• 한국수정란이식학회지
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• 제28권2호
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• pp.113-119
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• 2013

The study focuses on the quality assessment of Black Bengal buck semen preserved at chilled condition. In this in vitro trial, collected semen from Black Bengal bucks was preserved at chilling temperature ($4{\sim}5^{\circ}C$) in tris-glucosecitrate yolk medium of 1:5 ratios for four days. Artificial Vagina (AV) method was utilized to collect semen from buck. General evaluation of semen includes the color, mass activity and density were measured by direct visual examination. However, computer-assisted sperm analysis (CASA) and phase contrast microscopy were used to figure out the motility (%), hyper-activated (HYP) motility (%) and number of abnormal spermatozoa (%) initially, and at every 24 h intervals. The result revealed that spermatozoa preserved at chilling temperature showed significantly (P<0.05) lower motility and HYP motility with the progression of preservation. The number of phenotypically abnormal spermatozoa significantly (P<0.05) increased following preservation. Although significant positive correlation (r=0.945; P<0.05) was existed between % motile and % HYP motile spermatozoa however, the % of morphologically abnormal spermatozoa was negatively correlated with % motile (r=-0.997; P<0.05) and % HYP motile spermatozoa (r=-0.946; P<0.01). Therefore, we concluded that the quality of chilled semen progressively losses its viability and doesn't remain useable after certain period of preservation with respect to its motility and morphology.

• 한국임상수의학회지
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• 제11권2호
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• pp.545-552
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• 1994

This experiment was carried out to investigate the renditions to maintain good post-thaw motility and viability of canine spermatozoa when the semen was frozen using methanol. The semen from two male dogs which had been proven to be fertile in the previous one year was treated with different compositions of semen diluent and was frozen at different freezing temperatures, When canine semen was frozen at-2$0^{\circ}C$, -6$0^{\circ}C$, or -8$0^{\circ}C$, the spermatozoa frozen and stored at -2$0^{\circ}C$ showed very low post-thaw motility and viability from day 2 to 7 and showed no viability since day 15 after freezing. The spermatozoa frozen and stored at -6$0^{\circ}C$ or -8$0^{\circ}C$ showed higher post-thaw motility and viability on day 2, 1, 15 and 30 after freezing than that frozen and stored at-2$0^{\circ}C$(p<0.01), with no difference between two groups. Among different composition groups of the semen diluents of control(tris + egg yolk + glycerol), egg yolk-free, 히ycerol-free, and tris-free, Prior to freezing, the egg yolk-free diluent showed significantly love. motility and viability than the other diluents(p<0.05). On each thawing day (from day 2 to 15 after freezing), control group showed considerably higher motility and viability than the other groups(p<0.01). The canine spermatozoa frozen and stored at -6$0^{\circ}C$ and -8$0^{\circ}C$ showed gradual decrease of motility from day 2 to 30 after freezing and the spermatozoa of these two groups thawed on day 30 showed considerably love. motility than those thawed on day 2 after freezing, respectively(p<0.01). These results indicate that the freezing temperature of either -6$0^{\circ}C$ or -8$0^{\circ}C$ can be applicable to the freezing method using methanol and also all of the components of the semen diluent including cryoprotectant, buffer and cold-shock buffer are very important to maintain motility and viability of canine spermatozoa in the freezing and thawing procedure.

• 한국가축번식학회지
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• 제23권2호
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• pp.127-132
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• 1999

