Preservation of rooster post-thawed sperm epigenetic modifications, fertility potential and other quality parameters in different extenders using reduced glutathione.

Although rooster semen cryopreservation is an efficient procedure to spread qualified semen samples for reproductive goals, some post-thawed qualified semen samples resulted in poor fertility rate that could be related to epigenetic modifications during the cryopreservation process. This research was conducted to investigate the effect of reduced glutathione (GSH) in different cryopreservation extenders (Lake and Beltsville) on preservation of epigenetic modifications, fertility potential and other quality parameters of rooster sperm after thawing. Semen samples were collected and diluted in Lake and Beltsville extenders as follows: L-0: Lake without GSH, L-G: Lake with GSH, B-0: Beltsville without GSH, and B-G: Beltsville with GSH. After freeze-thawing process, sperm motility, membrane functionality, mitochondrial activity, acrosome integrity, viability, apoptosis status, lipid peroxidation, DNA fragmentation, ROS concentration, epigenetic modifications and fertility potential were evaluated. In results, the type of extender had no effect (P > 0.05) of post-thawed sperm quality. The treatments containing GSH presented higher (P ≤ 0.05) total motility, progressive motility, membrane functionality, mitochondrial activity, acrosome integrity, viability, DNA methylation, fertility as well as lower (P ≤ 0.05) lipid peroxidation, apoptosis, DNA fragmentation and ROS concentration than other treatments. Extender supplementation with GSH had no effect (P > 0.05) on histone methylation, histone acetylation and hatching rate. In conclusion, supplementation of rooster sperm cryopreservation extender with GSH could be an effective strategy to preserve post-thawed sperm DNA methylation, fertility and other quality parameters during reproductive programs.

Flow cytometry study of post-thawed buck spermatozoa: Mito-TEMPO improves cryopreservation performance by controlling apoptosis rate, DNA fragmentation and ROS production.

Sperm cryopreservation is used to spread qualified semen for artificial insemination, but the freezing process reduces sperm quality. This study assessed the efficacy of Mito-TEMPO on post-thawed goat sperm quality. Semen samples divided to five equal groups and after dilution, received different doses of Mito-TEMPO (0, 1, 10, 100 and 1000 µM), and cryopreserved in liquid nitrogen. After thawing, flow cytometry analysis was performed to evaluate sperm mitochondria membrane potential, viability, apoptotic-like changes, DNA fragmentation and ROS concentration. According to the results, Mito-TEMPO (10 and 100 µM) improved (P ≤ 0.05) sperm viability and decreased (P ≤ 0.05) apoptotic-like changes and ROS concentration compared to the other groups. Mitochondria membrane potential was higher (P ≤ 0.05) in groups received 1, 10 and 100 µM Mito-TEMPO. The lowest (P ≤ 0.05) DNA fragmentation was observed in group received 10 µM Mito-TEMPO. In conclusion, mitochondria-targeted antioxidant Mito-TEMPO could be an efficient cryo-additive to enhance flowcytometric quality parameters of post-thawed buck semen.

Cysteamine enhances quality and fertility potential of rooster semen in cooled storage.

This study investigated the effects of supplementing Lake extender with cysteamine (CYS) on rooster semen quality in cold storage and it's fertility performance. Semen samples were diluted with Lake extender supplemented with different concentrations of CYS (0, 1, 2, 4 and 8 mM) and were cooled and stored at 5 °C for a period of 46 h. Motility, membrane functionality, viability, lipid peroxidation, and mitochondria membrane potential were evaluated at 0, 23 and 46 h of storage. Fertility was assessed at 23 h of storage. Although at the beginning time (0 h), parameters were not affected, 1 mM of CYS improved (P ≤ 0.05) total motility, progressive motility and mitochondria membrane potential during 23 and 46 h storage. Moreover, 1 and 2 mM CYS improved (P ≤ 0.05) membrane functionality and viability compared to other groups. Lipid peroxidation was lower (P ≤ 0.05) in samples diluted with 1 and 2 mM CYS compared to the others. Artificial insemination with 23-hrs cooled-stored semen produced the higher (P ≤ 0.05) fertility rate in groups received 1 and 2 mM CYS compared to the control group. In conclusion, addition of 1 and 2 mM CYS to the extender could be helpful to protect rooster semen against structural and functional damages of cooling storage process.

