• Journal of Embryo Transfer
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• v.20 no.2
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• pp.169-176
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• 2005

Loop-mediated isothermal amplification (LAMP) is novel DNA amplification methods that amplifies a target sequence specifically under isothemal condition. The present study was to assess the in vitro viability afier biopsy and sexing rate of different types of embryo biopsied. In vivo compact morulae and blastocyst embryos were obtained from Korean Native Cow (KNC) superovulated with FSH (Antorin, R-10) on 7 Day after artificial insemination. in vitro compact morulae and blastocyst embryos were obtained with KNC or Holsteins that were gained on 6, 7 or 8 day after in vitro fertilization(IVF) with frozen semen. Biopsy of bovine embryo was carried out in a $80{\mu}l$ drop with $Ca^{2+}-Mg^{2+}$ free D-PBS and the viability of biopsied embryos were evaluated in IVMD (IFP, Japan) medium at 12 hrs culture time. The sex ratio of biopsied Hanwoo embryos were male vs. female of $43.5\%\;vs.\;56.5\%$ in vivo and $33.9\%\;vs.\;49.2\%$ in vitro respectively, and male rate of biopsied Holstein embryos were significantly higher than female $(70.8\;vs.\;29.2\%)$. and indefinite rate of in vitro embryos was $16.9\%$ and in vivo was not. The degeneration rate of biopsied embryo, in vitro embryos were significantly higher than in vivo $(13.2\%\;vs,\;0.0\%,\;p<0.05)$. The survivability of in vivo embryo were between biopsied following punching method was significantly (P<0.05) higher than bisection method produced embryos $(100\%\;vs.\;83.3\%)$ and in vitro had no difference. However, the degeneration rate of biopsied embryo by bisection method was significantly higher than punching methods between in vivo and in vitro $(16.7\;vs.\;22.6\%,\;respectively,\;p<0.05)$. In conclusion, these results indicate that punching method was optimal and survivability after embryo biopsy was useful for reducing the damage caused by the embryo biopsy procedure for LAMP-based embryo sexing.

• Journal of Embryo Transfer
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• v.20 no.2
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• pp.185-190
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• 2005

This study was performed to investigate the effects of multiple superovulation and parity on embryo production in Hanwoo cows. Donors were superovulate 4 times $1\~2$ months interval and inserted CIDR plus (with the capsule of estradiol benzoate 10mg) on Day 10 from standing heat for 9 days and injected 2.5ml FSH (Antorin R-10) 2 times in a day on 6th day to 10th day from insertion of CIDR and the doses of FSH were decreasing 0.5ml on every 2 times. On 3th day of FSH injection, 25ml $PGF_2{\alpha}$ were injected i.m. and on 4th day, CIDR was removed. After 2 days from removing CIDR, AI was performed 2 times 12 hour apart with 2 straws of Korean Proved frozen Semen and simultaneously 200ug/ml GnRH was injected and embryos were recovered on 7th day from Al. The response rates of superovulated donors were $85.7\%,\;90.5\%,\;62.5\%,\;100\%$ from 1 to 4 times of superovulation, respectively. There were significant differences among No. of superovulation times (P<0.05). The results of transferable embryos were 3.7, 3.4, 3.4, 5.7 from 1 to 4 times of superovualtion, respectively. There were no differences among No. of superovulation times. The results of transferable embryos were 2.5, 3.0, 5.3, 3.0, 3.4 form heifer, first born to 4 the born, respectively. There were significant differences among the parities of donors (P<0.05). These results suggested that even 4 times of superovulations of Hanwoo donors could be able to recover transferable embryos, it might be used the donors maximally and improved the adaptation of embryo transfer to farms safely.

