The aim was to discuss the optimal storage environment and proper insemination time after thawing of 0.25 mL straw frozen semen. Straw frozen semen was thawed at 40 ℃ for 20 s, and then stored at 0 -4 ℃, 14 - 16 ℃,...The aim was to discuss the optimal storage environment and proper insemination time after thawing of 0.25 mL straw frozen semen. Straw frozen semen was thawed at 40 ℃ for 20 s, and then stored at 0 -4 ℃, 14 - 16 ℃, 25 -27 ℃ for 2, 4, 6, 8 and 10 h, respectively. The sperm motility was detected. After thawing, semen was stored at 0 - 4 ℃ and 14 - 16 ℃ for 10 h. Their sperm motilities （0.434 ±0. 016 7 and 0.423 ±0.019 6） had no significant differences （P 〉 0.05） with initial thawing motility （0.441 ± 0.030）. Sperm motility reduced as the storage time prolonged at 25 -27 ℃. Sperm motility after 6 h had signifi- cant differences with that of initial thawing motility （P 〈 O. 05 ）, and sperm motilities after 8 and 10 h showed extremely significant differences （P 〈 0.01 ）. Thus, sperm motility after thawing was still very high after stored at 0 -4 ℃ and 14 - 16 ℃ within 10 h, which met the requirements for insemination. Under this temperature and time ranges, sperm could be carried over long distances, which had small effects on sperm quality and reached the expected insemination effects. However, under the temperature of 25 - 27 ℃, semen should be used for insemination within 6 h after thawing.展开更多
The aim of this study was to assess sperm quality (motility, viability and acrosomal integrity) sperm from commercially frozen semen straws two breeds of bulls Bos taurus (holstein Frisian) and Bos indicus (Brahman). ...The aim of this study was to assess sperm quality (motility, viability and acrosomal integrity) sperm from commercially frozen semen straws two breeds of bulls Bos taurus (holstein Frisian) and Bos indicus (Brahman). 9 commercial straws 0.5 ml of Holstein bull semen and 9 Brahman bull were thawed, they were kept for two hours at room temperature and motility, viability and acrosomal integrity (NAR) was assessed. The results were 30% motility, viability 40% and 30% of NAR in the Holstein breed. Brahma race for motility 40%, 50% and 40% viability was obtained NAR. In conclusion, according to the results of the variables analyzed, the Brahman breed in sperm quality was better than the Holstein breed;however, the results of both races meet minimum standards of quality sperm for use in artificial insemination (AI) field level.展开更多
[Objectives]This study was conducted to develop a molecular marker immunomagnetic bead sorting technology method that can specifically identify dead spermatozoa.[Methods]This study first confirmed the specific binding...[Objectives]This study was conducted to develop a molecular marker immunomagnetic bead sorting technology method that can specifically identify dead spermatozoa.[Methods]This study first confirmed the specific binding of Annexin V to dead bovine spermatozoa,and tried to remove dead spermatozoa in semen combining with the immunomagnetic bead technology,so as to improve the separation efficiency of target spermatozoa in the process of sex-controlled semen preparation on a flow cytometer.[Results]The spermatozoon motility,membrane integrity and mitochondrial activity after sorting and the rate of dead spermatozoa during the on-machine X/Y separation were all improved to different degrees(P<0.05),indicating that the technical process design could effectively remove some dead spermatozoa,and there was no significant effect on frozen sexed semen prepared from the separated X or Y spermatozoa(P>0.05),indicating that the technical process did not cause additional damage to the spermatozoa.[Conclusions]Combining the specificity of Annexin V with the immunomagnetic bead method could effectively remove dead spermatozoa from bovine spermatozoa,and significantly reduce the rate of dead spermatozoa in bovine permatozoa during sex-controlled separation(P<0.05).The method developed can effectively improve the production efficiency of frozen sexed semen of dairy cows,reduce the production cost,and promote the industrial application of the product.展开更多
The objective of the present study was to determine the effects of docosahexaenoic acid （DHA）-enriched hen egg yolks and L-cysteine supplementation on the qualities of the cryopreserved boar semen. A total of 15 eja...The objective of the present study was to determine the effects of docosahexaenoic acid （DHA）-enriched hen egg yolks and L-cysteine supplementation on the qualities of the cryopreserved boar semen. A total of 15 ejaculates from 5 Pietrain boars were divided into 4 groups according to the compositions of the freezing extenders used, that is, normal hen egg yolk （group Ⅰ）, DHA-enriched hen egg yolk （group Ⅱ）, normal hen egg yolk with 5 mmol L^-1 of cysteine supplementation （group Ⅲ） and DHA-enriched hen egg yolk with 5 mmol L1 of cysteine supplementation （group Ⅳ）. The semen was cryopreserved using controlled rate freezer and was thawed at 50℃ for 12 s. Progressive motility, sperm viability, acrosome integrity and functional integrity of sperm plasma membrane of the post-thawed semen were evaluated. The supplementation of L-cysteine in the freezing extender alone （group Ⅲ） improved progressive motility （P 〈 0.05）, and the supplementation of L-cysteine in combination with DHA-enriched hen egg yolk （group Ⅳ） improved both progressive motility （P 〈 0.05） and acrosome integrity （P 〈 0.01）. The use of DHA-enriched hen egg yolk alone （group Ⅱ） did not enhance any of the post-thawed semen qualities （P 〉 0.05）. In conclusion, the supplementation of antioxidant L-cysteine alone or in combination with DHA-enriched hen egg yolk significantly improved the post-thawed semen qualities, especially progressive motility and acrosome integrity.展开更多
