• Korean Journal of Animal Reproduction
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• v.21 no.4
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• pp.405-409
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• 1997

This study was carried out to investigate the general characteristics such as semen volume, pH, sperm motility and sperm concentration of the semen collected from Korean Jindo dogs by the mothod of Digital manipulation of penis, and the effect of temperature and preservation time on motility of fresh semen. Multiple ejaculates were collected from four male Korean Jindo dogs. The results obtained in this experiment were as follows : 1. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the second fraction and the small volume of third fraction from the ejaculate were 3.29ml, 6.30, 96.70% and 1.64$\times$108 cells/ml, respectively. 2. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the first fraction from the ejaculate were 1.16ml, 6.10, 6.67% and 5.07$\times$105cells/ml. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the second fraction from the ejaculate were 2.30ml, 6.33, 97.66% and 1.92$\times$108cells/ml. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the third fraction from the ejaculate were 3.24ml, 6.51, 93.33% and 3.13$\times$107cells/ml. 3. Motility of fresh semen during preservation were higher at 17$^{\circ}C$ than at 5$^{\circ}C$ or 36$^{\circ}C$. When preservation temeprature was 17$^{\circ}C$, motility were 95.75% at 1 h, 90.00% at 6 h, 84.25% at 12 h, 68.00% at 18 h, 36.25% at 24 h and 28.75% at 30 h, respectively.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.2
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• pp.149-157
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• 2002

Objectives: To assess the scavenging effect of carnitine derivatives on oxidative damage to sperm during sperm processing, cryopreservation and thawing. Materials and Methods: Fresh semen samples from 20 normal healthy volunteers were collected by masturbation after at least 48 hours abstinence. After liquefaction of semen samples at room temperature, the specimens were diluted with sperm wash media (Ham's F-10, Life technologics) to a uniform density of $20{\times}10^6/ml$. L-carnitine or acetylcarnitine were added with various concentration of $0{\mu}M$, $10{\mu}M$, $30{\mu}M$ in semen sample or cryoprotectant. All specimens were cryopreservated at $-196^{circ}C$ $LN_2$ for 3 days. Sperm motility, vitality, fertilizing capacity, reactive oxygen species formation and the level of lipid peroxidation were analyzed by computer assisted semen analyzer, eosin-nigrosin stain, hypoosmotic swelling test, chemiluminescence and thiobarbituric acid method, respectively, during sperm processing, cryopreservation and thawing. Results: The sperm motility was only increased in proportion to the concentration of acetylcarnitine with no statistical significance (p>0.05). The sperm vitality was also significantly improved in proportion to the concentration of acetylcarnitine with statistical significance (p<0.05). The sperm fertilizing capacity was significantly increased in proportion to the concentration of L-carnitine and acetylcarnitine and reactive oxygen species generation and lipid peroxidation were significantly decreased with same fashion (p<0.05). On comparison of effects between L-carnitine and acetylcarnitine, acetylcarnitine was superior to L-carnitine on the improvement of sperm motility and vitality as well as the suppression of reactive oxygen species generation and lipid peroxidation. Conclusions: These results suggest that carnitine derivatives have a scavenging effect against oxidative damages during sperm processing, cryopreservation and thawing. Therefore, carnitine derivatives may be useful as an oral antioxidant in patients with male infertility due to increased ROS generation.

• Clinical and Experimental Reproductive Medicine
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• v.18 no.1
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• pp.23-34
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• 1991

In order to study the sperm-egg interaction during fertilization process in mouse, the effects of sperm concentration, the duration of capacitation and insemination, the stages of maturation and development of eggs, and sperm extracts and BSA on sperm binding to egg were examined. Sperm-egg binding was increased depending on sperm concentration within the range of $10^3-10^6$ sperm/ml. It showed the most numbers of sperm-egg binding at 60min from the beginning of preincubation(capacitation) and insemination, respectively. During sperm capacitation, sperm-egg binding inhibitor was released from sperm into the incubation medium. Sperm extracts containing trypsin-like enzyme which is secreted through the acrosome reaction increased the binding. BSA in the culture medium showed a positive effect on the binding. It is suggested that physicochemical alterations of zona pellucida in the process of maturation and fertilization of eggs leaded to inhibition of sperm-egg binding.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.4
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• pp.500-503
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• 2002

This study was carried out to investigate the effects of season influencing semen characteristics, frozen-thawed sperm viability and testosterone concentration in Duroc boars. There were no significant differences in the semen volume and sperm concentration of Duroc boars among spring, summer, autumn and winter. However, the pH of sperm-rich and sperm-poor fractions in autumn and winter season was higher than in spring and summer season in Duroc boars. Sperm motility and normal acrosome of raw semen in Duroc boars did not differ significantly among spring, summer, autumn and winter. However, motility and normal acrosome of frozen-thawed sperm were higher in spring season than in summer, autumn and winter. Serum testosterone concentrations in Duroc were higher in spring than summer, autumn and winter. In conclusion, when serum testosterone concentrations were higher in seasons, frozen-thawed sperm viability in Duroc boars were higher.

