To the best of our knowledge, this study is the first to determine the effect of ginseng as an antioxidant supplement in freezing extenders on the quality of cryopreserved rooster semen. Semen samples were collected from 40 Thai native roosters (Pradu Hang Dum) using the dorso-abdominal massage method and then pooled and divided into five groups according to the concentrations of ginseng supplementation (0, 0.25, 0.50, 0.75, and 1 mg/mL) in a freezing extender. The semen suspensions were loaded into a medium straw and cryopreserved using the liquid nitrogen vapor method. The post-thaw semen was evaluated for sperm quality (sperm motility and membrane integrity), seminal plasma characteristics (lipid peroxidation, superoxide dismutase [SOD], catalase [CAT], and glutathione peroxidase [GPx]), and fertility. The results showed that ginseng extract supplementation at 0.25 mg/mL yielded the highest total motility, progressive motility, and membrane integrity (59.47%, 30.82%, and 48.30%, respectively; p < 0.05) in cryopreserved rooster semen. Higher malondialdehyde concentrations were observed in the control group than in the other groups (p < 0.05). SOD, CAT, and GPx increased compared with those in the control group (p < 0.05). The results showed that the fertility rate with 0.25 mg/mL of ginseng was higher than that of the control group (62.80% vs. 46.28%: p < 0.05). In conclusion, supplementation with 0.25 mg/mL of ginseng is recommended as an alternative component to the freezing extender to improve rooster semen cryopreservation.

The cooling of semen storage at 5 °C from a Thai native rooster (Pradu Hang Dum), supplemented with herbs possessing antioxidant properties, provided limited research. This study was conducted to evaluate the efficiency of Aloe vera (AV) gel supplementation at various levels on the quality of cooled semen and subsequent fertility after artificial insemination. Sixty-four chickens had semen pooled, diluted, and supplemented with different levels of AV gel (0% as control, 0.25%, 0.50%, 1.0%, 2.5%, 5.0%, 10%, and 20%), and then stored for 72 h. In Experiment 1, semen quality, malondialdehyde (MDA) levels, and pH values were assessed at 0, 24, 48, and 72 h after storage. Experiment 2 assessed fertility potential using the most effective cooled storage semen from Experiment 1. Results showed a decrease in semen quality with prolonged storage time (p < 0.001). The highest semen quality was observed in the group supplemented with 1.0% AV gel (p < 0.001), whereas the lowest was noted in the 20% AV gel group (p < 0.001). Furthermore, the 1.0% AV gel group exhibited the highest semen quality at 24, 48, and 72 h of storage. The evaluation of fertility and hatchability rates revealed a statistically significant improvement in fertility potential (p < 0.05) in the group supplemented with 1.0% AV gel. In summary, this study represents the first investigation of stored Thai native rooster semen using a semen extender supplemented with Aloe vera gel at 5 °C, demonstrating its efficacy for storage up to 72 h. The addition of 1% AV gel was recommended as an antioxidant supplementation during the semen storage process at 5 °C to enhance semen quality and fertility rates.

Cryopreservation of native Thai bull semen often results in significant post-thaw quality reduction, underscoring the need for effective cryoprotective strategies. This study investigated the effect of Moringa oleifera leaf extract (MOLE) as an antioxidant supplementation by incorporating four MOLE concentrations (0–1.5% [w/v]) into a standard semen extender, followed by cryopreservation using liquid nitrogen vapor freezing. Data were analyzed using a randomized complete block design with Tukey’s post hoc test (p < 0.05). Post-thaw analysis of semen revealed that 1 mg/mL MOLE significantly enhanced total sperm motility, progressive sperm motility, sperm viability, and sperm plasma membrane integrity compared to the control and other MOLE concentrations (p < 0.05). This concentration also improved the amplitude of lateral head displacement and curvilinear velocity and reduced malondialdehyde levels in semen samples (p < 0.05), indicating reduced lipid peroxidation. Higher MOLE concentrations negatively impacted semen quality. In conclusion, supplementation with 1 mg/mL MOLE markedly improved post-thaw semen quality and reduced lipid peroxidation, suggesting its potential as an antioxidant for enhancing reproductive outcomes in native Thai bulls.