• Journal of Food Hygiene and Safety
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• v.25 no.2
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• pp.170-179
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• 2010

This study was conducted to investigate for mineral contents, total polyphenol compounds, betaine, choline and DPPH free radical scavenging activity of halophyte. The mineral concentrations of Salicornia herbacea (top part) were Na 100,006 mg/kg, K 1,385 mg/kg, Mg 6,263 mg/kg, Ca 2,750 mg/kg, Fe 90.4 mg/kg, Mn 98.9 mg/kg, Zn 33.3 mg/kg, Cu 3.4 mg/kg respectively. And Suaeda Japonica (top part) were Na 85,332 mg/kg, K 710 mg/kg, Mg 7,005 mg/kg, Ca 4,344 mg/kg, Fe 1,434.9 mg/kg, Mn 119.1 mg/kg, Zn 19.2 mg/kg, Cu 2.7 mg/kg respectively. The betaine contents of Salicornia herbacea (top part) were 15.09 mg/g and Suaeda Japonica (top part) were 14.64 mg/g. The choline contents estimated by the DBAP-choline derivatives of Salicornia herbacea (top part) were 20.9 mg/100 g, Salicornia herbacea (root) were 23.4 mg/100 g, Suaeda Japonica (top part) were 23.1 mg/100g and Suaeda Japonica (root) were 23.8 mg/100 g. Total polyphenol compounds of Salicornia herbacea (top part) were high 36.0 mg/g in growth phase. The DPPH radical scavenging activities of methanol extract Salicornia herbacea (top part) were high 90.1% in growth phase. The frozen dried powder of Salicornia herbacea (top part) 1 g was equal to Quercetin 30.26 mg, Rutin 42.65 mg, TBHQ 20.32 mg, BHA 25.86 mg, BHT 40.75 mg, Ascorbic acid 22.86 mg in DPPH radical scavenging activities.

• Journal of Embryo Transfer
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• v.33 no.1
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• pp.17-22
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• 2018

This study was conducted to determine the effect of pentoxifylline levels on sperm motility, survival rate, sperm membrane integrity of frozen semen and fresh-extended equine semen in Jeju cross-bred horses. As a result of sperm characteristic comparison depending on pentoxifylline levles at 30 minutes post-thaw, the progressive motilities were $53.25{\pm}2.87$ (4mM pentoxifylline) and $50.28{\pm}2.14$ (8mM pentoxifylline) and significantly higher compared to the control group($40.09{\pm}5.15$) and other treatment group (16mM pentoxifylline, $41.27{\pm}2.82$). The progressive fast motility were $22.44{\pm}1.62$ (4mM pentoxifylline,) and $22.74{\pm}3.07$ (8mM pentoxifylline) and significantly higher compared to the control group ($13.47{\pm}1.48$) and other treatment group (16mM pentoxifylline, $14.66{\pm}3.68$) (p<0.05). As a result of sperm characteristic comparison depending on pentoxifylline levles at 30 minutes post-thaw were $68.96{\pm}1.64$ (4mM pentoxifylline) and $67.90{\pm}6.72$ (8mM pentoxifylline) and significantly higher compared to the control group ($53.48{\pm}4.84$) and other treatment group (16mM pentoxifylline, $58.14{\pm}2.65$) (p<0.05). In conclusion, these results suggest that treatment groups with 4mM and 8mM pentoxifylline were higher compare to equine seperm mobility and the control group and treatment groups with more than 16mM pentoxifylline has a negative effect on sperm characteristics. After thawing, the total motility in post-thawed equine sperm has increased by 10 percent for 1 hour. these results suggest that pentoxifylline contributes to the improvement of the equine sperm motility and characteristics in post-thawed semen.

• Korean Journal of Food Science and Technology
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• v.26 no.2
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• pp.110-116
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• 1994

