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
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
[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
Aim: This study was undertaken to compare the reduction in color of two dyes methylene blue (MBRT) and resazurindye (RRT) with other tests of bull semen quality and to examine their relationship to fertility. Methods:...Aim: This study was undertaken to compare the reduction in color of two dyes methylene blue (MBRT) and resazurindye (RRT) with other tests of bull semen quality and to examine their relationship to fertility. Methods: One hundredsixty-four ejaculates from 59 bulls were examined, processed, and used for 30 016 inseminations. Results: Bulls usedin artificial insemination have been selected for high semen quality and fertility, and semen from these bulls averaged80.6 % unstained sperm, only 11% had abnormalities, and fertility ranged from 64 %to 76 %. The MBRT and RRTwere run with standardized sperm numbers to prevent sperm concentration from influencing the dye reduction time. Ashort RRT was correlated with higher fertility ( r = -0.26) but MBRT was not correlated ( r = -0.05, P >0.0,5).The correlations were low, but axe typical and reflect the statistical effect of the large binomial variance associated withpregnancy or nonpregnancy being coded as 1 or O. Conclusion: The fact that the RRT was significantly correlated withfertility when sperm numbers were standardized, and published reports that resazurin is useful for monitoring other semencharacteristics, indicate that this dye may provide valuable information not given by other simple laboratory tests. (AsianJ Androl 1999 Sep; 1: 109 - 114 )展开更多
Objective:To explore the effect of Diospyros kaki on cattle spermatozoa during chilling and cryopreservation.Methods: Five milliliter of blended Persimmon (Diospyros kaki) flesh was added to 45 mL TCF to obtain 10% st...Objective:To explore the effect of Diospyros kaki on cattle spermatozoa during chilling and cryopreservation.Methods: Five milliliter of blended Persimmon (Diospyros kaki) flesh was added to 45 mL TCF to obtain 10% stock solution. Kaki enriched extender (KEE) was prepared by adding to TCF in concentrations 0.0/5.0 mL (control, 0%), 0.5/4.5 mL (1%), 1/4 mL (2%), 1.5/3.5 mL (3%), 2.0/3.0 mL (4%), 2.5/2.5 mL (5%), 3.0/2.0 mL (6%), 3.5/1.5 mL (7%), 4.0/1.0 mL (8%), 4.5/0.5 mL (9%) and 5.0/0.0 mL (10%) to obtain a final volume 5 mL in each tube. Whole egg yolk was added to each tube to obtain KEE with 20% egg yolk (KEEY), all tubes were centrifuged to get rid of debris. Semen was added to the supernatants in other tubes. Extended semen was subjected to evaluation (motility, alive sperm and intact sperm membrane (HOST) %) in both chilled and cryopreserved semen. Conception rate was carried out.Results:Sperm motility was significantly (P<0.0001) kept high after 11 d of chilling with the concentration 1%, 2%, 3%, 4%, 5% as compared to the control (41.67±1.67, 41.67±1.67, 40.00±0.00, 41.67±1.67 and 41.67±1.67, respectively) and also non-significantly kept high at the other concentrations up to 9 d of chilling. Addition of KEE had significantly (P<0.0033) improved post thawing sperm motility % with the concentrations 1, 2, 3, 4, 5 and 6% as compared to the control (51.67±5.27, 55.00±3.16, 48.33±1.05, 45.00±3.96, 57.00±2.50, 55.00±5.00 and 43.33±5.11 respectively).While the other concentrations exhibit no effect. Addition of KEE maintained alive sperm%, abnormalities% and % of intact spermatozoa membranes (HOST %) as good as the control with all concentrations of kaki used in our study. The conception rate upon using frozen semen in insemination showed higher conception rate in concentrations of 2%, 4% and 6 % KEE in cattle.Conclusion: It could be concluded that some concentrations ofDiospyros kaki improved bull semen quality post-chilling and post-freezing.展开更多
