Preservation of the quality and fertility potential of post-thawed rooster sperm using MitoQ.

Cryopreservation of rooster spermatozoa is an efficient procedure to spread qualified semen samples for reproductive goals in commercial flocks, but the freeze-thawing process reduces the quality and fertility potential of post-thawed sperm cells. This study was aimed to investigate the effect of the mitochondria-targeted antioxidant MitoQ on rooster sperm quality and fertility potential preservation during freeze-thawing process. Semen samples were collected and diluted in the Lake medium, assigned into five equal aliquots, supplemented with 0, 1, 10, 100 and 1000 nM MitoQ, and cryopreserved in liquid nitrogen. After thawing, sperm motility, membrane functionality, abnormal morphology, mitochondria active potential, acrosome integrity, viability, apoptosis status, lipid peroxidation, DNA fragmentation, ROS concentration and fertility potential were evaluated. According to the results, freezing extender supplementation with 10 and 100 nM MitoQ presented higher (P ≤ 0.05) total motility, progressive motility, average path velocity, membrane functionality, mitochondria active potential, acrosome integrity and viability compared to the other groups. On the other hand, lipid peroxidation, apoptosis rate, DNA fragmentation and ROS concentration were lower (P ≤ 0.05) in groups received 10 and 100 nM MitoQ compared to other groups. Moreover, fertility rate was higher in groups received 10 and 100 nM MitoQ compared to control group. Therefore, MitoQ is able to preserve quality parameters and fertility potential of post-thawed spermatozoa in rooster and it could be an effective additive for supplementation of rooster's semen cryopreservation medium during reproductive programs.

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.

Conservation of Buck's Spermatozoa during Cooling Storage Period through Cooling Medium Supplementation with L-Carnitine.

This research examined the influence of the addition of L-carnitine (LC) to cooling medium on buck's semen quality during cooling storage periods at 4oC. Semen samples were collected, diluted, assigned into four groups, and received LC (0, 1, 5, and 10 mM LC). The samples were then chilled to 4oC and stores for 48 h. Sperm total motility, progressive motility, viability, lipid peroxidation, membrane integrity, and mitochondrial activity were examined at 0, 24, and 48 h of cooling storage. At time 0 of cooling storage, different treatments showed no impact on the quality of sperm samples (P>0.05). During 24 and 48 h of chilling periods, the supplementation of cooling medium with 5 mM LC presented greater motility, viability, membrane integrity, and mitochondrial activity (P≤0.05), compared to the other groups. Moreover, the treatment of 5 mM LC caused lower lipid peroxidation (P≤0.05) than the other treatments at 24 and 48 h storage times. In conclusion, the supplementation of buck's cooling storage medium with 5 mM LC is a suitable way to protect buck spermatozoa during 24 and 48 h storage against cold-induced structural and functional damages.

Effects of in combination antioxidant supplementation on microscopic and oxidative parameters of freeze-thaw bull sperm.

The present study was conducted to determine the effect of reduced glutathione (GSH) and superoxide dismutase (SOD) supplementation in the bull semen freezing extender on post-thaw parameters of Holstein and Simmental bull sperm. Semen were collected from seven bulls (four Holstein and three Simmental) and cryopreserved in the Tris-egg-yolk based extender as control group and supplemented with various concentrations of GSH × SOD (5 × 100, 7.5 × 100, 5 × 150, and 7.5 × 150 mM × IU mL(-1)) in treatment groups. Microscopic parameters were evaluated in terms of total motility parameters using computer assisted semen analysis and viability and membrane integrity were assessed using Eosin-Nigrosin stains and hypo-osmotic swelling test (HOST), respectively after thawing the semen. Malonaldialdehyde (MDA) level, SOD and glutathione peroxides (GPx) activities were assessed immediately after thawing. Results showed that supplementation of the cryopreservation medium with various concentrations of GSH × SOD improved total motility (TM) and progressive motility parameters for Holstein (P < 0.05) semen, and values of TM and HOST for Simmental semen compared to the control group (P < 0.01) after semen thawing. Addition of antioxidant to Holstein semen samples decreased the level of MDA and increased GPx activities compared to control groups (P < 0.05). SOD activities increased in Simmental bull samples compare to the control group (P < 0.01), but not differ in Holstein, while these activities. In conclusion, supplementation of antioxidant to the semen extender as combination (GSH × SOD) improved the semen post-thaw qualities which may be associated with a reduction in lipid peroxidation as well as an increase in the antioxidant enzyme activities.

Effects of flushing and hormonal treatment on reproductive performance of Iranian Markhoz goats.

Forty-eight Iranian Markhoz goats were allocated to six groups (n = 8) to study the effect of flushing and hormonal treatments on reproductive performance. Treatments were divided into two categories including, hormonal treatments and flushing. The goats in each group were fed the same basal ration and received one of the following treatments: Groups A and B--injection of GnRH and equine chorionic gonadotropin (eCG) respectively; Groups C, D and E--a supplement of barley grain, soybean oil and sunflower oil in flushing diets, respectively, were offered and Group F--control (only received basal diet). In the flushing treatments, only the source of energy was different between rations. Both hormonal treatments and flushing treatments improved fertility and kidding rates. Treatment B with 16 and control with seven kids represented the highest and the lowest number of progeny respectively. Among flushing treatments, group C resulted in the highest number of kids being 15. Oestrogen levels in follicular phase increased with the injection of eCG and consumption of barley grain. GnRH injection and consumption of oil sources in the diet increased blood progesterone levels during ovulation and post-ovulation periods. Under current market conditions, using hormone or flushing can be profitable for Markhoz goats farmers.