• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.119-124
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• 2002

This study was carried out to investigate the effects of packing materials of frozen boar semen to improve reproductive performance efficiency in pig. Boars were raised at Swine Artificial Insemination Center in National Livestock Research Institute, Sunghwan, Chungnam, Korea. We compared packing protocols for frozen boar semen among 5$m\ell$ maxi-straw, 5$m\ell$ cryogenic-vial, and aluminum-pack. Cryogenic-vial packing material showed similar sperm characteristics compared with maxi-straw packing material when the sperm was frozen above 15cm from liquid nitrogen and thawed at 52$^{\circ}C$ for 190 seconds. We investigated different thawing times to find out the optimal condition of freezing and thawing protocol with cryogenic-vial. Freezing above 15cm from liquid nitrogen and thawing at 52$^{\circ}C$ for 190 seconds were the optimal protocol compared with 120 and 150 seconds. However, normal acrosome rates did not show any differences among thawing times. Post-thawing results of maxi-straw in water at 52$^{\circ}C$ for 45 seconds had better total motility and curve linear velocity than those of cryogenic-vial in water 52$^{\circ}C$ for 190 seconds. However, there were no differences on straightness and normal apical ridge of sperm between maxi-straw and cryogenic vial. Non-return rate, farrowing rate and litter size of sows inseminated with frozen boar semen of commercial farms were higher in the maxi-straw than cryogenic-vial, but there were no significant differences between maxi-straw and cryogenic-vial. In conclusion, there were no significant differences between maxi-straw and cryogenic-vial and so, we may replace cryogenic-vial packing method instead of maxi-straw packing method by improvement of freezing and thawing rate.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.77-77
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• 2001

본 연구는 돼지 동결정액의 번식성적을 개선코자 기존의 maxi-straw와 cryogenic-vial을 이용하여 포장방법에 따른 동결방법과 융해방법별 정액성상 및 번식성적을 비교하였다. 동결방법은 두 가지 포장방법 모두 정액의 양(5$m\ell$)과 농도(5.0$\times$$10^{9}$/dose)가 동일한 조건으로 처리하였으며, LYE(Lactose egg york extender) 보존액으로 희석하여 액체질소 상단 15cm에서 20분간 동결하였다. 융해방법은 maxi-straw는 52$^{\circ}C$에서 45초간 cryogenic-vial은 52$^{\circ}C$에서 190초간 융해하여 $25^{\circ}C$로 가온 된 80$m\ell$ BTS (Beltsville thawing solution) 보존액과 혼합하였다. 정액성상검사는 정자자동분석기(SAIS : Sperm Analysis Image System, Korea)를 이용하였다. 총활력(TM : Total motility)과 정자의 빠르기(VCL : Curve linear velocity)는 maxi-straw가 54.3%와 46.6%로 cryogenic-vial의 35.6%와 36.6%보다 우수하였다(P<0.05). 정자의 직진성(STR : Straightness)과 NAR은 maxi-straw가 53.2%와 32.6%로 cryogenic-vial의 47.3%와 29.8%와 비슷한 경향을 나타내었다. 수태율과 분만율 및 총산자수는 maxi-straw가 77.3%, 68.2% 및 8.0두로 조사되어 cryogenic-vial포장방법의 66.7%, 61.9% 및 7.4두보다 다소 우수하였으나 통계적인 유의차는 인정되지 않았다. 이상의 결과로 볼 때 cryogenic-vial방법이 새로운 돼지 동결정액 포장방법의 가능성을 나타낸다고 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.4
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• pp.507-515
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• 2009

A series of experiments were conducted to determine the suitable freezing and thawing temperatures for the freezing of boar semen in 5 ml maxi-straws. The ultrastructure, in vitro fertilization (IVF) and artificial insemination (AI) of frozen-thawed semen were also be evaluated. The 5 cm freezing height gave the best results not only in post-thaw motility rate (54.00%), but also in normal acrosome morphology rate (NAR) (80.23%). There was no significant difference in the post-thaw motility between different thawing temperatures and corresponding thawing times (p>0.05); the group of $52^{\circ}C$ and 25 s gave the highest motility rate (45.00%). As a whole, not only from the motility but also the NAR, thawing at $42^{\circ}C$ was better than the other two treatments. In the freezing packages, 5 ml maxi-straw gave a little lower mobility (40%), viability rate (49.58%), plasma membrane integrity rate (53.91%) and NAR (52.65%) than the 0.25 ml straw, but there was no significant difference between the two straw volumes (p>0.05). The IVF capacity of frozen-thawed semen in this experiment was similar to fresh semen. From ultrastructure observation, the main damage to boar spermatozoa after freezing was seen in the acrosome, such as swelling and formation of vesicles. After AI in recipient Shanghai White sows, frozen-thawed semen from 5 ml maxi-straws and pellets produced 72.2% and 80% conception rate and 7.8 and 8 litter sizes, respectively, and there was no significant difference between the 5 ml maxi-straw and the pellet (p>0.05).

