• Journal of Embryo Transfer
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• v.20 no.1
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• pp.43-48
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• 2005

This experiment was carried out to investigate the extender, cooling rate and concentration of glycerol for freezing of boar semen. The result obtained were summarized as follows: 1. Optimal cooling rate was $0.17\~022^{\circ}C/min$ from 25 to $5^{\circ}C$ in LEY extender on the viability and normal acrosome after thawed. 2. The LEY extender was effective in protecting frozen boar semen from cold shock among the extenders(p<0.001, respectively). 3. The sperm viability and normal acrosome rates after thawing was showed greater in the 3 or $4\%$ of glycerol concentration than $2\%$ in LEY extender. 4. Viability of sperm was higher when both 15mM of fructose and 3 or $4\%$ glycerol were added to the LEY extender compared with other concentrations of fructose and glycerol were added it(p<0.001).

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2011.10a
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• pp.745-748
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• 2011

현재 양돈산업에서의 인공수정(Artificial Insemination, AI) 기술은 1994년 이후 본격적으로 국내 양돈농가에 보급되어 양돈 산업 발전에 많은 영향을 주었다. 현재 국내 비육돈 임신의 80% 이상이 인공수정 센터에 공급하는 정액에 의해 이뤄지는 등 양돈 산업에서 차지하는 비중과 중요성이 매우 중요하며, 또한 국내 양돈 산업분야의 인공수정 기술은 번식과 육종개량 분야에서 절대적인 영향을 미치고 있다. 이러한 중요성에도 불구하고 전국 AI센터의 정액 생산 및 공급에 관한 기록은 53%이상 수기에 의존하고 있다. 그나마 이에 대한 수집된 현장자료는 전국 AI센터의 개별시스템에 수작업으로 입력 관리되어 원시 데이타의 오류 및 통합정보 활용이 매우 어렵다. 이에 XML을 기반으로 전국 AI센터의 통합정보 관리시스템 모델을 제안한다. 제안된 정보관리 모델은 웹을 기반으로 전국 AI센터의 정보를 통합관리 활용할 수 있으며, 이를 통해 통합된 정보의 통계분석, 미래 예측분석 자료 등으로 활용되어 효율적인 돼지개량 체계를 구축할 것으로 기대한다.

• Korean Journal of Animal Reproduction
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• v.23 no.4
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• pp.333-335
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• 1999

This study was carried out to find out the effect on fertilizing capacity according to sperm concentration of liquid boar semen. Four different doses with various motile sperm cells of 3.0$\times$10$^{9}$ , 2.5$\times$10$^{9}$ , 2.0$\times$10$^{9}$ , and $1.5\times$10$^{9}$ per 80$m\ell$ plastic bottle were inseminated twice 12 h interval after standing estrus in 6,818 sows. Farrowing rate and total piglets per litter were 82.2% and 10.9, respectively, with no significant differences among the other treatments. The presumption of optimal concentration of motile sperm cells in the liquid boar semen was best at 2.0~2.3$\times$10$^{9}$ per dose.

• Journal of Animal Science and Technology
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• v.49 no.6
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• pp.729-736
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• 2007

The objective of this study was to investigate the anti-oxidative effects of pyruvate, taurine and melatonin on sperm characteristics(motility, membrane integrity) and lipid peroxidation(LPO) for in vitro storage of boar semen. Semen was treated with various antioxidants such as pyruvate(1mM), taurine(50mM) and melatonin(100nM) with or without 100uM H2O2. Antioxidant treatments were significantly increased the sperm motility when compare to control group in all incubation periods(P≤0.05). Hypoosmotic swelling test(HOST), membrane integrity was similar to the result of motility. In lipid peroxidation measurement by TBA reactions of spermatozoal plasma membrane, malondialdehyde(MDA) level in control and antioxidant treatments were lower than those of antioxidant plus H2O2 or H2O2 treatment for 3 to 6 h incubation period. Relationships of evaluation methods for sperm viability were investigated by motility, membrane integrity and lipid peroxidation. Among evaluation methods, LPO vs motility and membrane integrity vs LPO were negatively correlated(－0.23~－0.92 and －0.68~－0.85), but membrane integrity vs motility was positively correlated (0.53~0.94) in all treatments. These experiments indicate that supplementation of antioxidant to the semen extender can increase the sperm motility and membrane integrity and decrease the lipid peroxidation of spermatozoal plasma membrane. The HOST might be utilized to evaluate the sperm quality instead of lipid peroxidation or motility.

