• Clinical and Experimental Reproductive Medicine
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• v.33 no.4
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• pp.237-243
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• 2006

Objective: The aim of this study was to evaluate the efficacy of frozen-thawed ET in poor prognosis patients such as the old age (38~44 years; OA group) and the patients who did not achieve clinical pregnancy with the first fresh ET cycle (non-pregnant patients; NP group). Methods: Laboratory and clinical data were collected from fresh and frozen-thawed ET cycles of OA and NP group. Controlled ovarian hyperstimulation (COH) and conventional insemination or ICSI, in vitro culture and ET were performed by routine procedures. Supernumerary embryos were frozen by the slow freezing method, and frozen embryos were thawed by the rapid thawing method. Embryo development, pregnancy and implantation rates were statistically analyzed by Student t-test and chi square test Results: Mean ages were similar between fresh ET ($40.0{\pm}1.8$ years, n=206) and frozen-thawed ET ($39.9{\pm}1.9$ years, n=69) cycles in OA group. However, the clinical pregnancy and implantation rate of subsequent frozen-thawed ET significantly higher than those of fresh ET cycles (29.0% and 11.2% vs. 16.5% and 7.0%, p<0.05). In NP group, there was no difference in the mean age between fresh ET ($31.2{\pm}2.3$ years, n=40) and frozen-thawed ET ($31.9{\pm}3.1$ years, n=119) in subsequent cycles. The clinical pregnancy and implantation rates were similar between the subsequent fresh ET (42.5% and 22.6%) and the frozen-thawed ET (40.3% and 18.8%). Conclusion: In old age patients, higher pregnancy rate of frozen-thawed ET compared to fresh ET cycles in this study. It may be related that better uterine environments for implantation in frozen-thawed ET cycles than that of non-physiological hormonal condition in uterus of fresh COH cycles.

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.97-108
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• 2000

The objectives of this study were performed to increase the efficiency of the culture conditions of embryos produced in vitro, and to assess the developmental potential after transfer of those embryos into recipients. The mean number of folliclular oocytes recovered from an ovary was 10.7. The rates of maturation and fertilization in Grade I oocytes were significantly (P＜0.05) higher than Graden and III. Developmental rate into blastocyst in the culture group of TCM-199 with BOEC were significantly higher (P＜0.05) than the groups of TCM-199 and conditioned medium (24.7% vs. 12.4% and 18.2%). The survivability of post-thawed blastocysts equilibrated for 3 min in EFS solution was significantly (P＜0.05) lower than l0 for 1 and 2 min (32.1% vs. 82.9% and 73.3%). Significantly higher (P＜0.05) survival rate in blastocysts was seen after freezing than in morulae stage embryos. Out of all 105 recipients, 49 (46.7%) were confirmed in pregnant. On pregnancy of cattle, 48 calves were born from 40 recipients. The ratio of twin and single calves was 30.5% (32/40 and 7.6% (8/40), respectively. However, the others composed of abnormal, as judging as 6 (12.2%) for abortion and 3 (6.1%) for stillbirth during the pregnant period.

• Korean Journal of Plant Resources
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• v.18 no.1
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• pp.1-7
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• 2005

This study was conducted to investigate the possibility of cultivating tea plants planted in the mountain area of west Gyeongnam province, Korea for 2 years from March 27th, 2001 to July 30th 2003. Ninety each plants collected from 5 different sife were cultivated in nursery cup pot$(\phi\;16cm)$. in the greenhouse condition and transplanted in 5 different location and monitored their survival growing state etc. The results obtained are as follow : 1. Survival rate tea plants after transfer to soil was relatively high Gaya-myeon Hanyang-gun with $90.0\%$ and the lowest in Buksang-meyon Geochang-gun with $80.0\%$. The tea plant collected from Gaya-myeon showed the best growing activity at early stage. 2. For the second year of harvesting time, survival rate was the highest in Machun-myeon Hamyang-gun with $92.2\%$ and the lowest was Buksang-myeon Geochang-gun with $76.7\%$. 3. For the 3rd year of harvesting time, it was impossible to data collection because of the most upper parts of plants were killed by severe freezing weather condition. In the Baekjeon-myeon, Hamyang-gun Buksang-myeon, Ungyang-myeon Geochang-gun, which are severly cold$(below\;-10^{\circ}C)$ in winter season, seems not a suitable places for tea plant cultivation since it is very different to harvest the young loaves in growing season. In conclnsion we could select two sites Gaya-myeon, Hapcheon-gun, Machun-myeon, Hamyang-gun, as tea plant cultivation in the mountain area of west-Gyeongnam province, korea.

