• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.49 no.6
• /
• pp.772-778
• /
• 2016

To develop a value-added product from individually quick-frozen oysters Crassostrea gigas (IQFO), we prepared a retort pouched oyster soup (RPOS) from IQFOs and characterized its processing conditions and quality metrics. We found that the most appropriate manufacturing process for the RPOS consisted of half-thawing and washing raw IQF oysters, blanching, adding them to the retort pouch along with other ingredients (base soup stock, IQF oyster extract, radish, bean sprouts, garlic, and red pepper), sealing, retort sterilization ($120^{\circ}$, F0-value 10 min.), cooling, and packaging inspection. The moisture, crude protein, pH and salinity of the RPOS were 91.0%, 2.8%, 6.20 and 0.9%, respectively. The total amino acid content of the RPOS was 2,163.8 mg/100 g, and the main amino acids were glutamic acid, aspartic acid, leucine, proline, lysine and arginine. The primary inorganic ions were Na, K, S and Zn. In taste compounds, total free amino acid content was 313.4 mg/100 g, and the main free amino acids were glutamic acid, taurine, proline, hydroxyproline, aspartic acid, glycine, alanine, valine, lysine and arginine. This RPOS has good storage stability and organoleptic qualities compared with commercial retort pouched shellfish soup, and is suitable for commercialization as a value-added instant seafood soup.

• Journal of Embryo Transfer
• /
• v.9 no.1
• /
• pp.65-71
• /
• 1994

This study were carried out to investigate the effective concentration of cryoprotectant agents and sucrose by one-step straw method, and to determine the optimum thawing temperature and equilibration time of frozen bovine embryos. The bovine embryos following dehydration by cryoprotective agents and a various concentration of sucrose were directly plunged into liquid nitrogen and thawed in 3O$^{\circ}C$ water. Survival rate was defined by FDA test. The results are summarized as follows : 1. The survival rate of bovine embryos thawed after rapid freezing in the freezing medium containing a various kinds of cryoprotective agents added 0.25M and O.50M sucrose were 28.6% and 25.0%, 35.1% and 31.6%, 32.4% and 24.4%, 34.2% and 28.2%, 18.9% and 17.6%, 14.7.% and 21.6%, respectively. 2. The survival rate of bovine embryos thawed after rapid freezing in the freezing medium containing a various concentration of sucrose added 1.5M and 2.OM glycerol, i.5M and 2.OM DMSO and 1.5M and 2.OM propanediol were 22.9~37.8%, 2O.7~31. 3%, 19.2~30.0% and 17.2~25.0%, respectively. 3. The temperature thawed at 2$0^{\circ}C$ after rapid freezing of bovine embryos resulted in a significantly higher embryos survival rate than did at 3$0^{\circ}C$ and 35$^{\circ}C$. 4. The equilibration time on the survival rate of bovine embryos was attained after short period of time(2.5~5 min.) in the freezing medium higher than long period of time (1O~20 min.).

• Korean Journal of Veterinary Research
• /
• v.35 no.3
• /
• pp.635-641
• /
• 1995

The purpose of this study was to investigate the effects of developmental stage and equilibration time on survival of rabbit embryos following freezing by vitrification. Adult New Zealand White female rabbits were superovulated with PMSG and hCG. The 8-cell stage embryos were collected from 40 to 45 hours after hCG injection by flushing oviducts with Dulbecco's phosphated buffered saline and in vitro cultured in TCM-199 containing 10% fetal calf serum(FCS). Each embryos developed in vitro to 16-cell, compact morula and blastocyst was cryopreserved and cultured following thawing to examine their developmental potential to expanded blastocyst stage in vitro. The frozen-thawed-cultured embryos were stained with Hoechst 33342, and their nuclei were counted using a fluorescence microscope. On the toxicity test of EFS solution as cryopreservation, the survival rates of 8-cell stage embryos was decreased in reverse to increasing of exposure time over 5 minutes. The post-thaw survival rates of embryos on equilibration times was significantly(P<0.05) higher for 2 min. than for 5 or 10 minutes. From morula to blastocyst of rabbit embryos was more suitable than 8-cell stage for cryopreservation by vitrification. The higher post-thaw survival rate of embryos can be achieved by keeping the cryoprotectant at $4^{\circ}C$ than at $20^{\circ}C$. The mean number of nuclei per embryo following freezing by vitrification and in vitro culture to expanded blastocyst at compacted morula and blastcyst was not significantly differ from fresh blastocyst.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.51 no.6
• /
• pp.632-639
• /
• 2018

