• Korean Journal of Animal Reproduction
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• v.23 no.2
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• pp.127-132
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• 1999

This study was carried out to investigate the general characteristics such as volume, sperm concentration, sperm motility, sperm abnormality on whole semen, removed seminal plasma(RSP) semen and fractional semen of small dogs, and the effect of temperature and preservatio time and cryopreservation on motility of whole and removed seminal plasma semen. Multiple ejaculates were collected from small dogs by the digital manipulation of penis. 1. Average sperm concentration, sperm motility and abnormal sperm rates of whole semen and RSP semen were 5.07$\pm$2.32$\times$10$^{6}$ cells/$m\ell$, 95.42$\pm$2.65%, 4.42$\pm$0.157% and 4.69$\pm$3.27~4.25$\pm$3.65$\times$10$^{6}$ cells/$m\ell$, 91.17$\pm$3.85~88.52$\pm$3.85%, 6.57$\pm$0.43~5.54$\pm$0.52%, respectively. 2. Average semen volume per ejaculate, sperm concentration, sperm motility and abnormal sperm rate of 1st fractional semen were 0.92$\pm$0.7$m\ell$, 4.57$\pm$0.78$\times$10$^{6}$ cells/$m\ell$, 10.72$\pm$3.21% and 5.50$\pm$0.70%. Average semen volume per ejaculate, sperm concentration, sperm motility and abnormal sperm rate of 2nd fractional semen were 2. 14$\pm$0.19$m\ell$, 2.01$\pm$0.12$\times$10$^{6}$ cells/$m\ell$, 95.44$\pm$4.21% and 4.31$\pm$0.53%. Average semen volume per ejaculate, sperm concentration, sperm motility and abnormal sperm rate of 3rd fractional semen were 2.66$\pm$0.23$m\ell$, 2.35$\pm$0.21$\times$10$^{6}$ cells/$m\ell$, 90.71$\pm$2.63%, 6.33$\pm$0.91%, respectively. 3. Motility of whole semen and RSP semen were higher at 2$0^{\circ}C$ than at 4$^{\circ}C$ or 37$^{\circ}C$. When preservation temperature was 2$0^{\circ}C$, sperm motility were 98.32% at 1 hr, 92.15% at 5 hrs, 90.23% at 10 hrs 82.08% at 15 hrs 70.07% at 20 hrs 60.02% at 20 hrs 37.19% at 40 hrs respectively. 4. Average sperm motility of frozen 2nd fraction semen and RSP semen were 33.3$\pm$8.7, 54.7$\pm$9.5%, respectively. Sperm motility was significantly higher in frozen 2nd fraction semen and RSP semen compared with control group.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.81-81
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• 2003

돼지 정액의 동결보존은 돼지 정자가 내동성이 약한 특성을 가지고 있어 아직 만족할만한 결과를 얻지 못하여 액상정액을 이용하여 인공수정을 실시하고 있는 실정이다. 돼지 정액의 동결보존을 효과적으로 이룩한다면 돼지 인공수정의 효율성을 증대시킬 수 있고, 우수한 종돈의 보호 및 국제적 돼지 품종의 교류를 증진시킬 수 있을 것이다. 본 실험은 돼지 정액 동결시 동결온도와 시간, 항산화제로 알려진 Taurine의 농도별 첨가, 동해보호제 및 융해조건이 돼지 정액의 동결 융해후 생존성에 미치는 영향을 검토하고자 수행되었다.

• 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 Aquaculture
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• v.13 no.3
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• pp.187-191
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• 2000

The effects of osmolality on the sperm motility in black seabream (Acanthopagrus schlegeli) were studied. Sperm motility of black seabream was suppressed when the osmolality was equal to the seminal fluid. But sperm became motile when the osmolality increased in electrolyte solution (NaCl, KCl, $CaCl_2$, $MgCl_2$) and non-electrolyte solution (mannitol, glucose, fructose, sucrose). The changes of sperm motility index (SMI) by osmolality of diluents described a parabola. In all of the diluents, SMI was the highest at ca. 1,000 mOsm/kg, which is similar to the osmolality of seawater. Sperm motility was induced by osmolality of diluents, but exposure to hypotonic or hypertonic diluents was harmful to the sperm.

