• Journal of Life Science
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• v.27 no.5
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• pp.517-523
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• 2017

This study was to investigate the role of antioxidants on the characteristics of arsenite-damaged boar semen. Collected sperm was diluted with semen extender, and $100{\mu}M$ arsenite was used for sperm damage. Then melatonin, silymarin, curcumin, and vitamin E were applied for 3, 6, and 9 hr in arsenite-treated boar sperm. Sperm characteristics were then analyzed for motility, plasma membrane integrity, mitochondrial activity, and lipid peroxidation. In the results, sperm motility (control, $77.3{\pm}1.8%$) was decreased by arsenite ($33.3{\pm}1.5%$), while the antioxidant treatment groups (100 nM melatonin, $55.8{\pm}3.4%$; $2{\mu}M$ silymarin, $48.8{pm}3.4%$; $10{\mu}M$ curcumin, $53.9{\pm}2.8%$; and $500{\mu}M$ vitamin E, $54.5{\pm}3.1%$) showed increases compared to the arsenite group (p<0.05). $100{\mu}M$ arsenite decreased the sperm plasma membrane integrity ($24.5{\pm}1.6%$) and mitochondrial activity ($58.2{\pm}2.6%$), and increased lipid peroxidation ($5.3{\pm}0.2%$) at 3 hr (p<0.05). However, arsenite-treated samples with 100 nM melatonin, $2{\mu}M$ silymarin, $10{\mu}M$ curcumin, and $500{\mu}M$ vitamin E increased the plasma membrane integrity and mitochondria activity, and decreased lipid peroxidation compared to the arsenite-treated samples. In summary, arsenite may induce sperm damage and oxidation stress, while antioxidants such as melatonin, silymarin, curcumin, and vitamin E are useful for maintaining sperm characteristics. Therefore, antioxidants can protect sperm against damage by arsenite in fresh boar semen.

• Journal of Embryo Transfer
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• v.26 no.1
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• pp.71-78
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• 2011

This study was undertaken to find out the effect of cholesterol and serum albumin on sperm ability and lipid peroxidation levels period to the liquid storage of miniature pig sperm. Ejaculated semen from miniature pigs was collected by gloved-hand method into a pre-warmed ($37^{\circ}C$) thermos bottle, and extended with Modena solution {with and without BSA, methyl-beta-cyclodextrin (-cholesterol) and cholesterol loaded cyclodextrin (+cholesterol)}. Each semen was assessed for viability (SYBR-14/PI staining) and acrosome intactness, intensity and capacitation status by chlorotetracycline (CTC) staining at 1, 3, 5, 7 and 10 days of storage. At for the effects of cholesterol and serum albumin on lipid peroxidation, semen were incubated with $H_2O_2$ ($10\;{\mu}M$), and lipid peroxidation level were measured by flow cytometry using the lipid peroxidation reporter probe $C_{11}-BODIPY^{581/591}$. The result, lipid peroxidation level in sperm added with cholesterol were lower in $10\;{\mu}M$ $H_2O_2$ compared to the added sperm with serum albumin. Also, added cholesterol to sperm had significant (p<0.05) higher viability when storage for 7 and 10 days and lower when 10 days of storage percentage of acrosome-reacted sperm (AR pattern) in acrosome state as say result compared to other treated groups. In conclusion, role of cholesterol during lipid storage in miniature pig spermatozoa was protected boar spermatozoa from lipid peroxidation prior to lipid storage. Addition serum albumin during lipid storage in sperm may be induce sperm membrane damage by lipid peroxidation. Therefore, addition of cholesterol to miniature pig sperm will be lead to extension of liquid storage periods.

• Journal of Embryo Transfer
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• v.29 no.2
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• pp.141-148
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• 2014