To develop the high-value added seafood products from a regional speciality seafood, the seasoned dried pen shell Atrina pectinata adductor (SDPA) and seasoned smoke-dried pen shell adductor (SSPA) samples were prepared, and their optimal processing conditions, quality metrics, and shelf-life characteristics were examined. SDPA and SSPA samples were produced by thawing of frozen pen shell adductor, and cutting it into 6-7 mm slices, hot-air drying ($60^{\circ}C$, 20 min) or smoking ($110^{\circ}C$, 20 min), seasoning ($4^{\circ}C$, 12 h) with seasoning powder (60% sorbitol, 15% sucrose, 16% salt and 9.0% monosodium glutamate), hot-air drying ($60^{\circ}C$, 3 h), torching, vacuum-packaging in a laminated plastic film bag, heat treating with hot-water ($85^{\circ}C$, 15 min), and cooling. The moisture content of SDPA and SSPA samples was 44.5 and 43.0%, respectively, and the water activity was 0.845 and 0.842. The total amino acids in SDPA and SSPA samples were 20,986.8 and 21,312.4 mg/100 g, respectively, and the major amino acids in both products were aspartic acid, serine, glutamic acid, proline, glycine, alanine, valine, leucine, phenylalanine, lysine and arginine. The primary minerals were Na, S, K and P. Incubating tests indicated that the quality of SDPA and SSPA samples was maintained for 30 days of storage.

• Journal of Animal Reproduction and Biotechnology
• /
• v.37 no.2
• /
• pp.130-135
• /
• 2022

Epididymal sperm cryopreservation provides a potential method for preserving genetic material from males of endangered species. This pilot study was conducted to develop a freezing method for tiger epididymal sperm. We evaluated post-thaw sperm condition using testes with intact epididymides obtained from a Siberian tiger (Panthera tigris altaica) after castration. The epididymis was chopped in Tyrode's albumin-lactate-pyruvate 1x and incubated at 5% CO₂, 95% air for 10 min. The Percoll separation density gradient method was used for selective recovery of motile spermatozoa after sperm collection using a cell strainer. The spermatozoa were diluted with modified Norwegian extender supplemented with 20 mM trehalose (extender 1) and subsequent extender 2 (extender 1 with 10% glycerol) and frozen using LN₂ vapor. After thawing at 37℃ for 25 s, Isolate^® solution was used for more effective recovery of live sperm. Sperm motility (computerized assisted sperm analysis, CASA), viability (SYBR-14 and Propidium Iodide) and acrosome integrity (Pisum sativum agglutinin with FITC) were evaluated. The motility of tiger epididymal spermatozoa was 40.1 ± 2.0%, and progressively motile sperm comprised 32.7 ± 2.3%. Viability was 56.3 ± 1.6% and acrosome integrity was 62.3 ± 4.4%. Cryopreservation of tiger epididymal sperm using a modified Norwegian extender and density gradient method could be effective to obtain functional spermatozoa for future assisted reproductive practices in endangered species.

• Proceedings of the Korean Society of Embryo Transfer Conference
• /
• 2002.11a
• /
• pp.83-83
• /
• 2002

The purpose of this study was to investigate the warming temperature and exposed time on the post-thaw survival rate and viability of bovine blastocyst cryopreserved by GMP vitrification. Groups of three bovine IVP blastocysts were sequentially placed into vitrification solution before being loaded into the GMP straws and immersed into LN$_2$within 20 to 25 sec. The warming rate was increased 2 times of warming temperature for improvement of post-thaw survival rates. The frozen embryos were warmed either at 35 or 70$^{\circ}C$ for 1 or 2 sec and then diluted in sucrose solution. Post-thaw blastocysts were serially washed in 0.25 and 0.15 M sucrose in holding medium (HM: TCM199 supplemented with 10% FCS) and TCM-199 for each 5 min, respectively, and then cultured in TCM199 for 24 h. The rate of re-expanded blastocyst was significantly different fer 35 and 70$^{\circ}C$ warming temporature (76.4 vs. 89.3%; P<0.05). The rate of re-expanded blastocyst at 70$^{\circ}C$ for 1 sec was significantly higher than that for 2 sec (91.1 vs. 70.9%; P<0.05). The number of nuclei counted were significantly different among control, 35 and 70$^{\circ}C$ (121${\pm}$8.5 vs. 104${\pm}$11.7 vs. 114${\pm}$10.3; P<0.05). These results indicated that the increasing of warming rate can provide high survival rates of bovine IVP blastocysts. Especially, the best viability of post-thaw blastocyst could be thaw at 70$^{\circ}C$ for 1 sec. The warming temperature and exposed time far warming was considered to be limiting factors to the viability of bovine IVP embryos. he purpose of this study was to investigate the warming temperature and expose.

