• Development and Reproduction
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• v.22 no.4
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• pp.369-378
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• 2018

We determined the morphologic characteristics (body weight and degree of abdomen inflation) of the red spotted grouper, Epinephelus akaara, mother fish producing healthy eggs. Experimental fish were chosen from fish reared in a sea cage. The fish were divided into four size groups by body weight: 400~600, 600~800, 800~1,000, and 1,000~1,200 g and four stages (I~IV) of the degree of abdomen inflation. After hormone treatment, we observed the amount of ovulation-induced eggs, and rates of buoyancy, fertilization, embryonic survival, and hatching. As a result, mother fish with a body weight of 600 g or more spawned, and the fertilization rate, embryonic survival rate, and hatching rate were high in the 800~1,000 g range, thus showing effective ovulation induction. As a result of dividing the degree of abdomen inflation based on the anal fin of the mother fish into I-IV stages and determining hormone treatment time, the GSI was $0.9{\pm}0.2%$ at stage I, $2.3{\pm}0.2%$ at stage II, $5.6{\pm0.2%$ at stage III, and $7.9{\pm}0.9%$ at stage IV. The flotation rate and hatching rate were highest at stage III, and the fertilization rate and embryonic survival rate were highest at stage IV. Therefore, in terms of egg quality, the amount of eggs collected per mother fish, maturation, and histology were different depending on the degree of abdomen inflation. At stage III, where the abdomen inflation degree of the mother fish was based on the basal part of the dorsal fin relative to the height of the anal fin was 1, the egg quality was highest.

• Journal of Embryo Transfer
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• v.12 no.1
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• pp.27-36
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• 1997

The studies were carried out to investigate the effects of co-culture with cumulus cells and oviduct epithelial cells on the in vitro fertilization and cleavage rate of bovine follicular cocytes and to determine the optimum thawing temperature and equilibration time on in vitro developmental rate of frozen bovine embryos. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes were cultured in TGM-199 medium containing 10 IU /ml의 PM SG, 10 IU /ml의 hCG, ip g/ml의 $\beta$-estradiol and 10% FCS for 24~48 hrs in incubator with 5% $CO_2$ in air at 38.5$^{\circ}C$. The bovine embryos following dehydration by cryoprotective agents and a various concentration of sucrose were directly plunged into liquld nitrogen and thawed in 3$0^{\circ}C$ water. Survival rate was defined as developmental rate on in vitro culture or FDA-test. The results are sunanarized as followes :1. The in vitro fertilization and in vitro developmental rates of bovine oocytes co-cultured with cumulus cells in TCM499 medium were 75.0~76.8% and 17.3~27.6%, respect-ively. And in-vitro fertilization rates of cumulus-enclosed oocytes(55.4%)were significantly(p<0.05) higher than cumulus-denuded oocytes (23.1%). 2. The in vitro fertilization and in vitro developmental rates of bovine oocytes co-cultured with l$\times$ l04cells /ml, 1 x l06cells /ml, lx l08cells /ml and 1 x l015cells /ml oviduct epithelial cells in TCM-199 medium were 74.5~77.8% and 15.7~21.20 respectively.3. The in-vitro fertilization and in vitro developmental rates of bovine oocytes cocultured in '1CM-199 media containing PMSG, hCG, PMSG+hCG. PMSG+$\beta$-estradiol, hCG+$\beta$-estradiol 0 to 40 hrs after insemination were 74.0~77.4% and l8.9~23.l%, re-spectiv ely.4.The survival rates of bovine embryos thawed after rapid freezing in the freezing medium containing a various concentration of sucrose added 1.5M and 2.OM glycerol,DMSO and propanediol were 23.5~31.4% and 20.6~34.l%, respectively. 5. The temperature thawed at 3$0^{\circ}C$ after rapid freezing of bovine embryos resulted in a significantly higher embryos survival rate than did at 2$0^{\circ}C$ and 35$^{\circ}C$.6. The equilibration time on the survival rates of bovine embryos was attained after short period of time(2.5~5 min.) in the freezing medium higher than long period of time (10~20min.). (Key words : bovine embryos, co-culture, freezing, in vitro development)

• Horticultural Science & Technology
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• v.33 no.1
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• pp.24-30
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• 2015

