• Title/Summary/Keyword: fertilization times

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Studies on the Improvement of Fertilization Rates Using Intracytoplasmic Sperm Injection with In Vitro Matured Oocytes (소 체외성숙 난자의 세포질내 정자주입에 의한 수정율 향상에 관한 연구)

  • 유상식;김용섭;이봉구;김상근
    • Korean Journal of Animal Reproduction
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    • v.22 no.3
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    • pp.213-219
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    • 1998
  • This study was carried out to investigate on the improvement of fertilizing ability of in vitro matured oocytes from sperm density, motility and polyvinylpyrrolidine (PVP) concentration, by intracytoplasmic sperm injection(ICSI) into the bovine oocytes. 1. The in vitro fertilization and cleavage rates of oocytes from 1.0, 2.0, 3.0, 5.0 ($\times$106/ml) sperm concentration by IVF and ICSI of bovine oocytes were 45.0%~65.0%, 65.0%~90.0% and 10.0%~30.0%, 35.0%~70.0%, respectively. 2. The in vitro fertilization and cleavage rates of oocytes from 20, 40, 60, 80% of sperm motility by IVF and ICSI of bovine oocytes were 47.8%~75.0%, 78.3%~90.0% and 8.7%~25.0%, 34.8%~70.0%, respectively. 3. The in vitro fertilization and cleavage rates of oocytes from 0.01, 0.02, 0.03, 0.05% of PVP concentration by microinjection of single into the bovine oocytes were 72.7%, 90.9%, 83.3%, 76.9% and 45.5%, 72.7%, 58.3%, 61.5%, respectively and these values of 0.02% addition of PVP were higher than other concentrations of PVP. 4. The in vitro fertilization and developmental rates of oocytes by IVF and ICSI methods were 63.3%~64.6%, 26.7%~29.2% and 88.2%, 47.1%, respectively. This ICSI method was improved high fertilization rates of bovined oocytes.

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Nuclear Maturation and In Vitro Fertilization of Mouse Eggs Recovered at Various Times after Superovulation (과배란처리후의 경과시간이 생쥐 난자의 핵성숙과 체외수정에 미치는 영향)

  • 이상진;정길생
    • Korean Journal of Animal Reproduction
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    • v.13 no.2
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    • pp.70-78
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    • 1989
  • Mouse eggs recovered from oviducts at one hourly intervals between 10 and 20 hours after administration of hCG were fixed, stained and then investigated the rate of in vitro fertilization and nuclear maturation. In case of out- bred ICR mice, ovulations were occured between 11 and 13 hours after hCG injection. The stages of in vitro maturation of eggs recovered from female mice at various times after hCG injection were metaphase I, anaphase I, telophase I and metaphase II. However the majority was metaphase I(17.6 to 44.4%) and metaphase II(42.9 to 80.0%) stage. When the eggs were inseminated with epididymal spermatozoa, the fertilization rate was declined as the egg recovery time after hCG administration was delayed. That is, the proportion of eggs undergoing fertilization became higher(68.1 to 77.4%) in the eggs at 12 to 15hr after injection of hCG than those(17.5 to 56.4) at 16 to 20 hr after injection of hCG. Also, when nuclear maturation of the unfertilized eggs were observed at 8 hours after insemination, the majority was in metaphase I and metaphase II and no anaphase I and telophase I were observed.

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Ecological and Biogeochemical Response of Antarctic Ecosystems to Iron Fertilization and Implications on Global Carbon Cycle

  • Bathmann, Ulrich
    • Ocean and Polar Research
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    • v.27 no.2
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    • pp.231-235
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    • 2005
  • The European Iron Fertilization Experiment EIFEX studied the growth and decline of a phytoplankton bloom stimulated by fertilising $10km^2$ in the core of a mesoscale $(80{\times}120km)$ cyclonic eddy south of the Antarctic Polar Front with about 2 times 7 tonnes of iron sulphate. The phytoplankton accumulation induced by iron fertilization did not exceed $3{\mu}g\;chl\;a\;l^{-1}$ despite a draw down of $5{\mu}M$ of nitrate that should have resulted in at least double to triple the amount of phytoplankton biomass assuming regular Redfield-ratios for draw down after phytoplankton growth in the Southern Ocean. During EIFEX the fertilized core of the mesoscale eddy evolved to a hotspot for a variety of small and medium sized mesozooplankton copepods. In contrast to copepods, the biomass of salps (Salpa thompson)) that dominated zooplankton biomass before the onset of our experiment decreased to nearly extinction. Most of the species of the rnosozooplankton community showed extremely hiか feeding rates compared to literature values from Southern Ocean summer communities. At the end of the experiment, massive phytoplankton sedimentation reached the sea floor at about 3800m water depth.

Effects of pH on Fertilization and the Hatching Rates of Far Eastern Catfish Silurus asotus

  • Gao, Yang;Kim, Sun-Gyu;Lee, Jeong-Yeol
    • 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.