Objective:To explore the effect of silymarin on bull spermatozoa during cooling and cryopreservation. Methods: Pooled bull semen were diluted by Tris-Citrate-Fructose egg yolk diluents, purified silymarin powder (obta...Objective:To explore the effect of silymarin on bull spermatozoa during cooling and cryopreservation. Methods: Pooled bull semen were diluted by Tris-Citrate-Fructose egg yolk diluents, purified silymarin powder (obtained from the milk thistle silybum marianum), purchased from Unipharma, Al Obour city, Egypt, was soaked in Tris-citric acid-fructose diluent for 48 h at 10℃ making a stock solution (70 mg/mL), from this stock solution we obtained concentrations of 0.18 mg/mL, 0.36 mg/mL, 0.54 mg/mL, 0.72 mg/mL, 0.90 mg/mL in addition to the control (0.0 mg/mL) reaching a final volume of 5 mL in each tube. Egg yolk was added to each tube to obtain silymarin enriched semen extender (SEE) with 20% egg yolk, cooled slowly up to 5℃ and equilibrated for 4 h. Semen was packed into 0.25 mL polyvinyl French straws (IMV, France). After equilibration periods, the straws were placed horizontally on a rack and frozen in a vapor 4 cm above liquid nitrogen for 10 min and were then dipped in liquid nitrogen. Extended semen was subjected to evaluation (motility, alive %, abnormality %, intact sperm membrane (HOST)% and conception rate) in both chilled and frozen semen.Results: Table 1 revealed that Sperm motility of the concentrations 2, 3 and 4 after 8 d of chilling were significantly (P<0.02) higher than control. Sperm motility of the concentration 2 (45.00%±2.89%) after 9 d of chilling was higher than control and the other concentrations. Addition of SEE in concentration 1 and 2 gave post thawing sperm motility as high as the control (47.50±2.81 and 45.00±2.58, respectively) while other concentration have lower effects on motility as compared to the control. Addition of silymarin improved post thawing alive% and was significantly higher (P<0.0001) than the control. SEE decreased significantly (P<0.0001) the % of post thawing abnormal sperm in concentration 3 and 4 (11.83±0.65 and 16.00±0.58, respectively). SEE improved significantly (P<0.018) the % of post thawing intact spermatozoa membranes (HOST%) in concentrations 2, 4 and 5 (71.17±0.83, 71.83±0.91 and 75.00±3.42 respectively) (Table 2).Conclusion:It could be concluded that silymarin as a natural additive to semen extenders improved preservability in both chilled and frozen bull semen.展开更多
Objective:To evaluate effect of tris-extender supplemented with various concentrations of strawberry (Fragaria spp.) on bull semen preservability.Methods: Pooled bull semen were extended with tris-citrate-fructose egg...Objective:To evaluate effect of tris-extender supplemented with various concentrations of strawberry (Fragaria spp.) on bull semen preservability.Methods: Pooled bull semen were extended with tris-citrate-fructose egg yolk diluent (control, 0% strawberry) and various concentrations of tris strawberry (TSB) (1%-6%) to achieve 60 million motile spermatozoa per milliliter. Extended semen were subjected to semen freezing protocol. Semen assessment including motility, alive%, abnormality%, intact sperm membrane (hypo-osmotic swelling test) and conception rate were carried out for both chilled and frozen semen. Results: Results showed that sperm motility after chilling was enhanced in groups treated with various concentrations of TSB from 1% to 5% and exhibited higher significance (P<0.0001) at 6-day post-chilling. In frozen semen, 3%, 4%, 5% and 6% concentrations gave the best significance (P<0.0001) on sperm motility in comparison with the control. Concentration 1% revealed the highest significance (P<0.0001) on alive% as compared to the control. Hypo-osmotic swelling test was maintained as the control. Concentration 3% gave the lowest significance (P<0.0001) considering abnormality%. The conception rate upon using frozen semen in insemination showed higher conception rate in concentrations of 5% and 6% in cattle.Conclusions: It is concluded that 1%-5% concentrations of TSB ameliorate bull semen characteristics after chilling, and 3%-6% concentrations of TSB improve bull semen characteristics after freezing. Higher conception rate exists in 5% and 6% concentration of TSB.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
