Simple SummaryArtificial insemination (AI) is widely used in goats, stimulating the development of semen preservation techniques. Oxidative stress is considered to be the main cause of sperm quality decline over time. In this study, we explored the effect of grape seed procyanidin extract (GSPE) during the liquid preservation of goat semen. The results showed that adding GSPE into the basic diluent enhanced sperm quality by eliminating oxidative stress. The most suitable concentration for the preservation of goat semen at 4 °C was 30 mg/L. This work suggests that promotes the viability of goat semen stored at low temperatures and provides a theoretical foundation for the development of more efficient diluents.AbstractGrape seed procyanidin extract (GSPE) has been shown to possess antioxidative effects. This experiment was designed to study the effect of GSPE during the liquid storage of goat semen. Semen samples were collected from six sexually mature goats. The samples were treated with different concentrations of GSPE (10, 30, 50, and 70 mg/L) in basic diluent and stored at 4 °C for 120 h; samples without GSPE were used as the control group. The results showed that sperm motility, acrosome membrane integrity, mitochondrial activity, plasma membrane integrity, total antioxidative capacity (T-AOC), catalase (CAT) activity, and superoxide dismutase (SOD) activity in the treatment groups were significantly higher than in the control group, whereas malondialdehyde (MDA) content was lower than in the control group (p < 0.05). In the treatment group, sperm quality in the 30 mg/L GSPE group was significantly higher than the other groups (p < 0.05). Furthermore, artificial insemination (AI) results showed that litter sizes were higher in the 30 mg/L GSPE group than in the control group (p < 0.05). In summary, this experiment showed that adding GSPE to the basic diluent improved sperm quality and that 30 mg/L of GSPE was the most suitable concentration for the liquid preservation of goat semen at 4 °C.

Adding a certain amount of antioxidants to semen extender has been shown to improve semen quality. The aim of present study was to elucidate whether the supplementation of melatonin to the Tris-based extender (CTR) could enhance the quality of ram spermatozoa during storage at 4°C. Ram semen samples were collected and diluted with CTR extender containing different concentrations (0, 0.05 (M 0.05), 0.1 (M 0.1), 0.2 (M 0.2) or 0.4 (M 0.4) mM) of melatonin. Sperm routine indicators, mitochondrial activity, total antioxidant capacity (T-AOC) and malondialdehyde (MDA) content were analysed in control and melatonin treatment groups. The higher per cent of motility, plasma membrane integrity, mitochondrial activity and T-AOC activity was observed in M 0.05, M 0.1 and M 0.2 groups compared to control group at 5 days of storage (p < 0.05), while lower percentage of MDA content was observed among these groups (p < 0.05). In addition, there were no significant differences in acrosome integrity among the control and M 0.05, M 0.1 and M 0.2 groups during the experiment. The above results show that the addition of 0.05, 0.1, 0.2 mM of melatonin is beneficial to the preservation of ram semen during liquid storage at 4°C mainly through antioxidative stress. K E Y W O R D S antioxidant, melatonin, Ram, semen storage How to cite this article: Dai G-C, Meng Y, Zhang L, et al. Effect of addition of melatonin on liquid storage of ram semen at 4°C.

Background Goat milk is very similar to human milk in terms of its abundant nutrients and ease of digestion. To derive greater economic benefit, farmers require more female offspring (does); however, the buck-to-doe offspring sex ratio is approximately 50%. At present, artificial insemination after the separation of X/Y sperm using flow cytometry is the primary means of controlling the sex of livestock offspring. However, flow cytometry has not been successfully utilised for the separation of X/Y sperm aimed at sexing control in dairy goats. Results In this study, a novel, simple goat sperm sexing technology that activates the toll-like receptor 7/8 (TLR7/8), thereby inhibiting X-sperm motility, was investigated. Our results showed that the TLR7/8 coding goat X-chromosome was expressed in approximately 50% of round spermatids in the testis and sperm, as measured from cross-sections of the epididymis and ejaculate, respectively. Importantly, TLR7/8 was located at the tail of the X-sperm. Upon TLR7/8 activation, phosphorylated forms of glycogen synthase kinase α/β (GSK3 α/β) and nuclear factor kappa-B (NF-κB) were detected in the X-sperm, causing reduced mitochondrial activity, ATP levels, and sperm motility. High-motility Y-sperm segregated to the upper layer and the low-motility X-sperm, to the lower layer. Following in vitro fertilisation using the TLR7/8-activated sperm from the lower layer, 80.52 ± 6.75% of the embryos were XX females. The TLR7/8-activated sperm were subsequently used for in vivo embryo production via the superovulatory response; nine embryos were collected from the uterus of two does that conceived. Eight of these were XX embryos, and one was an XY embryo. Conclusions Our study reveals a novel TLR7/8 signalling mechanism that affects X-sperm motility via the GSK3 α/β-hexokinase pathway; this technique could be used to facilitate the efficient production of sexed dairy goat embryos.

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BackgroundGoat milk is most similar to human milk because of its abundant nutrients and ease of digestion. To derive more economic benefits, farmers need to obtain offspring with more does; however, the buck to doe sex ratio of offspring is approximately 50%. At present, artificial insemination after separation of X/Y sperm using flow cytometry is the primary means to control the sex of livestock offspring. However, flow cytometry has not been successful in separating X/Y sperm for sexing control in dairy goats.ResultsIn this study, a novel, simple goat sperm-sexing technology that activates the toll-like receptor 7/8 (TLR7/8), inhibiting X-sperm motility, was investigated. Results showed that TLR7/8 coding goat X-chromosome was expressed in approximately 50% of round spermatids in testis' cross-sections and sperm in the epididymis' cross-sections and ejaculate. Importantly, TLR7/8 was located at the tail of X-sperm. Upon TLR7/8 activation, phosphorylated forms of glycogen synthase kinase α/β (GSK3 α/β) and nuclear factor kappa-B (NFκB) were detected in the X-sperm, reducing mitochondrial activity, ATP levels, and sperm motility. High-motility Y-sperm segregated to the upper layer and low-motility X-sperm to the lower layer. Following in vitro fertilisation using the lower layer TLR7/8-activated sperm, 80.52 ± 6.75% of the embryos were XX females. TLR7/8-activated sperm was used for in vivo embryo production via the superovulatory response, and 88.89% of the embryos were XX females.ConclusionsOur study reveals a novel TLR7/8 signalling mechanism that affects X-sperm motility via the GSK3 α/β-hexokinase pathway, facilitating the efficient production of sexed dairy goat embryos.