Studies have explored the assisted reproductive technology(ART)outcomes of Y-chromosome azoospermia factor c(AZFc)microdeletions,but the effect of sperm source on intracytoplasmic sperm injection(ICSI)remains unknown....Studies have explored the assisted reproductive technology(ART)outcomes of Y-chromosome azoospermia factor c(AZFc)microdeletions,but the effect of sperm source on intracytoplasmic sperm injection(ICSI)remains unknown.To determine the ART results of ICSI using testicular sperm and ejaculated sperm from males with AZFc microdeletions,we searched Embase,Web of Science,and PubMed to conduct a systematic review and meta-analysis.The first meta-analysis results for 106 cycles in five studies showed no significant differences in the live birth rate between the testicular sperm group and the ejaculated sperm group(risk ratio:0.97,95%confidence interval[CI]:0.73-1.28,P=0.82).The second meta-analysis of 106 cycles in five studies showed no difference in the abortion rate between the testicular sperm group and ejaculated sperm group(risk ratio:1.06,95%Cl:0.54-2.06,P=0.87).The third meta-analysis of 386 cycles in seven studies showed no significant difference in clinical pregnancy rates between the testicular sperm group and the ejaculated sperm group(risk ratio:1.24,95%Cl:0.66-2.34,P=0.50).Inevitable heterogeneity weakened our results.However,our results indicated that testicular sperm and ejaculated sperm yield similar ART outcomes,representing a meaningful result for clinical treatment.More properly designed studies are needed to further confirm our conclusions.展开更多
Objective To investigate the empty zona pellucida for use in the cryopreservation of human sperm. Materials & Methods Human and hamster zona pellucidae were evacuated and injected with testicular, epididymal ...Objective To investigate the empty zona pellucida for use in the cryopreservation of human sperm. Materials & Methods Human and hamster zona pellucidae were evacuated and injected with testicular, epididymal and ejaculated sperm. The zona pellucidae with sperm were cryopreserved. Results After thawing, zona pellucidae were easily found, and sperm inside zona pellucidae were also easily observed. There were no differences in post-thaw motility and vitality between ejaculated and epididymal sperm groups (P>0.05), but these two parameters were lowered in testicular sperm group compared to both ejaculated and epididymal sperm (P<0.01). No significant difference was observed in post-thaw motilities among 6%, 7.5%, 9% glycerol concentrations (P>0.05). In addition, obvious differences in post-thaw motilities were not found between human and hamster empty zona pellucidae (P>0.05). Conclusion An evacuated zona pellucida is an ideal vehicle for the cryopreservation of a small number of human sperm.展开更多
Couples with a spinal cord injured male partner require assisted ejaculation techniques to collect semen that can then be further used in various assisted reproductive technology methods to achieve a pregnancy. The ma...Couples with a spinal cord injured male partner require assisted ejaculation techniques to collect semen that can then be further used in various assisted reproductive technology methods to achieve a pregnancy. The majority of men sustaining a spinal cord injury regardless of the cause or the level of injury cannot ejaculate during sexual intercourse. Only a small minority can ejaculate by masturbation. Penile vibratory stimulation and electroejaculation are the two most common methods used to retrieve sperm. Other techniques such as prostatic massage and the adjunct application of other medications can be used, but the results are inconsistent. Surgical sperm retrieval should be considered as a last resort if all other methods fail. Special attention must be paid to patients with T6 and rostral levels of injury due to the risk of autonomic dysreflexia resulting from stimulation below the level of injury. Bladder preparation should be performed before stimulation if retrograde ejaculation is anticipated. Erectile dysfunction is ubiquitous in the spinal cord injured population but is usually easily managed and does not pose a barrier to semen retrieval in these men. Semen analysis parameters of men with spinal cord injury are unique for this population regardless of the method of retrieval, generally presenting as normal sperm concentration but abnormally low sperm motility and viability. When sperm retrieval is desired in this population, emphasis should be placed on initially trying the simple methods of penile vibratory stimulation or electroejaculation before resorting to more advanced and invasive surgical procedures.展开更多
