• Korean Journal of Animal Reproduction
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• v.23 no.4
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• pp.313-321
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• 1999

The purpose of these study was to investigate the use of a glass micropipette (GMP) as a vessel for vitrification of bovine IVP blastocysts, to compare the post-thaw survival rates of bovine blastocysts frozen in GMP with those frozen in OPS that have been previously investigated, and to improve the hatching rate following vitrification with GMP method. The GMP vessel permits higher freezing and warming rate than the OPS due to the higher heat conductivity of the glass and lower mass of the solution that contains the embryos. Groups of three bovine IVP blastocysts were sequentially placed into vitrification solution before being loaded into either the OPS or GMP vessels and immersed into L$N_2$within 20 to 25 sec. Post-thaw blastocysts were serially washed in 0.25 and 0.15 M sucrose in HM and TCM-199 for each 5 min, respectively, and then cultured in TCM 199 supplemented with 10% FCS for 24 h. The rate of blastocyst re-expanding did not significantly different for OPS (75.9%) and GMP (80.0%) methods (P＞0.05). The hatching rates in OPS (34.1%) and GMP (37.5%) methods were significantly lower than that in control group (54.3%) (P＞0.05). In addition, the rate of blastocyst re-expanding was significantly lower if blastocysts were vitrified in the wide portion of the micropipette rather than the narrow portion of the micropipette (83.3 vs 56.7%) (P＞0.05), even though three blastocysts were loaded per vessel. The hatching rate in 0.05% pronase solution treatment for 30, 60 and 90 see (45.9, 54.7 and 57.5%) were significantly higher than that in control (35.0%), even though there was not significantly different between 30 see and control. These results indicate that both vitrification vessels can provide high survival rates of bovine IVP blastocysts. However, the GMP vessel has the advantage over the OPS, in that the former does not need a cap to protect the vessel from floating after immersion in L$N_2$. The location of the embryos (narrow or wide portion of immersion) were considered to be limiting factors to the viability of bovine IVP embryos. The exposing in 0.05% pronase solution for 60 or 90 see can increase hatching rates of post-thaw bovine IVP blastocysts.

• Journal of the Korean Geotechnical Society
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• v.16 no.1
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• pp.179-189
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• 2000

The purpose of this research is to complement the existing researches on landfill bottom liners behavior during the periods of freeze and thaw. Landfill-related researches have been typically focused on small-scale soil samples that are often compacted under conditions different from those used in the field. Although these tests have been invaluable in clarifying the problem of freeze and thaw, extending the results of such experimental studies to prototype landfills are questionable. In this investigation, the author utilized a large scale laboratory simulation allowing inclusion of the field depth of the cover systems, layered soil profiles, rainfall simulation, a cold climate and boundary conditions similar to those encountered in the landfill. The soil materials were stabilized soils (mixed clays, cements, and minerals) instead of clays. The bottom liners are made up of drainage layer (30 cm), stabilized layer (75 cm), and leach collection layer (60 cm). The stabilized layers are made up of supporting layer (45 cm) and low permeable layer (30 cm) - consisting of $P_A\; and\; P_B$ layer. As a results, depths of penetration increased by about 2~5 more centimeters at rainfall simulated designs than those at no rainfall simulated designs (that is design 3, design 5 and design 7) - it increased by about 20mm/day in the bottom liners and frost heaves also increased it by a few millimeters. Also, a few cracks appeared partly. According to these results, we can surmise that the compacted stabilized soil is more reliable than the compacted clay liners for construction of the landfill liners.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.9
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• pp.251-258
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• 2020

