Cryopreservation of ram semen results in low post-thaw survival. Thus, studies were conducted to optimize liquid-storage of chilled semen. In Experiment 1, semen collected from rams (n = 5) was extended using either m...Cryopreservation of ram semen results in low post-thaw survival. Thus, studies were conducted to optimize liquid-storage of chilled semen. In Experiment 1, semen collected from rams (n = 5) was extended using either milk, TRIS (tris[Hydroxymethyl]aminomethane), TEST (N-Tris (hydroxymethyl)-methyl-2-aminoethane sulfonic acid), or CJ-2 (choline-based extender) and stored at either 4℃ and 15℃. All extenders were supplemented with 5% (v/v) egg yolk. In Experiment 2, semen collected from 9 rams was distributed across TRIS or milk extenders supplemented with 5% or 20% (v/v) egg yolk, and 0 or 1% ethylene glycol (EG), during storage for 72 hours at 4℃. The effect of penicillamine, hypotaurine, and epinephrine (PHE) on motility parameters was also evaluated following storage. In Experiment 1, most of the sperm motility parameters were higher after extension, and storage at 4℃ compared with 15℃ (P P in vivo sperm viability and conception rates when using extended, stored semen for artificial insemination of ewes.展开更多
Aim： To evaluate two extenders and two cryoprotectant agents （CPA） for alpaca semen cryopreservation. Methods： Semen samples were obtained from four adult alpacas （Lama pacos） and frozen using extender Ⅰ （TRIS...Aim： To evaluate two extenders and two cryoprotectant agents （CPA） for alpaca semen cryopreservation. Methods： Semen samples were obtained from four adult alpacas （Lama pacos） and frozen using extender Ⅰ （TRIS, citrate, egg yolk and glucose） or extender Ⅱ （skim milk, egg yolk and fructose）, each containing either glycerol （G） or ethylene glycol （EG） as CPA. Consequently, four groups were formed： 1） extender Ⅰ-G; 2） extender Ⅰ-EG; 3） extender Ⅱ-G; and 4） extender Ⅱ-EG. Semen was diluted in a two-step process： for cooling to 5 ℃ （extenders without CPA）, and for freezing （extenders with CPA）. Viability and acrosome integrity were assessed using trypan blue and Giemsa stains. Results： When compared, the motility after thawing was higher （P 〈 0.05） in groups Ⅱ-EG （20.0 %±6.7 %） and Ⅱ-G （15.3 %±4.1%） than that in groups Ⅰ-G （4.0 %±1.1%） and Ⅰ-EG （1.0 %±1.4 %）. Viable spermatozoa with intact acrosomes in groups Ⅱ-EG （18.7 %±2.9 %） and Ⅱ-G （12.7 %±5.9 %） were higher than that in groups Ⅰ-G （5.7 %±1.5 %） and Ⅰ-EG （4.0 %±1.0 %）. Conclusion： The skim milk- and egg yolk-based extenders containing ethylene glycol or glycerol to freeze alpaca semen seems to promote the survival of more sperm cells with intact acrosomes than the other extenders. （Asian J Androl 2005 Sep; 7： 303-309）展开更多
The identification of superior sire is largely dependent on the semen quality evaluations, currently through computer-aided sperm class analyser (CASA). Immunoassays present a viable method for analysis of proteins as...The identification of superior sire is largely dependent on the semen quality evaluations, currently through computer-aided sperm class analyser (CASA). Immunoassays present a viable method for analysis of proteins associated with fertility, such as osteopontin. We have targeted osteopontin at an upstream epitope at position 249 - 267 amino acids and a downstream epitope at position 59 - 74 amino acids of full osteopontin protein in raw, BTS, Kobidil, Tris and Citrate liquid preserved Large white and Kolbroek boar semen. The Genetool analysis software was used to quantify the amount of osteopontin detected. Three boars per breed were used and experiments were repeated five times. Our results revealed that the downstream 15 amino acid epitope was more sensitive to osteopontin antibody. Between total motility and osteopontin, Kolbroek boar semen preserved in Kobidil, Citrate, BTS and Tris extenders revealed positive correlations of 0.564, 0.471, -0.960 and 0.330, respectively. Large white boars semen showed correlations of -0.043, 0.655, 0.778, and 0.202 Kobidil, Citrate, BTS and Tris, respectively. For rapid motility and osteopontin, Kolbroek boar semen revealed positive correlations of 0.842, 0.601, 0.164 and 0.202, respectively while large white boars semen showed correlations of -0.909, -0.144, -0.210, and 0.089, respectively. Only BTS and Kobidil showed high negative correlations between osteopontin and viability in Kolbroek (-0.975) and Large white (-0.746) boar semen, respectively. No significant correlations with osteopontin were observed when Tris extender was used in both breeds. In conclusion, Kolbroek boar semen requires osteopontin in Kobodil extender while large white boar semen requires osteopontin in BTS extender.展开更多