Background Protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo development.In some mammals,like pigs,ejaculates are emitted in three separa...Background Protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo development.In some mammals,like pigs,ejaculates are emitted in three separate fractions:pre-sperm,sperm-rich(SRF)and post sperm-rich(PSRF).These fractions are known to vary in volume,sperm concentration and quality,as well as in the origin and composition of seminal plasma(SP),with differences being also observed within the SRF one.Yet,whether disparities in the DNA integrity and chromatin condensation and pro-tamination of their sperm exist has not been interrogated.Results This study determined chromatin protamination(Chromomycin A3 test,CMA_(3)),condensation(Dibromobi-mane test,DBB),and DNA integrity(Comet assay)in the pig sperm contained in the first 10 m L of the SRF(SRF-P1),the remaining portion of the sperm-rich fraction(SRF-P2),and the post sperm-rich fraction(PSRF).While chromatin protamination was found to be similar between the different ejaculate fractions(P>0.05),chromatin condensation was seen to be greater in SRF-P1 and SRF-P2 than in the PSRF(P=0.018 and P=0.004,respectively).Regarding DNA integrity,no differences between fractions were observed(P>0.05).As the SRF-P1 has the highest sperm concentra-tion and ejaculate fractions are known to differ in antioxidant composition,the oxidative stress index(OSi)in SP,calcu-lated as total oxidant activity divided by total antioxidant capacity,was tested and confirmed to be higher in the SRF-P1 than in SRF-P2 and PSRF(0.42±0.06 vs.0.23±0.09 and 0.08±0.00,respectively;P<0.01);this index,in addition,was observed to be correlated to the sperm concentration of each fraction(Rs=0.973;P<0.001).Conclusion While sperm DNA integrity was not found to differ between ejaculate fractions,SRF-P1 and SRF-P2 were observed to exhibit greater chromatin condensation than the PSRF.This could be related to the OSi of each fraction.展开更多
South African indigenous bucks are raised under extensive conditions and are more likely to reject artificial vagina (AV) due to their limited contact with human beings, as they are less handled. The purpose of the st...South African indigenous bucks are raised under extensive conditions and are more likely to reject artificial vagina (AV) due to their limited contact with human beings, as they are less handled. The purpose of the study was to compare goat semen collection techniques used for goats based on semen traits of South African indigenous bucks during the natural breeding season. A total of eight South African indigenous bucks were used, four bucks per semen collection technique due to their scarcity. Before semen collection with AV group, ten bucks were trained and exposed to AV. Semen was then collected from both groups over a period of six weeks per individual buck. Semen volume, pH and sperm concentration were evaluated immediately. The Computer Aided Sperm Analysis (CASA) assessed the sperm motility rates. Sperm morphology was evaluated using a fluorescence microscope with the aid of eosin-nigrosin staining. Analysis of variance was used to test the differences among the semen collection groups following assessment by CASA. Semen collected with EE resulted in higher semen volume (1.1 mL) compared with AV (0.5 mL) technique. However, AV technique resulted in higher total sperm motility and rapid movement (91.9% and 48.7%), live sperm (72%) and sperm concentration (635.6 × 106 sperm/mL) compared with EE (86.3% and 30.4%;69% and 463.7 × 106 sperm/mL, respectively). There were no significant differences observed in pH and sperm morphology characteristics between two semen collection techniques. Moreover, EE had higher sperm moving in a progressive (53.6%) and medium rate compared with AV (41.1% and 21.9%) technique. It was concluded that the AV yielded higher total sperm motility rate, sperm concentration and rapid sperm movement compared with EE. Therefore, AV was more suitable for semen collection of South African indigenous bucks, even though training was problematic to such indigenous bucks.展开更多