• Journal of the Korean Geographical Society
• /
• v.43 no.3
• /
• pp.296-311
• /
• 2008

In order to examine the rates and factors of path widening in Mount Halla, the retreat of path sidewalls was monitored at 32 sites of Seongpanak Hiking Trail located between 875 m and 1,400 m in elevation. The mean rate of sidewall retreat for the period 2002-2008 is 50.6 mm, equivalent to 10.0 mm/yr. The retreat rate of frozen period is 19.3 mm/yr, while the rate of unfrozen period is 4.3 mm/yr. The latter is divided into the rainy and dry periods that exhibit the retreat rates of 5.9 mm/yr and 2.9 mm/yr, respectively. The retreat rate of sidewalls is also varied with seasons; winter shows the maximum rate of 42.2 mm/yr, while summer exhibits the minimum rate of 1.3 mm/yr. Spring and fall show the intermediate rates of 13.9 mm/yr and 6.4 mm/yr, respectively. Soil hardness and elevation are not closely related to the retreat rate of sidewalls, even though the retreat rate is larger at the north-faced sidewalls than the south-faced sidewalls during the frozen period. Pipkrake is likely to be the most important factor contributing to the path widening in that the retreat of winter months accounts for 76.7% of the total retreat. The hiking trail is placed under the climatic conditions which develop pipkrake in 85 days annually. In addition, it is usual to observe the path sidewall covered with pipkrake in the freezing month of December and the thawing months of March and April. On the other hand, deflation and rainsplash erosion are not important due to the weak wind speed and the forested trail. Rainwash is also insignificant in that the path has been almost paved to mitigate trampling effects. Although biological activity is not dominant, hikers cause a large retreat of sidewalls in the thawing months since they would walk on the sidewalls to avoid snow-melting pools on the path.

• Journal of Veterinary Clinics
• /
• v.30 no.6
• /
• pp.442-448
• /
• 2013

The aim of the present study was to develop glycerol-free TRIS extender using glucose for dog sperm cryopreservation. We determined the appropriate concentration of glucose in glycerol-free TRIS and the exposure time in glycerol-free TRIS containing 0.3 M glucose at $4^{\circ}C$. Ejaculates of six dog sperm were cooled in glycerol-free TRIS through $4^{\circ}C$ for 100 min, cooled at $4^{\circ}C$ in TRIS with different glucose concentrations 0 M, 0.04 M, 0.1 M, 0.2 M and 0.3 M, respectively for 30 min followed by cryopreservation. After thawing at $37^{\circ}C$ for 25 sec, membrane and acrosome integrities of dog sperm were evaluated. In addition, the effect of exposure time (10, 30, 50 and 70 min) of sperm to glycerol-free TRIS containing 0.3 M glucose at $4^{\circ}C$ on progressive motility, viability, and DNA integrity following sperm cryopreservation was studied. Membrane integrity and acrosome integrity were assessed by 6-carboxyfluoresceindiacetate (6-CFDA)/propidium iodide (PI) fluorescent staining and Pisum sativum agglutinin conjugated to fluorescein isothiocyanate, respectively. DNA integrity was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling, using flow cytometry. Sperm frozen in glycerol-free TRIS supplemented with 0.2 M or 0.3 M glucose have an intact plasma membrane (CFDA+/PI-) after cryopreservation than sperm frozen in the extenders with lower glucose concentrations (p<0.05). Acrosome integrity was significantly higher in the 0.3 M group than less than 0.1 M groups (p<0.05). The sperm DNA fragmentation index did not differ according to exposure time, although progressive motility was significantly higher in the 50 min exposure group than the other groups (p<0.05). These results indicate that cryopreservation of dog sperm is feasible and yields more motile sperm following freezing and thawing in glycerol-free TRIS containing 0.3 M glucose with the exposure time for 50 min at $4^{\circ}C$.