본 연구는 소형견 정액의 원정액과 정장제거 정액의 일반성상, 채취분획별 및 단기보존시 정자의 생존성과 아울러 동결보존시의 정자의 생존성에 대해 조사하고자 수행하였다. 1. 원정액과 정장제거 정액에 saline 및 Tris-buffer를 희석 후 각각 일반성상 검사를 실시했을 때 원정액의 경우 정자농도는 5.07$\pm$2.32$\times$$10^{6}$cells/$m\ell$, 정자의 운동성은 95.42$\pm$2.65%, 기행정자수는 4.42$\pm$0.15%로 나타났으며, RSP(saline 및 Tris-buffer)군의 경우 정자농도는 4.69$\pm$3.27~4.25$\pm$3.65$\times$$10^{6}$cells/$m\ell$, 정자의 운동성은 91.17$\pm$3.85~88.52$\pm$3.85%, 기형정자수는 6.57$\pm$0.43~5.54$\pm$0.52%로 나타났다. 2. 분획별 채취 전정액중 제 1 분획에서는 정액량이 0.92$\pm$0.7$m\ell$, 정자농도는 4.57$\pm$0.78$\times$$10^{6}$cells/$m\ell$, 정자활력은 10.72$\pm$3,21%, 기형정자수는 5.50$\pm$0.70%로 나타났으며, 제 2분획에서는 정액량이 2.14$\pm$0.19$m\ell$, 정자농도는 2.01$\pm$0.12$\times$$10^{6}$cells/$m\ell$, 정자활력은 95.44$\pm$4.21%, 기형정자수는 4.31$\pm$0.53%로 나타났다. 또한, 제3 분획에서는 정액량이 2.66$\pm$0.23$m\ell$, 정자농도는 2.35$\pm$0.21$\times$$10^{6}$cells/$m\ell$, 정자활력은 90.71$\pm$2.63%, 기형정자수는 6.33$\pm$0.91%로 나타났다. 3. 원정액과 정장제거 정액을 4$^{\circ}C$ 와 2$0^{\circ}C$ 및 37$^{\circ}C$ 에서 각각 보존했을 때 보존시간별 정자의 활력은 2$0^{\circ}C$ 의 경우 1, 6, 13, 24, 30 및 40 시간에서 각각 98.51%와 98.32%, 86.32%와 92.15%, 83.71% 와 89.20%, 74.29% 와 82.08%, 52.98% 와 72.07%, 15.45% 와 60.02%, 2.41% 와 37.19% 의 정자활력을 나타내어 4$^{\circ}C$와 37$^{\circ}C$에 비해 높은 정자운동성을 나타냈다. 4. 제 2 분획 정액과 정장제거 정액을 각각 제 1차 및 제 2차 희석액으로 희석 평형 시킨 후 동결 융해했을 때 정자의 생존율은 각각 33.3$\pm$8.7, 54.7$\pm$9.5%로서 대조군의 15.4$\pm$5.2% 에 비해 높게 나타났다.

• 대한수의학회지
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• 제40권2호
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• pp.403-414
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• 2000

To improve the semen production, the selenium(Se) and vitamin E(Vit. E) were administrated into Hanwoo young sire for intensification an antioxidant system and the taurine were supplemented into semen extender for improving the semen characteristics. The 16 heads ranging from twenty to thirty two months of age were randomly assigned to control group, Se-admistrated group(Se-group), Vit. E-administrated group(Vit. E-Group) and Se and Vit. E administrated group(Se and Vit. E-group). Se and Vit. E dministrated 3 times every 30 days by intramuscular injection. The administration of Se, Vit. E, and Se and Vit. E didn't affect on semen volume, sperm concentration, and total sperm number among all groups(p>0.05). Adiministration of Se improved sperm motility and viability. Motility of Se-group and control were 26.01% and 19.20%, respectively(p<0.05). Viability of Se-group(47.07%) was higher than control group(35.73%), Vit. E group(36.55%)(p<0.05). The administration of Se and Vit. E didn't affect sperm capacitation and acrosome reaction. The 100mM taurine supplement into semen extender increased the motility of frozen/thawed semen in the Vit. E-group(p<0.05) and had a beneficial effects on decreasing abnormality of frozen/thawed semen in all groups(p<0.05). These results indicate that Se administration improve sperm motility and viability, and the taurine supplement into semen extender decrease abnormality in Hanwoo young sire.

• Reproductive and Developmental Biology
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• 제35권1호
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• pp.55-60
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• 2011