Objective studies of men＇s reproductive function are hindered by their reliance on： （i） self-reporting to quantify sexual activity and （ii） masturbation to quantify sperm output rendering both types of estimate ...Objective studies of men＇s reproductive function are hindered by their reliance on： （i） self-reporting to quantify sexual activity and （ii） masturbation to quantify sperm output rendering both types of estimate vulnerable to unverifiable subjective factors. We therefore examined whether detection of spermatozoa and measurement of prostate-specific antigen （PSA） in urine could provide objective semiquantitative estimates of sperm output and recent ejaculation, respectively, using widely available laboratory techniques. Of 11 healthy volunteers who provided urine samples before and at intervals for 5 days after ejaculation, sperm was present in 2111 men before, and in all 11111 samples immediately after ejaculation, but by the second and subsequent void, spermatozoa were present in -10%. PSA was detectable at high levels in all urine samples, peaking at the first post-ejaculatory sample but returning to baseline levels by the second post-ejaculatory void. We conclude that urinary spermatozoa and PSA are objective biomarkers for sperm production and sexual activity, but only for a short-time window until the first post-ejaculatory urine void. Hence, for a single urine specimen, the presence of spermatozoa and PSA are valid biomarkers, reflecting sperm production and recent ejaculation only until the next micturition, so their measurement should be restricted to the first morning urine void.展开更多
Objective To investigate sperm parameters of infertile men with retrograde ejaculation. Methods Twelve infertile men with retrograde ejaculation （group A） were enrolled into this study. Sperm samples were obtained f...Objective To investigate sperm parameters of infertile men with retrograde ejaculation. Methods Twelve infertile men with retrograde ejaculation （group A） were enrolled into this study. Sperm samples were obtained from the postejaculation urine. After sperm recovery and washing procedure, sperm parameters were assessed. Twelve semen samples from normospermic donors were Used as the control （group B）. Results In all retrograde cases, motile sperm with forward movement were observed in the medium. Motility of group A was significantly lower than that of group B （P〈0.01）. In group A, sperm motility ranged from 11% to 56%, sperm with intact both head and tail membranes was 42.2 ±12.3%, sperm count ranged （13-85）×^ 106/ml, and the sperm survival time was highly shortened. Sperm with normal morphology and intact acrosome were observed in retrograde specimens. Conclusion Sperm parameters recovered from retrograde specimens were highly variable between subjects. The toxicity of urine caused deleterious to sperm functions. Motile sperm could be collected by sperm recovery procedure. Sperm parameters could meet the requirement for the use of assisted reproductive techniques for treating infertile men with retrograde ejaculation.展开更多
Objective:To evaluate the effects of semen collection methods and different cooling rates on cooled and frozen-thawed Saanen goat semen.Methods:Twenty bucks were divided into two groups of 10 animals per group based o...Objective:To evaluate the effects of semen collection methods and different cooling rates on cooled and frozen-thawed Saanen goat semen.Methods:Twenty bucks were divided into two groups of 10 animals per group based on semen collection methods:artificial vagina and electro-ejaculator.Samples from each collection method were pooled and diluted with Tris-based extender.The pooled semen was divided into two cooling methods:slow cooling and fast cooling.Sperm motility and velocity,sperm plasma membrane integrity,sperm viability,mitochondrial membrane potential and DNA integrity were evaluated before dilution,immediately after cooling at 4℃and at 24 h after freezing by using computer assisted sperm analyser method and epifluorescence microscope.Results:Mean values for total motility,rapid-speed and progressive motile spermatozoa were significantly higher in semen collected with artificial vagina than electro-ejaculator(P<0.001).Slow cooling resulted in higher percentages of total motile,rapid-speed and progressive motile spermatozoa as compared to fast cooling(P<0.05).The post-thaw percentage of rapid-speed spermatozoa collected by artificial vagina in slow cooled sperm was significantly higher as compared to electro-ejaculator method(P<0.05).The mean values of curvilinear velocity and straight-line velocity were significantly higher in slow cooled semen obtained by artificial vagina(P<0.001).Similarly,the combination of artificial vagina and slow cooling resulted in significantly higher post-thaw percentages of straight-line velocity,average-path velocity,sperm plasma membrane integrity,viability and acrosome integrity(P<0.01).No significant differences were observed in slow cooling between artificial vagina and electro-ejaculator in terms of post-thaw sperm mitochondrial membrane potential and DNA integrity(P>0.05).Conclusions:The use of artificial vagina and slow cooling rate in cryopreservation protocol improves the cooled and post-thaw sperm quality of Saanen buck semen.展开更多