In order to study the effects of enzyme modification on the physico-chemical and functional properties of myofibrillar protein prepared from the frozen sardine, Sardinops melanostica, the protein was hydrolyzed with pepsin under the enzyme-substrate ratio 1:100 at $37^{\circ}C$ and pH 1.65 for 1, 4, 8, 12, and 24 hr, respectively. The properties of pepsin-modified sardine myofibriliar protein were determined. The extents of proteolysis with pepsin as a fuction of time was showed a typical enzyme hydorlysis curve with an initial region of 4 hour period followed by plateau region. The SDS-acrylamide slab gel electrophoresis patterns of pepsin-modified proteins showed mainly disappearances of minor protein bands, but no changes of main protein bands. The gel filtration patterns through Sephadex G-75 of sardine myofibrillar protein showed two big peaks and three small peaks. All the small peaks were disappearanced by proteolysis with pepsin in one hour. and during the period of proteolysis the fast big peak became gradually smaller and the late big peak eluted more slowly. By proteolysis, the emulsifying activity and emulsifying capacity of sardine myofibrillar protein were all decreased. The effects of pepsin-modification on emulsifying capacity were greater than those on emulsifying activity of protein. The aeration capacity of the protein was increased about 1.9 folds and the foam stability decreased to 0.6 folds of control by pepsin-modification. The pepsin-modified sardine myofibrillar proteins showed about 0.6 folds of heat coagulation and 1.4 folds of viscosity of control. The pH dependence of solubilities of sardine myofibrillar protein showed two isoelectric areas of pH 5 and 9. The pepsin-modified protein showed more clear pH dependences at the early stage but not at the late stage of proteolysis.

• Journal of Aquaculture
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• v.16 no.3
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• pp.190-195
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• 2003

Dietary optimum ratio of frozen raw fish and commercial compound meal (CCM) in moist pellet (MP) were investigated to improve the growth rate, and feed and economical efficiency in the flounder, Paralichthys olivaceus. Experimental fish (average body weight, about 48 g) were divided into 6 groups and each group was fed with raw fish (FRF), and MPs (ratio of raw fish and CCM=9:1, 8:2, 7:3, 6:4 and 5:5) for 10 weeks. The 9:1, 8:2, 7:3, 6:4, 5:5 groups showed no significant difference in weight gain as 203.5~217.3%, while the FRF group showed significantly low growth rate as 183.1%(P<0.05). The feed efficiency gradually increased with the increase in the ratio of CCM and was the highest in the 5:5 group as 89.7%. As a result of analysis of body composition after the experiment, moisture was significantly low in the 7:3 group (P<0.05) and crude protein was significantly low in the 9:1 group (P<0.05). The crude lipid increased as the ratio of raw fish increased, and it was the highest in the raw fish group (8.3%) and the lowest in the 5:5 group (4.6%). There were no significant difference in hepatosomatic index (HSI) and condition factor(CF) among the experimental groups. Visceralsomatic index (VSI) increased with the increase in the ratio of raw fish and was significantly high in the raw fish group as 5.49 (P<0.05). For the unit cost of feed, it was found that raw fish was economical when mackerel, a source of raw feed, was 400 won/kg, while the ratio of 5:5 was economical when it was more than 500 won/kg. Results of this study concluded that sole use of raw feed is not desirable in view of growth, environment and economy, and the 5:5 group showed highest effect under the least use of raw feed.

• Korean journal of food and cookery science
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• v.23 no.5
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• pp.626-634
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• 2007

This study investigated the effects of 3% ground fresh paprika (FP) and 5% freeze-dried paprika powder (FDP) on lipid oxidation inhibition and warmed-over flavor (WOF) development in cooked ground pork (CGP; meat:fat = 70:30), using two packaging methods (atmosphere packaging and vacuum packaging) during 8 days at $4^{\circ}C$ and 4 months at $-26^{\circ}C$. In the CGP containing FP with atmosphere packaging, at both $4^{\circ}C\;and\;-26^{\circ}C$, peroxide formation increased sharply, and was similar to that of the CGP without paprika. Peroxide formation, in both the CGP without paprika and with FP and packaged with vacuum packaging, repectively, was much lower than that found with atmosphere packaging. Vacuum packaging was superior to atmosphere packaging for lipid oxidation inhibition. In the CGP containing FP with vacuum packaging and stored at $-26^{\circ}C$, peroxide formation almost didn't occur, which was similar to the CGP containing FDP. The peroxide value and thiobarbituric acid (TBA) value did not increase in the CGP containing FDP over the storage periods ($4^{\circ}C\;and\;-26^{\circ}C$) for both the atmosphere and vacuum packaging. Therefore, FDP was the most effective for lipid oxidation inhibition during refrigerated storage, regardless of the packaging method. Both FP and FDP with vacuum packaging during frozen storage showed similar antioxidant activities. The development of WOF in the CGP containing FDP with vacuum packaging was delayed until 7 days at $4^{\circ}C$ and 3 months at $-26^{\circ}C$, respectively. WOF was highly correlated with TBA value in the CGP stored at $-26^{\circ}C$ with vacuum packaging (r = 0.88, p<0.05). The oxidative stability of the lipid in the CGP containing FDP with vacuum packaging was excellent.