Objective:To evaluate the effect of cooling rates,selenium supplementation,and three semen extenders on cooled and frozen-thawed Saanen buck sperm.Methods:Twenty Saanen bucks were divided into two groups:the selenium ...Objective:To evaluate the effect of cooling rates,selenium supplementation,and three semen extenders on cooled and frozen-thawed Saanen buck sperm.Methods:Twenty Saanen bucks were divided into two groups:the selenium supplemented group and the control group.Ejaculates were collected once weekly by artificial vagina.The first experiment examined the effects of cooling rates and selenium supplementation on semen characteristics of Saanen bucks.Pooled semen was diluted with triladyl extender and split into two aliquots for slow and fast cooling.The second experiment explored the effect of selenium supplementation and semen extenders on post-cryopreserved sperm quality.Ejaculates from each group were divided into three aliquots and diluted with three extenders(i.e.clarified egg-yolk,whole egg yolk,Tris without egg-yolk extenders).All samples were cooled for 2 h at 4℃and frozen at-196℃for 24 h.The sperm characteristics such as sperm motility,acrosome integrity,normal morphology,and viability were evaluated by using a phase-contrast microscope.Results:In the first experiment,all sperm characteristics were significantly increased in selenium supplemented samples when slow cooling was used(P<0.05).In the second experiment,in cooled semen,sperm motility and viability were significantly increased in both clarified egg yolk and Tris without egg yolk extenders in selenium supplemented samples as compared with whole egg yolk extender,respectively(P<0.05).After freezing-thawing,all sperm parameters of selenium supplemented samples in clarified egg yolk extender were significantly greater than those in Tris without egg yolk extender(P<0.05).However,normal morphology and acrosome integrity of selenium supplemented samples in whole egg yolk extender were similar to those of selenium supplemented samples in clarified egg yolk extender.Conclusions:The characteristics of cooled and post-cryopreserved sperm are greater when clarified egg yolk extender is used in semen from selenium supplemented bucks.展开更多
This article investigates general extenders in American English,using a more than one-billion-word corpus(COCA)classified by different genres.In this study,the differences and frequencies of general extenders in diffe...This article investigates general extenders in American English,using a more than one-billion-word corpus(COCA)classified by different genres.In this study,the differences and frequencies of general extenders in different genres are displayed.The study also illustrates the tendency of the use of general extenders in American English from 1990 to 2020.Moreover,the salient features of general extenders used in different genres are observed.The study can provide some practical implications for the use of general extenders for second language learners.展开更多
文摘Cryopreservation of ram semen results in low post-thaw survival. Thus, studies were conducted to optimize liquid-storage of chilled semen. In Experiment 1, semen collected from rams (n = 5) was extended using either milk, TRIS (tris[Hydroxymethyl]aminomethane), TEST (N-Tris (hydroxymethyl)-methyl-2-aminoethane sulfonic acid), or CJ-2 (choline-based extender) and stored at either 4℃ and 15℃. All extenders were supplemented with 5% (v/v) egg yolk. In Experiment 2, semen collected from 9 rams was distributed across TRIS or milk extenders supplemented with 5% or 20% (v/v) egg yolk, and 0 or 1% ethylene glycol (EG), during storage for 72 hours at 4℃. The effect of penicillamine, hypotaurine, and epinephrine (PHE) on motility parameters was also evaluated following storage. In Experiment 1, most of the sperm motility parameters were higher after extension, and storage at 4℃ compared with 15℃ (P P in vivo sperm viability and conception rates when using extended, stored semen for artificial insemination of ewes.