Promiscuous mating systems provide the opportunity for females to bias fertilization toward particular males.However,distinguishing between male sperm com-petition and active female sperm choice is difficult for speci...Promiscuous mating systems provide the opportunity for females to bias fertilization toward particular males.However,distinguishing between male sperm com-petition and active female sperm choice is difficult for species with internal fertilization.Nevertheless,species that store and use sperm of different males in different storing struc-tures and species where females are able to expel all or part of the ejaculates after cop-ulation may be able to bias fertilization.We report a series of experiments aimed at pro-viding evidence of female sperm choice in Euxesta eluta(Hendel),a species of ulidiid fly that expels and consumes ejaculates after copulation.We found no evidence of greater reproductive success for females mated singly,multiply with the same male,or mated multiply with different males.Female E.eluta possesses two spherical spermathecae and a bursa copulatrix for sperm storage,with a ventral receptacle.There was no significant difference in storing more sperm in spermathecae 24 h after copulation than immediately after copulation.Females mated with protein-fed males had greater reproductive success than similar females mated to protein-deprived males.Protein-fed females prevented to consume the ejaculate,retained more sperm when mated to protein-fed males than when mated to protein-deprived males.Our results suggest that female E.eluta can exert control of sperm retention of higher quality males through ejaculate ejection.展开更多
Cervids show a high degree of abnormalities in their sperm cells.Thus,this study aimed to select high-quality spermatozoa using magnetic-activated sperm sorting(MASS)compared to density gradient centrifugation(DGC)by ...Cervids show a high degree of abnormalities in their sperm cells.Thus,this study aimed to select high-quality spermatozoa using magnetic-activated sperm sorting(MASS)compared to density gradient centrifugation(DGC)by assessing the post-selection cell quality.Semen from six Mazama deer was collected by electroejaculation after chemical restraint.The semen was analyzed in four samples:Fresh,DGC,SEMgood-non-apoptotic fraction,and SEMpoor-apoptotic fraction.The material was analyzed for motility and vigor(light microscopy),concentration(Neubauer chamber),semen morphology(phase contrast),and supravital staining test(eosin/nigrosine).The DGC method used 20 x 106 cells in 90%and 45%percoll®gradient.The MASS used 10 x 106 cells with 20μl of iron nanoparticles attached to Annexin V and filtration in a magnetic separation column.Both processing methods(DGC and MASS)were effective in producing high-quality sperm samples,with a marked reduction in abnormalities from 41.83±10.25(fresh)to 14.83±3.17(DGC)and 12±3.01(SEMgood),with 80.3%±2.06 livings cells.These findings suggest that this nanotechnological method,using nanoparticles,effectively produces high-quality semen samples in cervids for use in assisted reproduction.展开更多
基金the Project of Medical Science and Technology Research Foundation of Guangdong Province(No.A2019336)Guangzhou Science and Technology Plan Project(No.201707010394).
文摘Studies have explored the assisted reproductive technology(ART)outcomes of Y-chromosome azoospermia factor c(AZFc)microdeletions,but the effect of sperm source on intracytoplasmic sperm injection(ICSI)remains unknown.To determine the ART results of ICSI using testicular sperm and ejaculated sperm from males with AZFc microdeletions,we searched Embase,Web of Science,and PubMed to conduct a systematic review and meta-analysis.The first meta-analysis results for 106 cycles in five studies showed no significant differences in the live birth rate between the testicular sperm group and the ejaculated sperm group(risk ratio:0.97,95%confidence interval[CI]:0.73-1.28,P=0.82).The second meta-analysis of 106 cycles in five studies showed no difference in the abortion rate between the testicular sperm group and ejaculated sperm group(risk ratio:1.06,95%Cl:0.54-2.06,P=0.87).The third meta-analysis of 386 cycles in seven studies showed no significant difference in clinical pregnancy rates between the testicular sperm group and the ejaculated sperm group(risk ratio:1.24,95%Cl:0.66-2.34,P=0.50).Inevitable heterogeneity weakened our results.However,our results indicated that testicular sperm and ejaculated sperm yield similar ART outcomes,representing a meaningful result for clinical treatment.More properly designed studies are needed to further confirm our conclusions.
