• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.42 no.1
• /
• pp.63-67
• /
• 2009

The effects of adhesive substrates and bottom materials on the release of fertilized eggs by the Pacific cod, (Gadus macrocephalus) were examined to find an effective release method. Palm twin as an adhesive substrate displayed a significantly elevated adhesive rate compared to gulf weed. However, the survival rate of the fertilized eggs attached to palm twin was as low as 17.5% 8 days after fertilization (DAF). In contrast, fertilized eggs in the absence of adhesive substrate displayed the highest survival rate (47.0%). Concerning bottom materials, the survival rate of fertilized eggs on sand was significantly high (approximately 51.1%) on 8 and 9 DAF. The observations indicates that adhesive substrates are not needed and spawning ground bottom materials such as sand or mud should be considered when releasing fertilized eggs of Pacific cod.

• Journal of Aquaculture
• /
• v.12 no.1
• /
• pp.39-45
• /
• 1999

In order to obtain the basic biological data for artificial seedling production of sea cucumber, Stichopus japonicus, the influence of water temperature and air dry for spawning induction, egg development and larvae rearing was investigated during the period from April, 1995 through September, 1995. Spawning induction rate by the water temperature was 6.0~17.5% and air dry was responsed 1.4~4.0%. Number of eggs spawned of Stichopus japonicus were $50~500\times10^4$ individuals, the fertilization and hatching rate were ranged 84.0~96.0%, 71.4~84.6% respectively. The fertilized egg of Stichopus japonicus appeared mean diameter of $154{\mu}m$. At a constant water temperature of $23^{circ)C$, it become 4 tell stage from fertilization after 2 hours 10 minutes, hatching larvae after 14 hours half, auricularia larvae after 3 days, doliolaria larvare after 11 days and pentactula larvae after 15 days ready for settlement. The suitable food in the larvae reared for 17 days after fertilization were shown the best growth and survival in the larvae food of Chaetoceros calcitrans. Optimum density for larvae rearing were maintained of the larval density lower than 2 individuals/ml.

• Clinical and Experimental Reproductive Medicine
• /
• v.19 no.2
• /
• pp.143-152
• /
• 1992

The present experiments were focussed to modify a short slow-cooling protocol used for freezing of early stage embryo(Testart et al., 1986) and also to apply the modified method for the cryopreservation of hamster oocytes with Zona or without. The protocol was modified by changing the 4-step equilibration into 1-step and the 1-step thawing into 2-step. The oocytes were added in 1.5M PROH and 0.1M Sucrose, seeded at $-7^{\circ}C$, slow cooled($0.3^{\circ}C$/min) to $-30^{\circ}C$ before plunging to $-196^{\circ}C$. The oocytes were thawed at $23-25^{\circ}C$ air(20sec/150sec) and/or 33-35 water(10sec). The survival of the frozen-thawed oocytes was determined by morphologic criteria and their fertilizing ability was also estimated by Sperm Penetration Assay(SPA) system(Chang et al, 1990) using fertile men semen sample. One-step equilibration showed slightly higher survival rate(83.9% vs. 71.0%) and fertilization rate(83.9% vs. 71.0%) compared with four-step(p>0.05). And two-step thawing(air & water exposing) of oocytes frozen after 1-step equilibration showed significantly higher survival rate(96.3%) than one-step thawing at air(85.2%) or water(65.0%) only(p<0.05). Therefore, by the modified method(l-step equilibration & 2-step thawing), Zona-intact(ZI) and Zona-free(ZF) oocytes were frozen and thawed. ZI-oocytes showed significantly higher survival rate(95.4%, 308/323 vs. 67.6%, 240/355) than ZF-oocytes(P<0.01). But the survival of ZF-oocytes was as high as ZI-oocytes in fourteen of twenty-four replicates. ZI-oocytes was also significantly higher fertilization rate($92.4{\pm}8.9%$ vs. $63.7{\pm}18.5%$) and higher mean number of penetrated sperm($6.2{\pm}4.2$ vs. $3.9{\\pm}3.3$) than ZF-oocytes, but not higher than control(fresh oocytes;$99.3{\pm}2.4%$, $8.4{\pm}4.2$)(P<0.001). Conclusively, this modified method will contribute to freeze effectively the hamster oocytes for simplifing of the logical consideration of performing SPA and also to freeze the human and other animal oocytes.