文摘Aim： To evaluate two extenders and two cryoprotectant agents （CPA） for alpaca semen cryopreservation. Methods： Semen samples were obtained from four adult alpacas （Lama pacos） and frozen using extender Ⅰ （TRIS, citrate, egg yolk and glucose） or extender Ⅱ （skim milk, egg yolk and fructose）, each containing either glycerol （G） or ethylene glycol （EG） as CPA. Consequently, four groups were formed： 1） extender Ⅰ-G; 2） extender Ⅰ-EG; 3） extender Ⅱ-G; and 4） extender Ⅱ-EG. Semen was diluted in a two-step process： for cooling to 5 ℃ （extenders without CPA）, and for freezing （extenders with CPA）. Viability and acrosome integrity were assessed using trypan blue and Giemsa stains. Results： When compared, the motility after thawing was higher （P 〈 0.05） in groups Ⅱ-EG （20.0 %±6.7 %） and Ⅱ-G （15.3 %±4.1%） than that in groups Ⅰ-G （4.0 %±1.1%） and Ⅰ-EG （1.0 %±1.4 %）. Viable spermatozoa with intact acrosomes in groups Ⅱ-EG （18.7 %±2.9 %） and Ⅱ-G （12.7 %±5.9 %） were higher than that in groups Ⅰ-G （5.7 %±1.5 %） and Ⅰ-EG （4.0 %±1.0 %）. Conclusion： The skim milk- and egg yolk-based extenders containing ethylene glycol or glycerol to freeze alpaca semen seems to promote the survival of more sperm cells with intact acrosomes than the other extenders. （Asian J Androl 2005 Sep; 7： 303-309）
文摘The identification of superior sire is largely dependent on the semen quality evaluations, currently through computer-aided sperm class analyser (CASA). Immunoassays present a viable method for analysis of proteins associated with fertility, such as osteopontin. We have targeted osteopontin at an upstream epitope at position 249 - 267 amino acids and a downstream epitope at position 59 - 74 amino acids of full osteopontin protein in raw, BTS, Kobidil, Tris and Citrate liquid preserved Large white and Kolbroek boar semen. The Genetool analysis software was used to quantify the amount of osteopontin detected. Three boars per breed were used and experiments were repeated five times. Our results revealed that the downstream 15 amino acid epitope was more sensitive to osteopontin antibody. Between total motility and osteopontin, Kolbroek boar semen preserved in Kobidil, Citrate, BTS and Tris extenders revealed positive correlations of 0.564, 0.471, -0.960 and 0.330, respectively. Large white boars semen showed correlations of -0.043, 0.655, 0.778, and 0.202 Kobidil, Citrate, BTS and Tris, respectively. For rapid motility and osteopontin, Kolbroek boar semen revealed positive correlations of 0.842, 0.601, 0.164 and 0.202, respectively while large white boars semen showed correlations of -0.909, -0.144, -0.210, and 0.089, respectively. Only BTS and Kobidil showed high negative correlations between osteopontin and viability in Kolbroek (-0.975) and Large white (-0.746) boar semen, respectively. No significant correlations with osteopontin were observed when Tris extender was used in both breeds. In conclusion, Kolbroek boar semen requires osteopontin in Kobodil extender while large white boar semen requires osteopontin in BTS extender.
文摘Objective:To evaluate the effect of cooling rates,selenium supplementation,and three semen extenders on cooled and frozen-thawed Saanen buck sperm.Methods:Twenty Saanen bucks were divided into two groups:the selenium supplemented group and the control group.Ejaculates were collected once weekly by artificial vagina.The first experiment examined the effects of cooling rates and selenium supplementation on semen characteristics of Saanen bucks.Pooled semen was diluted with triladyl extender and split into two aliquots for slow and fast cooling.The second experiment explored the effect of selenium supplementation and semen extenders on post-cryopreserved sperm quality.Ejaculates from each group were divided into three aliquots and diluted with three extenders(i.e.clarified egg-yolk,whole egg yolk,Tris without egg-yolk extenders).All samples were cooled for 2 h at 4℃and frozen at-196℃for 24 h.The sperm characteristics such as sperm motility,acrosome integrity,normal morphology,and viability were evaluated by using a phase-contrast microscope.Results:In the first experiment,all sperm characteristics were significantly increased in selenium supplemented samples when slow cooling was used(P<0.05).In the second experiment,in cooled semen,sperm motility and viability were significantly increased in both clarified egg yolk and Tris without egg yolk extenders in selenium supplemented samples as compared with whole egg yolk extender,respectively(P<0.05).After freezing-thawing,all sperm parameters of selenium supplemented samples in clarified egg yolk extender were significantly greater than those in Tris without egg yolk extender(P<0.05).However,normal morphology and acrosome integrity of selenium supplemented samples in whole egg yolk extender were similar to those of selenium supplemented samples in clarified egg yolk extender.Conclusions:The characteristics of cooled and post-cryopreserved sperm are greater when clarified egg yolk extender is used in semen from selenium supplemented bucks.
文摘This article investigates general extenders in American English,using a more than one-billion-word corpus(COCA)classified by different genres.In this study,the differences and frequencies of general extenders in different genres are displayed.The study also illustrates the tendency of the use of general extenders in American English from 1990 to 2020.Moreover,the salient features of general extenders used in different genres are observed.The study can provide some practical implications for the use of general extenders for second language learners.