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.1
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• pp.40-47
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• 2024

Background: Sperm quality and the number of sperm introduced into the uterus during artificial insemination (AI) are pivotal factors influencing pregnancy outcomes. However, there have been no reports on the relationship between sperm concentration at AI and sperm quality in Hanwoo cattle. In this study, we examined sperm quality and pregnancy rates after AI using sperm inseminated at different concentrations. Methods: We evaluated the motility, viability, and acrosomal membrane integrity of sperm at different concentrations (10, 15, 18, and 20 million sperm/straw) in 0.5-mL straws. Subsequently, we compared the pregnancy rates after AI with different sperm concentrations. Results: After freeze-thawing, sperm at the assessed concentrations showed similar viability and acrosomal membrane integrity. After AI, cattle in the 10 million group had significantly lower pregnancy rates compared to those in the 18 and 20 million groups. Conversely, there were no statistically significant variances observed between cattle in the 10 and 15 million groups. Conclusions: Sperm at concentrations of 10, 15, 18 and 20 million per straw exhibited comparable motility, viability, and acrosomal membrane integrity. However, a concentration of at least 18 million sperm per straw is required to achieve a consistent rate of pregnancy rate in Hanwoo cattle after AI.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.6
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• pp.755-759
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• 2004

The present investigation was carried out to assess the effect of Sephadex (G-15) filtration on the post thaw bull semen quality and conception rate. Post thaw unfiltered (control) and Sephadex filtered semen from four healthy bulls (three cross bred and one pure bred Holstein Friesian) were subjected to microscopic examination viz. sperm concentration, individual motility, live sperm count and sperm morphology. Sixty-two healthy, normal cycling crossbred cows were inseminated with post thaw unfiltered (n=32) and filtered semen (n=30). Sephadex filtration of post thaw semen significantly (p<0.05) decreased total sperm concentration and sperm with abnormal head, mid piece and tail. The overall average total sperm concentration, head and tail defects in filtered semen decreased significantly (53.4, 1.2 and 6.4 million) than in the unfiltered semen (80.4, 2.4 and 15.7 million, respectively). However, after filtration significant (p<0.05) increase in overall average motile and live sperm concentration were observed (38.8 and 38.0) as compared to unfiltered semen (29.2 and 32.0 million, respectively). The overall conception rate recorded was 21.9% with post thaw unfiltered semen and 56.7% with filtered semen. It was concluded that Sephadex filtration of post thaw semen improved its quality and conception rate.

• Transactions of the Korean Society of Mechanical Engineers A
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• v.32 no.12
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• pp.1115-1122
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• 2008

This paper presents a new microchip which can separate motile sperm by chemotaxis. The microchip was developed to create longitudinal concentration gradient in the microchannel due to diffusion. Linearly good concentration gradient of chemoattractant was generated without any fluid control devices. In sperm separation experiment with the developed microchip, mouse sperm was used as sample and acetylcholine was selected as chemoattractant. Human tubal fluid (HTF), buffer solution, was introduced into the microchannel of the microchip and attractants diluted in ratio of 1, 1/2, 1/4, 1/8, 1/16, 1/32 and 1/64 including control (DI water) were dropped in each outlet by $2\;{\mu}l$ volume with micropippet. After 5min, $1\;{\mu}l$ sperm solution was dropped into inlet of the chip. After 10 min, when sperms reached to the outlet by chemotaxis, we counted sperms in each outlet by using microscopy. Consequently, we could separate progressive motile sperm with the new microchip. In the experiment, the most sperms were isolated at the outlet dropped with 1/16 diluted solution. The optimal concentration gradient to induce chemotaxis was about 0.625 mg/ml/mm.

• Korean Journal of Animal Reproduction
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• v.15 no.3
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• pp.189-193
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• 1991

