• Horticultural Science & Technology
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• v.19 no.3
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• pp.373-377
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• 2001

This study was conducted to find optimum pollination methods for overcoming pre-fertilization incompatibility in interspecific crosses between L. longiflorum 'Gelria' (LG) and L. cernuum (LC). In cross combination of $LC{\times}LG$, the survival rate of the ovaries was higher in the cut style pollinations than in normal stigmatic pollination. Among the various cut style pollination methods, pollination after cutting the styles 2 3mm length above the ovary and top of the ovary were the most effective ones. No ovaries and ovules survived in cross pollination of $LC{\times}LG$. In self-pollination of LG and cross pollination of $LC{\times}LG$, pollens germinated on the stigma by 12 hours and reached to the ovules by 90 hours after pollination. However, in the cross pollination of $LC{\times}LG$, pollen tubes elongated very slowly until 48 hours after pollination and a lot of deformed tubes were observed in the styler canal having swelled tube tip and irregular heavy callose deposition. The pollination time after anthesis also affected on the survival rate of ovaries and ovules showing the best result when pollinated on three days after anthesis. Stigma exudate promoted slightly the enlargement of ovaries, but the survival rate of ovaries and ovules was not increased.

• Journal of Embryo Transfer
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• v.23 no.1
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• pp.19-24
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• 2008

The purpose of this study was to compare the efficiency of slow freezing with that of vitrification method for the cryopreservation of human embryos. Human embryos were derived from in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) and the mixed solution of propanedial (1.5, 1.0, 0.5M PROH) and sucrose (0.1M), ethylene glycol (7.5, 15%), dimethyl sulfoxide (7.5, 15% DMSO), sucrose (0.5, 1.0M) and SPS (Serum Protein Substitute) was used for a cryoprotectant for slow freezing and vitrification solution, respectively. Rates of recovery after thawing, morphological normality, post-thaw viability, arrest, morphological abnormality and preimplantation development were compared between two protocols. After freezing-thawing, recovery and survial rate of slow freezing was (88.6% and 73.4%), whereas vitrification was (99.2% and 96.2%) (p<0.05). The arrest rate of slow freezing was significantly lower compared with those of vitrification(8.7% vs 29.9%) (p<0.05). Preimplantation development to the 2-cell (83.8% vs 67.7%), 4-cell (69.0% vs 47.2%) and 8-cell (62.4% vs 37.8%) stages 24, 48 and 72 h after thawing, respectively, were higher in the slow freezing than the vitrification. After slow freezing and vitrification of human embryo at 2-8cell stage, the rate of recovery rate, survival rate and partial damage rate were 92.0% vs 100%, 80.4% vs 96.2% and 52.2% vs 19.0%, respectively. And partial damage rate was significantly lower than those of slow freezing method (p<0.05). These results demonstrate that a slow freezing using PROH is more efficient than a vitrification for cryopreserving the human zygotes, although the vitrification yielded better recovery, survival and partial damage of frozen-thawed 2-8 cell stage embryos than slow freezing method.

• Journal of Korean Society of Forest Science
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• v.108 no.1
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• pp.29-39
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• 2019

The objective of this study was to suggest a suitable amount of fertilizer using the changes in growth performances and soil properties for improving survival and quality of Zelkova serrata trees in a harvested Pinus rigida plantation. One-year-old containerized seedlings of Z. serrata were planted with the density of 3000 seedlings $ha^{-1}$ in end of March 2011 at Gwangneung experimental forest, Pocheon. Solid compound fertilizer (N:P:K=3:4:1) were applied yearly in three amounts (control: no fertilization, F1: $180kg\;ha^{-1}$, and F2: $360kg\;ha^{-1}$) every May from 2011 to 2013. We analyzed soil properties before (2011) and after (2012 and 2017) fertilization. And we measured the root collar diameter and height of Z. serrata trees from 2011 to 2016, and then calculated H/D ratio and stem volume. Soil properties at Z. serrata plantation did not show difference according to fertilization level in every investigation year. As time passed after planting, however, concentrations of total nitrogen and available phosphorus were increased from decreased. The growth of root collar diameter, height and stem volume of Z. serrata trees at F2 plot were significantly higher those at the other plots after only 2 years of fertilization. Because Z. serrata tree demand to more nutrient during the early growing period. The survival rate of Z. serrata trees at control plot was significantly lower than that at the other plots. This might be due to Z. serrata trees at control plot had not the upper hand from competition with vegetation at the early in planting. However, the growth of height and stem volume of Z. serrata trees between F1 and F2 plots did not show difference over 6 years after planting. Consequently, we could suggest that Z. serrata trees need to F1 fertilization level for considering improving survival and quality of Z. serrata trees and economical efficiency of plantation managements after harvesting P. rigida plantation.