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
Objective:To explore the effect of BHT on cattle spermatozoa during cooling and cryopreservation.Methods: Pooled bull semen were diluted by Tris-Citrate-Fructose egg yolk (TCFY) diluent considered as control (0 BHT) a...Objective:To explore the effect of BHT on cattle spermatozoa during cooling and cryopreservation.Methods: Pooled bull semen were diluted by Tris-Citrate-Fructose egg yolk (TCFY) diluent considered as control (0 BHT) and different concentrations of BHT (1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 mM were prepared in ethanol in prewarmed (37℃) test tubes. The ethanol was allowed to evaporate so that, a thin crystallized layer of BHT was deposited on the inner surface of the tubes. Then extended semen was added into the tubes and incubated at 37℃ for 5 min to allow uptake of BHT by spermatozoa. The tubes were cooled slowly (approximately for 2 h) up to 5℃ and equilibrated for 4 h. After equilibration, semen freezing process was carried out. Extended semen was subjected to evaluation (motility, alive sperm, intact sperm membrane (HOST) % and acrosome integrity) in both cooled and cryopreserved semen. Results:The result revealed that sperm motility of post-cooled spermatozoa improved (P<0.05) by the use of BHT concentrations (1, 2 and 3 mM) in Tris semen extender if compared to the control (85.00±1.09), (83.33± 0.63), (81.67± 0.63) and (78.33± 0.63), respectively. Alive sperm percent was significantly higher in all concentrations of BHT. Sperm abnormalities percent were significantly lower in concentrations of BHT 1 and 2 (11.2±0.2), (11.8±0.2)and (13.4±0.4), respectively. Sperm membrane integrity were significantly higher in BHT concentrations (1, 2, 3, 4 and 5 mM). It is exhibited that improved sperm motility in post-thawed frozen semen in the concentrations of BHT (1, 2, 3 and 4 mM) if compared to the control. The sperm membrane integrity were significantly improved at all concentrations of BHT. Acrosome integrity was significantly higher at BHT concentration 1 mM (81.80±0.57) and (76.00±2.05), respectively.Conclusions: It could be concluded that some concentrations of BHT improved bull semen quality post-cooling and post-freezing.展开更多
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 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.
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Aim: This study was undertaken to compare the reduction in color of two dyes methylene blue (MBRT) and resazurindye (RRT) with other tests of bull semen quality and to examine their relationship to fertility. Methods: One hundredsixty-four ejaculates from 59 bulls were examined, processed, and used for 30 016 inseminations. Results: Bulls usedin artificial insemination have been selected for high semen quality and fertility, and semen from these bulls averaged80.6 % unstained sperm, only 11% had abnormalities, and fertility ranged from 64 %to 76 %. The MBRT and RRTwere run with standardized sperm numbers to prevent sperm concentration from influencing the dye reduction time. Ashort RRT was correlated with higher fertility ( r = -0.26) but MBRT was not correlated ( r = -0.05, P >0.0,5).The correlations were low, but axe typical and reflect the statistical effect of the large binomial variance associated withpregnancy or nonpregnancy being coded as 1 or O. Conclusion: The fact that the RRT was significantly correlated withfertility when sperm numbers were standardized, and published reports that resazurin is useful for monitoring other semencharacteristics, indicate that this dye may provide valuable information not given by other simple laboratory tests. (AsianJ Androl 1999 Sep; 1: 109 - 114 )
