The trade creation and conservation of wild species in Amazon, including collared peccaries （Pecari tajacu）, may be favored by artificial insemination, but the time of semen storage may lead to reduced fertility in ...The trade creation and conservation of wild species in Amazon, including collared peccaries （Pecari tajacu）, may be favored by artificial insemination, but the time of semen storage may lead to reduced fertility in sperm of some wild animals. Thus, the aim of this research was to evaluate the influence of semen storage at 17 ℃ over time on seminal features of peccaries in captivity. Eight adult males were sedated and underwent electroejaculation. The ejaculates （n = 65） were evaluated for volume, concentration, pH, sperm motility, vigor, and cell with intact plasma membrane intact （IPM） and sperm morphology. Selected ejaculates （n = 21） were diluted （1：1） in Beltsville Thawing Solution and kept during 48 hours under controlled temperature （17 ℃）. Assessments were made after dilution （TO）, after 24 hours （T24）, and 48 hours after the onset of cooling （T48）. The storage impacted on sperm survival （P 〈 0.05）. Semen characteristics changed throughout the storage period studied and after 48 hours storage. The decline of sperm motility was of 55.2% for 10.9%, vigor was 2.3 for 0.5 and IPM cells were of 59.0% for 15.8%. Primary defects sperm increased of 19.8% for 32.2%, secondary defects of 9.8% for 10.4% and total defects of 29.4% for 42.7%. However, within 24 hours of preservation, chilled semen peccaries presented sperm motility average rate and IMP cells levels indicative to use in assisted reproductive events. These results indicate chilled semen for 24 hours could be used in experimentally artificial insemination of peccaries, technology that still has not been performed before.展开更多
Resveratrol(3,5,4’-trihydroxystilbene,RSV) has been widely used in mammalian cells,but whether it can be used during freezing boar semen is still unknown.The effects of RSV treatment during boar semen freezing on its...Resveratrol(3,5,4’-trihydroxystilbene,RSV) has been widely used in mammalian cells,but whether it can be used during freezing boar semen is still unknown.The effects of RSV treatment during boar semen freezing on its anti-freezing ability,apoptosis,and possible apoptotic pathways were observed in this study.Sperm motility,mitochondrial membrane potential(ΔΨm),adenosine triphosphate(ATP) content,terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick-end labeling(TUNEL)-positive apoptotic state,and messenger RNA(mRNA) expression levels of apoptotic genes involved in different apoptotic pathways after freezing with or without RSV treatment were tested.The results showed that:(1) Compared with fresh sperm,the motility,normal acrosome rate,and plasma membrane integrity rate of frozen boar sperm decreased significantly(P<0.05),and RSV did not significantly increase the sperm motility(0.44 vs.0.40,P>0.05),but it did significantly improve the normal acrosome rate(57.65% vs.47.00%,P<0.05) and plasma membrane integrity rate(46.67% vs.38.85%,P<0.05).(2) After freezing,most boar sperm showed low mitochondrial ΔΨm.RSV treatment could increase the rate of high mitochondrial ΔΨm of boar sperm.(3) RSV treatment significantly decreased reactive oxygen species(ROS) levels(58.65% vs.88.41%,P<0.05)and increased the ATP content(0.49 μmol/L vs.0.25 μmol/L,P<0.05) of boar sperm during freezing.(4) The apoptotic rate of the freezing group(80.41%) with TUNEL detection increased significantly compared to the fresh group(9.70%,P<0.05),and RSV treatment greatly decreased the apoptotic rate(68.32%,P<0.05).(5) Real-time polymerase chain reaction(RT-PCR) showed that not only the genes from the death receptor-mediated apoptotic pathway(tumor necrosis factor-α(TNF-α),Fas ligand(FasL),and Caspase-8),but also the genes from the mitochondria-mediated apoptotic pathway(manganese superoxide dismutase(MnSOD),B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),and Caspase-9) were both significantly changed after freezing.RSV treatment during freezing greatly changed their expression levels.Although RSV treatment during boar semen freezing did not significantly increase motility after thawing,it still played an efficient antioxidant role,which could enhance the mitochondrial function and decrease the apoptotic level induced by both the death receptor-and mitochondria-mediated apoptotic pathways.展开更多