The aim of the present study was to evaluate effect of 3 levels of butylated hydroxytoluene （BHT） supplemented in extender for fighting bull semen. Ejaculates were obtained from 10 healthy fighting bulls by artifici...The aim of the present study was to evaluate effect of 3 levels of butylated hydroxytoluene （BHT） supplemented in extender for fighting bull semen. Ejaculates were obtained from 10 healthy fighting bulls by artificial vagina. Semen with motility of more than 70% was included in the experiment. Tris-fructose-glycerol-egg yolk extender containing 0, 1, 3 and 7 mM BHT was used to dilute semen to final concentration of 200 × 10^6 cells/ml. Diluted semen was frozen in 0.25 mL straws and frozen semen was thawed in a 37 ℃ water bath for 15 sec before being evaluated at 0 and 1 hour. Sperm motility at 0 and 1 hour after thawing did not differ among groups tested （ranged 47.86-53.57 and 21.79-24.29 respectively）. At 0 hour after thawing, percentage of live sperm of 3 mM BHT （37.39% ± 2.91%） was lower than those of 0 and 1 mM BHT （43.71%± 2.76% and 41.46% ±2.59% respectively, P 〈 0.05） but not different from 7 mM BHT （40.89%± 2.50%）. The effect was more significant at 1 hour after thawing and 3 mM BHT （32.07% ± 2.45%） suppressed live cells more than other concentrations （ranged 35.07%-37.46%, P 〈 0.05）. At 0 hour after thawing, percentage of membrane integrity （hypo-osmotic swelling test） was not affected by BHT concentration （ranged 23.89%-28.54%）. However, at 1 hour after thawing, percentage of membrane integrity of 3 mM BHT （19.75% ± 1.41%） was lower that of 7 mM BHT （23.29% ± 1.88%, P 〈 0.05） but not different from those of 0 and 1 mM BHT （20.32% ±1.81% and 22.07% ± 2.27% respectively）. No significant effect was found on percentage of abnormal sperm. There was no effect of BHT supplementation in extender on most of the Hamilton-thorn motility analyser parameters. It may be concluded that 3 mM BHT can be harmful to fighting bull spermatozoa and lower concentrmion can be used without detrimental effect. Further study may be needed to verify use of BHT in lower range of concentrations.展开更多
基金Supported by the Technology Research and Demonstrational Popularization Project of Beijing Vocational College of Agriculture(XY-YF-15-07)(XY-YF-14-21)
文摘The aim was to discuss the optimal storage environment and proper insemination time after thawing of 0.25 mL straw frozen semen. Straw frozen semen was thawed at 40 ℃ for 20 s, and then stored at 0 -4 ℃, 14 - 16 ℃, 25 -27 ℃ for 2, 4, 6, 8 and 10 h, respectively. The sperm motility was detected. After thawing, semen was stored at 0 - 4 ℃ and 14 - 16 ℃ for 10 h. Their sperm motilities （0.434 ±0. 016 7 and 0.423 ±0.019 6） had no significant differences （P 〉 0.05） with initial thawing motility （0.441 ± 0.030）. Sperm motility reduced as the storage time prolonged at 25 -27 ℃. Sperm motility after 6 h had signifi- cant differences with that of initial thawing motility （P 〈 O. 05 ）, and sperm motilities after 8 and 10 h showed extremely significant differences （P 〈 0.01 ）. Thus, sperm motility after thawing was still very high after stored at 0 -4 ℃ and 14 - 16 ℃ within 10 h, which met the requirements for insemination. Under this temperature and time ranges, sperm could be carried over long distances, which had small effects on sperm quality and reached the expected insemination effects. However, under the temperature of 25 - 27 ℃, semen should be used for insemination within 6 h after thawing.
文摘The aim of this study was to assess sperm quality (motility, viability and acrosomal integrity) sperm from commercially frozen semen straws two breeds of bulls Bos taurus (holstein Frisian) and Bos indicus (Brahman). 9 commercial straws 0.5 ml of Holstein bull semen and 9 Brahman bull were thawed, they were kept for two hours at room temperature and motility, viability and acrosomal integrity (NAR) was assessed. The results were 30% motility, viability 40% and 30% of NAR in the Holstein breed. Brahma race for motility 40%, 50% and 40% viability was obtained NAR. In conclusion, according to the results of the variables analyzed, the Brahman breed in sperm quality was better than the Holstein breed;however, the results of both races meet minimum standards of quality sperm for use in artificial insemination (AI) field level.
