• Korean Journal of Animal Reproduction
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• v.25 no.3
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• pp.201-206
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• 2001

This study was carried out to investigate the effects of spring (March~May) and summer (June~August) influencing semen characteristics, frozen-thawed sperm viability and serum testosterone concentration in Yorkshire boars. Results of this study were as follows: 1. There were no significant differences in the semen volume, pH and sperm concentration of sperm-poor fraction of Yorkshire boars between spring and summer. However, sperm concentrations of sperm-rich fractions in spring were higher than those in summer (P＜0.05). 2. Sperm motility and normal acrosome of raw semen in Yorkshire boars did not differ significantly between spring and summer, However, motility and normal acrosome of frozen-thawed sperm were higher in spring season than in summer season (P＜0.05). 3. Serum testosterone concentrations in Yorkshire boars were 4.04 ng/$m\ell$ in spring and 2.85 ng/$m\ell$ in summer. Serum testosterone concentrations in spring were higher than those in summer (P＜0.05). 4. In conclusion, when serum testosterone concentrations in Yorkshire boars were higher, frozen-thawed sperm viability was higher.

• Development and Reproduction
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• v.15 no.2
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• pp.151-158
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• 2011

The comparison of the chemical properties of semen of black porgy Acanthopagrus schlegelii long-term acclimated reared in freshwater (BFW) and seawater (BSW) with sperm activity of salinity and ion composition. The chemical properties of seminal plasma on BFW of the factors that most there was not significant difference in the BSW. However, osmolality in seminal plasma of BFW and BSW was $307.0{\pm}4.6$ and $337.3{\pm}10.1$ mOsm/kg, respectively, where BFW showed significant lower concentration in contrast to BSW. Salinity effect on sperm motility of BFW and BSW in 0 psu solution, no sperm motility was observed, whereas in 10 psu solution, both BFW and BSW sperms showed low motility and short time post sperm activation. However, diluted in 20 and 32 psu solutions, highest motility and long time post sperm activation were observed in BFW and BSW sperm. SAI of BFW and BSW varied in depend on the osmolality regardless of ion kind and it showed the highest value in the similar osmolality of artificial seawater (956 mOsm/kg). Accordingly, even in sperm released from BFW, factors initiating sperm motility are determined by osmolality.

• Korean Journal of Ichthyology
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• v.19 no.2
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• pp.168-172
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• 2007

In the present study, attempts were made to find out the physico-chemical properties of milt and the sperm motilities in various osmotic conditions using Panther puffer, Takifugu pardalis. The average concentration of sperm in the milt was $12.1{\pm}3.2{\times}10^9/mL$. pH and osmolality of seminal plasma were $8.2{\pm}0.3$, $385.5{\pm}12.5mOsm/kg$, respectively. Spermatozoa were immotile when the milt was mixed with solutions (electrolyte or non-electrolyte) of lower osmolality than the average seminal plasma osmolality ($385.5{\pm}12.5mOsm/kg$), but became motile after mixing milt with hyperosmotic solutions.

• Journal of Veterinary Clinics
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• v.28 no.1
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• pp.13-19
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• 2011

Semen cryopreservation induces the formation of reactive oxygen species (ROS), and the ROS cause sperm damage. We aimed to investigate the effects of the antioxidative enzyme catalase (CAT) on sperm quality and ROS during cryopreservation. Sperm rich fractions collected from five Duroc boars were cryopreserved in freezing extender with (200 or 400 U/mL) or without CAT (control). After thawing, sperm motility, viability, normal morphology, plasma membrane integrity, mitochondrial function and intracellular ROS were evaluated. CAT significantly improved total sperm motility at a concentration of 400 U/mL (P < 0.05), but didn't improve progressive sperm motility, viability, morphological defects, plasma membrane integrity and mitochondrial function in frozen-thawed boar sperm. In evaluation of ROS, CAT had no effect on reduction in ${\cdot}O_2$, but scavenged $H_2O_2$ in viable frozen-thawed boar sperm at concentrations of 200 and 400 U/mL (P < 0.05). In conclusion, CAT was not enough to improve quality of frozen-thawed sperm, but can reduce $H_2O_2$ generation in viable boar sperm during cryopreservation.

