• Asian-Australasian Journal of Animal Sciences
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• v.19 no.12
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• pp.1716-1721
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• 2006

The present study was conducted to observe the effect of osmolality of glycerolated TEST-yolk glycerol extenders on post-thawing sperm kinematics of ram spermatozoa of the native Malpura breed maintained in a semi-arid tropical environment. Good quality semen obtained from adult rams was pooled, split and diluted to 1,000 million spermatozoa per ml in complete TEST-yolk-glycerol extenders of 900, 1,200, 1,500 and 1,800 mOsm/kg osmolality. Diluted semen samples were loaded in 0.25 ml straws and cooled down to $-125^{\circ}C$ freezing temperature at the rate of $-25^{\circ}C$ per minute under controlled conditions before plunging into liquid nitrogen for storage. The thawing of straws was performed at $50^{\circ}C$ in a water bath for 10 seconds and sperm kinematics of the frozen-thawed spermatozoa were assessed by a computer-assisted sperm analysis technique. Osmolality of diluent had no significant effect on post-thawing % motility, % rapid, % medium and % slow moving frozen-thawed spermatozoa but significantly (p< 0.05) affected the % linearity and % straightness. The post-thawing % motility and % rapid motile spermatozoa were highest in samples extended in diluent of 1,500 mOsm/kg osmolality and lowest in 900 mOsm/kg. The curvilinear velocity of spermatozoa was significantly (p<0.05) higher for samples extended in 1,800 mOsm/kg, compared to those in 900 and 1,200 mOsm/kg, but the effect was not significantly different to those extended in diluent of 1,500 mOsm/kg osmolality. The study indicated that ram spermatozoa could tolerate a wide osmolality range for dilution in the complete TEST-yolk-glycerol extender for their cryosurvival. The highest recovery of motile spermatozoa following thawing was achieved in samples extended in the TEST-yolk-glycerol diluent of 1,500 mOsm/kg osmolality.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2002.11a
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• pp.116-117
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• 2002

This study was conducted to investigate the effects of dilution rate and stored time of semen at 5, 25 and 35$^{\circ}C$ on mobility in liquid rooster semen. At 5$^{\circ}C$ cold temperature, no significant difference were found in sperm mobilities on dilution rate and stored time among treatments stored at 5$^{\circ}C$. Sperm mobility for the conservation of 1 hours at 25$^{\circ}C$ and 3 hours at 35$^{\circ}C$ were significantly higher for 1：2 and 1：3 dilution rate(semen： diluent) groups than for 1：1 dilution rate group(P〈0.05), In Fertility results after artificial insemination with different number of sperm per dole, fertilization rate of liquid rooster semen diluted with skim milk-glucose solution were 90.67, 94.00, 96.00, and 98.67% for 0.2${\times}$10$\^$8//dose, 0.4${\times}$10$\^$8//dose, 1.0${\times}$10$\^$8//dose, and 2.0${\times}$10$\^$8//dose groups, respectively. To have more than 90% fertility, 0.2${\times}$10$\^$8/ sperm per dose for the artificial insemination (AI) could be used, And to have more 94% fertility, 0.4${\times}$10$\^$8/ sperm per dose AI could be used practically.

• Journal of Embryo Transfer
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• v.27 no.1
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• pp.51-55
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• 2012

The objective of this study was to investigate the motility and kinematics of boar sperm that while stored at 4C. The samples of fresh boar semen were place into an extender, Androhep, and stored at $4^{\circ}C$. In three of these samples, cryoprotectants were added. The sperm's motilities and kinematics were evaluated by using microscope (${\times}400$) and the viability status was evaluated by using with eosin staining method. The 5 sample groups are; Goup A:Androhep (extender), stored at $17^{\circ}C$. Group B:Androhep (extender), stored at $4^{\circ}C$. Group C:Androhep (extender), + 3% glycerol (cryoprotectant), stored at $4^{\circ}C$. Group D:Androhep (extender), + 3% DMSO (cryoprotectant), stored at $4^{\circ}C$. Group E:Androhep (extender), + 3% ethylene glycol (cryoprotectant), stored at $4^{\circ}C$. In group A, the sperm's motility was reduced. On day one the sperm's motility was ($85.7{\pm}2.3$) and day 5 the motility was ($43.9{\pm}3.3$). In group B, C and D the sperm's motility were reduced to 0 on day 5. In group E the sperm's percentage of motility decreased. On day one the sperm's motility was ($42.0{\pm}0.5$) and day 5 the motility was ($2.3{\pm}0.3$). When comparing cryoprotectant in samples of boar sperm there is a slight improvement in the results when the use of Androhep Lite (extender), + 3% ethylene glycol (cryoprotectant), stored at $4^{\circ}C$ are used. Based on these results, ethylene glycol can protect sperm from heat shock at $4^{\circ}C$, but not satisfactory level. However, it showed the possibilities of liquid semen preservation at $4^{\circ}C$ by using cryoprotectant.

