• Journal of Embryo Transfer
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• v.17 no.2
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• pp.91-100
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• 2002

The experiment was divided into two phases. In phase-I fresh embryos were transferred and in Phase-II frozen embryos were transferred. Embryos were collected by using Dulbecco's phosphate buffered saline. In phase-I total of 65 ova were collected out of 107 ovulation in 18 goats. Recovery of ova was 60.74％, of which 51 (78.46％) was fertilized. Sixteen embryos were transferred to 10 recipient goats and kidding was observed in 6 goats, that produced 10 kids. Thus, 62.50％ embryo survival and 60％ kidding were achieved in phase-I. In phase-II of the experiment, 17 regular cyclic Black Bengal goats were used. The main purpose was to study the viability of caprine embryos after cryopreservation. In this phase the embryos were collected and frozen using Bio-cool freezers. A two step addition of cryoprotectants (5％ glycerol and 10％ glycerol) and three-step dilution of cryoprotectants with 1mole (M) sucrose was used. Embryos were preserved for 10 to 45 days. Out of 27 embryos preserved, 18 were recovered after freezing and thawing (37$^{\circ}C$ water bath) with 33.33％ embryonic loss. Seventeen frozen and thawed embryos were transferred in 9 recipient goats, out of which kidding was observed in 6 goats and 7 kids were produced, giving a 66.66％ kidding and embryo survival of 41.17％. The technique utilized for fresh and frozen embryo transfer can be successfully utilized to produce goats of superior genetic merits. The protocol used for addition of cryoprotectant, freezing, thawing and dilution was found suitable for caprine embryo freezing.

• Korean Journal of Animal Reproduction
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• v.7 no.1
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• pp.19-23
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• 1983

Present studies were conducted to investigate the developmental stage and the location of embryos in the reproductive tract at various times after ovulation, the morphologically normal after thawing of embryos preserved in liquid nitrogen, and the survival after transferring frozen-thawed embryos. The results obtained were as follows: 1. Embryo stage and location in the reproductive tract after hCG administration. For the investigation of embryo stage and location in the reproductive tract after ovulation, rabbits were laparotomized at 24, 40, 48, 72 and 120 hrs post hCG injection, simultaneously with mating. the oviducts and uteri were flushed out with PBS medium containing 50% rabbit serum, respectively. 1) Most of embryos was remained in the oviduct within 48 hrs, with the lapse of time, embryos were started to move to uterus and shifted in uterus at 72 hrs after hCG injection. 2) The representatives of embryos stage collected at 24, 40, 48, 72 and 120 hrs were 1-cell(60.4%), 8-cell to early morula (52.3, 39.3%), late blastocyst (95.5%) stages, respectively. 2. Morphological normality and survival of the frozen-thawed embryos. For the evalution of the quality and viability on the frozen-thawed embryos, immediately after thawing, embryos were assessed by morphologically normal under a dissecting microscope, and a further test of frozen-thawed embryos was made by transferring the morphologically normal embryos to the uteri of recipient rabbit induced pseudopregnancy by the injection of hCG at the time of hCG injection in donor rabbits. 1) The propotions of embryos which a, pp.ared morphologically normal was higher when 8-cell (85.7%) and morula(90.5%) were used for freezing than when 4-cell (66.7%) and blastocyst (75.8%) were used. 2) Preganacies were observed at Day 15 after transfer of frozen-thawed 8-cell (7/13), morula (19/42) and blastocyst (3/19) but not after transfer of embryos at 4-cell stage.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.51 no.1
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• pp.91-94
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• 2018

We develop a time-temperature indicator (TTI) that can determine whether thawing of fish and other fishery products has occurred during frozen storage. A polypropylene tube with an internal diameter of 3 mm was prepared and cut to a length of 14 to 20 mm. One end of the tube was thermally sealed and 0.1% acetic acid was injected into the other end; the tube was then frozen at $20^{\circ}C$. Then the open side of the frozen tube was blocked by sinking the tube into a 10% gelatin solution. The tube was attached to a polyvinyl packaging bag along blue litmus paper and the bag was put into a freezer at $-20^{\circ}C$. After freezing, the bag was removed to an ambient temperature of $20^{\circ}C$, and the time dependence of the color change of the litmus paper was observed. The color changed from blue to red, with the length of the red region increasing with time. Our TTI can be used as a part of a visible detection system and the detection program can conduct the elapsed time analysis on the length of the red region of the litmus paper indicating the degree of thawing. Thus, the TTI is a useful tool in the temperature management of frozen fish and fishery products.