• Development and Reproduction
• /
• v.7 no.1
• /
• pp.9-13
• /
• 2003

A series of experiments were conducted to compare the effects of various diluents in cold storage on the sea urchin, Hemicentrotus pulcherimus sperm. Various diluents of glucose solutions, artificial sea water(ASW) and 50% ASW were used to store the sperm at 4$^{\circ}C$. The storage effect was evaluated using sperm activity index(SAI), survival rate of sperm and fertilization rate to egg. ASW and 1.2 M glucose were found to be better diluents which maintained high motility and survival rate of sperm f3r a storage period of 30 days. Optimal pH of diluent to store the sperm at 4$^{\circ}C$ is 7.0∼8.0. In order to keep high SAI and survival rate of sperm, addition of 400 ppm neomycin into the diluent revealed the best storage results.

• Journal of Korean Society of Forest Science
• /
• v.104 no.1
• /
• pp.90-97
• /
• 2015

The objective of this study was to find optimal amount of fertilization of container seedling production for Zelkova serrata, Ulmus parvifolia, Betula costata and Tilia amurensis. To reach our goal, we measured root collar diameter (RCD), height, biomass and seedling quality index (SQI) of container seedlings of four deciduous hardwood species grown at three different fertilization treatments ($0.5g{\cdot}L^{-1}$, $1.0g{\cdot}L^{-1}$ and $2.0g{\cdot}L^{-1}$). Z. serrata seedlings grown at $2.0g{\cdot}L^{-1}$ fertilization and B. costata seedlings grown at $0.5g{\cdot}L^{-1}$ fertilization showed the highest RCD, height, biomass and SQI. The RCD and height of U. parvifolia and T. amurensis seedlings showed no significant differences by fertilization treatments. Seedlings of two species at $0.5g{\cdot}L^{-1}$ fertilization showed the lowest SQI, however, SQI at 1.0 and $2.0g{\cdot}L^{-1}$ fertilization treatments were not significantly different. Based on these results, it is appeared that container seedlings of Z. serrata at minimum $2.0g{\cdot}L^{-1}$ fertilization, U. parvifolia and T. amurensis at $1.0g{\cdot}L^{-1}$ fertilization and B. costata at maximum $0.5g{\cdot}L^{-1}$ fertilization were optimal nutrient conditions. Practice of optimal fertilization rate will make us get better quality seedlings and reduction of production costs in the container nursery system as well as good field performances with higher survival rate after planting.

• Korean Journal of Ichthyology
• /
• v.28 no.4
• /
• pp.260-266
• /
• 2016

We determined the morphologic characteristics (body weight and abdomen inflation degree) of the mother fish of sevenband grouper, Epinephelus septemfasciatus producing the healthy eggs. Experimental fish were chosen from the reared fish in the sea cage for 6 years. The fish were divided into four size groups by body weight: 4.0~5.0, 5.0~6.0, and 6.0~7.0 kg and four stages (I~IV) by the abdomen inflation degree. After hormone treatment, we observed the ovulation amount of induced eggs, rate of buoyant, fertilization, embryonic survival, and hatching. Egg and oil globule diameter was measured. In order to observe gonadal development, we calculated gonadosomatic index (GSI) and conducted its historical analysis. The ovulation occurred from all experimental fishes over 5.0 kg. The rate of buoyant, fertilization, and embryonic survival was the highest in 6.0~7.0 kg. Hatching rate was the highest in 5.0~6.0 kg. Stage I and II did not induce ovulation. GSI was $0.31{\pm}0.10%$ in stage I, $0.74{\pm}0.25%$ in stage II, $4.68{\pm}0.40%$ in stage III and $6.86{\pm}0.12%$ in stage IV. The rate of buoyant, fertilization, embryonic survival and hatching was the highest in stage III.

• Clinical and Experimental Reproductive Medicine
• /
• v.43 no.1
• /
• pp.9-14
• /
• 2016