• Korean Journal of Poultry Science
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• v.45 no.4
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• pp.237-244
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• 2018

In this study, to increase the survival rate of frozen/thaw rooster semen, standard protocols of semen thawing procedures were tested by computer-assisted sperm assay (CASA). We tested 4 different thawing protocols for frozen semen, $5^{\circ}C$ for 2 min, $35^{\circ}C$ for 30 s, $54^{\circ}C$ for 13 s, and $70^{\circ}C$ for 7 s. The pooled semen from 5 to 8 Ogye rooster line was diluted in the HS-1 diluent and frozen in 8% methylacetamide (MA) in liquid nitrogen vapors. To determine standard thawing method, straws were plunged into different temperatures and times. The resulting motilities were recorded by the CASA system. The results of this study showed that the best viability of the spermatozoa was shown by exposure at $5^{\circ}C$ for 2 min. Moreover, the longevity test of thawed sperm at $5^{\circ}C$ for 2 min also supported the higher viability under low temperature preservation of $17^{\circ}C$ for 1 hr. Further research is needed to increase the motility of thawed rooster semen for field application. In addition, the in vivo tests for different rooster lines are also needed for the establishment of avian genetic resource bank.

• Korean Journal of Animal Reproduction
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• v.26 no.1
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• pp.1-7
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• 2002

The present study was carried out to investigate the effects of maturation media on penetrability of pig oocytes by liquid boar sperm coincubated with different sperm concentrations in a modified Tris­buffered medium (mTBM). Follicular oocytes collected from ovaries of prepubertal gilts were matured in a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. The sperm­ich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ far 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes in 5% $CO_2$in air at 38.5$^{\circ}C$, cumulus cells were removed and oocytes (30~40) were coincubated for 6 h in 0.5 $m\ell$ mTBM fertilization medium with five different (1$\times$10$^{6}$ , 2$\times$10$^{6}$ , 4$\times$10$^{6}$ , 6$\times$10$^{6}$, 10$\times$10$^{6}$ $m\ell$) sperm concentrations. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ NCSU-23 culture medium fur further culture of 6 h. At 12 h after IVF, sperm penetration, polyspermy and male pronuclear formation of oocytes were evaluated. Oocytes of NCSU-23 maturation medium decreased polyspermy and increased male pronuclear formation compared to those of mTCM­199 and mWaymouth MB 752/1 maturation media. Of oocytes matured in NCSU-23 medium and inseminated in mTBM medium with 2~4$\times$10$^{6}$ $m\ell$ sperm concentrations, 50.8~50.9% showed sperm penetration, 13.3~19.5% polyspermy and 43.9~45.4% male pronuclear formation. In conclusion, we found out that oocytes matured in NCSU­23 medium and inseminated in mTBM medium showed superior in­vitro fertilization compared to those matured in mTCM­199 and mWaymouth MB 752/1 maturation media and inseminated in mTBM medium. The optimum sperm concentrations for in-vitro fertilization of oocytes matured in NCSU-23 medium by liquid boar semen stored at 17$^{\circ}C$ for 5 days were 2~4$\times$10$^{6}$ $m\ell$.

• 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.

• 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.