This study investigated the effect of Zardaverine supplementation in freezing extender, on kinetic characteristics of post-thawed boar sperm. Cryopreservation of boar sperm is an important technique of assisted reproductive technology and genetic resource banking. Although this technique is particularly useful, freeze-thaw cycles associated with sperm cryopreservation significantly reduce sperm quality. Semen from mature Duroc boars were collected and cryopreserved in freezing extenders (LEY) treated with varying concentrations of Zardaverine (0, 20, 50, 75, 100 𝜇M). The time-dependent kinetic characteristics of post-thawed spermatozoa were determined after thawing by applying computer-assisted sperm analysis (CASA). We observed that the motility immediately after thawing was significantly higher in 20 𝜇M stocks than in control (0 𝜇M) and the other treatments (p<0.05). Curvilinear velocity (VCL) in 0 𝜇M and 20 𝜇M stocks were significantly higher than the other treatment groups, except 75 𝜇M (p<0.05). Higher average path velocity (VAP) was obtained at 20 𝜇M as compared to 100 𝜇M, whereas amplitude of head lateral displacement (ALH) was significantly higher at 20 𝜇M than 50 𝜇M and 100 𝜇M (p<0.05). No differences were obtained for Straight-line velocity (VSL) and Linearity (LIN). In conclusion, our results indicate that Zardaverine improves the motility, VCL, VAP, and ALH of post-thawed boar sperm.

• 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.

• Korean Journal of Animal Reproduction
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• v.23 no.1
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• pp.85-92
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• 1999

This study was designed to determine effect of cryoprotectant kinds and cell stages on OPP vitrification method in mouse embryos. The freezing speed, cryoprotectants and cell stage could affect of embryo viability following various vitrification methods. The vitrification solution used were consisting of 40% (v/v) ethylene glycol, 18% (w/v) Ficoll, 0.3 M sucrose solution in holding medium (D-PBS supplemented with 5% FCS: HM) (EFS) or 16.5% ethylene glycol , 16.5% dimethyl sulfoxide, 0.5 M sucrose in HM (EDS). The embryos were collected from oviduct at 18 h after hCG injection and then washed and cultured in mHTF medium until use. In experiment 1, the blastocysts were vitrified by OPP straw to determine the optimal vitrification solution of EFS or EDS. The post-thaw survival rates at re-expanded stage rates were significantly different between EFS and EDS (95.0 vs 100%), but at hatching stage was not different between EFS and EDS (90.0 vs 95.0%). respectively. In experiment 2, zygotes, 2-, 4-cell, morula and blastocysts were vitrified by OPP method to determine the acceptable of early stage embryos. The development rates to expanded blastocyst in zygote (70.0%) were significantly lower rather than those in 2-, 4- 8-cell, compacted morula or blastocyst (89.7, 90.0, 92.8, 97.6 or 97.5%), respectively. However, the cell number of post-thaw developed to expanded blastocyst in blastocyst and control blastocyst stage (39.6$\pm$2.81, 35.7$\pm$2.98) were significanty higher than those in zygote, 2-, 4-, 8-cell, compacted morula (29.8$\pm$3.21, 31.3$\pm$3.83, 29.3$\pm$3.58, 28.9$\pm$3.21 or 30.8$\pm$2.93). In experiment 3, the zygotes were exposed in VSl for 1, 2, and 3 min to the optimal exposed time. The cleavage rates (91.6, 88.5, 88.9%) and develop mental rates to blastocyst (83.3, 74.3 and 69.4%) depends on the exposed time in VSl were not significantly different among 1, 2, or 3 min, respectively. The cell number also were not significantly different among exposed time in VS1. respectively. These results indicate that OPP method could be useful for vitrification either EFS or EDS vitrification solution. The post-thaw survival rates at zygote were significantly lower than those at 2-, 4-, 8-cell, morula or blastocyst, respectively. The zygote stage were more sensitive rather than late stage embryos. The exposing time in VS1 for 1 min was better than that for 2 or 3 min, even it was not significantly different. The OPP vitrification method could be useful of mouse embryos either with EFS or EDS vitrification solution.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.12
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• pp.2019-2027
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• 2013

