• Korean Journal of Animal Reproduction
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• v.23 no.3
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• pp.263-270
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• 1999

This study was conducted to investigate effects of L-ascorbic acid and selenium on maturation, fertilization, and development ablity of porcine follicular oocytes in vitro. When the follicular oocytes were cultured in the media containing 0, 62.5, 100 and 300 $\mu$M of L-ascorbic acid for 40~44h, the percentages of germinal vesicle breakdown were 86.8, 92.9, 91.7 and 92.6% respectively, and the nuclear maturation rates (M II) were 44.7, 57.1, 52.8 and 53.7%. The nuclear maturation rates of treated groups were significantly higher than those of non-treated group (p＜0.05). When the follicular oocytes were cultured at 0, 0.4, 0.8, and $1.5\mu$M of selenium for 40~44h, the nuclear maturation rates of treated groups were significantly higher than those of non-treated group (p＜0.05). The addition of L-ascorbic acid or selenium to the maturation medium, the incidence of male pronuclear formation was significantly increased (p＜0.05) and polyspermy rate was significantly decreased (p＜0.05). The addition of L-ascorbic acid or selenium to the maturation medium increased the clevage rate, morula and blastocyst rate (p＜0.05). These results suggested that the addition of L-ascorbic acid and selenium to maturation medium increase the nuclear maturation rates, male pronuclear formation and normal embryonic development: in porcine oocytes matured and fertilized in vitro.

• Journal of Embryo Transfer
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• v.21 no.4
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• pp.323-330
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• 2006

본 연구의 목적은 3주 동안 장기간 냉각상태로 보관된 개 정자의 기능적 특성을 알아보고자 두 종류의 정자 희석액, Glucose-BSA(G-BSA), Dimitropoulos-II(BIMI)가 정자의 기능적 특성에 미치는 영향을 알아보고자 한다. 개 정액을 수지법에 의해 채취하여 원심분리한 후 정장은 제거하였다. 정자를 G-BSA나 DIMI으로 희석하여 최종 농도를 $100\times10^6$ sperm/ml로 하였다. 희석된 정자를 정자 운송 시스템을 이용하여 루지애나 주립 대학에서 뉴올리언즈 실험실까지 운송하였다. 정자 운송 시스템은 $20^{\circ}C$에서 $9^{\circ}C$까지 분당 $0.08^{\circ}C$ 냉각율로 설정하였다. 희석된 정자는 $4^{\circ}C$에서 3주간 보관하였다. 보관 1일부터, 매주 단위로 정자의 활력, 정자 원형질막 고유성(생존성), 정자 첨체의 고유성을 검사하였다. 또한 체외조건하에서의 정자 번식 능력을 평가하기 위해 zona binding assay를 실시하였다. 실험 결과 냉각 상태로 3주 동안 보관된 개 정자는 여전히 정자의 기능적 특성을 나타냈다. 냉각기간이 길어질수록 정자의 활력 및 생존성은 감소하였으며(P<0.01), 첨체의 고유성은 냉각기간에 따라 감소하였으나 유의성은 없었다. 정자의 특성 중 활력이 다른 특성에 비해 냉각처리에 민감한 반응을 보였다. G-BSA배지에 보관된 정자의 활력 및 생존성이 DIMI에 비해 높게 나타났으며, 1 주 동안 보관된 정자의 생존성의 비교시 유의성이 높게 나타났다(P<0.05). 그러나 첨체의 고유성은 1일 동안 DIMI에 보관된 정자에서 높게 나타났다(P<0.05). 3주 동안 냉각상태로 보관된 개 정자는 여전히 난자의 투명대에 결합하는 능력을 보였으며, G-BSA에 보관된 정자의 경우 난자에 부착되는 평균 정자 수가 보관일수에 따라 유의적으로 감소하였다(P<0.05). 그러나 희석액 종류에 따른 난자에 부착되는 평균 정자 수의 유의적 차이는 없었다. 정자의 활력 및 생존성과 난자의 투명대에 결합하는 정자 수와의 상관관계를 본 결과, 활력 및 생존성이 높은 정자일수록 난자의 투명대에 많이 결합하여 정자의 잠재적 번식력이 증가함을 보였다. 결론적으로 G-BSA나 DIMI의 희석액으로 희석된 후 $4^{\circ}C$에서 냉각 보관된 정자는 3주 동안 정자의 기능적 특성을 유지하였으며 냉각 보관 2주까지 좋은 성적의 정자 특성을 나타냈다. 개 정자 냉각 보존에 대안적인 희석액으로 G-BSA도 사용 가능한 것으로 사료된다.

