Effect of sperm concentration on characteristics and fertilization capacity of rooster sperm frozen in the presence of the antioxidants catalase and vitamin E.

The objective of this study conducted was to determine the influence of different levels of sperm concentration, including catalase (CAT) and vitamin E (VitE) in rooster semen extender on postthawed quality and fertility of rooster semen. Semen was collected twice a week from six roosters (Arian) and diluted according to experimental treatments consisting of sperm suspensions containing different sperm concentrations (200, 400, and 600 × 106 sperm/mL) without antioxidant supplementation as control (Con) groups (Con200, Con400, and Con600, respectively), sperm suspensions containing different sperm concentrations (200, 400, and 600 × 106 sperm/mL) supplemented with 5-μg/mL VitE (VitE200, VitE400, and VitE600, respectively) and different sperm concentrations (200, 400, and 600 × 106 sperm/mL) supplementation with 100 IU/mL CAT (CAT200, CAT400, and CAT600, respectively). After thawing; sperm motility, membrane integrity, and mitochondrial function were assessed. Fertility and hatchability rates were determined by using 100 artificially inseminated hens. The percentage of total motility (TM) and activity of mitochondria decreased (P < 0.05) as the sperm concentration increased in control groups. So, the lowest percentage of the TM and activity of mitochondria were observed in the Con600 as compared with other treatment groups. Extenders containing 100 IU/mL CAT and 5-μg/mL VitE resulted in higher (P < 0.05) TM, progressive motility, membrane integrity, and activity of mitochondria compared with control groups. Adding VitE and CAT in different sperm concentrations, the percentage of TM, membrane integrity, and activity of mitochondria decreased (P < 0.05) as the sperm concentration decreased. The highest (P < 0.05) membrane integrity, TM, and progressive motility were recorded at VitE400 and CAT400. Including VitE and CAT in rooster extender with different level sperm concentrations had no effect (P > 0.05) on fertility and hatchability rates. In conclusion, although adding VitE and CAT in extender with different levels of sperm concentration improved postthawed quality of rooster semen, but adding VitE and CAT in the extender have no effect on fertility rate.

Moghbeli M ，Kohram H ，Zare-Shahaneh A ，Zhandi M ，Sharideh H ，Sharafi M ... -  《-》

Are the optimum levels of the catalase and vitamin E in rooster semen extender after freezing-thawing influenced by sperm concentration?

To date, there has no report to evaluate the interaction effects of antioxidant and sperm concentration in rooster semen cryopreservation. This study was aimed to investigate the effects of vitamin E (VitE) and catalase (CAT) at different sperm concentrations on the rooster post-thawed sperm quality. Semen samples were collected twice a week from ten roosters (ROS 308) and diluted according to experimental treatments. The treatments consist of different sperm concentrations (200, 400 and 600 × 106 sperm/mL) with supplementation VitE (5 μg/mL; VitE200, VitE400, and VitE600, respectively) or CAT (100 IU/mL; CAT200, CAT400, CAT600, respectively) and without antioxidants [Control (Con); Con200, Con400, Con600, respectively]. After thawing, motion characteristics were assessed using a CASA system. Plasma membrane integrity and malondialdehyde (MDA) level were evaluated with Hypoosmotic swelling test (HOST) and Thiobarbituric acid (TBA), respectively. The higher percentage of total motility, progressive motility, viability and membrane integrity were obtained in VitE400 (81.16 ± 1.21, 18.44 ± 1.19, 85.47 ± 1.07, 86.91 ± 1.16, respectively) and CAT400 (79.38 ± 1.21, 17.19 ± 1.19, 83.42 ± 1.07, 85.73 ± 1.16, respectively) compared to control groups. Moreover, the lowest percentage of MDA was measured in VitE400, VitE600 and CAT400 rather than other groups (1.489, 1.500, 1.510 ± 0.06, respectively). In conclusion, the results of the present study demonstrate that VitE (5 μg/mL) and CAT (100 IU/mL) independently at sperm concentration, 400 million sperm/mL could beneficial effect for preservation of rooster semen during cryopreservation.