In this study, we tried to develop a coating agent for the fresh-cut fruits used in cakes. First, the coating agent mixing ratios of sugar, pectin, sodium alginate, carrageenan, xanthan gum, vitamin C, and purified water were selected to be 55, 2, 2, 0.04, 0.1, 0.05, and 40.81% (w/w), respectively. In a freeze-thaw stability of the coating agent, the viscosity remained constant for 3 cycles of freezing and thawing repetition process, but showed a slightly decreasing trend in the 4th repetition process (P<0.05). On the other hand, the sugar content, pH, and chromaticity remained constant even in the 4th repetition process. Pineapple coated with the coating agent had smaller weight loss, hardness changes, and total bacteria distribution compared to the uncoated pineapple (P<0.05). In the chromaticity, both of the two pineapples experienced browning with increasing storage duration, as L value decreases and b value increases. However, when the color difference was compared, the progress of browning for the uncoated pineapple was faster than the coated pineapple. Also, the progress of browning at $4^{\circ}C$ was found to be slower than the progress of browning at $25^{\circ}C$. Therefore, the storage stability of the fresh-cut fruits could be improved by coating the fresh-cut fruits for cakes with the coating agent and storing at a low temperature, which would contribute to extending the shelf-life of cakes.

• Journal of the East Asian Society of Dietary Life
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• v.25 no.5
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• pp.893-901
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• 2015

This study investigated the effects of thawing conditions on physiological activities and quality of peeled garlic. Peeled frozen garlic was analyzed after thawing at low temperature ($4^{\circ}C$), room temperature ($20^{\circ}C$), tap water ($20^{\circ}C$), radio frequency of 27.12 MHz, and 2.45 GHz in a microwave. As a result, the time required to thaw garlic to $0^{\circ}C$ by various thawing methods was shortest at2.45 GHz in a microwave, followed by $20^{\circ}C$ tap water, radio frequency of 27.12 MHz, $20^{\circ}C$, and $4^{\circ}C$. Microwave thawing was faster than other methods, but it resulted in significant non-uniformity of heating. The hardness of peeled garlic significantly decreased upon freeze-thawing, whereas it showed improved hardness upon radio-frequency thawing. Total color difference in garlic increased upon freeze-thawing, and it was not improved by various thawing methods. Antioxidant activities were determined for DPPH radical scavenging ability, SOD-like activity, and reducing power. Total phenolic compounds and flavonoids in garlic extract were measured as $3.222{\pm}0.214{\mu}g$ GAE/g and $0.149{\pm}0.03{\mu}g$ QE/g, respectively. The content of total phenolic compounds was significantly reduced by 2.45 GHz microwave thawing ($1.90{\pm}0.02{\mu}g$ GAE/g); however, flavonoid contents were slightly reduced under freezing and thawing conditions. The DPPH radical scavenging ability of garlic extracts was not affected by thawing methods; however, SOD-like activity and reducing power were slightly reduced by freeze-thawing. These results indicate that physiological activities were not improved by radio-frequency thawing; however, thawing time and maintain hardness were reduced compared with conventional thawing methods.

• Journal of the Korean Geographical Society
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• v.41 no.6 s.117
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• pp.657-669
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• 2006

Rates and processes of bare patch denudation were observed at Janggumokoreum(1,710m) and Minoruem(1,600m) in order to clarify some characteristics of turf exfoliation in the subalpine grassland of Mt. Halla. The bare patches have marginal terrace fronts with a maximum height of 85 cm. The terrace risers usually develop an overhanging edge 2 to 38 cm long that eventually hangs down and protects the riser beneath. The patches are largely covered with angular pebbles and cobbles. The mean rate of riser retreat for the period 2002-2004 is 39.2 mm, equivalent to 19.6 mm/yr. However, there is a disparity of the rate of riser retreat at individual sites. The maximum rate is 131 mm measured at Janggumokoreum patch while the minimum rate is 0 mm at Minoreum patch. The rate of riser retreat also varies with seasons. The thawing season of April exhibits a maximum rate of retreat. The freezing season of October and November and the rainy season of June and July show relatively high rates of retreat. Several Processes such as frost action, aeolian deflation, rainwash, rainsplash and fauna activity cause the denudation of bare patches. In particular, the needle ire action which is combined with rainwash or deflation plays a primary role in turf exfoliation due to the diurnal freeze-thaw cycles occurred over 100 days, melted snow and strong wind in the subalpine zone of Mt. Halla. Rainwash is also an important contributing process in the rainy season because Mt. Halla has the highest precipitation in Korea. By contrast, rainsplash erosion has a minor effect on the bare patch denudation due to the overhanging edge of terrace risers. Recent increase in roe deer appears to be responsible for turf destruction.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.11
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• pp.1527-1533
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• 2001