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.11
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• pp.1553-1558
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• 2002

This study was carried out to obtain informations regarding the effect of N-acetyl-D-glucosamine in the LEY (lactoseegg yolk) diluent according to incubation time in 5 ml maxi-straw and the effects of freezing rate, thawing temperature and thawing time in the LEN (lactose-egg yolk and N-acetyl-D-glucosamine) diluent on acrosome morphology and motility of frozen-thawed boar sperm. The study showed that the LEN diluent was higher post-thaw NAR (normal apical ridge) acrosome than the LEY diluent for 0.5 h incubation at 37$^{\circ}C$. However, there were no differences between the LEN and LEY diluents on post-thaw sperm motility according to incubation time. The straws frozen from 5.0 cm (20$^{\circ}C$/min) to 17.0 cm (1$^{\circ}C$/min) above the liquid nitrogen surface did not show any significant differences on post-thaw sperm motility. However, the straws frozen above 5.0 cm from the liquid nitrogen surface were higher NAR acrosome than those frozen above 17.0 cm. The post-thaw percentages of motile sperm and NAR acrosome were significantly higher (p<0.05) for the maxi-straws submerged for 40 or 45 sec in a 52$^{\circ}C$ water bath than for 30, 35, 50 or 55 sec. The mean sample temperatures of maxi-straws after 40 or 45 sec submersion were 20.7 or 26.4$^{\circ}C$. In conclusion, the sample temperature of the thawed semen was very important for post-thaw sperm survival in the LEN diluent of 5 ml maxi-straw. When the temperature of the thawed semen was 20.7$^{\circ}C$, the percentages of motile sperm and NAR acrosome were highest.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.111-117
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• 2002

This study was carried out to investigate the effects of frozen boar semen on reproductive performance in swine artificial insemination (AI). Many factors, which were breeds, time of insemination, sperm concentration per dose, farm and year were investigated to improve reproductive performance efficiency. Boars were raised at Swine Artificial Insemination Center in National Livestock Research Institute, Sunghwan, Chungnam, Korea. This experiment was carried out from 1995 to 2000. There were no differences in swine AI with frozen boar semen using 5$m\ell$ maxi-straw among 3 breeds (Landrace, Yorkshire, Duroc), 2 or 3 times insemination per estrus, and 3 different sperm numbers of 3.0, 4.0, and 5.0$\times$10$^{9}$ per dose of insemination. However, non-return rate and litter size of sows inseminated with frozen boar semen of commercial farms were different according to farm management system and inseminator's skill. Conception rate, farrowing rate and number of pigs born alive per litter by artificial insemination with frozen boar semen (5$m\ell$ maxi-straw) from 1995 to 1999 was 68.3~74.6%, 61.7~67.6% and 8.1~8.7 heads.

• 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.

• Journal of Embryo Transfer
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• v.11 no.1
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• pp.81-83
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• 1996

This study was carried out to investigate the effect of sperm concentration of 5ml maxi-straw on farrowing rate and number of pigs born alive per litter in deep frozen boar semen. We did not find out the effect of sperm concentration on post-thaw sperm motility and NAR acrosome. However, farrowing rate and number of pigs born alive per litter of 7. 5 x 10˚ /5ml and 10.0 x 10˚ /5m1 sperm concentrations were higher than those of 5. 0 /10˚ /5ml sperm concentration.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.5
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• pp.612-616
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• 2004

This study was carried out to compare the semen characteristics, frozen-thawed sperm viability and testosterone concentration and in vitro fertilization (IVF) and development of in vitro matured pig oocytes between two Yorkshire boars. Semen and blood samples were collected once per week from October to November 2002 from two adult Yorkshire boars at 18 months of age with 170 kg body weight. Sperm were deep frozen in 5 ml maxi-straws with lactose-egg yolk and N-acetyl-D-glucosamine (LEN) diluent and stored in liquid nitrogen. Blood samples were obtained at 10 a.m. by inserting a 21 gauge, hypodermic needle attached to 10 ml syringe into surface veins in the ear. The concentration of testosterone was determined by Competitive Enzyme Immunoassay. Ovaries were collected from prepubertal gilts at a local slaughter house. Cumulus oocyte complexes were aspirated from antral follicles (3 to 6 mm in diameter). The medium used for oocyte maturation was modified TCM 199. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at $38.5^{\circ}C$, 5% $CO_2$ in air. For IVF, one frozen 5 ml straw was thawed at $52^{\circ}C$in 40 sec and was diluted with 20 ml Beltsville thawing solution at room temperature. Sperm were washed 2 times in mTLP-PVA and inseminated without preincubation after thawing. Oocytes were inseminated with $2{\times}10^7$/ml sperm concentration. Oocytes were coincubated for 6 h in 500 ${\mu}$l mTBM fertilization medium. At 6 h after IVF, oocytes were transferred into 500 ${\mu}$l NCSU-23 culture medium for further culture of 48 and 144 h. There were no significant differences in the semen volume, motility, normal acrosome morphology and sperm concentration of raw semen between A and B of Yorkshire boar. However, motility and normal acrosome of boar A were higher than those of boar B at 0.5, 2, 3, 4, 5 and 6 h incubations of frozen-thawed sperm. Testosterone concentration (3.75 ng/ml) of boar A was higher than that (2.34 ng/ml) of boar B. The rate of blastocyst formation (15.1%) of boar A was higher than that (10.4%) of boar B. In conclusion, serum testosterone concentration of boar showed very important role for the frozen-thawed sperm viability and the blastocyst formation of pig oocytes matured in vitro.