• Journal of Life Science
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• v.30 no.1
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• pp.77-81
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• 2020

The aim of this study was to overcome some of the limiting factors that the maxi cryopreservation straw of 5 ml presents in processing boar semen. Cryopreservation of semen samples was conducted in 0.5 ml and 5.0 ml straws at two freezing rates: -140℃ in 8 minutes and 30 seconds (FR-1) and -140℃ in 14 minutes (FR-2). The straws were then thawed and the semen parameters were compared by Computer Assisted Sperm Analysis, and sperm morphology and acrosome status were examined by Coomassie blue staining. The effects of different thawing temperatures and durations were also compared, namely 37℃ for 115 sec, 50℃ for 45 sec, or 70℃ for 25 sec. In general, the FR-1 group showed higher (p<0.05) sperm viability and motility than the FR-2 group in the 5.0 ml straws. Compared to other ranges, thawing at 50℃ for 45 sec showed the highest sperm viability and motility (68.4±3.6% and 69.5±2.2%, p<0.05), suggesting that thawing temperature should be adjusted concurrently with freezing rate. Sperm morphology and acrosome integrity did not significantly differ among the groups (p>0.05). The data obtained in this study suggest that improving the freezing-thawing protocol for one artificial insemination dose straws (5.0 ml) retains the sperm's parameters from 0.5 ml cryopreservation, and is more convenient to handle, which could result in enhanced reproductive performance.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.25-25
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• 2003

본 연구는 돼지 동결-융해 정자의 배양에 의한 체외수정능력과 glycosidase activity의 관계를 검토하였으며, 또한 돼지난자의 투명대내에서 발견된 당잔기에 대한 정자의 glycosidase 특이성을 확인하기 위하여 $\alpha$-L-fucosidase, $\alpha$-D-mannosidase, $\beta$-D-galactosidase 및 N-acetyl-$\beta$-D-glucosaminidase ($\beta$-GlcNAc'ase)의 activity를 분석하였다. 그 결과 glycosidase activity는 동결정자의 융해 후 배양하지 않았을 때보다 2시간 배양했을 때 더 높게 나타났다. $\beta$-GlcNAc'ase의 activity는 정자 배양 유무에 관계없이 다른 glycosidase 처리시보다 최소한 2배 이상 높게 나타났다. 또한 첨체반응이 유기된 정자의 비율은 glycosidase ($\alpha$-D-mannosidase; P<0.05)에 의해 영향을 받았으며 정자를 배양하지 않은 경우보다는 배양된 정자에서 높게 나타났다. 그러나 배양시간에 따른 정자의 생존성에 대해 glycosidase의 종류에 따른 유의차는 인정되지 않았다. 한편 투명대내 정자의 접착과 침입에 대한 또 다른 실험에서, 서로 다른 glycosidase가 첨가된 배양액내에서 수정된 정자가 배양시간이 길어짐에 따라 정자의 침입율은 낮아졌다($\beta$-GlcNAc'ase; P<0.05). 투명대내의 정자접착 정도는 glycosidase의 첨가시에 무첨가시보다 접착정도가 더 높았으며, 가장 높은 접착율은 $\beta$-GlcNAc'ase첨가시 나타났다. 또한 모든 glycosidase 처리시 2시간 배양한 정자보다는 배양하지 않은 정자에서 투명대에 대한 접착정도가 높게 나타났으며, $\alpha$-D-mannosidase의 처리시 유의적인 차이를 보였다(P<0.05). 본 연구의 결과, $\beta$-GlcNAc'ase가 주로 돼지정자의 원형질막내에 존재하는 것으로 추측되며, 배양된 정자에 의한 투명대 접착정도와 침입율이 낮았음에도 불구하고 glycosidase activity가 증가하는 것으로 나타났다

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.263-271
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• 2006