基金This study was supported by the Science & Technology Commission of Guangdong ProvinceP.R.China
文摘Objective To investigate the empty zona pellucida for use in the cryopreservation of human sperm. Materials & Methods Human and hamster zona pellucidae were evacuated and injected with testicular, epididymal and ejaculated sperm. The zona pellucidae with sperm were cryopreserved. Results After thawing, zona pellucidae were easily found, and sperm inside zona pellucidae were also easily observed. There were no differences in post-thaw motility and vitality between ejaculated and epididymal sperm groups (P>0.05), but these two parameters were lowered in testicular sperm group compared to both ejaculated and epididymal sperm (P<0.01). No significant difference was observed in post-thaw motilities among 6%, 7.5%, 9% glycerol concentrations (P>0.05). In addition, obvious differences in post-thaw motilities were not found between human and hamster empty zona pellucidae (P>0.05). Conclusion An evacuated zona pellucida is an ideal vehicle for the cryopreservation of a small number of human sperm.
文摘Couples with a spinal cord injured male partner require assisted ejaculation techniques to collect semen that can then be further used in various assisted reproductive technology methods to achieve a pregnancy. The majority of men sustaining a spinal cord injury regardless of the cause or the level of injury cannot ejaculate during sexual intercourse. Only a small minority can ejaculate by masturbation. Penile vibratory stimulation and electroejaculation are the two most common methods used to retrieve sperm. Other techniques such as prostatic massage and the adjunct application of other medications can be used, but the results are inconsistent. Surgical sperm retrieval should be considered as a last resort if all other methods fail. Special attention must be paid to patients with T6 and rostral levels of injury due to the risk of autonomic dysreflexia resulting from stimulation below the level of injury. Bladder preparation should be performed before stimulation if retrograde ejaculation is anticipated. Erectile dysfunction is ubiquitous in the spinal cord injured population but is usually easily managed and does not pose a barrier to semen retrieval in these men. Semen analysis parameters of men with spinal cord injury are unique for this population regardless of the method of retrieval, generally presenting as normal sperm concentration but abnormally low sperm motility and viability. When sperm retrieval is desired in this population, emphasis should be placed on initially trying the simple methods of penile vibratory stimulation or electroejaculation before resorting to more advanced and invasive surgical procedures.
基金This research was supported by the European Union’s Horizon 2020 research and innovation scheme under the Marie Skłodowska-Curie grant agreement No.801342(Tecniospring INDUSTRYGrant:TECSPR-19-1-0003)+4 种基金the Ministry of Science and Innovation,Spain(Grants:PID2020-113320RB-I00,PID2020-113493RB-I00,RYC2021-034546-I and RYC2021-034764-I)the Catalan Agency for Management of University and Research Grants,Regional Government of Catalonia,Spain(Grants:2017-SGR-1229 and 2021-SGR-00900)the Seneca Foundation,Regional Government of Murcia,Spain(Grant:21935/PI/22)La Marato de TV3 Foundation(Grant:214/857-202039)and the Catalan Institution for Research and Advanced Studies(ICREA).
文摘Background Protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo development.In some mammals,like pigs,ejaculates are emitted in three separate fractions:pre-sperm,sperm-rich(SRF)and post sperm-rich(PSRF).These fractions are known to vary in volume,sperm concentration and quality,as well as in the origin and composition of seminal plasma(SP),with differences being also observed within the SRF one.Yet,whether disparities in the DNA integrity and chromatin condensation and pro-tamination of their sperm exist has not been interrogated.Results This study determined chromatin protamination(Chromomycin A3 test,CMA_(3)),condensation(Dibromobi-mane test,DBB),and DNA integrity(Comet assay)in the pig sperm contained in the first 10 m L of the SRF(SRF-P1),the remaining portion of the sperm-rich fraction(SRF-P2),and the post sperm-rich fraction(PSRF).While chromatin protamination was found to be similar between the different ejaculate fractions(P>0.05),chromatin condensation was seen to be greater in SRF-P1 and SRF-P2 than in the PSRF(P=0.018 and P=0.004,respectively).Regarding DNA integrity,no differences between fractions were observed(P>0.05).As the SRF-P1 has the highest sperm concentra-tion and ejaculate fractions are known to differ in antioxidant composition,the oxidative stress index(OSi)in SP,calcu-lated as total oxidant activity divided by total antioxidant capacity,was tested and confirmed to be higher in the SRF-P1 than in SRF-P2 and PSRF(0.42±0.06 vs.0.23±0.09 and 0.08±0.00,respectively;P<0.01);this index,in addition,was observed to be correlated to the sperm concentration of each fraction(Rs=0.973;P<0.001).Conclusion While sperm DNA integrity was not found to differ between ejaculate fractions,SRF-P1 and SRF-P2 were observed to exhibit greater chromatin condensation than the PSRF.This could be related to the OSi of each fraction.