This study was conducted to select proper light intensity, pot media, and fertilization level for potted Hepatica asiatica Nakai native to Korea. The plants were grown under various light intensities (shading rate, 52, 82, 90, 97%) imposed by shading net. Plants grown with 52% shading showed a low survival rate (65%). Survival rate increased as shading increased, with over 80% survival in shading above 90%. Growth indexes such as fresh weight and leaf number did not show any significant difference between shading treatments. Plants grown in a soil mixture of decomposed granite:fertilizer-amended media:Kanumatsuchi (60:10:30, v/v/v) or river sand:fertilizer-amended media:bark (50:20:30) showed over 85% survival. However, plants grown in a soil mixture of river sand:fertilizer-amended media:Kanumatsuchi (50:30:20) or upland:river sand (40:60) showed very low survival, below 60%. Leaf number and plant height were the highest in a soil mixture of decomposed granite:fertilizer-amended media:Kanumatsuchi (60:10:30) as well. To select a proper fertilization level for H. asiatica, hyponex solution diluted 1,000- or 2,000-fold were applied weekly or biweekly. The survival rate was lowest at weekly application with 1,000-fold diluted solution, and no significant difference was observed between other treatments. In conclusion, H. asiatica exhibits preferences for very low light intensity and soil with air permeability, and is adaptable to a broad range of fertilization levels.

• The Korean Journal of Malacology
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• v.19 no.1
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• pp.33-40
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• 2003

To obtain the basic information on culture conditions for the larvae of Saxidomus purpuratus, experiments were conducted on the population from southern coast for (1) the success in fertilization and development from artificial fertilization among different months of a year, (2) the viability of sperms after exposure to seawater, (3) and the effects of temperature, salinity, and food organism on the survival and growth of larvae. Gametes obtained from dissection showed high rate of fertilization at all months. But the rate of development was higher only May-July. Developmental success seemed to be related with the quality of eggs at the time of fertilization. Developmental times for 2-cell, 4-cell, 8-cell, blastula, trochophore larva, and veliger larva at 20$^{\circ}C$ were 1.5, 2, 4, 18, 24, and 32 hr, respectively. Sperms could survive for more than 8 hr, however, actively swimming sperms could be found within 1 hr after exposure to seawater. It is recommended that sperms should be used for fertilization as soon as possible when they are exposed to seawater. At temperature of 35$^{\circ}C$, all the larvae died during 48 hr. Larval survival decreased when salinity was either lower than 20 psu or higher than 40 psu, and was 0% when salinity was 10 psu. Optimal range of temperature and salinity for rearing larvae of S. purpuratus were 20-25$^{\circ}C$ and 20-40 psu, respectively. Larvae grew from 111.5 to 235.3 ${\mu}$m during 21 days. Larvae fed mixed diets grew faster than unialgal diets. The fastest growth was observed when larvae were fed on the mixture of Isochrysis galbana and Nannochloris oculata.

• Environmental Analysis Health and Toxicology
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• v.20 no.1
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• pp.57-65
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• 2005

Effects of five polycyclic aromatic hydrocarbon (PAHs) constituents (naphthalene, fluorine, fluoranthene, benzo(a)pyrene, pyrene) on fertilization and early development of sea urchin egg, sperm and fertilized egg were investigated. The eggs, sperm and fertilized eggs were exposed to several concentrations of PAHs (1, 10, 100, 1000 and 10000㎍/L). The rate of fertilization and hatching decreased when the eggs and sperm were exposed to aqueous solution of PAHs. Also, Exposure of fertilized eggs with each PAHs did decrease survival and hatching rate. Concentration-dependent toxic effects on the rate of fertilization, hatching, survival and abnormality in A. crassispina were observed following exposure to PAHs (1, 10, 100, 1000 and 10000㎍/L). These data show that PAHs exposure decreased in fertilization success of sea urchin egg and sperm and producted abnormal embryo. It is plausible to suggest that PAHs had the potential to significantly reduce coastal recruitment of sea urchin.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.2
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• pp.191-200
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• 2000

Objective: This study was carried out to compare the effects of the stepwise exposure treatments on the morphological normality, fertilization and blastocyst formation rate of the frozen-thawed mouse mature oocytes by vitrification or ultra-rapid freezing and to use as a fundamental data for the cryopreservation of human oocytes. Materials and Methods: The morphological normality and fertilization rates of the vitrified and ultra-rapid frozen mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were observed. After choosing the 3step exposure treatment groups, we observed the morphological normality and fertilization, blastocyst formation rate of the vitrified and ultra-rapid frozen mouse mature oocytes. Results: The morphological normality and fertilization rates of the vitrified mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were 75%, 85%, 88% and 58%, 61 %, 54% respectively. There were no significant differences among treatments(p>0.05). The morphological normality and fertilization rate of the control was 92% and 65%. There were no significant differences in fertilization rate among control and treatments (p>0.05). The morphological normality and fertilization rates of the ultra-rapid frozen mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were 83%, 83%, 84% and 75%, 63%, 56% respectively. There were no significant differences among treatments (p>0.05). The morphological normality and fertilization rate of the control was 95% and 67%. There were no significant differences among control and treatments (p>0.05). The morphological normality and fertilization rate of the vitrified or ultra-rapid frozen mouse mature oocytes after 3step exposure treatment were 69% and 75%, respectively. The blastocyst formation rate was 60% and 57%. The results did not differ significantly between vitrification and ultra-rapid freezing (p>0.05). Conclusion: As known in the above results, there were no significant differences in the fertilization and blastocyst formation rate of the frozen-thawed mouse mature oocytes by vitrification or ultra-rapid freezing among the control and treatments. It is suggested that vitrification and ultra-rapid freezing method were effective for the cryopreservation of mouse mature oocytes.