• Korean Journal of Animal Reproduction
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• v.23 no.2
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• pp.127-132
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• 1999

This study was carried out to investigate the general characteristics such as volume, sperm concentration, sperm motility, sperm abnormality on whole semen, removed seminal plasma(RSP) semen and fractional semen of small dogs, and the effect of temperature and preservatio time and cryopreservation on motility of whole and removed seminal plasma semen. Multiple ejaculates were collected from small dogs by the digital manipulation of penis. 1. Average sperm concentration, sperm motility and abnormal sperm rates of whole semen and RSP semen were 5.07$\pm$2.32$\times$10$^{6}$ cells/$m\ell$, 95.42$\pm$2.65%, 4.42$\pm$0.157% and 4.69$\pm$3.27~4.25$\pm$3.65$\times$10$^{6}$ cells/$m\ell$, 91.17$\pm$3.85~88.52$\pm$3.85%, 6.57$\pm$0.43~5.54$\pm$0.52%, respectively. 2. Average semen volume per ejaculate, sperm concentration, sperm motility and abnormal sperm rate of 1st fractional semen were 0.92$\pm$0.7$m\ell$, 4.57$\pm$0.78$\times$10$^{6}$ cells/$m\ell$, 10.72$\pm$3.21% and 5.50$\pm$0.70%. Average semen volume per ejaculate, sperm concentration, sperm motility and abnormal sperm rate of 2nd fractional semen were 2. 14$\pm$0.19$m\ell$, 2.01$\pm$0.12$\times$10$^{6}$ cells/$m\ell$, 95.44$\pm$4.21% and 4.31$\pm$0.53%. Average semen volume per ejaculate, sperm concentration, sperm motility and abnormal sperm rate of 3rd fractional semen were 2.66$\pm$0.23$m\ell$, 2.35$\pm$0.21$\times$10$^{6}$ cells/$m\ell$, 90.71$\pm$2.63%, 6.33$\pm$0.91%, respectively. 3. Motility of whole semen and RSP semen were higher at 2$0^{\circ}C$ than at 4$^{\circ}C$ or 37$^{\circ}C$. When preservation temperature was 2$0^{\circ}C$, sperm motility were 98.32% at 1 hr, 92.15% at 5 hrs, 90.23% at 10 hrs 82.08% at 15 hrs 70.07% at 20 hrs 60.02% at 20 hrs 37.19% at 40 hrs respectively. 4. Average sperm motility of frozen 2nd fraction semen and RSP semen were 33.3$\pm$8.7, 54.7$\pm$9.5%, respectively. Sperm motility was significantly higher in frozen 2nd fraction semen and RSP semen compared with control group.

• Journal of Animal Science and Technology
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• v.47 no.6
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• pp.925-936
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• 2005

The present study evaluated whether pentoxifylline added to the freezing extender could improve the sperm characteristics and function in canine frozen semen. Also the conception rate following AI with frozen-thawed semen was investigated. The beneficial effects of pentoxifylline supplementation were visible in motility, viability, acrosome reaction, and tail swelling patterns. Especially, highest sperm viability and function were obtained in the forzen semen supplemented with 1mM pentoxifylline. The follicle size measured by ultrasonography was 6.48 mm, 11.52 mm and 8.9 mm on 11, 13 and 15 days after the onset of natural estrus, respectively and ovulation occurred on 13 and 15 days. The pregnancy rates in bitches inseminated with frozen semen on natural and induced estrus were 71.4% and 75.0%, respectively. There was no significant difference between the pregnancy rates in bitches inseminated with frozen semen following natural and induced estrus, but the litter size was slightly increased in natural cycle.

• Korean Journal of Ecology and Environment
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• v.37 no.1 s.106
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• pp.36-46
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• 2004