文摘South African indigenous bucks are raised under extensive conditions and are more likely to reject artificial vagina (AV) due to their limited contact with human beings, as they are less handled. The purpose of the study was to compare goat semen collection techniques used for goats based on semen traits of South African indigenous bucks during the natural breeding season. A total of eight South African indigenous bucks were used, four bucks per semen collection technique due to their scarcity. Before semen collection with AV group, ten bucks were trained and exposed to AV. Semen was then collected from both groups over a period of six weeks per individual buck. Semen volume, pH and sperm concentration were evaluated immediately. The Computer Aided Sperm Analysis (CASA) assessed the sperm motility rates. Sperm morphology was evaluated using a fluorescence microscope with the aid of eosin-nigrosin staining. Analysis of variance was used to test the differences among the semen collection groups following assessment by CASA. Semen collected with EE resulted in higher semen volume (1.1 mL) compared with AV (0.5 mL) technique. However, AV technique resulted in higher total sperm motility and rapid movement (91.9% and 48.7%), live sperm (72%) and sperm concentration (635.6 × 106 sperm/mL) compared with EE (86.3% and 30.4%;69% and 463.7 × 106 sperm/mL, respectively). There were no significant differences observed in pH and sperm morphology characteristics between two semen collection techniques. Moreover, EE had higher sperm moving in a progressive (53.6%) and medium rate compared with AV (41.1% and 21.9%) technique. It was concluded that the AV yielded higher total sperm motility rate, sperm concentration and rapid sperm movement compared with EE. Therefore, AV was more suitable for semen collection of South African indigenous bucks, even though training was problematic to such indigenous bucks.
文摘Objective studies of men＇s reproductive function are hindered by their reliance on： （i） self-reporting to quantify sexual activity and （ii） masturbation to quantify sperm output rendering both types of estimate vulnerable to unverifiable subjective factors. We therefore examined whether detection of spermatozoa and measurement of prostate-specific antigen （PSA） in urine could provide objective semiquantitative estimates of sperm output and recent ejaculation, respectively, using widely available laboratory techniques. Of 11 healthy volunteers who provided urine samples before and at intervals for 5 days after ejaculation, sperm was present in 2111 men before, and in all 11111 samples immediately after ejaculation, but by the second and subsequent void, spermatozoa were present in -10%. PSA was detectable at high levels in all urine samples, peaking at the first post-ejaculatory sample but returning to baseline levels by the second post-ejaculatory void. We conclude that urinary spermatozoa and PSA are objective biomarkers for sperm production and sexual activity, but only for a short-time window until the first post-ejaculatory urine void. Hence, for a single urine specimen, the presence of spermatozoa and PSA are valid biomarkers, reflecting sperm production and recent ejaculation only until the next micturition, so their measurement should be restricted to the first morning urine void.
基金This is a part of the project (A 04010449) supported by Natural Science Foundation of Guangdong Province, China.
文摘Objective To investigate sperm parameters of infertile men with retrograde ejaculation. Methods Twelve infertile men with retrograde ejaculation （group A） were enrolled into this study. Sperm samples were obtained from the postejaculation urine. After sperm recovery and washing procedure, sperm parameters were assessed. Twelve semen samples from normospermic donors were Used as the control （group B）. Results In all retrograde cases, motile sperm with forward movement were observed in the medium. Motility of group A was significantly lower than that of group B （P〈0.01）. In group A, sperm motility ranged from 11% to 56%, sperm with intact both head and tail membranes was 42.2 ±12.3%, sperm count ranged （13-85）×^ 106/ml, and the sperm survival time was highly shortened. Sperm with normal morphology and intact acrosome were observed in retrograde specimens. Conclusion Sperm parameters recovered from retrograde specimens were highly variable between subjects. The toxicity of urine caused deleterious to sperm functions. Motile sperm could be collected by sperm recovery procedure. Sperm parameters could meet the requirement for the use of assisted reproductive techniques for treating infertile men with retrograde ejaculation.