• Journal of Veterinary Clinics
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• v.34 no.2
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• pp.156-160
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• 2017

We investigated the effect of ethylene glycol and antioxidants such as taurine, hypotaurine and trehalose with extenders during cryopreservation of Korean Jeju Black Bull spermatozoa. The cryopreservation of freshly collected spermatozoa was conducted with four different conditions. As a control, spermatozoa were cryopreserved with Tris egg-yolk extenders added 5% ethylene glycol (EG). Taurine (20 mM), hypotaurine (20 mM) and trehalose (20 mM) were individually added into tris egg-yolk extenders with 5% EG. After thawing of frozen spermatozoa with four different conditions, sperm viability, motility, acrosomal integrity, and membrane integrity were investigated. The significant (p < 0.05) improvement of sperm viability showed in all antioxidant treated thawed spermatozoa (taurine; $68.1%{\pm}4.4$, hypotaurine; $69.2%{\pm}6.7$ and trehalose; $68.0%{\pm}4.4$) when compared to control ($63.4%{\pm}5.6$). Neither positive nor detrimental effects of three antioxidants were shown sperm motility after thawing. The results of hypo-osmotic swelling test showed that the membrane integrity of taurine, hypotaurine or trehalose treated thawed spermatozoa ($64.1%{\pm}5.4$, $61.5%{\pm}3.7$ and $59.0%{\pm}4.0$, respectively) had significantly (p < 0.05) higher rate of the swollen sperm compared to control ($53.7%{\pm}9.7$). Hypotaurine treated frozen-thawed spermatozoa had siginificantly higher (p < 0.05) F pattern ratio than taurine, trehalose and control treated frozen-thawed spermatozoa. Trehalose added frozen-thawed spermatozoa had significantly higher (p < 0.05) acrosome reaction pattern ratio than taurine and hypotaurine added frozen-thawd spermatozoa. In this study, we found that antioxidants such taurine, hypotaurine and trehalose treatments during cryopreservation process could reduce damage of spermatozoa of Korean Jeju Black Bull and improved sperm capability of fertilization.

• Journal of Fisheries and Marine Sciences Education
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• v.28 no.4
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• pp.1152-1158
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• 2016

This study was used samples, mackerel (Scomber japonicas), Japanese Spanish mackerel (Scomberomorus niphonius), alaska pollock (Theragra chalcogramma), yellow croaker (larimichthys crocea) known as some of the major species fisheries products in Korea We were investigated temperature change during thawing processing, water holding capacity, drip loss, extractive-nitrogen and viable cell count by various thawing methods, thawing at the room temperature (TRT), hot-air thawing (HAT), aeration thawing (AT), high-frequency thawing (HFT). The temperature changes have taken 2~3 hours in HFT and 24 hours by TRT. The expressible drip loss was 0.47~0.87 g/100 g in HFT, 1.91~4.42 g/100g in TRT, 1.31~4.93 g/100g in HAT, and 2.01~4.59 g/100g in AE. The water holding capacity was higher samples thawing by HFT than other thawing methods. Extractive-nitrogen was 276~452 mg/100 g in HFT, 177.21~420.27 mg/100 g in TRT. Viable cell count was $10^2$ to $10^3$ in HFT, $10^3$ to $10^5$ in other thawing methods. The processing speed and uniformity by HFT was minimized the risk of product degradations (drip losses, deterioration of sensorial, chemical and physical characteristics, bacteria growth, etc.), thus helping to preserve at its best the product quality than those by thawing methods. Therefore, HFT was expected to make high-quality thawing products anticipate future thawing technology

• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.103-109
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• 1997

The studies on the carried out to investigate to determine the optimum thawing temperature and equilibration time and 1 step straw method of frozen bovine embryos. The follicular oocytes were cultured in TCM-199 medium containing 10 IU/ml PMSG(Sigma, USA), 10 IU/ml hCG(Sigma, USA), 1 $\mu\textrm{g}$/ml $\beta$-estradiol(Sigma, USA) and 10% FCS for 24~48 hrs in incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18 hrs with motile capacitated sperm by preincubation of heparin. The bovine embryos following dehydration by cryoprotective agents and various concentration of sucrose were directly plunged into liquid nitrogen and thawed in 3$0^{\circ}C$ water. Survival and in vitro developmental rate was defined as developmental rate on in vitro culture or FDA-test. The results are summarized as followes : 1. The equilibration time on in vitro developmental rates of bovine embryos was attained after short period of time(2.5~5 min.) in the freezing medium higher than long period of time (10~20 min.). 2. The temperature thawed at 3$0^{\circ}C$ after rapid freezing of bovine embryos resulted in a significantly higher in vitro developmental rate than did at 2$0^{\circ}C$ and 35$^{\circ}C$. 3. The thawing time on in vitro developmental rates of bovine embryos was attained after short period of time(1~5 min.) in the freezing mediuim higher than long period of time(10min.). 4. The in vitro developmental rates of bovine embryos after rapid frozen-thawing by 1 step straw method in the freezing medium added 1.5M, 2.0M glycerol, DMSO, propanediol and 0.25M, 0.50M, 0.75M, 1.00M sucrose were 12.5~19.4%, 10.0~15.6%, 9.1~13.8% and 6.7~12.9%, respectively.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.2
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• pp.127-133
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• 2003