文摘Objective:To explore the effect of Diospyros kaki on cattle spermatozoa during chilling and cryopreservation.Methods: Five milliliter of blended Persimmon (Diospyros kaki) flesh was added to 45 mL TCF to obtain 10% stock solution. Kaki enriched extender (KEE) was prepared by adding to TCF in concentrations 0.0/5.0 mL (control, 0%), 0.5/4.5 mL (1%), 1/4 mL (2%), 1.5/3.5 mL (3%), 2.0/3.0 mL (4%), 2.5/2.5 mL (5%), 3.0/2.0 mL (6%), 3.5/1.5 mL (7%), 4.0/1.0 mL (8%), 4.5/0.5 mL (9%) and 5.0/0.0 mL (10%) to obtain a final volume 5 mL in each tube. Whole egg yolk was added to each tube to obtain KEE with 20% egg yolk (KEEY), all tubes were centrifuged to get rid of debris. Semen was added to the supernatants in other tubes. Extended semen was subjected to evaluation (motility, alive sperm and intact sperm membrane (HOST) %) in both chilled and cryopreserved semen. Conception rate was carried out.Results:Sperm motility was significantly (P<0.0001) kept high after 11 d of chilling with the concentration 1%, 2%, 3%, 4%, 5% as compared to the control (41.67±1.67, 41.67±1.67, 40.00±0.00, 41.67±1.67 and 41.67±1.67, respectively) and also non-significantly kept high at the other concentrations up to 9 d of chilling. Addition of KEE had significantly (P<0.0033) improved post thawing sperm motility % with the concentrations 1, 2, 3, 4, 5 and 6% as compared to the control (51.67±5.27, 55.00±3.16, 48.33±1.05, 45.00±3.96, 57.00±2.50, 55.00±5.00 and 43.33±5.11 respectively).While the other concentrations exhibit no effect. Addition of KEE maintained alive sperm%, abnormalities% and % of intact spermatozoa membranes (HOST %) as good as the control with all concentrations of kaki used in our study. The conception rate upon using frozen semen in insemination showed higher conception rate in concentrations of 2%, 4% and 6 % KEE in cattle.Conclusion: It could be concluded that some concentrations ofDiospyros kaki improved bull semen quality post-chilling and post-freezing.
文摘Objective:To explore the effect of silymarin on bull spermatozoa during cooling and cryopreservation. Methods: Pooled bull semen were diluted by Tris-Citrate-Fructose egg yolk diluents, purified silymarin powder (obtained from the milk thistle silybum marianum), purchased from Unipharma, Al Obour city, Egypt, was soaked in Tris-citric acid-fructose diluent for 48 h at 10℃ making a stock solution (70 mg/mL), from this stock solution we obtained concentrations of 0.18 mg/mL, 0.36 mg/mL, 0.54 mg/mL, 0.72 mg/mL, 0.90 mg/mL in addition to the control (0.0 mg/mL) reaching a final volume of 5 mL in each tube. Egg yolk was added to each tube to obtain silymarin enriched semen extender (SEE) with 20% egg yolk, cooled slowly up to 5℃ and equilibrated for 4 h. Semen was packed into 0.25 mL polyvinyl French straws (IMV, France). After equilibration periods, the straws were placed horizontally on a rack and frozen in a vapor 4 cm above liquid nitrogen for 10 min and were then dipped in liquid nitrogen. Extended semen was subjected to evaluation (motility, alive %, abnormality %, intact sperm membrane (HOST)% and conception rate) in both chilled and frozen semen.Results: Table 1 revealed that Sperm motility of the concentrations 2, 3 and 4 after 8 d of chilling were significantly (P<0.02) higher than control. Sperm motility of the concentration 2 (45.00%±2.89%) after 9 d of chilling was higher than control and the other concentrations. Addition of SEE in concentration 1 and 2 gave post thawing sperm motility as high as the control (47.50±2.81 and 45.00±2.58, respectively) while other concentration have lower effects on motility as compared to the control. Addition of silymarin improved post thawing alive% and was significantly higher (P<0.0001) than the control. SEE decreased significantly (P<0.0001) the % of post thawing abnormal sperm in concentration 3 and 4 (11.83±0.65 and 16.00±0.58, respectively). SEE improved significantly (P<0.018) the % of post thawing intact spermatozoa membranes (HOST%) in concentrations 2, 4 and 5 (71.17±0.83, 71.83±0.91 and 75.00±3.42 respectively) (Table 2).Conclusion:It could be concluded that silymarin as a natural additive to semen extenders improved preservability in both chilled and frozen bull semen.