文摘Objective:To evaluate the effects of semen collection methods and different cooling rates on cooled and frozen-thawed Saanen goat semen.Methods:Twenty bucks were divided into two groups of 10 animals per group based on semen collection methods:artificial vagina and electro-ejaculator.Samples from each collection method were pooled and diluted with Tris-based extender.The pooled semen was divided into two cooling methods:slow cooling and fast cooling.Sperm motility and velocity,sperm plasma membrane integrity,sperm viability,mitochondrial membrane potential and DNA integrity were evaluated before dilution,immediately after cooling at 4℃and at 24 h after freezing by using computer assisted sperm analyser method and epifluorescence microscope.Results:Mean values for total motility,rapid-speed and progressive motile spermatozoa were significantly higher in semen collected with artificial vagina than electro-ejaculator(P<0.001).Slow cooling resulted in higher percentages of total motile,rapid-speed and progressive motile spermatozoa as compared to fast cooling(P<0.05).The post-thaw percentage of rapid-speed spermatozoa collected by artificial vagina in slow cooled sperm was significantly higher as compared to electro-ejaculator method(P<0.05).The mean values of curvilinear velocity and straight-line velocity were significantly higher in slow cooled semen obtained by artificial vagina(P<0.001).Similarly,the combination of artificial vagina and slow cooling resulted in significantly higher post-thaw percentages of straight-line velocity,average-path velocity,sperm plasma membrane integrity,viability and acrosome integrity(P<0.01).No significant differences were observed in slow cooling between artificial vagina and electro-ejaculator in terms of post-thaw sperm mitochondrial membrane potential and DNA integrity(P>0.05).Conclusions:The use of artificial vagina and slow cooling rate in cryopreservation protocol improves the cooled and post-thaw sperm quality of Saanen buck semen.
基金supported by the Fund for Scientific and Technological Research(FONCyT PICT 2018-03521).
文摘Promiscuous mating systems provide the opportunity for females to bias fertilization toward particular males.However,distinguishing between male sperm com-petition and active female sperm choice is difficult for species with internal fertilization.Nevertheless,species that store and use sperm of different males in different storing struc-tures and species where females are able to expel all or part of the ejaculates after cop-ulation may be able to bias fertilization.We report a series of experiments aimed at pro-viding evidence of female sperm choice in Euxesta eluta(Hendel),a species of ulidiid fly that expels and consumes ejaculates after copulation.We found no evidence of greater reproductive success for females mated singly,multiply with the same male,or mated multiply with different males.Female E.eluta possesses two spherical spermathecae and a bursa copulatrix for sperm storage,with a ventral receptacle.There was no significant difference in storing more sperm in spermathecae 24 h after copulation than immediately after copulation.Females mated with protein-fed males had greater reproductive success than similar females mated to protein-deprived males.Protein-fed females prevented to consume the ejaculate,retained more sperm when mated to protein-fed males than when mated to protein-deprived males.Our results suggest that female E.eluta can exert control of sperm retention of higher quality males through ejaculate ejection.
文摘Cervids show a high degree of abnormalities in their sperm cells.Thus,this study aimed to select high-quality spermatozoa using magnetic-activated sperm sorting(MASS)compared to density gradient centrifugation(DGC)by assessing the post-selection cell quality.Semen from six Mazama deer was collected by electroejaculation after chemical restraint.The semen was analyzed in four samples:Fresh,DGC,SEMgood-non-apoptotic fraction,and SEMpoor-apoptotic fraction.The material was analyzed for motility and vigor(light microscopy),concentration(Neubauer chamber),semen morphology(phase contrast),and supravital staining test(eosin/nigrosine).The DGC method used 20 x 106 cells in 90%and 45%percoll®gradient.The MASS used 10 x 106 cells with 20μl of iron nanoparticles attached to Annexin V and filtration in a magnetic separation column.Both processing methods(DGC and MASS)were effective in producing high-quality sperm samples,with a marked reduction in abnormalities from 41.83±10.25(fresh)to 14.83±3.17(DGC)and 12±3.01(SEMgood),with 80.3%±2.06 livings cells.These findings suggest that this nanotechnological method,using nanoparticles,effectively produces high-quality semen samples in cervids for use in assisted reproduction.
