• Journal of Veterinary Clinics
• /
• v.30 no.5
• /
• pp.384-386
• /
• 2013

A two-year-old, female English bulldog was referred for breeding by artificial insemination with frozen semen of male English bulldog, a litter of female bulldog's grandfather. Intrauterine artificial insemination was done two days after the ovulation day. Sperm was evaluated after thawing by computer assisted sperm analyzer, and its motility was 89.8% with normal shape. Pregnancy bearing eight fetuses was diagnosed by ultrasonography and radiography. Cesarean section was performed sixty days after the artificial insemination. Eight pups were delivered with safe, but the entire pup had abnormalities including severe bow-legged malformations, cleft lip, cleft palate, and enlarged cranial part.

• Food Science of Animal Resources
• /
• v.27 no.2
• /
• pp.190-196
• /
• 2007

This study was carried out to investigate the effects of pressure assisted freezing(PAF) on physicochemical properties of pork meat. Pork meat was frozen under pressure up to 200 MPa at $-60^{\circ}C$, and compared with fresh control. Phase transition temperature decreased with increasing pressure level, while pressure level had no effect on supercooling extent. Increasing pressure level increased pH of meat significantly(p<0.05). Thawing losses of all treatments were significantly higher(p<0.05) than control with the exception of PAF at 200 MPa. Water holding capacity(WHC) was increased significantly(p<0.05) with increasing pressure level up to 100 MPa. Cooking loss tended to decrease with increasing pressure level. In color, CIE $L^*-\;and\;b^*-value$ increased with increasing pressure level, while CIE $a^*-value$ decreased significantly(p<0.05). Increasing pressure level up to 150 MPa increased shear force significantly(p<0.05), however, no significant difference between 150 and 200 MPa in shear force was found(p>0.05). Therefore, the results indicated that excessive pressure level in PAF caused several losses in meat qualities, while PAF at mild pressure level improved meat qualities compared to atmosphere freezing.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.21 no.4
• /
• pp.490-496
• /
• 2020

A styrofoam box is used as a simple and easy freezing method to preserve animal semen as a livestock genetic source. This study optimized the methods of freezing chikso brindled cattle semen. To test the freezing box, the motility of spermatozoa was compared between two box sizes (length×width×heigh) with the dimensions of 23.5×30.5×22.5 cm and 25.5×46.5×26.5 cm. The motility of thawed sperm from brindled Korean bulls was used to confirm the efficiency of the freezing boxes. The box having a larger inner space with larger horizontal and height measurements supported better motility after thawing (60.4±5.3% vs 67.2±3.1%) with 10 min of exposure time in liquid nitrogen vapor. The optimized freezing space is estimated to be an essential element for successful freezing results and the larger box could be used for production of more than 60 frozen semen straws. These properties are also helpful to optimize the cryopreservation techniques that would control the quality and quantity of semen straws according to different animal species.

• Korean Journal of Animal Reproduction
• /
• v.24 no.4
• /
• pp.439-449
• /
• 2000

This study was to investigate the expression of transgene after co-injection of spermatozoon and EGFP gene into mature oocytes in cattle. From frozen semen, spermatozoa were treated by DTT with 0.03% Tween-20, freezing and thawing or 0.02% Triton X-100 to disrupt their plasma membranes. The sperm injected oocytes were co-cultured with bovine oviduct epithelial cells in CRlaa, and expression of EGFP in embryos were observbed under epifluorescent microscope. Two pronuclei were formed in oocytes injected with sperm treated by DTT (44.6%), DTT-Tween-20 (48.4%), DTT-freezing (44.4%) and DTT-Triton X-100 (42.9%). Cleavage and blastocyst formation rates of bovine oocytes which injected with sperm treated by DTT, DTT-Tween-20, DTT-freezing, and DTT-Triton X-100 were 49.1, 58.5, 43.9, and 48.4% and 10.2, 13.0, 6.8, and 6.5%, respectively. Although the most of embryos were showing mosaicism, embryos expressing EGFP gene were observed in all treated groups. Therefore, these results indicate that membrane disrupted sperm could interact with exogenous DNA, and that this procedure may be useful to introduce foreign gene into bovine oocytes.

