• Journal of fish pathology
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• v.22 no.3
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• pp.335-342
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• 2009

The stability of immunoglobulin M (IgM) on different serum storage conditions and specific antibody response were tested using the serum collected from sevenband grouper Epinephelus septemfasciatus by enzyme-linked immunosorbent assay (ELISA). To test the effect of storage temperature and duration, sevenband grouper antiserum against bovine serum albumin (BSA) was stored at -80, -20 or 4${^{\circ}C}$ for 1, 34, 61 or 119 days. In addition, to test the effect of repeated freeze-thawing condition, the anti-BSA fish serum was frozen at -20 and -80${^{\circ}C}$ and then thawn and frozen for 1, 5 or 10 times repeatedly. Consequently, no significant difference was found in ELISA optical density (O.D.) values of sera for the above mentioned storage conditions: different temperatures (-80, -20 and 4${^{\circ}C}$), durations of storage (1, 34, 61 and 119 days), and repeated thaw-freeze cycles (1, 5, and 10 times), indicating that IgMs of test fish were stable. The specific antibody response of sevenband grouper was observed after BSA-immunization of the test fish reared at 20 ${^{\circ}C}$ or 25${^{\circ}C}$. At the rearing temperature of 20${^{\circ}C}$, the specific antibody against BSA first appeared at 14 days and maximum antibody titer was observed between 21 and 28 days, while at the rearing temperature of 25 ${^{\circ}C}$, specific antibody appeared at 7 days and maximum antibody titer was observed between 14 and 21 days. In conclusion, the rearing temperature at 25${^{\circ}C}$ gave a faster and higher specific antibody response than at 20${^{\circ}C}$ and the specific antibody response maintained for approximately 2 months at 20℃ and 25${^{\circ}C}$.

• Applied Biological Chemistry
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• v.13 no.1
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• pp.105-109
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• 1970

Thiamin in the various materials which used for Takja braving was determined and its quantitative changes in the Takju mashes which were mashed with polished rice and wheat flour respectively using Nuruk only for the enzyme source, and with wheat flour and corn powder respectively using Nuruk and Koji, were checked at 24 hours intervals during the whole brewing period. The results obtained were as follows. a) The thiamin contents in 100 grams of each materials were: Polished rice 107.8${\gamma}$ Wheat Pour 185.0${\gamma}$ Corn powder 410.2${\gamma}$ Nuruk 347.4${\gamma}$ Koji of wheat flour 170.1${\gamma}$ Koji of corn powder 257.3${\gamma}$ b) About 40 to 50 percent of thiamin in the polished rice, wheat flour and, corn powder were losed by steaming at 10 wounds for 50 minutes. c) The quantitative changes of thiamin in the Takju meshes showed a similar tendency. Thiamin contents were decreased markedly during 2 days after mashing but thereafter, almost unchanged till the mashes were ripened. Its decreasing rates in tile two plots uses Nuruk and Koji wereless thaw those in the two plots used Nuruk only. d) Approximately 6${\gamma}$ of thiamin was determined in 100 ml. of the ripened mash of the rice plot and 7.5${\gamma}$ in tire whet flour plot in the case of using Nuruk only. 12.4${\gamma}$ of thiamin was determined in 100 ml. of the ripened mash of the wheat flour plot and 15.4${\gamma}$in the corn powder plot in the case of using Nuruk and Koji. e) It was assumed that 2.4 to 3.5${\gamma}$ of thiamin were contained in 100 ml. of finished Takju of the two plots used Nuruk only and 5 to 7${\gamma}$ in the two plots used Nuruk and Koji. f) About 43.7${\gamma}$ of thiamin were determined in 100 grams of Takju cake of the rice plot and 56.1${\gamma}$ in the wheat flour plat used Nuruk only, and 81.4${\gamma}$ in the corn powder plot and 57.7${\gamma}$ in the wheat flour plot used Nuruk and Koji.

• Korean Journal of Poultry Science
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• v.40 no.3
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• pp.207-216
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• 2013

This study was conducted to establish the method for preserving PGCs that enables long-term storage in liquid nitrogen for developmental engineering or preservation of species. The purpose of this study is to clarify the effects of freeze-thaw treatment on viability of PGCs in chickens. PGCs were collected separately from a germinal gonad of an early embryo of 5.5~6 day (stage 28) of Isa brown, Korean Oge (KO), White Leghorn and Commercial breeds. PGCs separated from a germinal gonad of an early embryo of 5.5~6 day (stage 28) are suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and glycerol). The PGCs were then purified using magnetic activated cell sorting (MACS) method. The viability of PGCs after thawing was $87.4{\pm}0.4%$ and $89.4{\pm}0.2%$ with the 10% EG treatments with no significant difference between the Isa brown and Commercial breeds. The viability of PGCs after freeze- thawing was significantly higher for Isa brown ($87.4{\pm}0.4%$) and Commercial breeds ($89.4{\pm}0.2%$) than Korean Oge (KO) ($77.6{\pm}1.1%$) and White Leghorn ($76.2{\pm}0.9%$)(p<0.05) using 10% EG cryoprotectant. This study established a method for pre- serving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at agermplasm repository and ease of entry into a data base. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