Studies on in vitro Maturation and Fertilization of Porcine Follicular Oocytes (돼지 난포란의 체외성숙 및 수정에 관한 연구)

  • 김상근;이만휘;이명헌;신용호
    • Korean Journal of Animal Reproduction
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    • v.14 no.1
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    • pp.23-30
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    • 1990
  • These studies were carried out to investigate the effects of fetal calf serum(FCS), estrous porcine serum(EPS), porcine follicular fluid(PFF), hormone and matured cumulus cell(MCC) on in vitro maturation and fertilization of porcine follicular oocytes. The ovaries and testes were obtianed from slaughtered Landrace sow and boars, respectively. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluid from the visible follicles of diameter 3~5 mm and the semen were prepared from boar's epididymal cauda. The follicular oocytes were cultured in TCM-199 medium containing hormones, FCS, EPS, PFF and MCC for 48hrs. in a incubator with 5% CO2 in air at 36$^{\circ}C$ and then matured oocytes were again cultured for 18~20 hrs. with $1.5\times$106/ml motile capacitated sperm in the modified Tyroide solution containing 100$\mu\textrm{g}$/ml of heparin. The results obtained in these experiments were summarized as follows : 1. The maturation and fertilization rate of the follicular oocytes, cultured in the TCM-199 medium supplemented with 10% FCS and PMSG+HCG were 55.6~64.5% and 33.3~37.1%, respectively. 2. The maturation and fertilization rate of the follicular oocytes cultured in the TCM-199 medium supplemented with 20% EPS and PMSG+HCG were 50.0~55.0% and 30.3~33.3%, respectively. 3. The maturation rate(59.0~64.2%) and fertilization rate(34.8~39.3%) of follicular oocytes cultured in TCM-199 medium supplemented 20% FCS and 50% PFF were higher than those of follicular oocytes cultured in TCM-199 medium supplemented with 5%, 10% and 15% FCS and 10% and 50% PFF. 4. The maturation rate(60.0%) and fertilization rate(40.0%) of follicular oocytes cultured in TCM-199 medium supplemented with 20% FCS and granulosa cell (1$\times$106/ml) were significantly higher than those of fiollicular oocytes cultured in TCM-199 medium supplemented with 5%, 10% and 15% FCS and granulosa cell.

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Studies on the Fertilization Rates using Intracytoplasmic Sperm Injection with In Vitro Matured Porcine Oocytes (돼지 체외성숙 난자의 세포질내 정자주입에 의한 수정에 관한 연구)

  • 김상근;김민수;남윤이
    • Korean Journal of Animal Reproduction
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    • v.23 no.2
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    • pp.113-118
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    • 1999
  • This study was carried out to investigate on the improvement of fertilizing ability of in vitro matured oocytes from sperm density and motility by intracytoplasmic sperm injection(ICSI) into the porcine oocytes. 1. The in vitro fertilization and cleavage rates of oocytes from 1.0, 2.0, 3.0, 5.0 ($\times$10$^{6}$ $m\ell$) sperm concentration by IVF and ICSI of porcine oocytes were 46.7%~75.0%, 60.0%~85.7% and 10.6%~25.0%, 20.0%~64.3%, respectively. 2. The in vitro fertilization and cleavage rates of oocytes from 20, 40, 60, 80% of sperm mortilty by IVF and ICSI of porcine oocytes were 46.4%~71.4%, 67.9%~85.7% and 7.1%~21.4%, 28.6%~60.7%, respectively. 3. The in vitro fertilization and developmental rates of oocytes by IVF and ICSI methods were 55.6%~60.0%, 77.8%~80.0% and 17.8%~24.0%, 42.2%~56.0%, respectively. This ICSI method was improved high fertilization rates of porcine oocytes.

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Two Case Report of Spontaneous Pregnancy Treated with Gamiseogagjihwang-tang after Repeated Failure in In Vitro Fertilization or Intrauterine Insemination Treatment (보조생식술 실패 후 서각지황탕가미(犀角地黃湯加味) 복용치료를 통해 자연임신에 성공한 난임 환자 치험 2례)

  • Koh, Ji-Eun;Lyou, Myung-Sook
    • The Journal of Korean Obstetrics and Gynecology
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    • v.28 no.1
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    • pp.128-137
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    • 2015
  • Objectives: Spontaneous pregnancy is a rare event after repeated failure of in vitro fertilization (IVF) or Intrauterine insemination (IUI) treatment. So this paper is to report the natural pregnancy of Gamiseogagjihwang-tang treatments on two infertile patients who were repeatedly failed in IVF or IUI. Methods: One patient was a 35-year-old patient diagnosed with the adenomyosis and contralateral tubal obstruction, treated with 90 cc Gamiseogagjihwang-tang two times per day for 6 weeks after second failure of IVF. The other patient was a 34-year-old patient undergone induced abortion by three times, treated with 90 cc Gamiseogagjihwang-tang two times per day for 4 weeks after second failure of IUI. Results: Through taking Gamiseogagjihwang-tang without assisted reproduction techniques, two cases got pregnant naturally. Conclusions: These cases suggest that Gamiseogagjihwang-tang is effective in treating infertile female after failure in IVF or IUI and inducing spontaneous pregnancy. Therefore, there needs to be more trial on infertile patients treated with Seogagjihwang-tang.