文摘Objective:To evaluate effect of tris-extender supplemented with various concentrations of strawberry (Fragaria spp.) on bull semen preservability.Methods: Pooled bull semen were extended with tris-citrate-fructose egg yolk diluent (control, 0% strawberry) and various concentrations of tris strawberry (TSB) (1%-6%) to achieve 60 million motile spermatozoa per milliliter. Extended semen were subjected to semen freezing protocol. Semen assessment including motility, alive%, abnormality%, intact sperm membrane (hypo-osmotic swelling test) and conception rate were carried out for both chilled and frozen semen. Results: Results showed that sperm motility after chilling was enhanced in groups treated with various concentrations of TSB from 1% to 5% and exhibited higher significance (P<0.0001) at 6-day post-chilling. In frozen semen, 3%, 4%, 5% and 6% concentrations gave the best significance (P<0.0001) on sperm motility in comparison with the control. Concentration 1% revealed the highest significance (P<0.0001) on alive% as compared to the control. Hypo-osmotic swelling test was maintained as the control. Concentration 3% gave the lowest significance (P<0.0001) considering abnormality%. The conception rate upon using frozen semen in insemination showed higher conception rate in concentrations of 5% and 6% in cattle.Conclusions: It is concluded that 1%-5% concentrations of TSB ameliorate bull semen characteristics after chilling, and 3%-6% concentrations of TSB improve bull semen characteristics after freezing. Higher conception rate exists in 5% and 6% concentration of TSB.
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Objective:To explore the effect of BHT on cattle spermatozoa during cooling and cryopreservation.Methods: Pooled bull semen were diluted by Tris-Citrate-Fructose egg yolk (TCFY) diluent considered as control (0 BHT) and different concentrations of BHT (1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 mM were prepared in ethanol in prewarmed (37℃) test tubes. The ethanol was allowed to evaporate so that, a thin crystallized layer of BHT was deposited on the inner surface of the tubes. Then extended semen was added into the tubes and incubated at 37℃ for 5 min to allow uptake of BHT by spermatozoa. The tubes were cooled slowly (approximately for 2 h) up to 5℃ and equilibrated for 4 h. After equilibration, semen freezing process was carried out. Extended semen was subjected to evaluation (motility, alive sperm, intact sperm membrane (HOST) % and acrosome integrity) in both cooled and cryopreserved semen. Results:The result revealed that sperm motility of post-cooled spermatozoa improved (P<0.05) by the use of BHT concentrations (1, 2 and 3 mM) in Tris semen extender if compared to the control (85.00±1.09), (83.33± 0.63), (81.67± 0.63) and (78.33± 0.63), respectively. Alive sperm percent was significantly higher in all concentrations of BHT. Sperm abnormalities percent were significantly lower in concentrations of BHT 1 and 2 (11.2±0.2), (11.8±0.2)and (13.4±0.4), respectively. Sperm membrane integrity were significantly higher in BHT concentrations (1, 2, 3, 4 and 5 mM). It is exhibited that improved sperm motility in post-thawed frozen semen in the concentrations of BHT (1, 2, 3 and 4 mM) if compared to the control. The sperm membrane integrity were significantly improved at all concentrations of BHT. Acrosome integrity was significantly higher at BHT concentration 1 mM (81.80±0.57) and (76.00±2.05), respectively.Conclusions: It could be concluded that some concentrations of BHT improved bull semen quality post-cooling and post-freezing.