Objective: Autophagy contributes to the clearance and recycling of macromolecules and organelles in response to stress. We previously reported that vitrified mouse oocytes show acute increases in autophagy during warming. Herein, we investigate the potential role of Atg7 in oocyte vitrification by using an oocyte-specific deletion model of the Atg7 gene, a crucial upstream gene in the autophagic pathway. Methods: Oocyte-specific Atg7 deficient mice were generated by crossing Atg7 floxed mice and Zp3-Cre transgenic mice. The oocytes were vitrified-warmed and then subjected to in vitro fertilization and development. The rates of survival, fertilization, and development were assessed in the Atg7 deficient oocytes in comparison with the wildtype oocytes. Light chain 3 (LC3) immunofluorescence staining was performed to determine whether this method effectively evaluates the autophagy status of oocytes. Results: The survival rate of vitrified-warmed $Atg7^{f/f}$;Zp3-Cre ($Atg7^{d/d}$) metaphase II (MII) oocytes was not significantly different from that of the wildtype ($Atg7^{f/f}$) oocytes. Fertilization and development in the $Atg7^{d/d}$ oocytes were significantly lower than the $Atg7^{f/f}$ oocytes, comparable to the $Atg5^{d/d}$ oocytes previously described. Notably, the developmental rate improved slightly in vitrified-warmed $Atg7^{d/d}$ MII oocytes when compared to fresh $Atg7^{d/d}$ oocytes. LC3 immunofluorescence staining showed that this method can be reliably used to assess autophagic activation in oocytes. Conclusion: We confirmed that the LC3-positive signal is nearly absent in $Atg7^{d/d}$ oocytes. While autophagy is induced during the warming process after vitrification of MII oocytes, the Atg7 gene is not essential for survival of vitrified-warmed oocytes. Thus, induction of autophagy during warming of vitrified MII oocytes seems to be a natural response to manage cold or other cellular stresses.

• The Korean Journal of Malacology
• /
• v.21 no.2 s.34
• /
• pp.107-111
• /
• 2005

In the present study, the fecundation and fertilization behaviors of the trumpet shell, Charonia fampas sauliae were observed from November 2003 to March 2004. Female has multiple fecundation and multiple fertilization, and the fertilization continued for two days. The fertilized female stayed with the egg mass to protect the eggs, and the fecundation was made during the fertilization. The highest survival rate of egg capsule was 60% at $15^{\circ}C$. Charonia lampas sauliae fed with Chaetoceros calcitrans, Isochysis galbana, and Pavlova lutheri. Growth and survival rates of C. lampas sauliae in the conditions of various water temperature were observed. The highest survival rate was 23% at $15^{\circ}C$ reared for 60 days. The growth was presented in the $15^{\circ}C$ group, with shell length from $408{\pm}21.52{\mu}m\;to\;625{\pm}19.76{\mu}m$ during the experiment.

• Korean Journal of Animal Reproduction
• /
• v.16 no.3
• /
• pp.217-224
• /
• 1992

These studies were carried out ot investigate on the transferred embryo development following ultrarapid frozen for 8-cell and morula of in vitro fertilization mouse embryos. The post-thaw embryo survival was evaluated and compared by cell stage of embryos and by equilibration time before ultrarapid freezing. The results obatined were summerized as follows: 1. The effects of equilibration time of 3 vs. 6 minutes before ultrarapid freezing and after thawing on the morphological survival and the viability of 8-cell and morulas embryos were not significant. 2. When the ultrarapid frozen-thawed 32 eight-cell and 33 morula embryos, and 30 fresh blastocysts were transferred to pseudopregnant recipient mice, the number of normal offsprings produced were 9(28.1%), 14(42.4%) and 18(60.0%), respectively. From the above resutls, it was concluded that the optimal conditions of pH osmolality of the media for mouse IVF and embryo culture, and the period of sperm preincubation might be 7.1, 310 mOsm and 120 min., respectively,a nd somewhat high conception rate might be resulted from transfer of frozen embryos of morula stage and fresh embryos of blastocyst stage.

• Korean Journal of Environment and Ecology
• /
• v.21 no.6
• /
• pp.544-553
• /
• 2007

This study is focused on the cultivation of seedlings and cuttings of roundleaf chastetree(Vitex rotundifolia L. f.) according to the sorts of soil and fertilization methods. The results are summarized as follows: When roundleaf chastetree seedlings(aged 1-0) and cuttings(aged 1-0) were cultivated in three different kinds of soil- sand, granite soil, and yellow silt- without any fertilization, 70% of the seedlings survived in sand and granite soil, and 35% did in yellow silt; 90% of the cuttings survived in granite soil and 80% did in sand and yellow silt. The general growth of both seedlings and cuttings was good in granite soil, and sand but relatively inferior in yellow silt. Seedlings showed good stem growth while cuttings had fine root growth. In case of cultivation of seedlings with different kinds and quantities of fertilizer adjusted and with granite soil as basic soil, seedlings and cuttings survived in the experimental section where only the organic fertilizer was used as basic fertilization; in contrast, seedlings grown in the experimental section were all dead where both inorganic and liquefied fertilizer were used. Cuttings showed considerably higher survival rates than seedlings at the experimental fertilization section; particularly, the survival rate of cuttings was twice as much as that of seedlings at the experimental section with low fertilization. This study results revealed that cuttings are desirable than seedlings in general rootage, growth, adaptability to soil, and cuttings have lower demand for soil fertility in time of growing roundleaf chastetree saplings.