• Journal of Veterinary Clinics
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• v.24 no.4
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• pp.486-492
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• 2007

Seminal plasma(SP) is usually removed from semen that is to be cryopreserved. However, some reports indicate that SP has beneficial effects on spermatozoa during chilling and freezing. The purpose of this study was to determine the effect of SP on sperm survival by adding SP to the extender before cooling and freezing canine spermatozoa. In replicate experiments, ejaculates obtained from four healthy dogs(1-4 years old) of various breeds were pooled, centrifuged at $300{\times}g$ for 10 min at $25^{\circ}C$, and the supernatant of seminal plasma was decanted. Spermatozoa were suspended in egg yolk-Tris(EYT) buffer. The study comprised two experiments: [Exp 1] Sperm were suspended in EYT extender containing either 0, 20, 40, 80 or 100% SP and were slowly cooled to $4^{\circ}C$ for 2h or held at $25^{\circ}C$ as controls. Sperm concentration was adjusted to $2{\times}10^8/ml$. [Exp II] Sperm samples, each of which contained $1{\times}10^8/ml$, were assigned to nine groups to be frozen. In the first four groups, sperm in EYT containing either 20, 40, 80 or 100% SP were cooled to $4^{\circ}C$, then diluted to contain final concentrations of EYT+0.6M glycerol and then were frozen. The final concentrations of SP were 10, 20, 40 or 50%. In the other four groups, sperm in EYT alone were first cooled slowly to $4^{\circ}C$, then diluted to contain final concentrations of EYT+0.6M glycerol plus 10, 20, 40 or 50% SP and then were frozen. Spermatozoa, which chilled in EYT alone and diluted to contain final concentrations of EYT+0.6M glycerol without seminal plasma, and then frozen, was regarded as control. Spermatozoa were frozen at $25^{\circ}C/min$ of cooling rate in plastic straws that were suspended above liquid nitrogen and thawed in water at $38^{\circ}C$ for 1 min. Sperm survival was assayed by determining progressive motility and integrity of plasma and acrosome membranes. Progressive motility was determined by microscopic examination at $200{\times}$ magnification. Membrane integrity was assessed by use of a double fluorescent dye, and acrosome integrity by staining sperm with Pisum sativum agglutinin. The results of the first experiment showed that adding SP did not improve motility of spermatozoa compared to those incubated without SP regardless of temperature. The results of the second experiment showed that spermatozoa suspended in EYT+0.6M glycerol containing SP exhibited the higher progressive motility before being frozen(P<0.05). However, frozen-thawed spermatozoa that had suspended in EYT+0.6M glycerol containing SP showed the similar or lower viability(P<0.05). In summary, although seminal plasma did not affect spermatozoa that were chilled in EYT without cryoprotectant(CPA), addition of seminal plasma to EYT containing CPA did significantly improved progressive motility of canine spermatozoa that were chilled.

• Korean Journal of Animal Reproduction
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• v.25 no.1
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• pp.79-84
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• 2001

This study was carried out to investigate the effects of spring (March~May) and summer (June~August) influencing semen characteristics, frozen-thawed sperm viability and serum testosterone concentration in Duroc boars. Results of this study were as follows: 1. There were no significant differences in the semen volume, pH and sperm concentration of Duroc boars between spring and summer. 2. Sperm motility and normal acrosome of raw semen in Duroc boars did not differ significantly between spring and summer. However, motility and normal acrosome of frozen-thawed sperm were higher in spring season than in summer season(P＜0.05). 3. Serum testosterone concentrations in Duroc boars were 2.15 ng/$m\ell$ in spring and 0.65 ng/$m\ell$ in summer. Serum testosterone concentrations in spring were higher thin those in summer (P＜0.05). 4. In conclusion, when serum testosterone concentrations in Duroc boars were higher, frozen-thawed sperm viability were higher.

• Clinical and Experimental Reproductive Medicine
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• v.34 no.3
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• pp.159-166
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• 2007

Objective: To investigate whether semen parameters in infertile couples who undergone intrauterine insemination (IUI) change in the subsequent IUI cycle and the subsequent in vitro fertilization (IVF) cycle. Methods: Fifty-three infertile couples who had failed to become pregnant after the first IUI cycle with computer-assisted semen analysis (CASA) were included. After the first IUI, thirty-eight couples underwent the second IUI (Group 1), and fifteen underwent IVF-ET procedure (Group 2). All semen parameters including semen volume, concentration, motility and total motile sperm count were analyzed in the second IUI or IVF-ET procedure for comparison with the result of first IUI. Results: There were no significant differences in husband age, interval between the first and second procedure and cause of infertility. In Group 1, only sperm motility at the time of the latter IUI was significantly decreased when compared to the former IUI irrespective of the first semen parameters. In Group 2, sperm concentration, motility and total motile sperm count at the time of subsequent IVF were lower than the former IUI. By sub-analyses of Group 2, in the group of optimal semen parameter at IUI cycle, sperm concentration and total motile sperm count at the time of subsequent IVF were lower than the former IUI, while in the group of suboptimal semen parameter at IUI cycle, only sperm motility at the time of subsequent IVF were lower than the former IUI. Conclusion: The semen parameters in couples converted to IVF cycle were more adversely affected than those remained in IUI cycle. Further study on psychological stress should be necessary to explain the reason.