Objective: The aim of this study was to investigate the morphological aspects of testicular tissue before and after freezing-thawing by light and transmission electron microscopy. Methods: Tissue biopsies were carried out on mouse testis for freezing. Samples in medium containing 20% glycerol were frozen by computer-controlled freezing program. The effect of freezing-thawing on the structural change of testicular tissues were examined by light and electron microscopy. Results: The freezing-thawing procedure had no significant effect on tubular diameter. However, it caused folding of the lamina propria, and notable damage to Sertoli cells, spermatogonia and spermatocytes. The cells were detached, desquamated from the basal lamina and had increased vacuolization. Round spermatids, elongated spermatids and spermatozoa were less affected, and most of them maintained their normal structure. Conclusions: The structure of spermatogonia, spermatocyte and basal compartments in seminiferous epithelium was significantly altered by freezing-thawing procedure of mouse testicular tissues. Thus, we need to develop a more reliable method for the cryopreservation of testicular tissues.

• Journal of Veterinary Clinics
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• v.17 no.2
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• pp.420-430
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• 2000

These studies were preformed to investigate the freezing conditions to achieve good post-thaw viability of spend and the practical methods of artificial insemination frozen canine semen. Semen were collected from nine male dogs which had been proved to be fertile in the past and the semen were treated for freezing procedure. Post-thaw motility and viability of canine sperm were evaluated to investigate individual tolerance of freezing, difference among freezing extenders, dif-ference among freezing equipments and freezing conditions, difference between fast and slow cooling rate, difference according to different glycerol concentration, effect of seeding on post-thaw viability, difference according to cutting part of straw, difference according to thawing temperatures, and dif-ference according to media added to thawed semen. Thawed semen for insemination were added with equal volnme of canine capacitation medium (CCM) and the volume of semen and the number per insemination were adjusted as 2-3 ml and $20-30 {\times}10^7,$ respectively. The semen were inseminated in vagina using balloon catheter and en17ryos were cellected from 9 to 11 days after the second Al to d determine fertilization.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.12
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• pp.1756-1763
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• 2017

Objective: The study aimed at assessing the effects of frozen storage duration on quality characteristics, lipid oxidation and sensory quality of various horse muscles. Methods: Five representative muscles: longissimus dorsi (LD), gluteus medius (GM), semimembranosus (SM), biceps femoris (BF), and triceps brachii (TB) at 24 h post-mortem obtained from 28-mo-old Jeju female breed horses (n = 8) were used in the present investigation. The muscles were vacuumpackaged and frozen at $-20^{\circ}C$ for 120, 240, and 360 days. All the samples were analyzed for thawing and cooking losses, pH, Warner-Bratzler shear forces (WBSF), color traits, total volatile basic nitrogen (TVBN), thiobarbituric acid reactive substances (TBARS) and sensory traits. The muscle samples analyzed on day 0 of frozen storage (fresh, non-frozen) were used for comparison. Results: Results revealed that thawing and cooking losses significantly (p<0.05) increased in all the muscles after 120 days and then remained unchanged up to 360 days of frozen storage. The TBARS and TVBN contents significantly increased as increasing frozen storage time up to 360 days (p<0.05). While, significant decreases in WBSF values were observed for all the muscles with increased frozen storage time (p<0.05). Frozen storage variously affected the color traits of the muscles for instance; the redness of LD, GM, and BF muscles showed a decreasing tendency during frozen storage while it was not changed in TB and SM muscles. Furthermore, the frozen storage did not produce detrimental effects on sensory quality as it did not cause flavor and juiciness defects whereas it partially improved the tenderness of all the muscles studied. Conclusion: Based on the results obtained from our work, it is concluded that frozen storage could be applied to increase the long-term shelf life of horsemeat while still retaining its sensory quality.

• Korean Journal of Animal Reproduction
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• v.16 no.3
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• pp.217-224
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• 1992

These studies were carried out ot investigate on the transferred embryo development following ultrarapid frozen for 8-cell and morula of in vitro fertilization mouse embryos. The post-thaw embryo survival was evaluated and compared by cell stage of embryos and by equilibration time before ultrarapid freezing. The results obatined were summerized as follows: 1. The effects of equilibration time of 3 vs. 6 minutes before ultrarapid freezing and after thawing on the morphological survival and the viability of 8-cell and morulas embryos were not significant. 2. When the ultrarapid frozen-thawed 32 eight-cell and 33 morula embryos, and 30 fresh blastocysts were transferred to pseudopregnant recipient mice, the number of normal offsprings produced were 9(28.1%), 14(42.4%) and 18(60.0%), respectively. From the above resutls, it was concluded that the optimal conditions of pH osmolality of the media for mouse IVF and embryo culture, and the period of sperm preincubation might be 7.1, 310 mOsm and 120 min., respectively,a nd somewhat high conception rate might be resulted from transfer of frozen embryos of morula stage and fresh embryos of blastocyst stage.