• Journal of Embryo Transfer
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• v.27 no.1
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• pp.29-35
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• 2012

The purpose of this study was to examine the effect of growing stages of the Korean Native Striped Bull (KNSB) on the freezability and fertility of frozen-thawed semen. First, we investigated the total motility (TM) and progressive motility (PM) according to the diluent used for semen freezing. Second, we examined the effect of the age of KNSB on semen volume, TM and PM of fresh and frozen-thawed semen. Third, we examined the effect of frozen semen from the different age of KNSB on the $in-vitro$ fertilization rate, and the artificial insemination pregnancy rate. The diluents used in this experiment were Triladyl$^{(R)}$ and Tris-egg yolk extender (EYE). Semen was collected from 5 KNSB in the growing stage (15 months) and 5 adult KNSB (36 months). When Triladyl or Tris-EYE extender was used for semen freezing, there was no difference of the mean TM and the mean PM. However, the mean TM was significantly higher in Bull No. 1885 than Bull No. 4283 ($p$ <0.05). The mean volume of semen collected from the 15-month-old bulls (2.3 ml) was significantly lower ($p$ <0.05) than that from the 36-month-old bulls (5.0 ml). The mean semen concentration was similar for the 15-month-old ($2.1{\times}10^9$ spermatozoa/ml) and 36-month-old ($1.8{\times}10^9$ spermatozoa/ml) bulls. For the 15-month-old and 36-month-old bulls, the mean TM of fresh semen were 93.7% and 88.3%, respectively, and the mean PM were 97.0% and 88.3%, respectively; the 15-month-old bulls showed a particularly high PM ($p$ <0.05). For the 15-month-old and 36-month-old bulls, the mean TM (56.0% and 58.0%, respectively) and the mean PM (64.0% and 70.7%, respectively) of frozen-thawed semen did not differ. The development rates of embryos after $in-vitro$ fertilization and the pregnancy rate after artificial insemination using frozen-thawed semen did not differ according to the bull's age. In summary, semen volume differed according to the bull's age, but semen concentration and survival rate, the $in-vitro$ fertilization rate, and the pregnancy rate did not differ according to the stripe bull's age. Accordingly, semen from bulls in the growing stage can be collected and frozen for the preservation and multiplication of rare livestock.

• Korean Journal of Environmental Biology
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• v.38 no.2
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• pp.323-331
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• 2020

The method of natural spawning is very passive and inconvenient for the study of developmental engineering in marine medaka, Oryzias dancena. The optimum concentration of human chorionic gonadotropin (HCG) and carp pituitary extract (CPE) for ovulation and spawning, and the injection time for the artificial spawning of marine medaka were analyzed in this study. The success rate, survival rate, and hatching rate were highest with 100IU HCG kg^-1 BW and 5mg CPE L^-1 in both male and female marine medaka (p<0.05). After obtaining unfertilized eggs and sperm by the injection of HCG and CPE into the broodstock of marine medaka, artificial fertilization could be successfully achieved any time fertilized eggs are needed in this species. This result should be useful for developing a study program for marine medaka as an experimental animal.

• Journal of Aquaculture
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• v.12 no.3
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• pp.229-236
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• 1999

in the present study, the zinc toxicity to larval development and seed attachment of the abalone, Haliotis discus hannai was obtained under continuous flow through system. The zinc concentration melted from zinc coating pipe for 7 months ranged from $89.00\pm2.55 \mu\textrm{g}/\ell to 15.23\pm2.58\mu\textrm{g}/\ell(Y=0.85M^2-19.71+109.96)$. Treatments were carried out with zinc concentration $0~160 \mu\textrm{g}/\ell$. The maximum and minimum of fertilization rate were $87.7\pm5.3%$ in control, $83.7\pm7.6%$ in zinc concentration $160\mu\textrm{g}/\ell$, respectively. The maximum and minimum of hatching rate were $87.5\pm4.5%$ in zinc concentration $10\mu\textrm{g}/\ell$, $79.3\pm5.6%$ in zinc concentration $160\mu\textrm{g}/\ell$, respectively. Both of the results were not significantly different (P>0.05). But the normality rate, setting rate and survival rate of abalone larvae at over zinc concentration TEX>$20\mu\textrm{g}/\ell$ decreased rapidly and showed significantly different from those of the other group(P<0.05).

• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.281-292
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• 1997

The objective of this study was to compare retrospectively the survival and pregnancy rates(PR) of cryopresered-thawed embryos obtained from intracytoplasmic sperm injection (ICSI) or conventional in vitro fertilization (IVF). Ninety-six cycles of cryopresered-thawed embryo transfer (ET) were performed in 79 patients from June, 1996 to September, 1997 and grouped as followings: 20 cycles (16 patients) inseminated by ICSI (ICSI Group) and 76 cycles (63 patients) by conventional IVF (IVF Group). Slow-freezing and rapid-thawing protocol was used with 1.5M propanediol (PROH) and 0.1M sucrose as cryoprotectant. All embryos were frozen-thawed at the two pronuclear (2 PN) stage excluding four cycles in which the early cleavage stage embryos were frozen, and allowed to cleave in vitro for one day before ET. The duration from freezing to thawing was comparable in both groups ($mean{\pm}SD$, $112.1{\pm}80.0$ vs. $124.8{\pm}140.1$ days). The age of female ($31.2{\pm}3.4$ vs. $32.6{\pm}3.3$ years) and the endometrial thickness prior to progesterone injection ($9.4{\pm}2.0$ vs. $9.3{\pm}1.8$ mm) were also comparable in both groups. There was no significant difference in the outcomes of cryopreserved-thawed ET between two groups: survival rate ($85.2{\pm}16.1%$ vs. $82.2{\pm}19.7%$), cleavage rate ($96.9{\pm}6.7%$ vs. $94.7{\pm}13.0%$), cumulative embryo score (CES, $54.5{\pm}31.1$ vs. $49.0{\pm}20.0$), preclinical loss rate (5.0% vs. 5.3%), clinical miscarriage rate (0% vs 29.4%), clinical PR per transfer (35.0% vs. 22.4%), implantation rate (9.9% vs. 5.6%), and multifetal PR (42.9% vs. 17.6%). In conclusion, human embryos resulting from ICSI can be cryopreserved-thawed and transferred successfully, and the survival rate and PR are comparable to conventional IVF.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.9 no.1
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• pp.43-55
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• 1976

