• Journal of Animal Science and Technology
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• v.63 no.1
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• pp.46-57
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• 2021

Fruits with antioxidant enrichment can be an economically affordable supplement for mitigating oxidative damage prone spermatozoa membrane pathologies. Computer-assisted sperm analyzer and oxidative status were utilized to evaluate the impact of watermelon (Citrullus lanatus) fortification of dextrose saline as diluent for rooster semen and fertility response of hens inseminated. Watermelon juice and dextrose saline were used to formulate diluent of 7 treatments consisting of unextended semen (positive control), 10%, 20%, 30%, 40%, 50% and only dextrose saline (negative control) designated as Treatments 1-7. Pooled semen was obtained from fertile roosters and equilibrated with diluents at ratio 1:2 in the various treatments and were evaluated using computer software coupled microscope and seminal oxidative status assay. 168 laying hens randomly divided into 7 treatment of 8 replicates and 3 hen per replicate. Hen were everted, and semen (2 × 108 Spermatozoa) deposited intra-vagina and eggs collected over 8 weeks to assess fertility and hatchability of eggs laid. The result obtained revealed that watermelon-dextrose saline rooster semen diluent enhanced progressive motility, sperm kinetics and lowered non-progressive motility in T2-T6 compared to T7 over the 3 hours of evaluation. Watermelon addition to rooster semen diluent enhance the antioxidant capacity of rooster semen and lowered lipid peroxide generation. The percentage fertility was highest in T3 (81.01%) and T4 (81.24%) with lowest value obtained in T7 (73.46%). The hatchability of eggs set of hens inseminated with undiluted semen (71.46%) was lower than values for hens inseminated with watermelon inclusive extended semen (75.71%-80.39%). The optimal inclusion of 30%-40% watermelon in dextrose saline diluent enhance rooster semen kinetics, seminal oxidative stability and egg fertility.

• Animal Bioscience
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• v.36 no.11
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• pp.1647-1654
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• 2023

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.

• Journal of Animal Reproduction and Biotechnology
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• v.37 no.1
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• pp.55-61
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• 2022

The acrosome cap allows sperm to penetrate the egg membrane and produce male pronuclei within female chicken eggs, facilitating successful fertilization. Given this, it is important to establish practical methods for evaluating the integrity of the acrosome cap and thus the quality of the rooster's sperm. There are several established methods for evaluating the acrosomes of mammalian sperm, but none of these methods are suitable for evaluating the acrosome status of rooster spermatozoa. Therefore, a simplified method for evaluating the rooster acrosome is needed. Here we evaluated the usefulness of CBB (coomassie brilliant blue) staining of the acrosome at concentrations of 0.04%, 0.08%, and 0.3% CBB solutions. Our data revealed a clear staining pattern for intact acrosome caps at 0.04% and 0.08% CBB but not at 0.3% CBB. This protocol revealed differences in acrosome integrity between fresh and frozen rooster sperm smears suggesting that CBB staining may facilitate easier semen evaluation in roosters. This protocol allows for the accurate differential staining of acrosome cap in rooster spermatozoa.

• Korean Journal of Poultry Science
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• v.30 no.2
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• pp.129-134
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• 2003

This study was conducted to investigate the effects of liquid rooster semen on reproductive ability in chicken. Raw and diluted semens were stored at 5$^{\circ}C$ cold temperature for 6, 30, and 54 hours after semen collection. There was no statistically difference in sperm motility throughout the 6 hours period of storage among raw semen and diluted semen groups with skim milk glucose solution (SM), egg yolk glucose solution (EY), and saline. But there was decrease in those throughout the period of 30 and 54 hours of storage. Sperm motility and normal sperm for the period of 30 and 54 hours of storage were significantly better in SM and EY diluted groups (P<0.05). Fertilization rates of rooster semen diluted with SM were 90.77, 87.70, and 59.46% for 6, 30, and 54 hours stored groups, respectively, those proved to be higher in SM-diluted group than other groups.

• Animal Bioscience
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• v.36 no.10
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• pp.1530-1535
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• 2023

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.

• Korean Journal of Poultry Science
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• v.45 no.4
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• pp.237-244
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• 2018

In this study, to increase the survival rate of frozen/thaw rooster semen, standard protocols of semen thawing procedures were tested by computer-assisted sperm assay (CASA). We tested 4 different thawing protocols for frozen semen, $5^{\circ}C$ for 2 min, $35^{\circ}C$ for 30 s, $54^{\circ}C$ for 13 s, and $70^{\circ}C$ for 7 s. The pooled semen from 5 to 8 Ogye rooster line was diluted in the HS-1 diluent and frozen in 8% methylacetamide (MA) in liquid nitrogen vapors. To determine standard thawing method, straws were plunged into different temperatures and times. The resulting motilities were recorded by the CASA system. The results of this study showed that the best viability of the spermatozoa was shown by exposure at $5^{\circ}C$ for 2 min. Moreover, the longevity test of thawed sperm at $5^{\circ}C$ for 2 min also supported the higher viability under low temperature preservation of $17^{\circ}C$ for 1 hr. Further research is needed to increase the motility of thawed rooster semen for field application. In addition, the in vivo tests for different rooster lines are also needed for the establishment of avian genetic resource bank.

