• Reproductive and Developmental Biology
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• v.32 no.4
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• pp.237-243
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• 2008

The objective of this study was to examine the effect of caffeine and sodium bicarbonate in a fertilization medium on the fertilizability of boar spermatozoa that were frozen in straws. Boar spermatozoa were extended with Beltsville F5 extender and frozen in 0.25-ml straws. In vitro matured porcine oocytes were fertilized in vitro (IVF) with frozen-thawed boar spermatozoa for 6h in a modified tris-buffered medium (mTBM) or in its modified medium by substituting the tris with 25mM sodium bicarbonate (modified bicarbonate-buffered medium; mBBM). Some of inseminated oocytes were fixed and stained for examination of sperm penetration. IVF embryos were cultured in a North Carolina State University-23 medium for embryo development. The percentage of live sperm was $47{\pm}4%$ and morphological abnormality of acrosome was found in $14{\pm}3%$ of spermatozoa. Optimal sperm concentration for IVF was $0.75{\sim}1.0{\times}1.0{\times}10^6$ sperms/ml when mTBM containing 5mM caffeine was used as the fertilization medium. Sperm penetration was significantly (p<0.05) stimulated by increasing caffeine concentration in the IVF medium. In addition, mBBM significantly (p<0.05) increased sperm penetration (92%) compared to mTBM (65%). More (p<0.05) blastocysts (22% vs. 32%) developed from the oocytes that were fertilized in mBBM containing 1mM caffeine than from those fertilized in mTBM with 5mM caffeine. Our results indicate that boar spermatozoa can be frozen successfully in straws with holding their normal fertilizability and that caffeine and sodium bicarbonate stimulates sperm penetration in vitro.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.385-391
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• 1997

The purpose of this study was to examine the effects of anti-sperm antibody (ASA) on the fertilization processes using conventional IVF and ICSI procedure in human and hamster oocytes. In human IVF, we have observed restricted fertilization with sperm testing positive for ASA. ($23{\sim}90%$ IgA, 60-97 % IgG). However, if ICSI was perform in the next IVF cycle with the same patients, we could successfully fertilize the oocytes (37%; p<0.001), thus achieving pregnancy and delivery. When the sperm were cocultured in medium containing ASA, there were binding of ASA to sperm surface. In addition, the mean rate of the acrosomal reaction in an in vitro acrosome reaction test was lower for Ab-bound sperm (43.5%) than for Ab-free sperm group (51.3%, p<0.05). We used human sperm and hamster oocytes to confirm the negative effects of the ASA on fertilization. The sperm and/or oocytes have been expose to medium containing ASA before IVF and ICSI. In this experiment, the ASA was bound to the oocyte and sperm surface. The following results were obtain by using various combinations of ASA free or ASA bound sperm with ASA free or ASA bound oocytes for IVF. When ASA free sperm were inseminate with ASA free and ASA bound hamster oocytes, the fertilization rates are 89.6% and 74.3% respectively. However, when ASA bound human sperm were use the results were 62.5% and 55.6% respectively. These shows the fertilization rate was significantly decreased in both ASA bound and ASA free oocytes when using ASA bound sperm. No difference found when ASA are present on the oocyte surface. When the hamster oocytes was treated by ICSI with ASA free or ASA bound human spermatozoa, no significant difference was found. These results showed that ICSI is the most promising method for couples who fertilization was not possible by conventional IVF because of ASA.

• 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.

• Journal of Korean Society of Forest Science
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• v.108 no.1
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• pp.21-28
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• 2019

This study was conducted to determine the carbon (C) and nitrogen (N) response of litterfall components as affected by N addition in compound fertilizer in a Korean red pine (Pinus densiflora S. et Z.) stand in southern Korea. Litterfall in a mature red pine stand was collected for two years following compound fertilizer application ($N_3P_4K_1$; $P_4K_1$) and no fertilization (control). The C concentration of litterfall components was not significantly (P > 0.05) different between the $N_3P_4K_1$ and the control plots, whereas the N concentration of the litterfall components was significantly higher in the $N_3P_4K_1$ plot than in the control plot. The $N_3P_4K_1$ and $P_4K_1$ additions induced a lower C/N ratio of litterfall components compared with the control plot. Annual C and N fluxes via litterfall components were not affected by the $N_3P_4K_1$ addition over the study period, except for reproduction litter. Annual N fluxes via reproduction litter were significantly higher in the $N_3P_4K_1$ plot than in the control plot. Thus, the $N_3P_4K_1$ and $P_4K_1$ additions could modify differently nutrient distribution of the forest floor and mineral soils in a red pine stand. These results indicate that N concentration and C/N ratio in litterfall components are more susceptible to fertilizer application than the C response in litterfall components.

• Journal of Aquaculture
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• v.7 no.1
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• pp.55-61
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• 1994

Triploid fish were induced successfully in olive flounder (Paralichthys olivaceus) by cold shocking fertilized eggs 3 minutes post fertilization at $2^{\circ}C$ for 45 minutes. Percent incidence of triploid was $92.6\%$ in this treatment. Floating rate and fertilization rate of eggs were not significanlty different from that of diploid controls (P> 0.05). However, hatchability and abnormal larvae of triploids were significantly different from that of diploid controls (P< 0.05). Incidence of triploidy was confirmed by erythrocyte measurements and chromosome counts. The surface area of triploid erythrocytes and nucleus was 1.6 times larger than that of diploids. Diploids had 48 acrocentric chromosomes, while triploids had 72.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.12
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• pp.1675-1677
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• 2001

The effect of the presence or absence of corpus luteum in the ovaries of slaughtered buffaloes was studied for the oocytes recovery and their subsequent maturation and fertilization in vitro. On an average, 0.41 and 0.67 oocytes per ovary were recovered from ovaries with and without corpus luteum, respectively. Immature oocytes were matured in TCM-199 medium. Significant difference was observed in maturation rate between good (74%) and fair (37%) oocytes. However, there was no significant difference in cleavage rate between the two types. The results of this study show that although the presence of corpus luteum in the ovary at the time of recovery significantly affected availability of total oocytes and in-vitro maturation, but fertilization and cleavage remained unaffected under in vitro conditions.