• Fisheries and Aquatic Sciences
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• v.14 no.4
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• pp.417-420
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• 2011

In this experiment, we examined the survival, fertilization, hatching times, and hatching rates of Far Eastern catfish Silurus asotus at pH ranging from 2 to 13 under laboratory conditions. Eggs could be fertilized at pH 3-12. In a hatching experiment, mortality was first observed at pH 13, when all fertilized eggs died within 8 min, followed by pH 2 (30 min), pH 12 (60 min), pH 3 (4 h), and pH 11 (5 h). Hatching only occurred at pH 4-10, with the highest hatching rate at pH 7 (52%) and the lowest at pH 10 (24%). Hatching rates in acid solutions were higher than in alkaline solutions, although the difference was not significant. Hatching was first observed at pH 10, beginning 27 h after fertilization and ending at the 31 h. A clear difference was observed between hatching times, ranging from 31 to 64 h and increasing in order with descending pH.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.3
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• pp.201-207
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• 2002

Objective: To observe the capability of fertilization and embryo development including blastocyst formation of the oocytes in simple media after thawing of the cryopreserved cumulus-free mouse oocytes by vitrification method. Methods: Oocytes were collected from 5 to 6 weeks old ICR female mice, and were denuded from the cumulus cells by 0.1% hyaluronidase. Recovered mature oocytes in study group were cryopreserved by vitrification method using EM grid for $5{\sim}7$ days. In brief, oocytes were exposed in dPBS containing 1.5 M EG and 5.5 M EG+1 M sucrose for 2.5 minutes and 20 seconds each, and then executed vitrification by plunging in LN2 after loading on EM grid. Thawing treated by exposure of 1, 0.5, 0.25 and 0.125 M sucrose solution for 2.5 minutes each in order and used for experiments. Spermatozoa aspirated form the epididymis of 12 weeks old ICR male mice were used for insemination after capacitation. T6 media containing 0.4% BSA were used for fertilization and development. Results: Survival and fertilization rates after thawing were 76.9% and 79.6% respectively. Fertilization rate was lower (p<0.005) than that of control group (92.9%). There was no difference in embryo developmental rates from 2-cell to morula, however, the blastocyst formation rate and mean cell numbers of blastocysts in study group (63.3%, $58.9{\pm}9.2$) were lower compared with those of control group (76.1%, $63.5{\pm}8.9$). Conclusion: Vitrification is an effective method for mouse mature oocyte cryopreservation with high survival and fertilization rate after thawing. And in simple media, fertilization rates and embryo development of frozen-thawed mouse oocytes are satisfactory.

• Korean Journal of Animal Reproduction
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• v.22 no.3
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• pp.207-211
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• 1998

본 연구는 소 미성숙 난자의 동결에 있어서 미성숙 난자의 1, 2, 4, 8 시간 성숙배양한 다음 동결 융해 후 체외배양하였을 때 체외수정율과 생존율을 조사하고저 수행하였다. 미성숙 난자들 1∼8시간 성숙배양후 동결융해 및 체외수정시켰을 때 수정율은 12.8∼30.9%로서 단시간의 체외성숙 배양이 높게 나타났으며(23.8∼28.8%), 또한 체외배양시 생존율은 11.8∼29.9%를 나타냈다.

• Journal of Aquaculture
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• v.10 no.4
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• pp.395-402
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• 1997

The study has been conducted in order to understand the inheritance of body color in the wild type zebrafish (zebra danio), Danio rerio, and its golden mutant (golden danio). The body color was also studied to determine the effect of golden coloration on the survival rate of zebrafish eggs and larvae up to 15 days after fertilization. Reciprocal monohybrid crosses between the wild and the golden type of zebrafish indicated that golden coloration was controlled by a single gene which had two alleles. Transmission of these alleles from parents to their progenies followed the principles of dominance and segregation based on Mendelian inheritance. Similar results from the reciprocal crosses implied that a locus for golden coloration was located on an autosomal chromosome. On the other hand, average survival rates from four different types of mating between, and within, zebra and golden danio suggested that golden coloration seemed to be associated with the survival rate of zebrafish, especially in its early embryonic stage. This indicated that homozygous recessive golden mutation was likely to weaken the golden danio's chance of survival.