• Development and Reproduction
• /
• v.4 no.2
• /
• pp.161-173
• /
• 2000

The aim of this study was to assess the effect of the frequency of the L$N_2$ infusion on the ultrastructure, metabolism and development of the embryo after freezing and thawing by computerized cell freezer. Two-cell embryos of ICR mouse were randomly allocated into fresh (control), high-frequency freezing (group 1) and low-frequency freezing (group 2). For fresh and frozen-thawed intact 2-cell embryos, total ceil number in the blastocyst was counted by fluorescent microscope after Hoechst 33258 staining. Relative amount of $H_2O$$_2$ was measured by DCHFDA. Intracellular location and membrane potential of the mitochondria were evaluated by staining with rhodamine 123 and JC-1. The structure of actin filament was also evaluated by confocal microscope. DNA fragmentation was assessed by TUNEL method after development into blastocyst. The survival rate of intact embryo was higher in group 1 than group 2 (50.7％ vs. 34.6％ respectively, p<0.05). The blastocyst developmental rate was significantly low in group 2 (86.7％, 76.7％ vs. 44.0％ for control, group 1 and group 2 respectively, p<0.05). Total cell number in the blastocyst was also significantly lower in group 2 than control (79.5$\pm$12.9, 71.6$\pm$8.0, and 62.5$\pm$4.7 for control, group 1 and group 2 respectively, p<0.05). The relative amount of $H_2O$$_2$ was higher in group 2 than other groups (15.3$\pm$3.0, 16.6$\pm$1.6 vs. 23.4$\pm$1.8, p<0.05). After JC-1 staining, relative intensity of mitochondria with high membrane potential was significantly lower in group 2 than control and group 1 (17.2$\pm$3.8, 17.4$\pm$1.3 vs. 13.2$\pm$2.0, p<0.05). In group 2, partial deletion and aggregation of the actin filament was found. DNA fragmentation rate was also hieher for group 2 versus other groups (30.8％, 36.0％ vs. 65.6％, p<0.05). The frequency of the L$N_2$ infusion is an important factor for the development of frozen-thawed mouse embryo. High-frequency infusion may prevent damages of cytoskeleton and mitochondria in the embryo probably by preventing the temperature fluctuation during dehydration phase. We speculate that the application of high-frequency infusion method in human embryo may be promising.

• Korean Journal of Animal Reproduction
• /
• v.20 no.1
• /
• pp.35-42
• /
• 1996

This study was carried out to investigate the effect of straw loading method and thawing protocol on the in vitro development of in vitro produced mouse blastocysts cryopreserved by vitrification. Three loading types of straw I, Il and III on loading and sealing method were made for vitrification. The ability of the solution on straw loading methods to remain vitreous during warming was tested by exposed in air for 1 to 10 s sec. and then plunged the vitrified straws into water bath at 25°C. Embryos to be vitrified were equilibrated to the 20% EG for 5min. and exposed in EFS 40 for 1min. The plug ends of Straw I and Straw II were sealed with straw powder and Straw III was treated straw powder, followed by heat sealing and then plunged into LN$_2$. The results obtained in these experiments were summarized as follows; 1) Straw I embryo column mostly changed from transparent to opaque upon thawing without exposure in air for 3-6 sec. Straw II embryo column was I improved partially but was not remained completely vitreous during warming. However, when Straw lll loading method was used, the embryo column was remained vitreous completely. 2) High survival rates and development rates of each groups (middle blastocysts and hatching blastocysts) of vitrified embryos were obtained by using Straw III loading method than Straw I method (P<0.05). And the range of s standard error was low in Straw lll method. 3) When the embryos vitrified-frozen were placed in air for 3, 5 and l0sec. and then warmed rapidly in water bath at 25$^{\circ}C$, the survival rates after 24h of culture were 72.7-87.1% and the development rates to hatching stage after 48h of culture were 34.0-48.4%. There were no significantly differences according to exposure time in air during warming. In conclusion, the present results showed that highly survival and low standard error of vitrified-frozen mouse bIastocysts were obtained by using straw lll loading, double sealing and appropriate 2 step warming method.