• Korean Journal of Soil Science and Fertilizer
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• v.31 no.4
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• pp.330-333
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• 1998

This experiment was conducted to investigate the effect of long-term fertilization on the yield and quality of soybean. The yield of soybean was highest in the NPK+lime $1.5Mg\;ha^{-1}$ treatment. Carbohydrate content of soybean was high in the non-fertilizer treatment and crude protein content was high in the NPK+lime $1.5Mg\;ha^{-1}$ and NPK+silicate $1.5Mg\;ha^{-1}$ treatment, and crude lipid and ash content was high in the NPK+silicate $1.5Mg\;ha^{-1}$ treatment. The content of inorganic compounds were not different among treatments. Correlation between $P_2O_5$/T-N and carbohydrate, crude lipid was $-0.99^{**}$, $-0.94^{**}$ respectively. Correlation between Mn and carbohydrate was $0.95^*$, but others were not significantly different. Yield of soybean increased with lime application and protein content increased with silicate application.

• Asian Journal of Turfgrass Science
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• v.4 no.1
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• pp.5-11
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• 1990

Korean Lawn. Zoysia japonica Steud. is the type of a perennial warm-season turfgrass keeping green leaf color for 5 months from May to September in natural condition in Korea. This Studies were carried out to obtain the effects of N-P-K fertilization, regrowth capacity after mowing and chlorophyll contents on prolongation of the green leaf color period in Zoysia japonica Steud turfgrass areas. The results obtained are summarized as follows ; 1.The effects of nitrogen fertilizer application in Zoysia japonica Steud, turfgrass areas were affected on regrowth ability after mowing green color of leaf canopies and chlorophyll contents, respectively(P<.05) 2.A experimental plots which applied 51 g/m$^2$/10 weeks of N$_3$P$_1$K$_1$fertilizer, there were high significant effects on prolongation of the green leaf color period and the plots of phosphorus and potassium had additive effects. 3.Prolongation of the green leaf color period to the last of October was obviously possible by slow degrees in proportation to increment of the fertilizer. However, it is interpreted that more prolongation of the green period is impossible with N-P-K fertilization and irrigation treatments. unless the intermediate turfgrasses between $C_3$ and $C_4$plants must be found and bred.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.10
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• pp.1360-1366
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• 2001

The aims of this study were first to evaluate the effects of different levels (20, 40 and 100%) and sources (follicular size: large, >7 mm; medium, >5-7 mm; small, 3-5 mm) of porcine follicular fluid (pFF) on the in vitro maturation (IVM) of porcine oocytes, and the effects of fertilization treatments and different culture conditions on development of fertilized oocytes were also investigated. No differences in the maturation (63.6-76.6%) and cleavage (24.8-34.3%) rates were observed among the 20,40 and 100% pFF groups (p>0.05). The cleavage rates of oocytes cultured and fertilized in 40% and 100% pFF maturation media were significantly higher than those fertilized in m199-NBCS (51.0-61.2% vs. 12.8-31.8%. p<0.05), regardless of sources of the pFF. When oocytes were fertilized in m199-NBCS followed by culture in rabbit oviducts for 4 days, the cleavage rate in 40% pFF group was better than that in 100% pFF group (46.9% vs. 32.5%, p<0.05). Two oocytes recovered from the oviducts in the 40% pFF group developed to blastocysts after IVC. However, none developed to blastocysts when fertilized in the IVM medium after being transferred to rabbit oviducts. In conclusion, addition of pFF accompanied with gonadotropins (FSH, LH) in IVM medium enhanced maturation and cleavage rates of porcine oocytes. Direct addition of sperm suspension to IVM medium may be an alternative to simplify the fertilization procedures and to reduce the mechanical lesion during manipulation. Furthermore, rabbit oviducts provide a better environment for the in vitro fertilized oocyte developing to the morula and blastocyst stages.

• Korean Journal of Medicinal Crop Science
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• v.15 no.3
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• pp.148-151
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• 2007

This experiment was carried out to investigate the effect of several organic fertilizers on the growth and root yield of Liriope platyphylla was planted 30 cm ${\time}$ 10 cm in field from 1998 to 1999. Six treatments of organic fertilizers applied : N.P.K, NPKC (N,P,K+Compost), NPKCO (N.P.K+Compost+Oilcake),NPKCOF (N.P.K+Compost+Oilcake+Fowldropping), NPKCOFP (N.P.K+Compost+ Oilcake+Fowldropping+plant ash), CO (Compost+Oilcake), COF (Compost+Oilcake+fowldropping). Organic matter, available phosphate, and MgO are NaO of soil increased in all of plots with the addition of organic fertilizers. The leaf length, number of leaves and of tillers were increased by the use of organic fertilizers compared to N.P.K fertilizers compared with N.P.K fertilization. The number of tubers were increased by using the organic fertilization N.P.K fertilization and showed higher in the soil depth of 0${\sim}$20 cm. The dry root yield was showed higher in organic fertilizers than in N.P.K fertilizer and highest dry root yield resulted from NPKOF 39% and NPKCOFP 49%.