• Korean Journal of Poultry Science
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• v.48 no.4
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• pp.185-191
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• 2021

Chicken spermatozoa have the ability to survive in low-temperature environments; however, the effects of low temperature on sperm motility and acrosome damage have not been studied in detail. The present study investigated semen longevity following dilution of rooster semen with Beltsville Poultry Semen Extender (BPSE) and Lake extender in preservation vessels (1.5 mL e-tube and 0.5 mL straw). Spermatozoa motility in the closed-type vessel (0.5 mL straw) was higher than that in the 1.5 mL e-tube on day 3 of preservation (68.6±3.1% vs. 22.1±5.7%). The motility of rooster semen diluted with BPSE in 0.5 mL straw was also higher than that of the Lake extender on day 3 of preservation (57.7±5.6% vs. 37.7±5.4%). Furthermore, acrosome intactness was higher in 0.5 mL straw than in the 1.5 mL e-tube, and the rate of acrosome cap damage increased with preservation days. The present study demonstrates that a closed 0.5-mL straw vessel could be used for low-temperature semen preservation, with an increased motility rate and acrosome integrity in fresh rooster semen.

• Korean Journal of Poultry Science
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• v.44 no.1
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• pp.59-65
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• 2017

This study examined factors affecting the analysis of motility of chicken semen. The viability of spermatozoa was estimated using varying dilution ratios and supplementation with BSA or fatty acid free (FAF)-BSA as protein sources in semen diluent. Fresh semen was examined after preparing dilutions in beltsvile poultry semen extender (BPSE) of 1/8, 1/16 and 1/32 at $25^{\circ}C$. The motility of incubated semen at each dilution was observed at 3 min (89.9%, 69.9% and 53.2%), 30 min (86.7%, 71.4% and 51.7%), 1 h (89.5%, 74.0% and 53.5%) and 3 h (78.5%, 66.5% and 45.7%), respectively. The addition of BSA or FAF-BSA to BPSE diluent significantly increased the viability of semen in 1/32 dilution with results of 53.2% (control), 84.8% (BSA) and 92.9% (FAF-BSA) (p<0.05). This phenomenon was also observed in the dilution of frozen semen, where FAF-BSA treatment increased the viability of thawed semen from 17.6% to 34.0% in a 1/8 dilution (p<0.05). When the protein sources were used in the dilution, the survival rates of diluted chicken semen were also increased with time lapse. These results show that FAF-BSA may act to protect chicken semen and is suitable as a basic component of chicken semen diluent for the method of analyzing rooster semen after freezing.

• Korean Journal of Poultry Science
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• v.41 no.1
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• pp.21-27
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• 2014

To preserve chicken genetic materials like cryopreserved spermatozoa, various kinds of freezing agents like glycerol, dimethylsuloxide, dimethylformamide or dimethylacetamide have been used for rooster semen preparation. Recently, the usage of N-methylacetamide (MA) for Ogye rooster semen preservation resulted in hatched chicken successfully. In this study, we investigated the effects of 7, 9 and 11% of MA on the viability, fertility and hatchability of frozen-thawed rooster semen using artificial insemination. The results of viability, fertility and hatchability in frozen semen with 7%, 9% or 11% MA were $35.16{\pm}6.12%$, $67.83{\pm}15.3%$ and $66.2{\pm}16.3%$ of motile sperm rate, 21.5%, 34.7% and 25% of fertility rate, and 100%, 89.5% and 87.5% of hatchability rate. The results of control group with frozen semen were 96.0% of fertility rate and 92.2% of hatchability rate. With these results, the concentration range of MA as a freezing agent of rooster semen could be 7~9% of media. The higher concentration of 9 % MA could decrease the fertility rate of thawed semen not the rate of hatchability rate. So the use of MA without affecting fertility rate would be a key point of freezing method of rooster semen for poultry genetic resource preservation.

• Animal Bioscience
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• v.37 no.7
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• pp.1177-1184
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• 2024

Objective: Semen cryopreservation is an effective method of preserving genetic material, particularly in native chicken breeds facing a substantial decline. In this study, we evaluated the quality of frozen/thawed rooster semen treated with different concentrations of oral administrations of black ginger (Kaempferia parviflora: KP) extract and determined its fertility. Methods: Thirty-two Thai native roosters (Pradu Hang Dum, 42 weeks old) were used in this study. The treatments were classified into four groups according to the concentration of KP extract administered to the roosters: 0, 100, 150, and 200 mg/kg body weight. The quality of fresh semen was analyzed before cryopreservation. Post-thaw sperm quality and fertility potential were determined. Also, lipid peroxidation was determined. Results: The results showed that sperm concentration and movement increased in roosters treated with 200 mg/kg of KP extract (p<0.05). The malondialdehyde (MDA) in the roosters receiving 200 mg/kg KP extract was lower than that in the other but had an insignificant difference within the KP treatment groups (p>0.05). The highest MDA levels were observed in the control group (p<0.05). The percentage of motile sperm (total motility and progressive motility) after semen thawing was higher in roosters that received 150 and 200 mg/kg KP extract than in those that received 100 mg/kg KP extract and the control (p<0.05). MDA levels decreased significantly in roosters that received 150 and 200 mg/kg KP extract than in those that received 100 mg/kg KP extract and the control (p<0.05). Fertility and hatchability were greater in the KP150 and KP200 groups than in the KP100 and control groups (p<0.05). Conclusion: The optimal amount of KP extract influencing initial sperm quality was determined to be 200 mg/kg. However, 150 mg/kg was the optimal low dosage of KP extract administration that maintained sperm quality and fertility following semen cryopreservation.