Fertilization and early development of turbo cornutus was studied based on the samples which were collected in Yeosu area. Particular emphasis was paid on induction of artificial spawing, fertilization rate, preembryonic development, the growth of the early larva and larval survival to various salinity. Among the various methods for induction of artificial spawning which have been tested for the present study, drying by exposure to air is the. most efficient, and percentage fertilization rate was $83.8-96.4\%$. The diameter of fertilized eggs was $0.182{\pm}0.0028mm$; and the diameter of egg membrane was $0.245{\pm}0.093mm$. Under the temperature range of $20.6-25.4^{\circ}C$ the larvae hatched out after 11:05-11:15 hours of fertilization. After 3.0-3.5 days of fertilization the planktonic larvae begand to settle, and the settlement terminated within 5 days. During the period of 150 days of early culturing the diameter growth of shell(M) and the diameter of shell aperture(A) was formulated as follows: $$1972\;M=0.33e^{0.02070D}$$ $$A=0.19e^{0.02282D}$$ $$1973\;M=0.32e^{0.02282D}$$ $$A=0.16e^{0.02596D}$$ During the same period of early culturing the relative growth of shell diameter and the diameter of shell aperture was formulated as follows : 1972 A=0.6478 S-0.1575 1973 A=0.5897 S-0.0515 After 11 days of larval hatching $0.02-0.18\%$ of planktonic larvae settled. After 150 days of settlement the survival rate of the early shells was $7.4-21.6\%$. Under the temperature range of $21.0-22.7^{\circ}C$ the optimum salinity range for the development of egg and the planktonic larvae was $30-35\%_{\circ}$.

• Applied Microscopy
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• v.29 no.2
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• pp.241-253
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• 1999

Fertilization pattern of north temperate bats is known to be unique for their sperm storage in the female reproductive tract during hibernation (e.g. Korean greater horseshoe bats). They copulate in fall but their ejaculated spermatozoa survive until the next spring. In another words they can persist to survive during long hibernation under the cold condition $(8\sim13^{\circ}C)$ and are to be fertilized with the ovum ovulated in the next spring, so called delayed fertilization. The present study was designed to observe morphological and functional changes of spermatozoa plasma membrane of the bats, hamsters which are hibernators, and mice which are non-hibernators in the room and the cold (bat-hibernation) temperatures and to confirm influence of the temperature on spermatozoa; survival rate, acrosome reaction rate, protein distribution, $Na^+-K^+-ATPase$ activities and scanning electron microscopic histochemistry. Based on the experimental results obtained in the present study, there were no significant morphological and functional differences in the spermatozoa plasma membrane in both the room and cold (bat-hibernation) temperatures and such an absence of difference suggests that the spermatozoa plasma membrane might play a pertinent role as a protector for consistent fertilization during and after hibernation.

• Journal of Embryo Transfer
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• v.17 no.1
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• pp.61-65
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• 2002

This study was carried out to study the viability of oocytes when vitrified at various maturation stages. Bovine cumulus-oocyte complexes were recovered from ovaries at a slaughter and then divided into five groups: control group(unvitrified oocytes), 0 hr. group(composed of oocytes vitrified before the onset of maturation) and 10, 14, and 20 hrs groups(vitrified at 10, 14 and 20 hrs after the onset of maturation, respectively). The oocytes remained vitrified for 24 hrs, and then were thawed in 3$0^{\circ}C$. Survival and cleavage rates were investigated by results of in vitro culture and aceto-orcein staining or FDA test. No difference in the incidence of diploid oocytes was observed among the control, non-vitrified group(3.6%) and oocytes vitrified at 14 hrs(6.7%) or 20 hrs(1.7%). However, more diploid oocytes were detected after vitrification at 0 hr.(26.7%) and 10 hrs(21.7%) post maturation. The survival rate of all vitrified immature oocytes(12.0~38.0%) was low, 48.0% of unvitrified oocytes and oocytes vitrified before maturation or 0~ 10 hrs after the onset of maturation were higher than that of other groups. The overall fertilization and cleavage rates of vitrified immature oocytes (32.3 ~ 64.6% and 4.6 ~ 32.3%) were low, and 55.0% of unvitrified oocytes and the rate of immature oocytes were very higher than that of mature oocytes.