The objective of this study was carried out to evaluate the efficiency of sperm collection methods on the post-thaw viability of cat semen. The cat semen was collected by artificial virginal (AV) and electronic ejaculate (EE) methods. The composition of semen extender was consisted of Tris-buffer supplemented with 20% egg yolk and 1% P/S antibiotics in Ext I, and more added 8% glycerol, 1.0% Equex STM paste of total volume in Ext II. The collected semen was adjusted the concentration and then diluted in Ext I for optimal concentration. The diluted semen was cooling to $5^{\circ}C$ temperature in refrigerator for at least 2 hrs and then diluted stepwise with Ext II for at least 1 hrs. After an equilibration for 1 hrs, the cooled semen was packaged in 0.5 ml straw and then freezing on the $LN_2$ vapor over 5 cm above from $LN_2$ and then immersed directly in $LN_2$ for cryopreservation. The frozen semen was thawed in $38^{\circ}C$ water for 15 sec and then evaluated the motility, viability, and morphology. Post-thaw semen were calculated the motility by SMI (sperm motility index). The live-dead sperm was evaluated by Eosin-B and morphological evaluation was by Diff-quik kit staining. The post-thaw concentration ($89{\times}10^6$ /ml vs. $128{\times}10^6$ /ml), viability ($22.6{\pm}10.6%$ vs. $37.1{\pm}26.1%$), morphological normality ($27.0{\pm}50.2%$ vs. $45.6{\pm}123.0%$) of EE and AV groups were not significant different, but the post-thaw motility was significant lower in EE than that in AV group ($53.1{\pm}3.6$ vs. $73.6{\pm}5.7$) (p<0.05). In conclusion, semen collection methods did not significant different between EE and AV groups except of post-thaw motility and so both semen collection methods could be applied in feline semen collection methods.

• 한국수정란이식학회지
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• 제25권4호
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• pp.263-266
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• 2010

Prediction of semen's fertilizing ability used in artificial insemination (AI) is one of very important factors on pig reproductive performance. In vitro fertilization (IVF) has been used for indirect evaluation of sperm's fertilizing ability and it has been showed as highly correlated index. In swine industry, increasing interest in preservation of boar semen raises questions on the sperm motility from semen used in commercial AI centers. Mitochondria in sperm mid-piece generate the energy to support motility and could be an explanation of impaired fertility. Objective of this study was to suggest usable sperm motility to farms in measuring the effect of sperm motility and sperm abnormality on in vitro production of embryo in which sperm's fertilizing ability can be determined indirectly. Semen samples were provided from local AI center and used within 3 days after collection. Semen samples were divided by 4 different motile groups (>70%; 61~70%; 51~60%; <50%) using CASA (computer-assisted sperm analysis) on the days of IVF. Developmental rate to the blastocyst stage from over 61% motile sperm group showed significantly higher rate than below 60% motile sperm group ($16.5{\pm}0.7{\sim}18.4{\pm}0.8%$ vs $6.3{\pm}0.8{\sim}11.5{\pm}0.7%$, p<0.05). In experiment to determine the relationship between sperm motility and viability and abnormality, over 61% motile sperm groups showed significantly higher viability rate compared to below 60% motile sperm groups ($84.8{\pm}4.0{\sim}88.1{\pm}4.0%$ vs $69.1{\pm}4.0{\sim}74.2{\pm}4.0%$, p<0.05). On the other hand, morphological sperm abnormality showed significantly higher in over 70% motile sperm group ($10.2{\pm}2.2$ vs $16.0{\pm}2.2{\sim}21.0{\pm}2.2%$, p<0.05). In experiment to find the correlation between sperm motility of 4 different motile groups and amount of mitochondria, lower motility group also showed lower level of mitochondria (p<0.05). The mitochondria parameter used in this study showed another possibility to differentiate the sperm motility. Taken together, because below 60% motile semen used in AI reduce the fertility, AI centers should provide the over 60% motile sperm to the farms at the time of AI.

• Asian-Australasian Journal of Animal Sciences
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• 제19권4호
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• pp.486-494
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• 2006

In order to protect the spermatozoa against cold shock, hen egg yolk is widely used as a cryoprotective agent in semen freezing extenders for domestic animals. The protective action of yolk is largely presumed to be due to low density lipoproteins (LDL). The effects of LDL on sperm quality of bull and northern pike (Esox lucius) after freezing-thawing have been reported, but no study has been made to evaluate the effect of LDL on boar sperm motility and other characteristics. The experiment was carried out to investigate the effect of LDL on the freezing of boar sperm in 0.25 ml straws. The aim was to evaluate the quality of boar spermatozoa cryopreserved in the presence of LDL. Motility of semen cryopreserved in LDL was analyzed and compared to semen cryopreserved with Tris-citric acid-glucose (TCG) and Tris-citric acid-fructose (TCF), two basic freezing extenders containing egg yolk. Similarly, acrosome and plasma membrane integrity were also evaluated and compared to semen cryopreserved with TCG and TCF. Analysis of sperm quality after freeze-thaw showed that the motility, acrosome and plasma membrane integrity were improved with LDL in the extender, as compared to the TCG and TCF. The highest post-thaw integrity of acrosome and plasma membrane and motility were obtained with 9% LDL (w/v). Consequently, the optimum LDL concentration in the extender was 9%. It is also suggested that the concentration of LDL addition is important for the effect on boar sperm protection during freezing and thawing. The percentage of motile spermatozoa was significantly higher after freezing in 9% LDL than in TCG and TCF 54.4% versus 30.4% and 30.1% (p<0.05), respectively. The integrity of acrosome and plasma membrane were also significantly higher at 70.3% and 50.5% respectively with semen frozen in 9% LDL extender compared to TCG at 37.8% and 30.3% and TCF at 36.4% and 29.9%, respectively (p<0.05),. In conclusion, we propose that extender containing LDL extracted from hen egg yolk could be used as a cryoprotective media with a better efficiency than TCG and TCF. LDL improved boar semen quality, allowing better spermatozoa motility, acrosome and plasma membrane integrity after the freeze-thaw process. Furthermore, we found out that the extender with 9% LDL concentration significantly enhanced motility, acrosome and plasma membrane integrity of boar sperm after freezing and thawing.