基金Supported by Targeted Poverty Alleviation Special Project of Hetao College(HYZX201955)Introduced Talent Scientific Research Start-up Fund of Hetao College(HYRC2019002)。
文摘[Objectives]This study was conducted to develop a molecular marker immunomagnetic bead sorting technology method that can specifically identify dead spermatozoa.[Methods]This study first confirmed the specific binding of Annexin V to dead bovine spermatozoa,and tried to remove dead spermatozoa in semen combining with the immunomagnetic bead technology,so as to improve the separation efficiency of target spermatozoa in the process of sex-controlled semen preparation on a flow cytometer.[Results]The spermatozoon motility,membrane integrity and mitochondrial activity after sorting and the rate of dead spermatozoa during the on-machine X/Y separation were all improved to different degrees(P<0.05),indicating that the technical process design could effectively remove some dead spermatozoa,and there was no significant effect on frozen sexed semen prepared from the separated X or Y spermatozoa(P>0.05),indicating that the technical process did not cause additional damage to the spermatozoa.[Conclusions]Combining the specificity of Annexin V with the immunomagnetic bead method could effectively remove dead spermatozoa from bovine spermatozoa,and significantly reduce the rate of dead spermatozoa in bovine permatozoa during sex-controlled separation(P<0.05).The method developed can effectively improve the production efficiency of frozen sexed semen of dairy cows,reduce the production cost,and promote the industrial application of the product.
文摘The objective of the present study was to determine the effects of docosahexaenoic acid （DHA）-enriched hen egg yolks and L-cysteine supplementation on the qualities of the cryopreserved boar semen. A total of 15 ejaculates from 5 Pietrain boars were divided into 4 groups according to the compositions of the freezing extenders used, that is, normal hen egg yolk （group Ⅰ）, DHA-enriched hen egg yolk （group Ⅱ）, normal hen egg yolk with 5 mmol L^-1 of cysteine supplementation （group Ⅲ） and DHA-enriched hen egg yolk with 5 mmol L1 of cysteine supplementation （group Ⅳ）. The semen was cryopreserved using controlled rate freezer and was thawed at 50℃ for 12 s. Progressive motility, sperm viability, acrosome integrity and functional integrity of sperm plasma membrane of the post-thawed semen were evaluated. The supplementation of L-cysteine in the freezing extender alone （group Ⅲ） improved progressive motility （P 〈 0.05）, and the supplementation of L-cysteine in combination with DHA-enriched hen egg yolk （group Ⅳ） improved both progressive motility （P 〈 0.05） and acrosome integrity （P 〈 0.01）. The use of DHA-enriched hen egg yolk alone （group Ⅱ） did not enhance any of the post-thawed semen qualities （P 〉 0.05）. In conclusion, the supplementation of antioxidant L-cysteine alone or in combination with DHA-enriched hen egg yolk significantly improved the post-thawed semen qualities, especially progressive motility and acrosome integrity.
文摘The aim of the present study was to evaluate effect of 3 levels of butylated hydroxytoluene （BHT） supplemented in extender for fighting bull semen. Ejaculates were obtained from 10 healthy fighting bulls by artificial vagina. Semen with motility of more than 70% was included in the experiment. Tris-fructose-glycerol-egg yolk extender containing 0, 1, 3 and 7 mM BHT was used to dilute semen to final concentration of 200 × 10^6 cells/ml. Diluted semen was frozen in 0.25 mL straws and frozen semen was thawed in a 37 ℃ water bath for 15 sec before being evaluated at 0 and 1 hour. Sperm motility at 0 and 1 hour after thawing did not differ among groups tested （ranged 47.86-53.57 and 21.79-24.29 respectively）. At 0 hour after thawing, percentage of live sperm of 3 mM BHT （37.39% ± 2.91%） was lower than those of 0 and 1 mM BHT （43.71%± 2.76% and 41.46% ±2.59% respectively, P 〈 0.05） but not different from 7 mM BHT （40.89%± 2.50%）. The effect was more significant at 1 hour after thawing and 3 mM BHT （32.07% ± 2.45%） suppressed live cells more than other concentrations （ranged 35.07%-37.46%, P 〈 0.05）. At 0 hour after thawing, percentage of membrane integrity （hypo-osmotic swelling test） was not affected by BHT concentration （ranged 23.89%-28.54%）. However, at 1 hour after thawing, percentage of membrane integrity of 3 mM BHT （19.75% ± 1.41%） was lower that of 7 mM BHT （23.29% ± 1.88%, P 〈 0.05） but not different from those of 0 and 1 mM BHT （20.32% ±1.81% and 22.07% ± 2.27% respectively）. No significant effect was found on percentage of abnormal sperm. There was no effect of BHT supplementation in extender on most of the Hamilton-thorn motility analyser parameters. It may be concluded that 3 mM BHT can be harmful to fighting bull spermatozoa and lower concentrmion can be used without detrimental effect. Further study may be needed to verify use of BHT in lower range of concentrations.