We evaluated the effect of limiting nutrients and N/P ratio on the growth of phytoplankton in a small eutrophic reservoir from November 2002 to December 2003. Nutrient limitation was investigated seasonally using nutrient enrichment bioassay (NEB). DIN/DTP and TN/TP ratio (by weight) of the reservoir during the study period ranged 17${\sim}$187 and 13${\sim}$60, respectively. Most of nitrogen in the reservoir account for $NO_3$-N, but sharp increase of ammonia was evident during the spring season. Seasonal variation of dissolved inorganic phosphorus concentration was relatively small. DTP ranged 26.5${\sim}$10.1 ${\mu}g\;P\;L^{-1}$, and the highest and lowest concentration was observed in August and December, respectively. Chlorophyll a concentration ranged 28.8${\sim}$109.7 ${\mu}g\;L^{-1}$, and its temporal variation was similar to that of cell density of phytoplankton. Dominant phytoplankton species were Bacillariphyceae (Melosira varians) and Chlorophyceae (Dictyosphaerium puchellum) in Spring (March${\sim}$April). Cyanophyceae, such as Osillatoria spp., Microcystis spp., Aphanizomenon sp. dominated from May to the freezing time. TN/TP ratio ranged from 46 to 13 (Avg. 27${\pm}$6) from June to December when cyanobacteria (Microcystis spp.) dominated. p limitation for algal growth measured in all NEB experiments (17cases), while N limitation occurred in 8 out of 17 cases. The growth rates of phytoplankton slightly increased with decreasing of DIN/DTP ratio. Evident increase was observed in the N/P ratio of > 30, and it was sustained with DTP increase until 50 ${\mu}g\;P\;L^{-1}$. Under the same N/P mass ratio with the different N concentrations (0.07, 0.7and 3.5 mg N $L^{-1}$), Microcystis spp. showed the highest growth rate in the N/P ratio of< 1 with nitrogen concentration of 3.5 mg N $L^{-1}$). The responses of phytoplankton growth to phosphate addition were clearly greater with increase of N concentration. These results indicate that the higher nitrogen concentration in the water likely induce the stronger P-limitation on the phytoplankton growth, while nitrogen deficiency is not likely the case of nutrient limitation.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.369-375
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• 2011

The aim of this study was to evaluate the effect of ${\alpha}$-tocopherol on blastocysts development and subsequent cryosurvival of the vitrification. The ${\alpha}$-tocopherol(0, 100, 200, 400 ${\mu}M$) was added in to culture medium for the bovine embryos. The blasocysts from the ${\alpha}$-tocopherol and untreated control groups were then frozen-thawed, and their cryosurvival was assessed by in vitro culture for 48 h. There were no differences in the overall cleavage rate($56.14{\pm}4.66$, $58.18{\pm}4.70$, $62.97{\pm}6.86$ and $51.17{\pm}7.28$) among four treatment groups. However, in blastocyst development and total cell number were significantly higher in ${\alpha}$-tocopherol 200 ${\mu}M$ ($38.60{\pm}7.12$; $106.33{\pm}3.50$) to culture medium than other treatment groups($29.30{\pm}5.24$, $31.60{\pm}7.12$ and $26.37{\pm}4.18$; $101.36{\pm}5.12$, $97.27{\pm}2.87$, and $91.23{\pm}7.52$ respectively). Before and after vitrification, the total cell number and blastocyst development of embryo were significantly higher in July to August than September to October. In conclusion, addition of ${\alpha}$-tocopherol 200 ${\mu}M$ to in vitro bovine embryo culture medium was beneficial for improving embryo quality by decreasing the embryo damage blsstocysts cell number and improving the tolerance of the embryos to cryopreservation.

• Korean Journal of Animal Reproduction
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• v.20 no.2
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• pp.119-126
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• 1996

This study was carried out to determine the optimal condition for successful and efficient c cryopreservation of zygotes, 1-cell embryos, using EFS40 which was 40% (v/v) ethylene glycol diluted in DPBS medium containing 30% Fic-oll (w/v) and 0.3 M sucrose. After mouse zygote produced by IVF was vitrified by two freezing methods, the post-warming survival rates of 1-cell zygotes were assessed as cleavage to the 2-cell stage and development into the hatching blastocysts at 5 day. In the one-step method, when embryos were directly exposed to the vitrification solution at 25$^{\circ}C$ for 1 min., survival and development rates of zygotes were 85.5% and 31.9% In the two-step method, embryos were equilibrated with a dilute 20% EG for 1, 3, 5 min. before 1 min. exposure to EFS40, re-spectively. However, the rates of development (17.7, 3.3, 0%) were lower than that of one-step method. The highest survival rate (95.9%) was obtained by one-step method which exposes embryos in EFS40 for 30 sec. In this condition, 63. 8% of cleaved 2-cell developed into hatching blastocysts. In the cell number of Total and ICM using differential labelling technique, there are no significant differences in the cell number of Total and ICM between blastocysts devel oped in vitrified-thawed embryos (63.2${\pm}$16.9, 1 13.5${\pm}$4.0) and control balstocysts (54.0${\pm}$15.2, 1 12.3${\pm}$4.6). Therefore, these results show that mouse zygotes can be successfully cryopreserved by a simple vitrification method although developmental rates of vitrified embryos were reduced. In conclusion, this proposed vitrifi cation procedures can be useful in the cryopreservation of mouse IVF zygotes.

