryopreserved in liquid nitrogen. After thawing, sperm motility, lipid peroxidation, membrane functionality, normal morphology, mitochondria active potential, acrosome integrity, viability, apoptotic-like changes, DNA fragmentation, hydrogen peroxide concentration and fertilizing capacity were evaluated. Supplementation of Lake medium with 5 and 50 μM Mito-TEMPO resulted in greater (P ≤ 0.05) total sperm motility, progressive motility, average path velocity, membrane functionality, mitochondria active potential, acrosome integrity and viability compared with semen of the other groups. Lipid peroxidation, late apoptotic-like changes, DNA fragmentation and hydrogen peroxide content, however, were less (P ≤0.05) in semen samples supplemented with 5 and 50 μM Mito-TEMPO compared to other groups. Furthermore, fertility percentages were greater when there was supplementation with 5 and 50 μM Mito-TEMPO compared to the control group. Mitochondria-targeted antioxidant Mito-TEMPO could be included in semen extender before cryopreservation to improve quality and fertilization capacity of rooster semen after thawing of cryopreserved samples.

Masoudi R ，Asadzadeh N ，Sharafi M 《-》

Supplementation of sperm cooling medium with Zinc and Zinc oxide nanoparticles preserves rooster sperm quality and fertility potential.

Supplementation of sperm cooling medium with helpful additives is a reasonable method to conserve sperm fertility potential during cooling storage process. This study was aimed to determine the effect of sperm cooling medium supplementation with Zinc and Zinc oxide nanoparticles (NZn and NZnO) on rooster semen quality and fertility efficiency during storage periods. Semen samples were diluted in the Lake medium and assigned into five equal aliquots. The first was Control group and the other groups received 50 μg/ml NZn, 50 μg/ml NZnO, 100 μg/ml NZn and 100 μg/ml NZnO. Then, the samples were cooled at 5 °C and conserved up to 45 h. Total motility, progressive motility, mitochondrial activity, viability, membrane integrity and lipid peroxidation of samples were analyzed during 0, 22 and 45 h post-cooling. Artificial insemination was also performed using 22- hrs cooled semen. No difference was found among groups during quality evaluations at 0 h storage. Extender supplementation with 100 μg/ml NZn and NZnO presented higher (P ≤ 0.05) total motility, progressive motility, mitochondrial activity, viability, membrane integrity and lower lipid peroxidation compared to other groups during 22 and 45 h cooling storage. Fertility rate of 22-h cooled-stored semen samples was higher (P ≤ 0.05) in groups contained 100 μg/ml NZn and NZnO compared to the Control group. In conclusion, addition of 100 μg/ml NZn and NZnO to the sperm storage medium could be introduced as an effective method to preserve rooster semen quality during cooling storage period.

Khodaei-Motlagh M ，Masoudi R ，Karimi-Sabet MJ ，Hatefi A ... -  《-》

Supplementation of ram's semen extender with Mito-TEMPO I: Improvement in quality parameters and reproductive performance of cooled-stored semen.

Supplementation of cooling medium with some antioxidants could be a helpful way to improve sperm quality during chilling process. The current study was aimed to assess the influence of using Mito-TEMPO in cooling medium on quality parameters and reproductive performance of sheep semen during chilling process. In this study, diluted semen samples were assigned into 5 parts, and received 0, 0.5, 5, 50 and 500 μM Mito-TEMPO. The prepared samples were stored at 5 °C up to 48 h. Chilled sperm motility, viability, abnormal morphology, mitochondrial membrane potential, membrane functionality and malondialdehyde concentration were assessed during 0, 24 and 48 h. For evaluation of reproductive performance, artificial insemination was performed via 24 h-chilled semen. In results, at time 0, no difference was observed among groups. Using 5 and 50 μM Mito-TEMPO resulted in higher (P ≤ 0.05) cooled sperm total motility, progressive motility, membrane functionality, viability and lower malondialdehyde concentration than the other groups during 24 and 48 h storage. The rate of mitochondrial membrane potential was greater (P ≤ 0.05) in treated groups with 5, 50 and 500 μM Mito-TEMPO. Pregnancy, parturition and lambing rates were higher (P ≤ 0.05) when ewes were inseminated with 24 h-chilled semen samples containing 5 and 50 μM Mito-TEMPO compared to the control group. Therefore, supplementation of cooling medium with Mito-TEMPO (5 and 50 μM) could be an efficient method to improve the quality and reproductive efficiency of ram's cooled semen during storage period.

Zarei F ，Kia HD ，Masoudi R ，Moghaddam G ，Ebrahimi M ... -  《-》

Mitochondria-targeted antioxidant "MitoQ" improves rooster's cooled sperm quality indicators and reproductive performance.

The cell membrane of rooster sperm is sensitive to cold due to the high content of polyunsaturated fatty acids, which are very susceptible to lipid peroxidation. The present study was conducted to determine the effect of different concentrations of the mitochondrial-targeting antioxidant "MitoQ" on sperm quality and fertility potential of chilled semen in roosters. Semen samples were collected from 10 roosters, diluted in Lake extender, assigned into 5 groups according to MitoQ concentrations (0, 1, 10, 100 and 1000 nM MitoQ) and stored at 5 °C up to 48 h. Motility, mitochondrial activity, viability, membrane integrity, and lipid peroxidation were assessed at 0, 24, and 48 h of cold storage periods. In addition, the fertility potential was assessed using 24 h-cooled semen samples. Our results showed that extender supplementation with MitoQ had no effect (P > 0.05) on chilled semen samples quality parameters at time 0, while at times 24 and 48 h storage, samples contained 100 nM MitoQ presented higher (P ≤ 0.05) total motility, progressive motility, viability and membrane integrity compared to the other groups. In addition, semen samples containing 10 and 100 nM MitoQ showed higher (P ≤ 0.05) mitochondrial activity and lower (P ≤ 0.05) lipid peroxidation than other groups at 24 and 48 h storage. Fertility rate was higher (P ≤ 0.05) when the hens were artificially inseminated with 24 h-chilled semen samples containing 100 nM MitoQ. In conclusion, supplementing Lake Extender with 100 nM MitoQ could be a helpful strategy to preserve chilled semen quality and fertility potential in the rooster.

Alipour-Jenaghard P ，Daghigh-Kia H ，Masoudi R ，Moghaddam G ，Qasemi-Panahi B ... -  《-》