The purpose of this study was undertaken to compare ability of frozen-thawed sperm characteristics between two strains (miniature pig and Duroc). The semen was collected by gloved-hand method into a pre-warmed ($37^{\circ}C$) thermos bottle. The semen was diluted with same volume extender and added to LEY solution for freezing. The diluted semen was placed in 0.5 ml straws, and freezing was initiated by exposing the straws to liquid nitrogen ($LN_2$) vapours for 10 min before placing them into $LN_2$ for cryopreservation. The frozen-semen straw were thawed at 20, 37 and $50^{\circ}C$ for 1 min, 45 sec and 10 sec within water-bath. The semen sample were evaluated at 0, 3, 6, 9, and 12 h after incubation at $37^{\circ}C$ for analysis of sperm ability. Abnormality of spermatozoa in miniature pig was significantly (p<0.05) higher than that in Duroc at 0, 9 and 12 h of post-thawing incubation after frozen-thawing. The percentage of F-patterned spermatozoa in miniature pig was significantly (p<0.05) lower, while the percentage of AR (acrosome reacted spermatozoa) pattern was higher in the miniature than in the Duroc. On the other hand, there was no significant difference in the viability of spermatozoa thawed at different temperature ($20^{\circ}C\;and\;37^{\circ}C$) between two species, but the viability in miniature pig was higher (p<0.05) than in Duroc when sperm was thawed at $50^{\circ}C$. In conclusion, this study suggest that suitable freezing method for miniature pig semen is required for increasing post-thawing viability and fertilization capacity.

• ANNALS OF ANIMAL RESOURCE SCIENCES
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• v.29 no.4
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• pp.166-171
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• 2018

The effect of bisphenol S (BPS) on the viability and production of reactive oxygen species (ROS) was studied in boar sperm and ovarian granulosa cells. Boar semen was incubated in Beltsville thawing solution with either 0 or $5{\mu}M$ BPS for 3 and 6 h. The viability of sperm was analyzed by SYBR14/PI doubling staining, and production of ROS was detected. Ovarian granulosa cells were also treated with BPS for 24, 48, and 72 h. Then, cell viability (0, 5, 10, and $20{\mu}M$) and ROS production (only 0 and $5{\mu}M$ BPS) were assessed. The results showed that, BPS decreased sperm viability at 3 and 6 h, and that BPS increased ROS production (p<0.05). Also, BPS reduced the viability of ovarian granulosa cells (p<0.05), and stimulated ROS production (p<0.05). These results suggest that BPS damages sperm activation and ovarian granulosa cells in the reproductive system.

• 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.

• Journal of Embryo Transfer
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• v.19 no.3
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• pp.265-273
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• 2004

This study was carried out to examine the effects of fertilization media and sperm concentration on in vitro fertilization (IVF) and development (IVD) of porcine oocytes matured in vitro. Cumulus-oocyte complexes (COCs) were collected from antral follicles of porcine ovaries collected from abattoir, and were matured in vitro in modified NCSU-23 (mNCSU-23) supplemented with 10％ porcine follicular fluid (pFF). After the fertilization by experimental scheme, putative embryos were developed in vitro in NCSU-23. The results are as follows. When the oocytes were fertilized in vitro in modified TBM or modified TLP-PVA by 1 ${\times}$10$^{5}$ sperm/$m\ell$, all of the fertilization parameters were not significantly different between two media. Subsequently, as these putative embryos were developed in vitro in NCSU-23, the percentage of oocytes cleaved and of blastocysts were not different between two media, either. When the oocytes were fertilized in vitro in mTBM by 5${\times}$10$^4$, 1${\times}$10$^{5}$ or 5${\times}$10$^{5}$ sperm/$m\ell$, all of the fertilization parameters were significantly (P<0.05 or P<0.01) increased as sperm concentration was elevated. Subsequently, as these putative embryos were developed in vitro in NCSU-23, the percentage of oocytes cleaved and of blastocysts were significantly boosted (P<0.01) as sperm concentration at fertilization was elevated from 5${\times}$10$^4$ to 1${\times}$10$^{5}$ sperm/$m\ell$, but were not different between 1${\times}$10$^{5}$ and 5${\times}$10$^{5}$ sperm/$m\ell$.