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.

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. Keywords: antioxidant gene expression, melatonin, semen cold storage, 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.

Pregnancy Rate after Intrauterine Insemination with the Presence or Absence of Leukocytospermia in Sperms Prepared using Density Gradient Method

Vitrifikasi merupakan suatu teknik kriopreservasi tanpa disertai pembentukan kristal es, baik intraseluler maupun ekstraseluler. Tingkat keberhasilan vitrifikasi sangat dipengaruhi oleh jenis dan konsentrasi krioprotektan yang digunakan. Sukrosa merupakan krioprotektan ekstraseluler yang mempunyai peranan dalam menjaga kestabilan membran sel pada saat proses dehidrasi. Tujuan penelitian ini adalah mengkaji efek penambahan sukrosa dalam dua level konsentrasi yang berbeda pada morfologi dan persentase hidup oosit pascavitrifikasi dengan menggunakan oosit domba yang telah dimatangkan secara in vitro sebagai model. Penelitian dilaksanakan di Laboratorium Riset dan Bioteknologi, Fakultas Peternakan, Universitas Padjadjaran periode September 2016–Desember 2016. Penelitian menggunakan rancangan acak lengkap dengan dua kelompok perlakuan, yaitu penambahan 0,5 M sukrosa atau 0,65 M sukrosa pada media vitrifikasi. Parameter yang diamati adalah persentase oosit hidup dan morfologi oosit yang mati pascavitrifikasi-pencairan kembali. Hasil yang diperoleh menunjukkan bahwa persentase oosit hidup pascavitrifikasi meningkat 20% setelah penambahan sukrosa 0,65M. Fraktur zona pelusida merupakan kerusakan morfologi yang banyak ditemukan pada oosit yang divitrifikasi dengan penambahan 0,5 M sukrosa dalam larutan vitrifikasi. Hasil penelitian ini mengindikasikan bahwa penambahan sukrosa 0,65 M sebagai krioprotektan ekstraseluler meningkatkan peluang hidup oosit setelah proses vitrifikasi. Kata kunci: Krioprotektan, morfologi oosit, sukrosa, vitrifikasiComparison of Oocyte Viability after Vitrification with Two Different Sucrose Concentration LevelVitrification is a cryopreservation technique without the formation of ice crystals, both intracellular and extracellular. The success of vitrification depends on the type and concentration of cryoprotectants. Sucrose is one of the extracellular cryoprotectants that play a role in maintaining cell membrane during dehydration process. The purpose of this study was to examine the effect of the addition of two different sucrose concentration levels on the morphology and viability of oocyte after vitrification using matured sheep oocytes as a model. This study was conducted at the Research and Biotechnology Laboratory, Faculty of Animal Husbandry, Universitas Padjadjaran during September–December 2016. The oocytes were randomly assigned into groups with two different concentrations of sucrose in vitrification media: i.e. 0.5 M and 0.65 M. The parameters observed were the percentage of live oocytes and the dead oocyte morphology after vitrification-thawing. The results showed that the percentage of live matured oocytes post-vitrification increased by 20% after the addition of 0.65M sucrose. Fracture of pellucida zone was the major finding in the post-vitrification dead oocytes in 0.5 M sucrose group. This indicates that vitrification using 0.65M sucrose as extracellular cryoprotectant presents a higher oocyte survival after vitrification-thawing. Key words: Cryoprotectant, oocytes morphology, sucrose, vitrification

In vitro maturation is the crucial step for in vitro embryo production. It needs a large number of oocytes as source gamet cells recovered. The present study is aimed to assess the influence of corpus luteum on the average number oocytes harvested, COCs quality and subsequent maturation of immature oocytes recovered from sheep ovaries. Sheep ovaries were collected from local slaughterhouse and COCs were collected by using slicing method. Collected COCs were graded into three categories dependent upon cumulus cells surrounding them and the homogenous of cytoplasm. COCs were maturated in maturation media at 5% CO2 for 24 hours. Maturation of oocytes evaluated base on the expansion of cumulus cells and extrusion of the first polar body. There was significantly higher on average of COCs harvested from ovaries with corpus luteum compared without corpus luteum. The presence of Corpus luteum did not affect the COCs quality and ability to reach the maturation stage. However, there was a dramatic effect of cultured COCs quality on maturation rate both groups. Collectively, these results indicate that COCs quality is the main factor affecting the subsequent of oocytes matured in vitro. Keywords: Corpus luteum; cumulus oocyte complex; in vitro maturation; maturation rate; ovaries