• Korean Journal of Animal Reproduction
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• v.17 no.2
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• pp.113-120
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• 1993

To provide the optimal culture conditions for the developm,ent of in vit개 produced embryos, we have been investigated various culture media as well as co-cultrue systems using porcine cumulus cells or mouse fetal fibroblast cells. Porcine ovaries were brought to the laboratory from local slaughter house within 1 hour after slaughtering and cumulus oocytes complexes were recovered from antral follicles(3~5mm) with 23 gauge needle. To maturate follicular oocytes, cumulus oocytes complexes were washed three times with TCM-199 containing 25mM HEPES and incubated(39$^{\circ}C$, 5% CO2 in air) in various maturation media for 42 hrs. Ejaculated and liquid storaged boar spermatozoa capacitated with different sperm capacitation methods and media were rpepared for fertilizing of matured follicular oocytes in vitro. Fertilization was performed by adding 5~10${\mu}\ell$ fo capacitated spermatozoa containing 1~5$\times$105 sperm/ml to droplets. Eighteen to twenty-eight hours after sperm insemination, fertilized eggs were washed three times with culture media and transferred to the various culture media, to the culture media with a monolayer of somatic cells. The in vitro development rates of 1-cell embryos cultured with three times with culture media and transferred to the various culture media, to the culture media with a monolayer of somatic cells. The in vitro development rates of 1-cell embryos cultured with three different media, m-KRB, BECM and TCM-HEPES were 0~1.0%, showing extremely lower rates. Especially, most of embryos were observed to arrest the development beyond 4-cell stages. The rates of embryos developed to 2-, 4-, 8-, 16-, 32-cell and morula or blastocyst stage in co-culture with porcine cumulus cells and mouse fetal fibroblast cells were 61.1~67.0%, 59.0~58.0%, 42.5~43.1%, 28.4~30.2% and 20.4~21.0%, respectively. These development rates upto morula or blastocyst stages were significantly higher than those of the embryos cultured in the basic culture medium(P<0.01). These findings suggest that co-culture of in vitro fertilized eggs with porcine cumulus cells or mouse fetal fibroblast cells enhance the development of fertilized eggs to morula or blastocyst stage in vitro.

• The Korean Journal of Pesticide Science
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• v.15 no.2
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• pp.208-217
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• 2011

Pyrimisulfan is a herbicide. In order to register this new pesticide, the series of toxicity data on animal testing were reviewed to evaluate its hazards to consumers and also to determine its acceptable daily intake. Pyrimisulfan was excreted mostly by feces. It has low acute oral toxicity while it has no dermal, ocular irritation and skin sensitization (As the result of subchronic and chronic toxicity and carcinogenicity showed changes of hematology and liver.). Two-generation reproduction toxicity, genotoxicity, carcinogenicity and prenatal development toxicity were not proven. Therefore, the ADI for Pyrimisulfan is 0.1 mg/kg/ bw/day, based on the NOAEL of 10 mg/kg/ bw/day of 90-days repeated dose oral toxicity study in dogs while applying an uncertainty factor of 100.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.10
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• pp.181-186
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• 2020

