Objective:To evaluate the cryoprotective effect of butylated hydroxytoluene on buck frozen semen.Methods:Semen was collected from Boer(n=6)and Zaraibi(n=6)bucks by electroejaculator for 5 weeks.Semen aliquots were dil...Objective:To evaluate the cryoprotective effect of butylated hydroxytoluene on buck frozen semen.Methods:Semen was collected from Boer(n=6)and Zaraibi(n=6)bucks by electroejaculator for 5 weeks.Semen aliquots were diluted at 38℃in Tris-buffer with egg yolk 15.0%(vol/vol)(Tris-egg yolk extender)or soya lecithin 2.5%(weight/vol)(Tris-soya lecithin extender)supplemented with butylated hydroxytoluene at 0.0(as the control),0.5,1.0,2.0 and 4.0 mM.Post-thawing motility(at 400×magnification),plasma(hypo-osmotic swelling test),acrosome(Trypan blue/Giemsa dual staining)membranes,DNA(comet assay),and lipid peroxidation(by malondialdehyde concentration)were assessed.Results:Spermatozoa motility was enhanced by butylated hydroxytoluene in Tris-soya lecithin extender at 0.5 mM in the two breeds,and in Tris-egg yolk extender at 1.0 mM in Boer and at 2.0 mM in Zaraibi bucks for up to 3 h post-thawing.Plasma and acrosome membranes and DNA integrity of the two breeds were maximally high with butylated hydroxytoluene at 1.0-2.0 mM in Tris-egg yolk extender and at 0.5-1.0 mM in Tris-soya lecithin extender.Lipid peroxidation was minimal with butylated hydroxytoluene at 1.0-2.0 mM in Tris-egg yolk and soya lecithin extenders in the two breeds.Butylated hydroxytoluene at 4.0 mM deteriorated spermatozoa motility,and plasma and acrosome membranes.Conclusions:The consequence of butylated hydroxytoluene on buck frozen-thawed spermatozoa varies with the levels of supplementation,buck breed,and phospholipid source in the extender.Semen parameters of Boer buck are better in their response to butylated hydroxytoluene than Zaraibi buck.Butylated hydroxytoluene at 1.0 and 2.0 mM in Tris-egg yolk extender,and at 0.5 mM in Tris-soya lecithin extender represents the best concentrations and profitably improves the semen quality of buck semen.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
建立了同时测定特丁基对苯二酚(TBHQ)、丁基羟基茴香醚(BHA)和二丁基羟基对甲酚(BHT)的高效液相色谱法。考察了流动相组成、柱温等因素对分析效率的影响。样品经甲醇提取后,以E-clipse plus C18(250mm×4.6mm,5μm)色谱柱为分析柱,0...建立了同时测定特丁基对苯二酚(TBHQ)、丁基羟基茴香醚(BHA)和二丁基羟基对甲酚(BHT)的高效液相色谱法。考察了流动相组成、柱温等因素对分析效率的影响。样品经甲醇提取后,以E-clipse plus C18(250mm×4.6mm,5μm)色谱柱为分析柱,0.02mmol/L乙酸铵水溶液-甲醇为流动相,线性梯度淋洗,紫外二极管阵列检测器检测,波长280nm。结果表明:TBHQ、BHA与BHT在0.5～20mg/L范围内有良好的线性关系,方法的检出限(S/N=3)分别为0.03,0.03和0.05mg/kg,加标回收率为83.45%～108.42%,相对标准偏差不大于12.79%。展开更多
文摘Objective:To evaluate the cryoprotective effect of butylated hydroxytoluene on buck frozen semen.Methods:Semen was collected from Boer(n=6)and Zaraibi(n=6)bucks by electroejaculator for 5 weeks.Semen aliquots were diluted at 38℃in Tris-buffer with egg yolk 15.0%(vol/vol)(Tris-egg yolk extender)or soya lecithin 2.5%(weight/vol)(Tris-soya lecithin extender)supplemented with butylated hydroxytoluene at 0.0(as the control),0.5,1.0,2.0 and 4.0 mM.Post-thawing motility(at 400×magnification),plasma(hypo-osmotic swelling test),acrosome(Trypan blue/Giemsa dual staining)membranes,DNA(comet assay),and lipid peroxidation(by malondialdehyde concentration)were assessed.Results:Spermatozoa motility was enhanced by butylated hydroxytoluene in Tris-soya lecithin extender at 0.5 mM in the two breeds,and in Tris-egg yolk extender at 1.0 mM in Boer and at 2.0 mM in Zaraibi bucks for up to 3 h post-thawing.Plasma and acrosome membranes and DNA integrity of the two breeds were maximally high with butylated hydroxytoluene at 1.0-2.0 mM in Tris-egg yolk extender and at 0.5-1.0 mM in Tris-soya lecithin extender.Lipid peroxidation was minimal with butylated hydroxytoluene at 1.0-2.0 mM in Tris-egg yolk and soya lecithin extenders in the two breeds.Butylated hydroxytoluene at 4.0 mM deteriorated spermatozoa motility,and plasma and acrosome membranes.Conclusions:The consequence of butylated hydroxytoluene on buck frozen-thawed spermatozoa varies with the levels of supplementation,buck breed,and phospholipid source in the extender.Semen parameters of Boer buck are better in their response to butylated hydroxytoluene than Zaraibi buck.Butylated hydroxytoluene at 1.0 and 2.0 mM in Tris-egg yolk extender,and at 0.5 mM in Tris-soya lecithin extender represents the best concentrations and profitably improves the semen quality of buck semen.
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.
文摘建立了同时测定特丁基对苯二酚(TBHQ)、丁基羟基茴香醚(BHA)和二丁基羟基对甲酚(BHT)的高效液相色谱法。考察了流动相组成、柱温等因素对分析效率的影响。样品经甲醇提取后,以E-clipse plus C18(250mm×4.6mm,5μm)色谱柱为分析柱,0.02mmol/L乙酸铵水溶液-甲醇为流动相,线性梯度淋洗,紫外二极管阵列检测器检测,波长280nm。结果表明:TBHQ、BHA与BHT在0.5～20mg/L范围内有良好的线性关系,方法的检出限(S/N=3)分别为0.03,0.03和0.05mg/kg,加标回收率为83.45%～108.42%,相对标准偏差不大于12.79%。