Garole is a prolific, rare, less known and small size Indian sheep breed found in low and humid Sunderban region of West Bengal. Although information on stored Garole ram liquid semen upto 24 h is available, but there is a need to further investigate the short-term and long-term preservability of Garole ram semen for extensive utilization of this valuable germplasm by artificial insemination. The aim of the present study was to apply computer-assisted sperm analysis technique for assessing the motion characteristics of Garole ram semen stored (i) in liquid state at refrigeration temperature for short-term preservation upto 48 h and (ii) in frozen state at $-196^{\circ}C$ for long-term preservation after packaging in mini straws. Short-term preservation had a significant effect on motility (p<0.01) as the motility progressively decreased from 90.1% at 0 h to 85.5% and 73.2% after 24 and 48 h of storage, respectively. Although the decline in rapid moving sperms was also significant (p<0.01) on storage but the decrease was more pronounced at 48 h as compared to 24 h of storage period. Storage of chilled semen had also a significant effect on % linearity (p<0.05), % straightness (p<0.01), sperm velocities (p<0.01), amplitude of lateral head displacement (p<0.01) and beat frequency (pO.Ol) of spermatozoa. The replication had a significant effect for all the variables except average path and straight line velocity. However, the interactions of short-term storage and replication were non-significant for most of the variables except % of medium moving sperms, sperm velocities and beat frequency. On long-term preservation of Garole ram spermatozoa under controlled conditions the mean post-thaw recovery of 70.4 and 71.4% motile spermatozoa was achieved having 48.8 and 48.9% of rapidly motile spermatozoa, respectively in both the replicates. The effect of replication on cryopreservation was significant (p<0.05) on amplitude of lateral head displacement and beat frequency, but there was no significant effect on motility, rapidly motile spermatozoa, linearity, straightness and sperm velocities of frozen-thawed spermatozoa. It can be concluded from these results that an average 70% motility can be achieved on storage of Garole ram semen in chilled liquid state upto 48 h or in liquid nitrogen after freezing under controlled conditions in straws. However, further studies are required to evaluate the fertility of short-term and long-term preserved Garole ram semen for extensive use of this prolific sheep breed.

• Korean Journal of Animal Reproduction
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• v.22 no.2
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• pp.171-176
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• 1998

This study was carried out to confirm whether the in vivo developmental potential of mouse hatched blastocysts (HBs) can be obtained by vitrification method using the cryoprotectant EFS35. The HBs ($\theta$ 130$\mu\textrm{m}$) were cultured in vitro until day 5 from zygotes produced in vivo and were equilibrated in 10% ethylene glycol(EG) for 5 min, and then were exposed or vitrified in EFS35 (35% EG, 18% Ficoll and 0.3M sucrose). After 30 min thawing, re-expanding HBs were transferred into one or both uterine horns of pseudopregnant recipients on day 3 (4~6 embryos /horn). Pregnancy rates of recipients and implantation were assessed by autopsy on day 15 of gestation. The results obtained in these experiments were summarized as follows : After thaw-ing, in vitro survival of HBs was not significantly different between exposed (65.5%) and vitrified(54.5%) group. Also, when the in vivo development potential was examined, total implantation was not different between control (58.5% ) and vitrified (41.0%) group, although the live fetus formation of vitrified group (24.0%) was significantly lower than that of control (58.3%) group (p< 0.05). These results suggested that vitrification freezing method of mouse HBs using EFS35 can be used to make wide the utility of embryo transfer of the more embryos produced in vitro.