• ANNALS OF ANIMAL RESOURCE SCIENCES
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• v.29 no.4
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• pp.158-165
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• 2018

The present study was undertaken to investigate the reproductive performance of the female breeding pigs after artificial insemination (AI) using the frozen boar semen imported from Canada, thereby finding insights into improving the efficiency of AI using the frozen semen (FSAI). Analyzed in the present study were the records of a total of 626 FSAI in a great grandparent (GGP) farm beginning from May through November of the year of 2016 (Farm A) and 2,024 FSAI beginning from 2015 through 2017 from a second GGP farm (Farm B). Both the total number of piglets born (TNB) and the number born alive (NBA) were greater during May than during September within FSAI (p<0.05) in Farm A (p<0.01 for the effect of the month). In Farm B, no difference was detected between the years in any of the farrowing rate, TNB, and NBA. When the records from Farm A and Farm B were pooled, the farrowing rate was greater for Farm A vs. Farm B (p<0.01), with no difference between the two farms in TNB and NBA. Moreover, TNB and NBA were less for FSAI than for AI using the liquid semen (LSAI; $10.9{\pm}0.3$ vs. $13.4{\pm}0.1$ and $10.0{\pm}0.3$ vs. $12.0{\pm}0.1$ piglets, respectively, for FSAI vs. LSAI in TNB and NBA, respectively; p<0.01). In conclusion, these results suggest that the reproduction efficiency for FSAI, which is lower than that for LSAI, could be improved by selecting an optimal period of the year for the use of the former.

• Korean journal of food and cookery science
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• v.16 no.6
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• pp.557-567
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• 2000

An optional ingredient, jasoja(Perillae semen), was adopted to improve the quality of Dongchimi. The final weight percentage of jasoja in Dongchimi was adjusted to 0, 0.25, 0.5, 0.75, or 1.0%, per radish, and sensory and microbiological characteristics were determined during fermentation at 10 for 45 days. The effect was varied depending on the amounts of jasoja, but Dongchimi fermented with 0.5% jasoja was most favored for color, flavor, taste, texture, and overall acceptability in sensory evaluation. According to a quantitative descriptive analysis for the product, the liquid portion of Dongchimi steadily became clearer and less sour in proportion to the amount of added jasoja. However, a strong off-taste was detected from 1.0% treatment. The viable cell numbers of total and lactic acid bacteria drastically increased during the first 2 days, and then gradually increased to their maximum values during fermentation and slowly decreased at the later stage. Dongchimi with 0.5% treatment showed a distinctive high number of microorganisms at the 15th-day of fermentation and this trend was maintained until the completion of fermentation. The lactic acid bacteria isolated and identified from Dongchimi were; Lactobacillus brevis, Lactobacillus plantarum, Leuconostoc mesenteroides, Lactococcus faecalis, and Lactococcus lactis. The combined number of Lactobacillus brevis and Lactobacillus plantarum began to increase right after preparation to as much as 10$\^$7/CFU/㎖, then decreased to 10-10$^3$CFU/㎖ afterward. This study showed that the addition of jasoja retarded the initial fermentation of Dongchimi; however, too much jasoja at above 1% weight level per Chinese radish might accelerate fermentation at the later fermentation stage and shoud be avoided. A comparable fermentation pattern was observed among the samples; however, more acceptable Dongchimi could be prepared by fermenting for 11 to 30 days at 0.5% jasoja concentration per radish.

• Asian-Australasian Journal of Animal Sciences
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• v.10 no.3
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• pp.329-332
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• 1997

A study on the conception rate of 885 cows and heifers bred under artificial insemination (AI) and natural service was carried out at AI sub-centers. A high conception rate was observed at first service in all breeding methods. It was revealed that the conception rates in AI with liquid semen (LS) were found to be 25.08, 18.58, 12.69, and 2.48% after 1, 2, 3 and 4 services, respectively. Inseminations of cattle with frozen semen (FS) were found to achieve 42.15, 14.46, 5.85, and 2.46% conception rates after 1, 2, 3, and 4 services, respectively. In natural service, conception rates were found to be 62.45, 6.75, 5.91, and 4.64%, respectively, after 1, 2, 3, and 4 services. Insemination of cows and heifers at mid-cycle had significantly (p < 0.05) higher conception rate as compared to early and late cycles. The existing AI machineries and breeding management need to be improved to achieve higher conception rate of dairy cattle under AI.