Quercetin and genistein, plentifully present in fruits and vegetables, are flavonoid family members that have antioxidative function and plant-derived phytoestrogen activity. The antioxidative effects of quercetin and genistein on boar sperm characteristics and in vitro development of IVF embryo were investigated. The sperm motility was increased by addition of genistein $50{\mu}M$ for 6 hr incubation compared to control (p<0.05). The sperm viability was increased by addition of quercetin 1 and $50{\mu}M$ and genestein 1 and $50{\mu}M$ for 3 hr incubation. In addition, the sperm viability seemed to be increased dose-dependantly by addition of quercetin or genistein 1 and $50{\mu}M$, respectively (p<0.05). The membrane integrities were not increased by quercetin or genistein treatments for 3 hr or 6 hr incubation period except for quercetin $1{\mu}M$ for 3 hr incubation. In mitochondrial activities, addition of quercetin $50{\mu}M$ for 6 hr incubation increased mitochondrial activity but decreased at $100{\mu}M$ concentration compared with control (p<0.05). When porcine IVF embryos were cultured in PZM-3 medium supplemented with low concentrations of quercetin ($1{\sim}10{\mu}M$), the developmental rates to morula and blastocyst increased but significantly decreased at high concentrations of quercetin ($25{\sim}50{\mu}M$). The highest developmental rate to blastocysts among all concentrations of quercetin was shown at quercetin $10{\mu}M$ (p<0.05). The developmental rates to morula or blastocysts at low ($0.01{\sim}1{\mu}M$) and high ($5{\sim}10{\mu}M$) concentrations of genistein were not significantly different among all treatment group and genistein did not affect on IVF embryo development. These results suggest that quercetin and genistein seem to have positive effects at certain concentrations on sperm characteristics such as motility, viability and mitochondrial activity. In addition, low concentrations of quercetin (1, 5 and $10{\mu}M$) in this experiment, seem to have beneficial effect on porcine IVF embryo development but genistein did not affect on it at all given concentrations ($0.01{\sim}10{\mu}M$).

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.7
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• pp.923-927
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• 2008

The objective of this study was to compare random regression model and multiple trait animal model estimates of the (co) variance of total sperm cells over the active lifetime of AI boars. Data were provided by Smithfield Premium Genetics (Rose Hill, NC). Total number of records and animals for the random regression model were 19,629 and 1,736, respectively. Data for multiple trait animal model analyses were edited to include only records produced at 9, 12, 15, 18, 21, 24, and 27 months of age. For the multiple trait method estimates of genetic and residual variance for total sperm cells were heterogeneous among age classifications. When comparing multiple trait method to random regression, heritability estimates were similar except for total sperm cells at 24 months of age. The multiple trait method also resulted in higher estimates of heritability of total sperm cells at every age when compared to random regression results. Random regression analysis provided more detail with regard to changes of variance components with age. Random regression methods are the most appropriate to analyze semen traits as they are longitudinal data measured over the lifetime of boars.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.259-259
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• 2004

Porcine sperm extract (PSE) supporting Ca/sup 2+/ osillation was microinjected into the in vitro matured porcine oocytes. In the presence of the capacitative Ca/sup 2+/ entry mechanism which can activate MII oocytes, preparation methods of sperm extraction were studied by many researchers. Such as freeze-thaw cycle, homogenation, sonication of boar sperm was used for certification of their activity of calcium signals. (omitted)

• Mycobiology
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• v.44 no.1
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• pp.54-57
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• 2016

Davallialactone (DAVA) is a hispidin analogue derived from the medicinal fungus Phellinus baumii. We examined the effect of DAVA on in vitro fertilization (IVF) of pigs. Boar spermatozoa were incubated in fertilization medium with varying concentrations of DAVA, then sperm motility and reactive oxygen species (ROS) level were evaluated. Higher sperm motility was found following the addition of 0.5 or $1{\mu}M$ DAVA after incubation than addition of other concentrations or controls. ROS level decreased significantly with the addition of DAVA. The rate of normal fertilization was higher in the presence of $1{\mu}M$ DAVA (65.1%) than were those of other concentrations or controls (45.4~59.4%), and the highest total fertilization rate (mono- and polyspermic oocytes) was observed at $1{\mu}M$ DAVA (83%). In conclusion, addition of DAVA to fertilization medium improved sperm motility, and reduced ROS level so as to potentially improve sperm-oocyte binding in IVF, suggesting the potential of a compound isolated from mushrooms in assisted reproductive technology for humans and animals.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.3
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• pp.199-205
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• 2020

Cryopreservation of semen is useful for animal breeding via artificial insemination (AI). However, the use of frozen-thawed boar semen is limited due to cryodamage. The aim of this study was to investigate the effects of different concentrations of MitoTEMPO (a mitochondria-targeted antioxidant) in lactose-egg yolk (LEY) extenders on kinetic characteristics of frozen-thawed boar sperms. Semen samples were collected from mature Duroc boars (2~3 years old) and cryopreserved in LEY extenders containing 0, 0.5, 5, 50, and 500 μM MitoTEMPO. The kinetic characteristics of frozen-thawed sperms were determined 0 and 30 min after thawing using computer-assisted sperm analysis (CASA). Results indicated that sperm motility immediately after thawing was significantly higher with 5 and 50 μM (50.46±2.71% and 46.96±2.66%, respectively) than with 500 μM MitoTEMPO (35.40±2.95%) (P<0.05). However, there were no significant differences in other kinetic characteristics except motility. In conclusion, the addition of MitoTEMPO to the sperm freezing extender may have a beneficial effect on motility of post-thawed boar semen.

• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.85-91
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• 2011

The objective of this study was to evaluated the efficiency on sperm cryosurvival and ability of in vitro fertilization using Triladyl and Lactose Egg-Yolk(LEY) as extenders for cryopreservation of separated sperm by 65% percoll in miniature pig. Sperm viability was measured with SYBR-14/PI double stained sperm by flow cytometry. Ability on embryo cleavage rate and blastocyst development were observed by in vitro fertilization after frozen-thawing of sperm separated by 65% percoll. The experimental groups were designed that separated sperm by 65% percoll with Triladyl (ST) or LEY(SL) and unseparated sperm with Triladyl(UT) or LEY(UL) for cryopreservation. As a results, the viability was significantly(p<0.05) higher in ST(55.1%), SL(63.1%), UL(58.8%) than UT(38.2%) group. Sperm viability in SL(63.1%) group was significantly(p<0.05) higher than other experimental groups. On the other hand, embryo cleavage rate was significantly(p<0.05) higher in ST(79.1%), SL(83.2) than UT(74.1) and UL(75.7%) groups at 96h after in vitro fertilization. Blastocyst development was also significantly(p<0.05) higher in ST(21.5%), SL(20.9%) than UT(17.0%) and UL(18.8%) groups. In conclusion, cryopreservation of miniature boar sperm separated by 65% percoll were beneficial to viability and capacity on in vitro fertilization.

• Reproductive and Developmental Biology
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• v.39 no.1
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• pp.7-11
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• 2015

Preservation of liquid semen is an important factor for breeding management in swine industry. Oxidative stress of spermatozoa during liquid preservation has a detrimental effect on sperm quality and decreases fertility. Objective of this study was to determine the effect of antioxidant, Quercetin, on capability of porcine liquid semen preservation. Freshly collected porcine semen from boars (n=3), having proven fertility was counted, diluted to $3{\times}10^7/mL$ and divided into 5 different semen extenders. Aliquots of diluted semen with different extenders were subjected to measure the pH, motility, viability and sperm DNA structure status on elapse time after preservation for 10 days. For the first 3 days, semen preserved in all 5 different extenders maintained their initial pH and either gradually decreased or increased thereafter, indicating lipid peroxidation has started. Sperm motility (r=0.52, p=0.01) and viability (r=0.55, p=0.03) had positive correlation with semen pH. Sperm motility was maintained well (p<0.05) in especially 2 extenders containing Tris and antioxidant compared to other extenders, suggesting both Tris and antioxidant worked as pH regulator and had beneficial effects on sperm characteristic during preservation. Sperm DNA structure status accessed by sperm chromatin structure assay on elapsed time after preservation, tended to be higher in semen preserved without antioxidant. Taken together, addition of antioxidant to extender prevents the sperm from oxidative stress during storage in mechanism by which antioxidant slows the lipid peroxidation, and thus reduced the reactive oxygen species in preserved porcine semen resulted in maintaining semen pH, sperm motility and viability for 7~10 days.

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.81-87
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• 2008

Production of transgenic animals for studying specific gene has been limited due to a low efficiency, lack of skilled researchers and the need for expensive equipment. Currently, the boar spermatozoa as a vector to deliver exogenous DNA into the oocyte were used to improve the efficiency of transfection rate. In this study, we revealed that the optimal conditions for DNA uptake in spermatozoa by liposome were to 90 min of incubation, $17^{\circ}C$, $10^5$ spermatozoa, 4 ng/ml of exogenous DNA and 0.5% (v/v) liposome, without damage to fertility. In addition, the developmental rate to the blastocyst stage of embryo in control group was significantly higher than those embryos with exogenous DNA and liposome, whereas there were no significant differences in embryo development between the liposome and type of DNA. The transfection rates of embryo using treated spermatozoa with both liposome and circular DNA were higher than those using linear DNA. These findings raise the possibility thattreated spermatozoa with liposome/DNA complexes could be used in in vitro fertilization, and the exogenous DNA transferred into the oocytes. Taken together, we demonstrated that liposome a vector for the uptake of exogenous DNA in boar spermatozoa could improve the efficiency of sperm-mediated gene transfer in creating transgenic pig and the other domestic transgenic animals.