• Applied Biological Chemistry
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• v.13 no.3
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• pp.207-218
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• 1970

Studies were carried out to develope the most economical and practical methods of packaging and preservation of kimchi, so commercialization of kimchi manufacture could proceed rapidly. The results obtained may be summarized as following. (1) It is generally established that the acceptable range of lactic acid content of kimchi is between 0.4% and 0.75%. Based on sensory evaluation, kimchi having lactic acid content below 0.4% and above 0.75% was not edible, and the time of optimum taste corresponded to the vicinity of 0.5% of lactic acid content. For the refrigeration storage with or without preservatives, the packaging kimchi in plastic film must be done at the lactic acid content of 0.45%, for lactic acid fermentation will continue slowly after the packaging. However, for the heat sterilized kimchi the packaging should be done at the 0.5% of lactic acid content for the best because lactic acid fermentation is completely stopped after the packaging. (2) Polyethylene, polypropylene, and polycello were chosen as suitable packaging materials. Polyethylene is cheapest among them but kimchi packaged in this film was damaged frequently in handling process and gave off kimchi flavor. On the other hand polypropylene also gave off kimchi flavor, but its higher mechanical strength gave better protection to kimchi and it had superior display effect due to the transparancy. Therefore polypropylene made much better packaging material. Polycello proved to be the best packaging material from the standpoint of physical characteristics but its price is higher than that of other plastic films. To be effective, the thickness of plastic films for packaging kimchi must exceed 0.08mm. (3) Keeping property of kimchi appeared to be excellent by means of freezing. However, by the time the frozen kimchi was thawed out at room temperature, moisture loss due to drip was extensive, rendering the kimchi too stringy. (4) Preservation of kimchi at refrigerated temperatures proved to be the best method and under the refrigerated condition the kimchi remained fresh as long as 3 months. The best results were obtained when kimchi was held at $0^{\circ}C$. (5) In general, preservatives alone were not too elective in preserving kimchi. Among them potassium sorbate appeared to be most effective with the four fold extension of self-life at $20^{\circ}C$ and two fold extension at $30^{\circ}C$. (6) In heat sterilization the thickness of packaged kimchi product had a geat effect upon the rate of heat penetration. When the thickness ranged from 1.5 to 1.8cm, the kimchi in such package could be sterilized at $65^{\circ}C$ for 20 minutes. Kimchi so heat treated could be kept at room temperature as long as one month without apparent changes in quality. (7) Among combination methods, preservation at refrigerated and heat sterilization could be favorably combined. When kimchi was stored at $4^{\circ}C$ after being sterilized at $65^{\circ}C$ for 20 minutes, it was possible to preserve the kimchi for more than 4 months.

• Korean Journal of Poultry Science
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• v.42 no.2
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• pp.109-116
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• 2015

Ogye rooster sperm chromatin status can be detected using well established sperm assays. In this paper, a simple and fast method to monitor rooster sperm chromatin status could be employed in field for assessment of chicken sperm quality. Using standard bright field microscope, Diff-Quik stains can be reproducibly, easily and routinely monitored with simple staining. The presence of abnormal chromatin staining of rooster sperm was determined by darker stain in head. In the fresh semen, the viabilities of three tested Ogye spermatozoa were 93.53%, 82.42% and 90.63% and normal chromatin rates were 87.96%, 74.25% and 85.10% respectively. However, after freezing, the rates of viability of thawed semen were reduced to 69.58%, 61.98% and 72.20% and normal chromatin rate also reduced to 58.91%, 48.49% and 63.34%. A significant correlation between live sperm and normal sperm nuclei was 0.875 in fresh semen and 0.513 in frozen semen. After incubation of sperm at $37^{\circ}C$ for 5min, the rates of viability, chromatin normality and sperm head activity were shown as $90.63{\pm}1.28%$, $82.44{\pm}8.09%$ and $66.68{\pm}10.29%$ in fresh semen. However, the rates of thawed semen were reduced to $67.92{\pm}7.55%$, $56.92{\pm}12.15%$ and 47.32{\pm}5.02%, respectively. The relationship between chromatin normality and sperm head movements in fresh and thawed semen were 0.564 and 0.540, respectively. With these results, the chicken sperm normality could be assessed by the Diff-Quik staining that could be used for chromatin status of sperm head and activated morphology of live spermatozoa, as a simple and rapid staining method.