Flow cytometry study of post-thawed bulk spermatozoa: Mito-TEMPO improves cryopreservation performance by controlling apoptosis rate, DNA fragmentation and ROS production.

Sperm cryopreservation is used to spread qualified semen for artificial insemination, but the freezing process reduces sperm quality. This study assessed the efficacy of Mito-TEMPO on post-thawed goat sperm quality. Semen samples divided to five equal groups and after dilution, received different doses of Mito-TEMPO (0, 1, 10, 100 and 1000 µM), and cryopreserved in liquid nitrogen. After thawing, flow cytometry analysis was performed to evaluate sperm mitochondria membrane potential, viability, apoptotic-like changes, DNA fragmentation and ROS concentration. According to the results, Mito-TEMPO (10 and 100 µM) improved (P ≤ 0.05) sperm viability and decreased (P ≤ 0.05) apoptotic-like changes and ROS concentration compared to the other groups. Mitochondria membrane potential was higher (P ≤ 0.05) in groups received 1, 10 and 100 µM Mito-TEMPO. The lowest (P ≤ 0.05) DNA fragmentation was observed in group received 10 µM Mito-TEMPO. In conclusion, mitochondria-targeted antioxidant Mito-TEMPO could be an efficient cryo-additive to enhance flowcytometric quality parameters of post-thawed bulk semen.

Supplementation of chilling storage medium with glutathione protects rooster sperm quality.

This study was aimed to evaluate the effect of addition of reduced glutathione (GSH) to the extender on the rooster's semen quality parameters and fertility potential. Semen samples were diluted with Lake extender contained 0, 0.5, 1, 2, 4 and 8 mM GSH. Then, were chilled to 5 °C and stored for a period of 48 h. Sperm motion characteristics, viability, membrane integrity, lipid peroxidation, mitochondrial activity and fertility potential were evaluated. At the initiation of the experiment (0 h), GSH did not affect sperm parameters, while 2-4 mM GSH improved (P ≤ 0.05) quality indicators during storage periods. Moreover, the samples treated with 2-4 mM GSH have had a lower lipid peroxidation compared to other groups (P ≤ 0.05). Artificial insemination using the semen samples, which had been stored in groups treated with 2-4 mM GSH for a period of 24 h, led to greater (P ≤ 0.05) fertilizing potential compared to the control group.

Genetic diversity of the Dwarf honeybee (Apis florea Fabricius, 1787) populations based on microsatellite markers.

Apis florea is one of two species of small, wild honeybee. The present study was conducted to evaluate the genetic diversity of Apis florea honeybee from 48 nests (colonies) using microsatellite markers in the South of Iran. All honeybee samples were analyzed for six microsatellite loci (A88, A107, A7, B124, A113 and A35). The six loci had different numbers of alleles in the sampled colonies ranging from 7 (loci A107) to 3 (loci A7, A35). Gene diversity in Apis florea ranged from 0.491 to 0.595. This range probably reflects the spreading of nests in a large region with a varied climate. Phylogenetic tree showed two distinct clusters including a) Minab region samples and b) Bandar Abbas, Bandar Khamir and Qeshm Island regions. All of these regions are geographically rich, having varied vegetation and climate conditions. Our findings are an important contribution to the methods of studying distribution and conservation of Apis florea.

Genetic variation among native chicken breeds of Iran based on biochemical polymorphisms.

(1) Genetic distances and heterozygosity were determined in 5 Iranian native chickens: Dashtyary, Lary, Marandy, Naked neck and Common breed, using three blood group systems (A, B and D) and 4 serum protein loci (albumin, transferrin, alkaline phosphatase and esterase). (2) The highest D(A) and D(S) were obtained between Dashtyary and Lary breeds. (3) The average heterozygosity for each breed in all loci ranged from 0.330 for Dashtyary to 0.370 for Common breed. (4) Dendrograms based on an unweighted pair-group method using an arithmetic average (UPGMA) showed two distinct clusters. One cluster included Dashtyary and the other contained the remaining 4 breeds.