Effect of melatonin supplementation on sperm quality parameters and expression of antioxidant genes during cold storage of buck semen extenders.

Background and Aim: Semen storage is an important reproductive method used in artificial livestock breeding. However, oxidative stress during storage reduces the quality of sperm. Melatonin supplementation in semen storage medium has not been well studied, but it has been shown to protect cells from oxidative stress. Therefore, this study aimed to determine the effect of melatonin supplementation on sperm quality parameters and antioxidant gene expression levels in semen extenders during cold storage. Materials and Methods: Semen extenders with melatonin concentrations of 0 (control), 0.1, 0.2, and 0.3 mM were added as treatment. The treated semen was then stored at 5°C for 72 h using a cold storage method, and quality parameters, including percentage of progressive motility, membrane integrity, intact acrosome, and DNA integrity, were measured every 24 h. In addition, messenger ribonucleic acid abundance levels of glutathione peroxidase (GPx) and superoxide dismutase (SOD) genes were sampled after 0 and 72 h of cold storage. Results: All observed sperm quality parameters decreased with increasing cold storage time; however, 0.2 mM melatonin demonstrated superior protection of sperm quality during cold storage. Gene expression analysis showed that GPx levels decreased significantly (p < 0.05) after 72 h in semen without melatonin but not in the melatonin-treated groups. A similar trend was also observed in SOD, indicating that exogenous antioxidants effectively protected the sperms. Conclusion: Melatonin supplementation at 0.2 mM in semen extenders during cold storage maintains sperm quality parameters for up to 72 h because melatonin protects sperm from oxidative stress. These findings can be used to improve the semen storage protocol by combining semen extender and antioxidant.

Fertility testing of preserved epididymal sperm by microinjection: A model for the rescue and utilization of genetically superior animals.

Background: Epididymal sperm preservation is a simple conservation approach that can help prevent the loss of high genetic quality of farm animals. The chance of loss increases, especially during disease outbreaks or other interruptions to normal reproduction function. Aim: This study looked into the ability of preserved ram epididymal sperm to fertilize oocytes. Due to motility becoming an issue following sperm storage for fertilization, the sperm microinjection known as intracytoplasmic sperm injection approach was employed. Methods: The study was divided into two parts. First, involved the preservation of epididymal sperm at 5°C for 12 days. During preservation, sperm quality parameters namely motility, viability, intact membrane, acrosome, and Deoxyribonucleic acid (DNA) are evaluated every three days. For the fertility test in the second experiment, matured oocytes were injected with immotile sperm discovered in the last days of preservation. The presence of pronucleus development following in vitro culture is used as an indicator of sperm's ability to activate and fertilize oocytes. Results: All sperm quality parameters significantly (p < 0.05) declined during preservation time. On day 12, motility was discovered to be 0%, but viable sperm, sperm with intact membrane, acrosome, and DNA remained at 41.86% ± 9.30%, 31.18% ± 5.15%, 21.88% ± 1.93%, and 33.35% ± 8.74%, respectively. On the fertility test, we inject immotile sperm from day 12 of preservation, which has the lowest motility found, into matured oocytes. Those sperms are able to activate (52.05% ± 7.15%) and fertilize (31.37% ± 1.75%) the injected oocytes, but their fertilizing ability is significantly lower (p < 0.05) when compared to the sperm derived from the ejaculate. Conclusion: In this study, simple preservation of epididymal sperm reduces all sperm quality criteria, particularly motility. Using the microinjection approach preserved sperm which had no motility, still demonstrated its ability to activate and fertilize the oocytes. According to that, this study provides potential approaches and tools for using genetically superior animals that have lost their ability to execute regular fertilization, and also prolong reproduction function.

Impact of glucose and pyruvate on adenosine triphosphate production and sperm motility in goats.

OBJECTIVE: This study evaluates goat sperm motility in response to metabolic substrates and various inhibitors, aiming to assess the relative contribution of glycolysis and mitochondrial oxidation for sperm movement and adenosine triphosphate (ATP) production. METHODS: In the present study, two main metabolic substrates; 0 to 0.5 mM glucose and 0 to 30 mM pyruvate were used to evaluate their contribution to sperm movements of goats. Using a 3-chloro-1,2-propanediol (3-MCPD), a specific inhibitor for glycolysis, and carbonyl cyanide 3-chlorophenylhydrazone as an inhibitor for oxidative phosphorylation, cellular mechanisms into ATP-generating pathways in relation to sperm movements and ATP production were observed. Data were analysed using one-way analysis of variance for multiple comparisons. RESULTS: Sperm motility analysis showed that either glucose or pyruvate supported sperm movement during 0 to 30 min incubation. However, the supporting effects were abolished by the addition of a glycolysis inhibitor or mitochondrial uncoupler, concomitant with a significant decrease in ATP production. Although oxidative phosphorylation produces larger ATP concentrations than those from glycolysis, sperm progressivity in relation to these two metabolic pathways is comparable. CONCLUSION: Based on the present study, we suggest that goat sperm use glucose and pyruvate to generate cellular energy through glycolysis and mitochondrial respiration pathways to maintain sperm movement.

Deleterious effect of short-term gavage of an ethanol extract of cogon grass (Imperata cylindrica L.) roots on testis and epididymal sperm quality.

BACKGROUND AND AIM: Cogon grass (Imperata cylindrica L.) (CGG) is a herbal medicine that could be developed into a male antifertility agent. The present study aims to determine the effect of an ethanol extract of CGG roots on mice testicular activity, reproductive hormone levels, and epididymal sperm quality. MATERIALS AND METHODS: This study was designed as completely randomized with three different doses, such as an ethanol extract of CGG roots at 0 (control), 90, and 115 mg/kg body weight. In total, 21 male DDY mice strain were treated with the CGG extract (by gavage) for 14 days, followed by an evaluation of reproductive organs, epididymal sperm quality, testis histology, histomorphometry, and reproductive hormone assays. All quantitative data were analyzed by analysis of variance, followed by Tukey's post hoc test at α=0.05. RESULTS: The results showed that the administration of the CGG root ethanol extract disrupted the testis interstitial area and seminiferous tubules, resulting in decreased epididymal sperm quality as well as serum testosterone levels in a dose-dependent pattern. CONCLUSION: Oral administration of a CGG root ethanol extract induced testicular damage, decreased epididymal sperm quality, and impaired testosterone secretion.