• Clinical and Experimental Reproductive Medicine
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• v.35 no.3
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• pp.203-212
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• 2008

Objective: During cryopreservation process, cold shock and cryo-injury affect the fertilizing capacity of the sperm by damaging cell membranes with loss of functional integrity. A longstanding concept for preventing the cryo-damage is to stabilize the plasma membrane by incorporating cholesterol. This study was to determine the effects of cholesterol in freezing media on the motility and functional integrity of human sperm after cryopreservation. Methods: Control group (non-cholesterol treated) and different concentrations of cholesterol-treated sperm (14 healthy males) were frozen and thawed. After freezing and thawing of sperm, the quality of sperm was evaluated by sperm analysis, acrosome reaction test and sperm chromatin structure assay. Results: When human sperm were incubated in sperm freezing medium (SFM) containing $0.5{\mu}g$ cholesterol and then freezing/thawing, the motility of sperm have significantly improved compared to those untreated cholesterol ($33.46{\pm}1.48%$ vs. $30.10{\pm}1.07%$, p<0.05). The rate of calcium ionophore-induced acrosome reactions in post-thawed sperm was significantly higher than that ($53.60{\pm}1.60%$ vs. $47.40{\pm}1.86%$, p<0.05) in SFM containing cholesterol. Sperm chromatin structure assay revealed that DNA damage to the sperm in the cholesterol-treated group was lower than that of non-treated group. Conclusion: These results suggest that increased cholesterol content of sperm plasma membrane by supplementation of cholesterol in SFM improves sperm motility, capacitation status, and DNA integrity. Therefore, addition of cholesterol into SFM could be a useful for protecting human sperm from cold shock and cryo-injury during cryopreservation.

• 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.

• Korean Journal of Poultry Science
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• v.43 no.4
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• pp.197-206
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• 2016

This work was carried out to investigate effects of the freezing/thawing method on duck meat kept in a freezer for a month. The meats used were breast muscle collected from Korean native ducks (KND) that were fed for 8 weeks (2.8 kg of live weight). Forty-five samples were used after being frozen in storage for one month and were then divided into 5 treatments (3 replications/treatment, 3 samples/replication). Five treatments (CON, FFFT, FFST, SFFT and SFST) were control groups (CON) and four were experimental groups, using $2{\times}2$ complex factors with two freezing methods (fast freezing, FF, $-50^{\circ}C$ in a deep freezer; slow freezing, SF, $-20^{\circ}C$ in a common freezer) and two thawing methods (fast thawing, FT, 5 h $12^{\circ}C$ with flow water; slow thawing, ST, 24 h $5^{\circ}C$ in a refrigerator). Lightness of KND meat in FF and FT groups was lower than that of control (P<0.05). Yellowness of KND meat of the ST group was higher than that of control (P<0.05). Cooking loss (CL) and water holding capacity (WHC) of KND meat in the control were lower than those of the freezing and thawing groups (P<0.01, P<0.05), but shear force (SF) of the control was higher than that of other groups (P<0.01). Moisture content of the ST group was higher than that of the FT group (P<0.05), and protein content of the FF group was higher than that of control (P<0.05). Stearic acid (C18:0) of the SF group was higher than that of the FF group (P<0.05). Arachidonic acid (C20:4n6) of control was higher than that of the SF and ST groups (P<0.01, P<0.05). Alanine, aspartic acid, glutamic acid, serine, and tyrosine content of the control were lower than that of the freezing and thawing groups (P<0.05). These results show that freezing and thawing methods affect meat color, shear force, cooking loss, and WHC-related water content.