• Journal of Embryo Transfer
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• v.13 no.3
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• pp.277-289
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• 1998

To establish a system for sperm swim-up separation through sucrose layer, indiscreet sperm migration should sufficiently to block but movement of sperm shouldn't inhibit. Thus, the effects of sucrose levels in sucrose layer, incubation times and types of sucrose layer on sperm separation were examined. And the results obtained were as follows; 1. Layer of 10mM sucrose inhibited sperm swim-up migration through sucrose layer. 2. Incubation for 25 minutes without sucrose layer significantly increased sperm swim-up migration. However, incubation for 10 minutes to induce swim-up through sucrose layer significantly stimulated sperm migration and maintained sperm movement. 3. There was no significant difference between Type I and Type II in barrier effect of sucrose layer. However, sucrose layer of Type II with shorter distance of barrier was efficient for sampling. To elucidate a function of follicular fluid on sperm chemotaxis using in vitro system of sucrose layer of Type II and incubation for 10 minutes, the effects of dilution, heat treatment, and protein and lipid extracts of follicular fluid on sperm swim-up separation were examined. And the results obtained were as follows; 4. Follicular fluid stimulated sperm migration and movement, and significantly-attracted capacitated-sperm at 10% level. 5. Follicular fluid heated at 55$^{\circ}C$ for 30 minutes maintained the effect of follicular fluid stimulating sperm migration and movement. 6. Follicular protein stimulated sperm movement that was reduced by filtration of the protein. 7. Follicular lipid didn't significantly stimulate sperm migration and movement. 8. Both of follicular protein and lipid reduced the effect of follicular fluid stimulating sperm migration and movement. In conclusion, sucrose layer could be used for a barrier against indiscreet sperm migration by swim-up. And follicular fluid stimulated migration and movement of sperm and attracted capacitated-sperm through sucrose layer. Especially, heat-resistant protein of follicular fluid stimulated sperm migration.

• Korean Journal of Animal Reproduction
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• v.23 no.1
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• pp.29-36
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• 1999

This study were performed the effect of methylxanthine derivatives on hamster epididymal sperm motility. To assess the effect of methylxanthine derivatives on motility kinematics of epididymal sperm, pentoxifylline, 2-deoxy-adenosine and hypoxanthine were added to TALP medium. 1 mM of pentoxifylline, significantly increased VCL, VAP, VSL of spermatozoa obtained from corpus and cauda epididymis. With 1 mM of 2-deoxyadenosine, VCL, VAP, VSL of spermatozoa obtained from corpus and cauda epididymis were significantly increased, but 2 mM ADE for cauda spermatozoa was effective than 1 mM. In the case of hypoxanthine, various concentrations were not significantly effective, but 2 mM HX showed higher effect than other concentrations. pentoxifylline 1 mM and 2-deoxyadenosine 1 mM significantly increased VCL, VAP of corpus and cauda epididymal spermatozoa and VSL of cauda epididymal spermatozoa. From these results it could be concluded that the addition of methylxanthine increase motility kinematics of hamster spermatozoa collected from epididymis.

• Journal of Embryo Transfer
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• v.18 no.1
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• pp.51-59
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• 2003

This study was carried out to evaluate the effect of different freezing and thawing rates on the viability, motility and acrosomal changes of frozen canine spermatozoa. The ejaculated semen was extended with Tris-egg yolk buffer containing 8% glycerol and equilibrated for 60 min after cooled to 4$^{\circ}C$ for 58 min. The straws were cryopreserved gradually by slow-cooling at different distance(6, 10 and 17 cm, respectively) from the liquid nitrogen (L$N_2$) to achieve temperature rate of 3, 8.9 and 19$^{\circ}C$ /min. Thawing of the straws was performed in a water bath fur 2 min at 37$^{\circ}C$ and 55$^{\circ}C$ , respectively. The motility of frozen-thawed spermatozoa was assessed by phase-contrast microscopy. To assess their viability and acrosome content, spermatozoa were stained with a vital stain and Fluorescence conjugated lectin Pisum Savitum Agglutinin (FITC/PAS), respectively. Concentration of the ejaculated fresh semen was normal range of 3.44 $\times$ 10$^{8}$ /ml. Freezing temperature were reduced to -110, -70 and -35$^{\circ}C$, as higher distance from liquid nitrogen, 6, 10 and 17 cm, respectively. Freezing at 3$^{\circ}C$/min in distance of 17 cm from liquid nitrogen yielded better motility, viability and rate of intact acrosome than 8.9 or 19$^{\circ}C$/min and the optimal thawing was 37$^{\circ}C$ for 2 min.