Moghbeli M ，Kohram H ，Zare-Shahaneh A ，Zhandi M ，Sharafi M ，Nabi MM ，Zahedi V ，Sharideh H ... -  《-》

Effect of vitamin E and selenium nanoparticles on post-thaw variables and oxidative status of rooster semen.

This study was conducted to determine the combined effects of adding vitamin E (VitE) and selenium nanoparticle (Nano-Se) as antioxidant supplements to rooster semen extender for freezing. Semen samples were collected from 12 White Leghorn roosters and pooled. Subsequently, the samples were divided into nine equal groups using modified Beltsville extender. Extenders were supplemented with either two amounts of VitE (5 and 10μg/mL) or two amounts of Nano-Se (1% and 2%) or a combination of both VitE and Nano-Se, and no antioxidants extender (control group). Using 5μg/mL VitE and 1% of Nano-Se improved (P<0.05) total sperm motility (79.28±3.86%), progressive sperm motility (18.03±1.02%), sperm viability (81.46±2.16%) and integrity of the sperm membrane (77.21±2.12%) after the freeze-thawing process compared with control group (54.08±3.86%, 10.96±1.02%, 60.53±2.16%, and 54.47±2.12%; respectively). Also, extenders supplemented with 5μg/mL Vit E or 5μg/mL VitE and 1% Nano-Se had a lesser malondialdehyde (MDA) concentration compared to control extender (1.15±0.32, and 1.29±0.32, respectively). Total abnormal morphology of sperm was decreased (P<0.05) by adding 5 or 10μg/mL VitE alone or in combined with 1% or 2% Nano-Se. Moreover, extenders containing Nano-Se demonstrated greater glutathione peroxidase (GPx) activity compared to other extenders. Catalase (CAT) activity was greater in extender supplemented with 10μg/mL VitE and 2% Nano-Se. Moreover, higher TAC was observed in extenders supplemented with VitE and Nano-Se. It can be concluded that addition of 5μg/mL vitamin E in combined with 1% Nano-Se improved the post-thawing quality and oxidative variables of rooster semen.

Safa S ，Moghaddam G ，Jozani RJ ，Daghigh Kia H ，Janmohammadi H ... -  《-》

The effects of different levels of catalase and superoxide dismutase in modified Beltsville extender on rooster post-thawed sperm quality.

Oxidative damage of sperm by means of reactive oxygen species generated by the cellular components of semen is one of the main reasons for decreased sperm motility and fertility during the freeze-thawing process. This study was conducted to determine the influence of catalase (CAT) and superoxide dismutase (SOD) on rooster sperm motility, viability and MDA level after freezing and thawing. Semen samples from 10 sexually-mature Ross 308 breeder roosters were collected and pooled, divided into nine equal parts and diluted with modified Beltsville extender containing no antioxidants (control), or supplemented with 50, 100, 200 and 300 μg/mL CAT, or 50, 100, 200 and 300 U/mL SOD. After thawing, sperm motility and motion parameters were assessed using a CASA system. Sperm viability and MDA level were assessed by eosin-nigrosin and MDA test, respectively. The results of this experiment showed that the extender supplemented with 100 and 200 μg CAT, and 50 U SOD had the highest sperm motility (P<0.05) in sperm motility. Also, addition 100, 200 and 300 μg CAT, and 50 U SOD can improve significantly viability after freeze-thaw. Extender supplemented with 100 μg CAT had significantly lower MDA level compared to control and 300 μg CAT. In conclusion, the results of the present study demonstrate that addition of CAT (100 μg/mL) and SOD (50 U/mL) independently have beneficial effect on quality of post-thawed rooster semen.

Amini MR ，Kohram H ，Zare-Shahaneh A ，Zhandi M ，Sharideh H ，Nabi MM ... -  《-》

Fertility and flow cytometric evaluations of frozen-thawed rooster semen in cryopreservation medium containing low-density lipoprotein.

Shahverdi A ，Sharafi M ，Gourabi H ，Yekta AA ，Esmaeili V ，Sharbatoghli M ，Janzamin E ，Hajnasrollahi M ，Mostafayi F ... -  《-》