• Journal of Korean Tunnelling and Underground Space Association
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• v.19 no.6
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• pp.1059-1075
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• 2017

The objective of this study was to evaluate the freezing and thawing resistance of the existing drainage system for leakage treatment of underground concrete structures operating in cold regions. The freezing and thawing test was conducted on 4 types of drainage system specimens to evaluate the freezing and thawing resistance of the drainage system. The freezing and thawing resistance was evaluated on 4 types of Hotty-gel, as a waterproofing material, connection methods and on two methods to fix the drainage board with Hotty-gel on the surface of cement concrete specimen. One cycle of the freeze-thaw testing was 48 hours (24 hours of freezing and 24 hours of thawing), and the temperatures of freezing and thawing were at $-18^{\circ}C$ and $10^{\circ}C$, respectively. Among the 4 types of Hotty-gel connection methods, leakage occurred after 28 cycles (8 weeks) of freeze-thawing only in the Hotty-gel connection method with the 'V' groove applied to the corner of the drainage board. No leakage occurred in the 3 types of Hotty-gel connection methods. In two fixing methods, leakage occurred in the method of fixing the drainage board on the cement concrete specimen using the washer, screw and plastic wall plug. Leakage occurred at one point after 10 cycles (3 weeks) of freezing and thawing. After 28 cycles (8 weeks) of freezing and thawing, leakage point increased to 5 points. As time passed, the leak point was not increased, but the amount of leakage was increased at each leak point. The Hotty-gel connection method with cross-sectional diagonal shape was evaluated to be the highest in the production efficiency considering the production time and manufacturing method of the Hotty-gel connection shape. In the construction efficiency considering the construction time and construction method, the fixing method of air nailer, fixed nail and washer was superior to that of the washer, screw and plastic wall plug.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.1
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• pp.57-64
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• 2010

Objectives: Likewise fresh cycle, it is also important to select right blastocysts for transfer in purpose of improving the pregnancy and implantation rates in frozen-thawed embryo transfer (ET) cycles. To investigate the relationship between the developmental velocity at the time of cryopreservation and pregnancy rates, we compared pregnancy rates between the day 5 cryopreservation group and the day 6 cryopreservation group. Methods: Transfers of frozen-thawed blastocysts which had been cryopreserved by vitrification on day 5 or day 6 were performed between January 2006 and June 2007. Ethylene glycol, DMSO, and pull and cut straws were used for vitrification and artificial shrinkage was done in expanded blastocysts. Thawing was performed on the day before transfer and thawed blastocysts were cultured in for 15~18 hrs in Quinn's blastocyct media. Blastocyst survival was assessed before transfer and post-thaw survival was defined as >50% of cells remaining intact and blastocoele re-expansion by the time of transfer. Results: Transfers of thawed blastocyst had been cryopreserved on day 5 were 52 cycles and 41 transfer cycles were cryopreserved on day 6. Patient characteristics, the number of transferred embryos and the survival rate of thawed blastocysts were not different in each cryopreservation day. But the biochemical pregnancy, clinical pregnancy, ongoing pregnancy, and implantation rate were significantly high in transfer of frozen-thawed blastocyst which were cryopreserved on day 5. Conclusions: The clinical pregnancy and implantation rate of day-5 blastocyst showed significantly higher than those of day-6 blastocyst in frozen-ET cycles. This result indicated that developmental rate of blastocyst at cryopreservation time in frozen-thawed cycle is discriminative marker of pregnancy outcome as like in fresh cycle.