• Journal of Plant Biotechnology
• /
• v.7 no.3
• /
• pp.183-186
• /
• 2005

Cryopreservation has been recognized as a practical and efficient tool for the long-term storage of vegetatively propagated plants. This study was conducted to investigate the effects of slow-freezing techniques on the cryopreservation of potato. In vitro plantlets of the potato genotypes of 'Atlantic', 'Superior’, 'Namseo', 'J138', and 'CTO5-5' were cold acclimated, and the excised axillary buds were precultured, osmoprotected, exposed to plant vitrification solution, frozen slowly to $-40^{\circ}C$ and then rapidly plunged into liquid nitrogen, thawed and finally plated on the regeneration medium. It was found that the higher the sucrose concentrations in the subculture medium of donor plantlets, the higher the survival rates of shoot tips after cryopreservation, and the highest survival (20%) was observed in the medium added with 0.25 M sucrose. As for the effect of cooling, $0.3^{\circ}C/min$ cooling speed showed the highest survival (25%). Different varieties showed different responses over different cryopreservation treatments. Survival rate was increased by slow-freezing technique method as compared with that of the basic cryopreservation method of vitrification alone in the diverse potato genotypes. Leaf and tuber morphologies of potatoes regenerated after cryopreservation using slow freezing technique were similar to those derived from the in vitro stock plantlets.

• Korean Journal of Veterinary Research
• /
• v.33 no.3
• /
• pp.547-554
• /
• 1993

This study was carried out to investigate the best condition for in vitro and in vivo culture after freezing and thawing of nuclear transplant 2 cell embryos. When nuclear transplant embryos were submitted to electrofusion, the significantly higher fusion rates of 2 cell donor nuclei were achieved at the electric field strength of DC 1.5 kV/cm for 100 and $150{\mu}sec$, DC 2.0 kV/cm for 100 and $150{\mu}sec$ than DC 1.0 kV/cm for 100 and $150{\mu}sec$(p<0.01). The significantly higher fusion rates of 4 cell donor nuclei were achievecl at DC 2.0 kV/cm for 100 and $150{\mu}sec$ than DC 1.0 kV/cm for 100 and $150{\mu}sec$(p<0.01). The fusion rates in 8 cell donor nuclei were 94.2~99.3%. The developmental potency to blastocyst in 2 cell donor nuclei was significantly higher in DC 2.0 kV/cm for $150{\mu}sec$ treated group(p<0.01). The significantly higher developmental potency to blastocyst in 4 cell donor nuclei were achieved at the electric field strength of DC 2.0 kV/cm for $150{\mu}sec$ than DC 1.5 kV/cm for 100 and $150{\mu}sec$, DC 2.0 kV/cm for $100{\mu}sec$ treated group(p<0.01). The develop mental potency to blastocyst in 8 cell donor nuclei was significantly higher in DC 2.0 kV/cm for $100{\mu}sec$ treated group(p<0.01). The developmental potency to blastocyst after nuclear transplantation was significantly higher in 2 cell donor nuclei than in 8 cell donor nuclei(p<0.01). When the recovered embryos in normal morphology were cultured in vitro, there were no significant differences in the developmental potency to blastocyst between the freezing methods and the concentrations of cryoprotectant(p<0.01). The production rates of offspring after transfer of nuclear transplant embryos to recipient mouse were no significant difference in 2, 4 and 8 cell donor nuclei.

• Korean Journal of Animal Reproduction
• /
• v.17 no.4
• /
• pp.339-346
• /
• 1994

An understanding of the structure and function of mammalian spermatozoa requires the iso-lation of these components. In this study, frozen-thawed bovine spermatozoa were treated by physical treatments (vortexing, 26 gauge needle, strained 26 gauge needles and freezing-thawing) or chemical treatments (trypsin, dithiothreitol, sodium dodecylsulfate and $\beta$-mercaptoethanoJ) to yield free heads and tails. The most effective treatment was repeated pumping of sperm suspension through a strained 26 gauge needle conneted to a syringe. Spermatozoa by this treatment were mainly broken at the junction of the head and the tail, resulting in 90-100% yields. Also, sperm head surface did not modify during strained 26 gauge needle treatment when either spermatozoa or sperm heads were incubated in 250${\mu}\textrm{g}$/ml of FITC-UEA 1 for 1 h at room temperature to detect the modification of sperm surface components. Other physical treatments were less efficient for the breakdown of spermatozoa. The effects of chemical treatments on bovine spermatozoa are not noticeable. Dissected sperm heads and tails should be fractional leading to nearly pure components by sucrose gradient centrifugation at 1,000 rpm for 15 min. The result suggest that the established method may be useful for the biochemical study of spermatozoal components, and the understanding of oocyte activation mechanism either by spermatozoal components during fertilization or microinjection of isolated components.