• 한국수정란이식학회지
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• 제30권3호
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• pp.143-147
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• 2015

Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Various researches have been trying to improve the quality of semen Post-thawed in boar. Recently, polymorphism (g. 35756 T>C) of Estrogen Receptor 1 (ESR1) gene reported to be significant association with MOT. This study was conducted to evaluate the ESR1 gene as a positional controlling for motility and kinematic characteristics of post-thawed boar semen. To results, The g.35756 T>C SNP of ESR1 was significantly associated with frozen semen motility and kinematic characteristics. The g.35756 T>C SNP was high significantly associated with MOT, VCL, VSL and VAP (p<0.001). The SNP was also significantly associated with ALH (P<0.05). Therefore, we suggest that the g. 35756 T>C polymorphism in the intron 1 region of the porcine ESR1 gene could potentially be applied in frozen semen programs to improve MOT trait, but only after validation in other populations.

• 한국수정란이식학회지
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• 제30권3호
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• pp.137-142
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• 2015

Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Various researches have been trying to improve the quality of semen post-thawed in boar. Recently, polymorphism (g.158T>C) of phospholipase C zeta (PLCz) gene reported to be significant association with MOT. This study was conducted to evaluate the PLCz gene as a positional controlling for motility and kinematic characteristics of post-thawed boar semen. To results, The g.158 T>C SNP of PLCz was significantly associated with frozen semen motility and kinematic characteristics. g.158 T>C SNP was high significantly associated with MOT, VCL, VSL and VAP (p<0.0001, p=0.0002, p<0.0001 and p<0.0001, respectively). Therefore, we suggest that the intron region of the porcine PLCz, may be used as a molecular marker for Duroc boar post-thawed semen quality, although its functional effect was not defined yet. Whether the association is due to the candidate gene or not require further verification. Thus, it will be of interest to continue association studies in the regions surrounding those genes.

• Clinical and Experimental Reproductive Medicine
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• 제41권4호
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• pp.165-167
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• 2014

Objective: The objective of this study is to estimate the effects of Inclear, a feminine cleanser, on sperm motility. Methods: Semen samples were obtained from infertile male patients. Following liquefaction, the raw semen samples were diluted with Ham's F-10 nutrient mixture medium containing 0.4% human serum albumin solution at a ratio of 1:3. The semen samples were subsequently centrifuged to separate the seminal plasma from the serum. The supernatant was discarded, and the pellet was resuspended. The sample was again centrifuged to remove cell debris, and the supernatant was removed. The final pellet was gently loosened by resuspension and incubated in medium alone as a control, and in a 10% solution of the medium plus Inclear. A sampling time of 30 minutes was selected on the basis of sperm transport studies. Sperm motility was evaluated with computer-assisted sperm analysis. Results: A total of 20 samples were analyzed. The mean age of patients was $34.40{\pm}2.96years$. There was no difference in sperm concentration and motility in the two samples at 0 minute and 30 minutes of incubation. In both semen samples, the sperm concentration and motility decreased after an incubation period of 30 minutes. However, there was no statistical difference between the samples. Sperm concentration and motility were not significantly different between the control and Inclear samples after 0 minute and 30 minutes of incubation. Conclusion: Inclear has no negative effects on sperm motility. This product can be recommended to pregnancy planners for vaginal hygiene and as a vaginal lubricant.