Objective: Aging roosters typically exhibit subfertility with decreasing semen quality, furthermore Thai native roosters reared in rural areas are raised for a longer duration than their usual lifespan. The present study therefore aimed to assess the effect of selenium supplementation as an antioxidative substance in diets to improve the semen cryopreservation of aged roosters.Methods: Semen samples were collected from young (n = 20) and aged (n = 20) Thai native roosters (Pradu Hang Dum) at 36 and 105 weeks of age when starting the experiment, respectively. They were fed diets either non-supplemented or supplemented with selenium (0.75 ppm). Fresh semen quality and lipid peroxidation of fresh semen was evaluated before cryopreservation using the traditional liquid nitrogen vapor method. Post-thaw sperm quality and fertility potential were determined.Results: Advancing age is unrelated to decreasing fresh semen quality (p>0.05). However, lipid peroxidation in rooster semen depended on age, and the malondialdehyde (MDA) concentration increased in aged roosters (p<0.05). Selenium supplementation in diets significantly decreased the MDA concentration and increased the sperm concentration (p<0.05). In contrast, cryopreserved semen was affected by advancing rooster age, and selenium influenced sperm quality (p<0.05). Younger roosters had higher post-thaw sperm quality and fertility potential than aged roosters (p<0.05). Likewise, diet selenium supplements improved post-thaw sperm quality and fertility compared with the non-supplement group.Conclusion: Rooster’s age does not influence the rooster sperm quality of fresh semen, while sperm cryotolerance and fertility were greater in young roosters than in aged roosters. However, sperm of aged roosters could be improved by dietary selenium supplementation.

Objective: Semen cryopreservation result in decreased sperm parameters and fertilization ability. Sericin exhibits antioxidant activity by reducing lipid peroxidation resulting from free radicals, which can potentially improve cryopreservation outcomes. The present study aimed to examine the efficacy of various sericin concentrations supplemented with a rooster semen-freezing extender on post-thaw semen quality and fertilizing ability of sperm after cryopreservation.Methods: Semen samples were collected from 40 roosters (5 reps), then were pooled, and divided into four groups by the levels of sericin supplementation (0%, 0.25%, 0.50%, and 0.75%) in a freezing extender. Semen suspensions were loaded in medium straw (0.5 mL) and cryopreserved with the traditional liquid nitrogen vapor method. Post-thawed semen was evaluated for sperm motility, sperm viability, and lipid peroxidation. Also, the fertility test was determined.Results: The results showed that supplementation of the freezing extender with 0.50% to 0.75% sericin resulted in greater total motility and progressive motility and lower malondialdehyde levels than the other groups after cryopreservation (p<0.05). However, the viability of 0.75% decreased compared with the value of 0.50% sericin supplementation (p<0.05). Moreover, the fertility and hatchability of total eggs were significantly higher in the 0.50% sericin group than in the other groups (p<0.05).Conclusion: In conclusion, 0.50% sericin is recommended as an alternative component of the freezing extender to improve cryopreserved rooster semen.

This study evaluated the effect of semen extenders and antioxidant supplementation strategies on the quality and fertility of Thai native rooster semen during cold storage. In Experiment 1, four extenders (EK, IGGKPh, NaCl, and NCAB) were evaluated for their ability to preserve sperm motility, viability, lipid peroxidation (MDA), and oxidative stress (reactive oxygen species; ROS) over 24 h at 5°C. EK and IGGKPh extenders yielded superior motility (68.72% and 70.27%) and viability (91.96% and 91.97%) while minimizing MDA (0.93 and 1.12 µmol) and ROS (14.44% and 14.04%) compared to NaCl (motility: 53.14%, viability: 82.09±3.15, MDA: 1.59 µmol; P < 0.01). Experiment 2 examined the effects of dietary selenium (0.6 ppm), extender-based glutathione (GSH) supplementation (2mM), and their combination across all extenders. Antioxidant treatments, particularly GSH and its combination with selenium, improved motility, viability, and oxidative stability (P < 0.05). At 24 h, GSH-enriched groups using EK and IGGKPh exhibited enhanced motility (67.90% and 71.49%) compared to the controls (57.97% and 55.09%). In the NaCl group, GSH and combined treatments significantly improved motility at 12 h (50.83% and 50.01%, respectively) compared to the control (40.80%; P < 0.01). Selenium also increased sperm concentration (4.24 ± 0.55 × 10⁹ vs. 2.46 ± 0.22 × 10⁹ spz/mL; P < 0.01). Experiment 3 fertility outcomes using EK extender and the same antioxidant treatments from Experiment 2. Fertility rates were significantly higher in the GSH (70.38%) and combined (69.16%) groups than in the control (54.26%; P < 0.01), while hatchability remained unaffected (P > 0.05).In summary, the composition of the extender and antioxidant strategy has a significant influence on semen quality and fertility. The EK and IGGKPh extenders, especially when supplemented with GSH, offered superior protection, and combining dietary selenium with GSH yielded synergistic benefits for semen preservation and reproductive success.