• Clinical and Experimental Reproductive Medicine
• /
• v.25 no.2
• /
• pp.129-134
• /
• 1998

The objective of this study was to test whether the developmental capacity of human multi-pronuclear (PN) zygotes after ultra-rapid freezing using EM grid can be maintained. For this experiment, multi-PN zygotes which produced in human IVF program were used as an alternatives of normal 2PN zygotes, and they were separated into 3PN or $\geq4PN$ zygotes to compare their in vitro development and cryoinjury according to PN number. As freezing solution, EFS30 which consisted of 30% ethylene glycol, 18% bcoll, 0.5 M sucrose and 10% FBS added D-PBS was used. The result obtained in this experiment was summarized as follows; When the multi..PN zygotes were ultrarapidly frozen and thawed, the high mean percentages (85.5%) were survived. Also when the cleavage rates between control and freezing group were compared with PN number, there were not significantly different in each group (3PN; 81.3% & 85.4% and $\geq4PN$; 90.0% & 95.7%). When the in vitro development rates after thawing were examined, freezing 3PN group (22.0%) was not differed to control 3PN group (38.5%), although the development result of freezing $\geq4PN$ group (45%) was significantly lower than that of control $\geq4PN$ group (44.4%) (p<0.05). These results demonstrate that developmental capacity of human zygote can be obtained by ultra-rapid freezing method using EM grid and EFS30.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.36 no.2
• /
• pp.228-232
• /
• 2007

The purpose of the study was to investigate the quality characteristics of Jeju island's indigenous pork fed with citrus byproducts. Samples were the Jeju island's indigenous pork loin without citrus byproduct (JNP-0) and the Jeju island's indigenous pork loin fed with 8% and 15% citrus byproducts during growing and fattening periods (JNP-1). The pH, VBN content, bacterial counts, L* value, frozen loss, thawing loss, water boiling loss, hardness, springiness, cohesiveness, chewiness, and shear force value were not significantly different between JNP-0 and JNP-1 (p<0.05). The TBARS, a* value, b* value, water holding capacity, and pan boiling loss of JNP-0 were significantly higher than those of JNP-1 (p<0.05), but the gumminess of JNP-1 was significantly higher than that of JNP-0 (p<0.05). For sensory characteristics, taste, flavor, juiciness, and palatability were not significantly different between JNP-0 and JNP-1, but tenderness of JNP-1 was sig-nificantly higher than that of JNP-0 (p<0.05).

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.38 no.6
• /
• pp.766-772
• /
• 2009

In this study, the effects of feeding citrus byproduct on physicochemical and sensory characteristics of chicken meats were investigated. The samples consisted of chicken meats provided with only feed for laying hen without citrus byproduct (T-0), and the chicken meats fed with 1.0%, 1.5% and 2.0% citrus byproduct during the starter (initial period feed; $1{\sim}9th$ day), the grower (middle period feed; $10{\sim}24th$ day), and the finisher (latter period feed; $25{\sim}36th$ day), respectively. The $L^*$ value of thigh was significantly lower in the T-1 than in the T-0, the $a^*$ value was significantly higher in the T-1 than in the T-0 (p<0.05). The water holding capacity of thigh was significantly higher in the T-1 than in the T-0 and the cooking loss was significantly higher in the T-0 than in the T-1 (p<0.05). The acid value was significantly higher in the T-0 than in the T-1 (p<0.05). Antioxidant activity was higher in the T1 than in the T-0 (p<0.05). There was no significance between T-0 and T-1 regardless of feeding citrus byproduct, in terms of chicken's $b^*$ value, frozen loss, thawing loss, hardness, springiness, cohesiveness, gumminess, chewiness, shear force, free amino acid content of hot water extracts, taste, flavor, tenderness, juiciness and palatability.