This study examined the artificial fertilization efficiency of crossbred goats from a farmhouse using frozen semen. Electrostimulation was used to ejaculate and collect semen to assess the artificial fertilization efficiency of crossbred goats. The sperm concentration, vitality, and vitality after melting were investigated. The sperm volume was within 2.5~3 ml, and the concentration was 21~25 × 10⁸/ml for each male crossbred goat. The melted semen had high vitality (≥90%). An IDEXX Rapid Visual Pregnancy Test kit was used for an earlier diagnosis of the pregnancy and to determine the pregnancy rate of fertilization using frozen-thawed semen. The reproductive performance of the artificially fertilized crossbred goats had the highest delivery rate (68%) from Farm C and the lowest delivery rate (45%) from farm A. The delivery rate through artificial fertilization was equal to the fertilization rate according to early pregnancy diagnostic kits. The artificial insemination efficiency was 45~68%. These findings can be used as the basis for improvement and breeding goats in goat farms and livestock research institutes.

• Korean Journal of Animal Reproduction
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• v.27 no.2
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• pp.143-152
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• 2003

The objective of this study were to investigate the effects of Se and Vit. E administration on blood chemical values and Vit. E concentrations of serum in calves born from Hanwoo dams injected Se and Vit. E at late pregnancying periods. The effects of Se and Vit. E administration of blood chemical values in calves born from Hanwoo dams injected Se and Vit. E at the last month of pregnancying periods were examined. In albumin concentration of blood, the Vit. E 2000IU group were slightly higher than other groups and the cholesterol concentrations were also higher in Vit. E 15001u group than other groups. But, another components examined of blood chemical values were not significantly different in all experimental groups. The blood chemical values in calves born from Hanwoo dams administrated Se and Vit. E at two months before parturition were examined. However, there were not significantly different in all experimental groups in concentrations of albumin, cholesterol, BUN, creatinine, glucose, inorganic phosphorous, calcium, total protein and triglycerides. The concentrations of serum Vit. E in calves born from Hanwoo dams administrated Se and Vit. E at 30 and 15 days before parturition were examined. The concentrations of Vit. E in serum were higher in treatment groups than in control group(P>0.05). The concentrations of serum Vit. E in calves born from Hanwoo dams administrated Se and Vit. E at 60 days before parturition were examined, the concentrations of Vit. E in serum were slightly higher in groups administrated than in control group(P>0.05), but there were not significantly difference in all experimental groups.

• Korean Journal of Animal Reproduction
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• v.15 no.1
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• pp.49-57
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• 1991

This study was carried out to investigate the effect of equilibration time, precooling and straw loading method on the post-thaw survival rates of mouse embryos cryopreserved by vitrification method. The effect of the vitrification procedure on the damage of the embryos was also examined by the straining of nuclei using Hoechst 33342. The results obtained were summarized as follows; 1. The equilibration in Medium-1 for 10 minutes was considered to be the optimum equilibration time. Embryos equilibrated in Medium-1 for 10 minutes(81.0%) showed significantly(P<0.05) higher survival rates than those equilibrated for 5 minutes(40.0%) or 15 minutes(74.1%). 2. The survival rate of embryos cryopreserved using the double Medium-2 column(81.0%) was significantly(P<0.01) higher than that using the single Medium-2 column, whish was considered to be due to the double Medium-2 column method being more reliable for preventing contamination of diluent solution of 1M sucrose. 3. The survival rate of compacted morula stage embryos cryopreserved with the precooled and non-precooled Medium-2 was not significantly(P<0.05) different. 4. The number of blastomeres of embryos at late blastocyst stage after culture of mouse morulae for 24 to 28hours was significantly(P<0.05) decreased by freezing embryos using vitrification(53.3${\pm}$1.6 vs 41.4${\pm}$1.5), which was considered to be due to the damage of embryos during vitrification and the delay of embryo development by handling in vitro. 5. The vitrification procedure is considered to be a very simple and efficient method for cryopreservation of embryos at early developmental stage.