• Reproductive and Developmental Biology
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• v.29 no.1
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• pp.25-30
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• 2005

This study was carried out to investigate the effects of semen characteristics, frozen-thawed sperm viability and serum FSH, LH, estradiol-17β and testosterone concentrations between breeds and among seasons in boars. In all seasons, Yorkshire boars produced higher semen volume compared with Duroc boars, whereas sperm concentration did not differ significantly between Duroc and Yorkshire boars. Semen volume in spring was higher compared with summer, autumn and winter in both Duroc and Yorkshire boars, but sperm concentration did not differ significantly among seasons. Sperm motility and normal acrosome rate of frozen-thawed sperm produced in spring were higher than those in summer, autumn and winter in both Duroc and Yorkshire boars. Sperm motility of frozen-thawed sperm in Yorkshire boars was higher than that in Duroc boars regardless of seasons. However, normal acrosome rate did not differ significantly between Duroc and Yorkshire boars. Serum FSH concentration in Yorkshire boars was lower than that in Duroc boars in all seasons. However, there were no significant differences on serum FSH concentration of Duroc and Yorkshire boars among seasons. Serum LH and estradiol-17β concentrations did not differ significantly between Duroc and Yorkshire boars. Also, there were no significant differences in serum LH and estradiol-17β concentrations of Duroc and Yorkshire boars among seasons. Serum testosterone concentration in Yorkshire boars was higher than that in Duroc boars in all seasons. In both breeds, serum testosterone concentrations were higher in spring than in summer, autumn and winter. In conclusion, when serum FSH concentrations were low, semen volumes were high, and when serum testosterone concentrations were high, sperm motility and normal acrosome rate of frozen-thawed sperm were high.

• Korean Journal of Animal Reproduction
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• v.25 no.4
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• pp.409-419
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• 2001

This study was to examine whether the in vitro friability, motility and intact acrosome of frozen-thawed bovine and human sperm can be improved by adding Pentoxifylline (PF) or Fertilization Promoting Peptide (FPP). Human semen was frozen ultra-rapidly using Test yolk-buffer (TYB) freezing medium. Additive (PF, FPP) effects in frozen-thawed bovine and human sperm were analyzed by microscopic count for sperm motility and coomassie brilliant blue staining method f3r sperm acrosome intact. The in vitro motility of frozen-thawed bovine sperm with 5 mM PF treatment group (50.0%) was significantly higher than that of control (34.0%) (P＜0.05). In the frozen-thawed bovine sperm was examined, the intact acrosome rate of 50 nM FPP treatment (49.0%) was significantly higher than those of control (30.0%) and 25 nM FPP (38.0%) treatment groups (P＜0.01). In human semen, when in vitro motility of sperm with PF addition prior to freezing was examined, the result of 5 mM treatment group (51.0%) was significantly higher than those of control and 2.5 mM treatment group (39.0, 40.0%) (P＜0.01). In addition, 50 nM (75.5%) FPP adding in all treatment procedures for human semen freezing (before freezing, freezing and after thawing) was significant effect on maintenance of the sperm intact acrosome percentage (control: 45.0; 25 nM: 53.0; 100 nM: 68.0%) (P＜0.01). Also, the intact acrosome rate of human sperm with FPP (65.0%) was significantly higher than that with PF (43.0%) (P＜0.05), although sperm motility was slightly higher in PF treatment group. These results suggest that improved sperm motility and intact acrosome of frozen thawed bovine and human sperm can be obtained by addition of PF or FPP, and that the enhanced in vitro viability, motility and intact acrosome can be obtained by addition of FPP in all semen freezing procedures.

• Journal of Life Science
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• v.29 no.4
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• pp.395-401
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• 2019

Phthalate is a chemical endocrine disrupter and interfere with the action of hormones, estrogens, androgens and thyroid hormones. It also affect cardiovascular, metabolic, immune and reproductive system in the human and animals. Curcumin is antioxidant, anti-inflammatory activity and -cancer properties in the human. We studied whether phthalates damage viability, mitochondrial activity and membrane integrity of sperm in boar semen. We also treated curcumin with/without phthalates in the boar semen. Fresh boar semen was treated with phthalates and/or curcumin for examining sperm characteristics. Sperm characteristics, sperm motility, viability, mitochondrial activity, and membrane integrity were determined during storage of boar semen. Sperm motility and viability in dose-dependent manner decreased by di-n-butyl phthalate (DBP), mono-n-butyl phthalate (MBP) and di-2-ethylhexyl phthalate (DEHP, p<0.05). Phthalates also decreased mitochondrial activity and membrane integrity of sperm (p<0.05). However, sperm motility and viability were higher than untreated-curcumin when DBP, MBP and DEHP treated with a curcumin in boar semen (p<0.05). Mitochondrial activity and membrane integrity of sperm were higher in DBP- and MBP-treated semen with curcumin (p<0.05). In conclusion, phthalates can damage sperm viability and quality during the boar semen storage, and curcumin may protect the boar sperms from phthalates during storage term.