• Journal of Embryo Transfer
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• v.32 no.1
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• pp.1-8
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• 2017

Cryopreservation of germ cells from genetically proven animals could be a source of restoration tools from the risk of extinction or disappearance of wanted characteristics. Using frozen semen, the genetic gains of Korean native cattle have been increased greatly for 70 years. The preservation of genetic resources as a form of frozen semen straw has limited availability due to the numbers. To circumvent this weakness of frozen semen, we tested two re-freezing methods with different initial thawing temperatures using frozen Korean proven semen and rare breed semen from albino, black and chikso breeders. It has been known that human sperm could resist to cryo-damages by repeated freeze-thaw cycles, but not for Korean proven bulls number (KPN) or for rare breeds. Total 7 frozen semem from brindled(2), black(1), Korean Albino(2) and KPN(1) bulls were used for our research. After thawing straws under $5^{\circ}C/2min$ or $37^{\circ}C/40sec$ with low temperature water bath and thermo jug, spermatozoa were re-diluted with triladyl diluents after first thawing and re-frozen. Sperm motilities were compared between animals and treated groups after re-thawing. Mean values of motility and viability of refrozen/thawed sperm for expansion of the number of straws were significantly higher in $5^{\circ}C$ than in $37^{\circ}C$ (P < 0.05). However, the activity of viable sperm thawed at $5^{\circ}C$ was significantly decreased before refreezing. It is estimated that re-freezing of frozen semen from rare Korean native cattle is possible with resistant properties of survived spermatozoa.

• Journal of Practical Agriculture & Fisheries Research
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• v.21 no.1
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• pp.29-40
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• 2019

In Korea kiwifruit growing area is limited to southern coastal region and Jeju island, partly due to the lack of information on their cold hardiness in winter. This study was carried out to investigate cold hardiness of Korean kiwifruit cultivars in a period of dormancy for using it as preliminary data to expand the cultivation area of kiwifruit in Korea. A total of five kiwifruit cultivars in two species and hybrid, Actinidia deliciosa ('Hayward' and 'Garmrok'), A. chinensis ('Goldone') and A. arguta hybrid ('Bangwoori' and 'Skinny Green') were subjected to five freezing treatments of -12℃, -15℃, -18℃, -21℃ and -24℃. Cell membrane damage in all cultivars initiated in -18℃/32h and cell membrane stability was lost in -24℃ in most cultivars, except for 'Skinny Green'. Cold hardiness was estimated by 50% lethal temperature (LT₅₀) which was determined by triphenyl tetrazolium chloride (TTC) reduction. In branches, LT₅₀ was -15℃ in 'Hayward' and 'Garmrok', -18℃ in 'Bangwoori' and -21℃ in 'Goldone.' The LT₅₀ of buds on 'Hayward' and 'Garmrok' was 56 and 42 hours in -15℃ and 4 and 11 hours in -18℃, respectively; however, LT₅₀ of buds on 'Goldone' was 51 hours in -18℃ and that on 'Bangwoori' was 3 hours in -24℃. Cold hardiness results imply that it may be difficult for cultivars in A. deliciosa such as 'Hayward' and 'Garmrok' to be grown in the north of southern coastal region in Korea; however, it can be possible for several cultivars in A. chinensis and A. arguta hybrid to be grown in the northern part of Korean kiwifruit belt if cold tolerance in the thaw is confirmed.

• Journal of Embryo Transfer
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• v.26 no.1
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• pp.19-25
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• 2011

This study was carried out to evaluate the effect of early pregnant cow as donor for Ovum Pick-Up (OPU) derived oocyte aspiration and embryo production in Holstein heifers. Four non-pregnant and 2 pregnant Holstein heifers were used as donor and then carried out total 17 OPU session for 10 weeks (2 times per week). Recovered cumulus-oocyte-complexes (COCs) were classified into 4 grade by oocyte cytoplasm and cumulus cells and matured in vitro in TCM-199 supplemented with 10% FBS, 10 mg/ml FSH and 1 mg/ml estradiol in 5% $CO_2$ and over 99% humidity for 24 h. After 24 h co-incubation with post-thaw sperm, the presumed zygotes were cultured in CR1aa medium with 4 mg/ml BSA for 3 days and then changed CR1aa medium with 10% of FBS for another 3~4 days. The Mean number of aspirated follicles and collected oocytes in the early stage pregnant and non-pregnant heifers were $13.0{\pm}4.3$ and $10.6{\pm}3.9$, $5.4{\pm}3.4$ and $7.7{\pm}3.6$ per session, respectively. Rate of collected oocyte from aspirated follicles were 59.2% and 50.5%, respectively. The average number of good quality oocytes (Grade I and II) in the early stage pregnant and non-pregnant heifers was $3.7{\pm}2.7$ and $4.9{\pm}2.6$ (Mean${\pm}$SD). Cleavage and blastocyst developmental rates in Grade I and II were 22.2% and 25.5%, and then $1.7{\pm}0.9$ and $1.4{\pm}1.1$ blastocyst per session, respectively. In conclusion, OPU technology can be used in early stage pregnant and non-pregnant heifers without any problem and so applied OPU derived embryo production to maximize the ability of genetically valuable females.