• Korean Journal of Animal Reproduction
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• v.15 no.1
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• pp.41-47
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• 1991

The ability of fertilization in vitro and subsequent development of superovulated oocytes was assessed in a controlled environment using an in vitro fertilization technique. The in vitro fertilization percentage of oocytes with cumulus mass declined significantly(P<0.05) with increased doses from 4~10 to 16~40IU of PMSG for superovulation. However, the porportion of polyspermic penetration varied from 2.3 to 9.7% and there was no significant difference between treatments in incidence of polyspermy. When morphologically normal ova with cumulus mass were cultured for 66~72h in a plastic mini-straw to undergo fertilization in vitro, the mean percentage of embryos developed to 2-16 and 4-16cell stage was 61.8 and 17.6%; it was slightly(P<0.05) superior to the corresponding results from petri dish. A total of 52 two-cell embryos fertilized in a mini-straw were transferred to seven pseudopregnant rat. Among these recipients, two normal young were born from one recipient which received a total of six embryos. These results suggest that superovulated oocytes are proportionately less competent than normally ovulated oocytes to undergo fertilization in a controlled environment using an in vitro fertilization technique. Also, a plastic mini-straw designed was slightly superior to petri dish as a culture vessel for fertilization and embryo development in vitro.

• Journal of The Korean Society of Grassland and Forage Science
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• v.39 no.1
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• pp.1-8
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• 2019

This study was conducted to predict the energy requirements for maintenance and growth of female Korean black goats during their growth and pregnancy phases. Fifty female goats ($18.7{\pm}0.27kg$) in their growth phase with an average age of 5 months were stratified by weight and randomly assigned into 5 groups. They were fed 5 diets varying in metabolic energy (ME) [2.32 (G1), 2.49 (G2), 2.74 (G3), 2.99 (G4), and 3.24 (G5) Mcal/kg] until they were 9-month-old. After natural breeding, 50 female goats ($30.7{\pm}0.59kg$) were stratified by weight and randomly assigned into 5 groups. They were fed 5 diets varying in ME [2.32 (P1), 2.43 (P2), 2.55 (P3), 2.66 (P4), and 2.78 (P5) Mcal/kg]. The average feed intake ranged between 1.5 and 2.0% of the body weight (BW), and there was no significant difference between the treatment groups with goats in growth or pregnancy phases. Average daily gain (ADG) in diet demand during the growth phase increased with an increasing ME density and ranged from 46 to 69 g/d (p<0.01). Feed conversion ratio (FCR) improved with the ME density during the growth phase (p<0.01). The intercept of the regression equation between ME intake and ADG indicated that energy requirement for maintenance of goats during growth and pregnancy phases was $103.53kcal/BW^{0.75}$ and $102.7kcal/BW^{0.75}$, respectively. These results may serve as a basis for the establishment of goat feeding standards in Korea. Further studies are required to assess the nutrient requirement of goats using various methods for improving accuracy.

• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.219-224
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• 2006

The objective of this study was to investigate the toxic effects of Bisphenol A(BPA) and di-2 ethyhexyl phthalate (DEHP) as endocrine disrupters on sperm motility, viability, membrane integrity and abnormality during in vitro incubation of boar semen. Semen were randomly divide into 24 groups and healed with different concentrations of BPA md DEHP($1{\sim}100{\mu}M$) for 3, 6 and 9 hrs, respectively. The percentages of sperm motility and viability decreased by treatment time with both BPA and DEHP, and obiously differ from the controls. The percentages of sperm motility and viability significantly decreased by incubation with both $100{\mu}M$ of BPA and DEHP compared to control and other treatment groups(p<0.05). Sperm membrane integrity was significantly reduced by incubation with 10 and $100{\mu}M$ of BPA and DEHP, respectively(p<0.05), but sperm abnormality were not significantly affect both BPA and DEHP. These results indicate that high concentration of BPA and DEHP($>10{\mu}M$ can affect noxiously the sperm characteristics.