文摘The trade creation and conservation of wild species in Amazon, including collared peccaries （Pecari tajacu）, may be favored by artificial insemination, but the time of semen storage may lead to reduced fertility in sperm of some wild animals. Thus, the aim of this research was to evaluate the influence of semen storage at 17 ℃ over time on seminal features of peccaries in captivity. Eight adult males were sedated and underwent electroejaculation. The ejaculates （n = 65） were evaluated for volume, concentration, pH, sperm motility, vigor, and cell with intact plasma membrane intact （IPM） and sperm morphology. Selected ejaculates （n = 21） were diluted （1：1） in Beltsville Thawing Solution and kept during 48 hours under controlled temperature （17 ℃）. Assessments were made after dilution （TO）, after 24 hours （T24）, and 48 hours after the onset of cooling （T48）. The storage impacted on sperm survival （P 〈 0.05）. Semen characteristics changed throughout the storage period studied and after 48 hours storage. The decline of sperm motility was of 55.2% for 10.9%, vigor was 2.3 for 0.5 and IPM cells were of 59.0% for 15.8%. Primary defects sperm increased of 19.8% for 32.2%, secondary defects of 9.8% for 10.4% and total defects of 29.4% for 42.7%. However, within 24 hours of preservation, chilled semen peccaries presented sperm motility average rate and IMP cells levels indicative to use in assisted reproductive events. These results indicate chilled semen for 24 hours could be used in experimentally artificial insemination of peccaries, technology that still has not been performed before.
基金Project supported by the Jiangsu Agri-animal Husbandry Vocational College Academy Research Project(No.NSF201509)the Jiangsu University Brand Speciality Construction Work Founded Project(No.PPZY2015C230),China。
文摘Resveratrol(3,5,4’-trihydroxystilbene,RSV) has been widely used in mammalian cells,but whether it can be used during freezing boar semen is still unknown.The effects of RSV treatment during boar semen freezing on its anti-freezing ability,apoptosis,and possible apoptotic pathways were observed in this study.Sperm motility,mitochondrial membrane potential(ΔΨm),adenosine triphosphate(ATP) content,terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick-end labeling(TUNEL)-positive apoptotic state,and messenger RNA(mRNA) expression levels of apoptotic genes involved in different apoptotic pathways after freezing with or without RSV treatment were tested.The results showed that:(1) Compared with fresh sperm,the motility,normal acrosome rate,and plasma membrane integrity rate of frozen boar sperm decreased significantly(P<0.05),and RSV did not significantly increase the sperm motility(0.44 vs.0.40,P>0.05),but it did significantly improve the normal acrosome rate(57.65% vs.47.00%,P<0.05) and plasma membrane integrity rate(46.67% vs.38.85%,P<0.05).(2) After freezing,most boar sperm showed low mitochondrial ΔΨm.RSV treatment could increase the rate of high mitochondrial ΔΨm of boar sperm.(3) RSV treatment significantly decreased reactive oxygen species(ROS) levels(58.65% vs.88.41%,P<0.05)and increased the ATP content(0.49 μmol/L vs.0.25 μmol/L,P<0.05) of boar sperm during freezing.(4) The apoptotic rate of the freezing group(80.41%) with TUNEL detection increased significantly compared to the fresh group(9.70%,P<0.05),and RSV treatment greatly decreased the apoptotic rate(68.32%,P<0.05).(5) Real-time polymerase chain reaction(RT-PCR) showed that not only the genes from the death receptor-mediated apoptotic pathway(tumor necrosis factor-α(TNF-α),Fas ligand(FasL),and Caspase-8),but also the genes from the mitochondria-mediated apoptotic pathway(manganese superoxide dismutase(MnSOD),B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),and Caspase-9) were both significantly changed after freezing.RSV treatment during freezing greatly changed their expression levels.Although RSV treatment during boar semen freezing did not significantly increase motility after thawing,it still played an efficient antioxidant role,which could enhance the mitochondrial function and decrease the apoptotic level induced by both the death receptor-and mitochondria-mediated apoptotic pathways.