• Journal of Aquaculture
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• v.11 no.1
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• pp.67-75
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• 1998

Experiments were performed to obtain cryopreservation techniques of black seabream (Acanthopagrus schlegeli) sperm. For sperm collection, brood stock reared in recirculating seawater system and fed with the commercial feed during experimental period. The results indicated that following cryopreservation method in block seabream sperm could be employed. Post-thaw survival rate of sperm revealed the highest value ($80{\pm}1.4$%) in 3% sodium citrate as a diluent for the cryopreservation. Cryopreserved sperm diluted with 5.4% glucose showed the highest fertilization rate to the ovulated eggs. Glycerol was a better cryoprotectant than dimethyl sulfoxide in sperm cryopreservation : survival rate and fertilizing capacity of cryopreserved sperm were decreased according to increase of glycerol concentration and varied in renges of 0.8~59.3% and 32.5~69.4% with 5~30% glycerol, respectively. A few of cryopreserved spermatozoa showed the enlarged head with granulated chromatin and ruptured plasma membrane by freezing and thawing injuries compared with unfrozen normal spermatozoa.

• Journal of Animal Science and Technology
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• v.54 no.2
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• pp.95-101
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• 2012

The objective of this study was to examine the effect of amides as a cryoprotectant for semen cryopreservation in Korean Jeju Black Bull. The semen was cryopreserved with extenders containing 5% dimethyl acetamide (DMA), 5% dimethyl formamide (DMF), 5% methyl formamide (MF) or 7% glycerol. Post-thawed sperm were evaluated for sperm motility, viability, acrosome integrity and membrane integrity. Post-thawed sperm motility was significantly higher (p<0.05) in glycerol and DMF ($64.00%{\pm}9.62$ and $59.00%{\pm}5.48$, respectively) than DMA and MF ($50.00%{\pm}3.24$ and $44.00%{\pm}4.18$, respectively). Sperm viability wassignificantly higher (p<0.05) in glycerol and DMF ($58.25%{\pm}7.35$ and $53.05%{\pm}3.77$, respectively) than others. However, for sperm motility and viability, there were no differences among glycerol and DMF. Also, swelling sperm ratio by hypo-osmetic selling test (HOST) was significantly increased (p<0.05) in glycerol and DMF treatments ($45.12%{\pm}25.08$ and $44.95%{\pm}8.58$, respectively). The percentage of capacitated sperm assessed by CTC staining, F pattern was lower (p<0.05) in DMF than others. B pattern was increased (p<0.05) in DMA, DMF and MF when compared with glycerol. AR pattern ratio was decreased (p<0.05) in glycerol and DMF when compared with DMA and MF. These results suggested that amides performed better and could be used as a cryoprotectant for semen freezing of Korean Jeju Black Bull.

• Journal of Embryo Transfer
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• v.18 no.3
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• pp.195-201
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• 2003

This study was carried out to establish most suitable freezing condition, to evaluate the different condition of freezing and thawing rates on the viability and motility of frozen canine spermatozoa. The ejaculated semen was added to obtained 200∼400 ${\times}$ 10$^{6}$ /$m\ell$ with extender I and was cooled to 4$^{\circ}C$ over 30, 60 and 120 minutes. And then, semen was diluted with extender II containing 4, 6 and 8％(v/v) glycerol for 60 min, respectively and packaged in 0.5$m\ell$ straw, equilibrated far 30, 60 and 120 min at 4$^{\circ}C$ and cryopreserved in liquid nitrogen vapor at different distance(3, 5, 7 and 9 cm, respectively), plunged into nitrogen tank. Samples were thawed by placing straws into 27, 37, 47, 57$^{\circ}C$ water bath for 120, 20 and 12 sec, respectively. The results were as follows; 1. The survival and motility rate immediately post-thawing was significantly higher in samples frozen in 4％ glycerol than 6 or 8％ glycerol(P< 0.05). 2. According to equilibration time at 4$^{\circ}C$, the survival and motility rate immediately post-thawing was significantly higher in samples frozen after 60 min equilibration than 30 or 120 min equilibration(P<0.05). 3. Freezing in distance of 5 cm from liquid nitrogen yield better survival and motility rate than the others(3, 7 or 9 cm)(P<0.05). 4. The effect of thawing rate on sperm survival were higher when the thawing was done at 37$^{\circ}C$ for 120 sec(P<0.05).