This study was carried out ot investigate effects of X-537A on hydrogen ion concentration insperm washed solution and sperm acrosome reaction. The results obtained were as follows. 1. When bovine sperm was twice washed with SHP solutions of pH 6.8 and 7.4 and again washed with SHP solution containing 4$\mu$M of X-537A, in case of pH 6.8 the sperm washed with 4$\mu$M of X-537A showed signifciantly(p<0.01) higher hydrogen ion concentration in sperm washed solution than the sperm washed without X-537A. 2. When the sperm was twice washed with SHP solution and then washed with SHP solution containing 4$\mu$M of X-537A, sperm acrosome rection rate was signifciantly(p<0.01) increased from 12min after incubation in the sperm washed without X-537A, but was signifciantly(p<0.01) increased from 8 min after incubation in the sperm washed with 4$\mu$M of X-537A. 3. When the sperm was twice washed with SHP solution and then washed with SHP solution containing 0, 4 and 40$\mu$M of X-537A, and then incubated in m-TALP for 120 min, sperm acrosome reaction rate was significantly(p<0.01) increased from 15 min after incubation in 0, 4 and 40$\mu$m OF X-537A. However at 60 min incubation 40$\mu$M of X-537A showed significantly(p<0.01) higher sperm acrosome reaction rate than 0 and 4$\mu$M and at 120 min incubation 4 and 40$\mu$M of X-537A showed signifciantly(p<0.01) higher acrosome reaction rate than 0$\mu$M of X-537A.

• Journal of Embryo Transfer
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• v.9 no.3
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• pp.269-277
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• 1994

The objective of this study was to develop an effective in vitro production system capable of obtaining more porcine embryos from immature oocytes These experiments were conducted to examine the effect of sperm factor on the IVF and IVD, and the effect of coculture with somatic cells on the IVD of embryos. Although the concentration of epididymal sperm for IVF did not affect on cleavage rate, but 5 x 105 sperm/mi showed the highest cleavage rate(48.7%) and the developmental potential of IVF oocytes from this concentration was also greatly higher (P$^{\circ}C$-stored sperm for l2hrs and the cleavage rate from fresh sperm was significantly higher (P<0.05) than that from frozen sperm, but the developmental potential after IVF was slightly high from the frozen sperm. The cleavage rate of IVF oocytes cocultured with oviductal epithelial cells and cumulus cells was 76.3% and 72.9%, respectively. There was no difference between two coculture systems but this rate was significantly higher(P<0.05) than that of medium alone(42.0%).

• Korean Journal of Veterinary Service
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• v.18 no.3
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• pp.1-12
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• 1995

To investigate effect of seeding on post-thaw motility and viability of canine spermatozoa, the semen from male dogs which had been proved to be fertile in the past were frozen and seeded during freezing process. Post-thaw motility and viability of canine sperm which were frozen and seeded were investigated according to different seeding temperatures of $-5^{\circ}C$, $-10^{\circ}C$, or $-l5^{\circ}C$ and also according to different concentration of glycerol of 2%, 5% and 10%. In addition, post-thaw motility of canine sperm frozen by direct freezing in a deep freezer or programmed freezing in a programmed cell freezer was investigated. Post-thaw motility of canine sperm was compared according to different seeding temperatures : The sperm seeded at $-5^{\circ}C$ showed considerably higher post-thaw motility than that of non-seeding, and that seeded at $-10^{\circ}C$, or $-l5^{\circ}C$, respectively, in 2% and 5% glycerol groups on both 2 and 7day after freezing(p<0.05). In 10% concentration of glycerol, the sperm seeded at each seeding temperature showed considerably higher post-thaw motility than that of non-seeding group on day 7 after freezing(p<0.01). Post-thaw viability of canine sperm was compared according to different seeding temperatures : The sperm seeded at $-5^{\circ}$ showed significantly higher post-thaw motility than that of non-seeding, and that seeded at $-10^{\circ}C$, or $-l5^{\circ}C$, in 5% and 10% glycerol groups on day 7 after freezing(p< 0.05). In comparison of post-thaw motility of canine sperm seeded according to different concentration of glycerol, 5% glycerol group and 10% glycerol group showed considerably higher post-thaw motility than 2% glycerol group without difference between those two groups in all seeding temperatures($-5^{\circ}C$, $-10^{\circ}C$ and $-l5^{\circ}C$) on day 2 and 7 after freezing(p<0.01). In comparison of post-thaw viability of canine sperm seeded according to different concentration of glycerol, 5% glycerol group and 10% glycerol group showed the same considerably higher post-thaw viability than 2% glycerol group on each thawing day(p<0.01). The canine sperm frozen and seeded by programmed freezing method showed considerably higher post-thaw motility than that frozen by direct freezing method in all different seeding temperatures($-5^{\circ}C$, $-10^{\circ}C$ and $-l5^{\circ}$). These results indicated that the higher post-thaw motility and viability was obtained in the spermatozoa seeded than that of non-seeding, that among different seeding temperatures of $-5^{\circ}C$, $-10^{\circ}C$ and $-l5^{\circ}C$, the sperm seeded at $-5^{\circ}C$ showed higher post-thaw motility and viability than the other temperatures, also among different concentrations fof glycerol of 2%, 5% and 10%, the sperm frozen and seeded in 5% and 10% concentration of glycerol showed higher post-thaw motility and viability than that in 2% of glycerol, and that the sperm frozen and seeded by programmed freezing method showed higher motility than that by direct freezing method.