Development of Techniques for the Production of Selenium and Germanium-enriched Chinese Cabbage and Pepper (셀레늄과 게르마늄 강화 배추와 고추 생산기술)

  • Yun, Hyung-Kwon;Zhang, Cheng-Hao;Seo, Tae-Cheol;Huang, Hua-Zi
    • Journal of Bio-Environment Control
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    • v.16 no.3
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    • pp.180-185
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    • 2007
  • The effects of selenium (Se) and germanium (Ge) fertilization on the growth and quality of Chinese cabbages cultivated in spring and autumn and peppers cultivated in spring were investigated. $Se\;(Na_2SeO_4)\;and\;Ge\;(GeO_2)$ were supplied 5, 10, or 20 times in an aqueous solution of 0, 2, 4, or $8mg{\cdot}L^{-1}$ during the cultivation of Chinese cabbages and peppers. The fresh weight of Chinese cabbages increased by Ge fertilization with high concentration. But it was not affected by Se fertilization. The content of vitamin C increased by 10 times application with $4mg{\cdot}L^{-1}$ of Se or Ge. The concentration of Se in Chinese cabbage increased according to increasing concentration of Se fertilization. Se concentration was higher in the outer leaves than in the inner leaves. Se concentration in the mesophyll was higher than that in the midrib. Ge fertilization increased the uptake and concentration of Ge in autumn-cultivated Chinese cabbages. Se and Ge fertilization did not affect the fresh weight of peppers. The content of vitamin C in pepper increased by 20 times application of $2mg{\cdot}L^{-1}$ of Se. Vitamin C content in red peppers was twice as much as in green peppers.

Effects of Harvesting Frequency and Fertilization Levels on Botanical Composition and Forage Productivity of Alpine Grassland at 800m Altitude (산지초지를 위한 대관령 표고 800m에서 예취횟수와 시비수준이 목초의 식생구성비율, 건물수량 및 사료성분에 미치는 영향)

  • Oh, Seung Min;Kim, Ji Yun;Lee, Bae Hun;Peng, Jinglun;Chemere, Befekadu;Nejad, Jalil Ghassemi;Sung, Kyung Il;Kim, Byong Wan
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.36 no.2
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    • pp.124-128
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    • 2016
  • The objective of this research was to determine the effects of harvesting frequency and fertilization levels on botanical composition, dry matter yield, and forage feed compositions of Alpine grassland at 800 m altitude. This research lasted for three years at National Alpine Agricultural Research Institute in Pyeongchang with two harvesting frequency schedules (two and three times annually) and two levels of fertilizer application (conventional level of fertilizer at 280-200-240 kg/ha and a lower level of fertilizer at 200-200-200 kg/ha for N, $P_2O_5$, and $K_2O$). Mixture combinations with seeding rate (kg/ha) were as follows: Orchardgrass 18, Tall fescue 9, Timothy 8, Kentucky bluegrass 3, and Ladino Clover 2. The gramineae ratio ranged from 93.2 to 95.3%. Therefore, gramineae forage was considered as the dominant plant in this experiment. No significant (p>0.05) difference was observed in forage dry matter yield between the two harvesting frequency treatments (two times at 9.8 ton/ha and three times at 8.6 ton/ha). However, forage dry matter yield in the two times of harvesting frequency tended to be greater than that in the three times of harvesting frequency. Significantly (p<0.05) higher forage dry matter yield in the standard fertilization level group than the lower fertilization level group (9.8 ton/ha vs. 8.7 ton/ha) was observed. However, there was no significant (p>0.05) difference in forage crude protein concentration between the two harvesting frequency treatment groups, although the concentration in the group with three times of harvesting frequency tended to be higher. In contrast, crude fiber concentration in the group with two times of harvesting frequency tended to be higher, although the difference was not statistically significant (p>0.05). Crude protein, ether extract, crude fiber, and organic matter concentrations were not significantly (p>0.05) different between the two groups with different fertilization levels. Based on these results, it was concluded that the group with two times of harvesting frequency with conventional fertilization level might be proper for obtaining better forage productivity for Alpine grassland at 800 m altitude.

Bovine Oocytes Can Be Penetrated in Modified Tris-buffered Medium

  • Park, Kwang-Wook;Niwa, Koji
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.4
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    • pp.500-506
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    • 2009
  • A modified Tris-buffered medium (mTBM) has been widely used as an insemination medium for porcine in vitro fertilization (IVF). We examined whether mTBM could be used for bovine IVF. Bovine cumulus-oocyte complexes (COCs) were cultured in a serum-free medium containing 30 ng/ml EGF for 22 h. After culture, COCs were inseminated with spermatozoa for 12 h in mTBM containing 5 mM caffeine and 10 g/ml heparin. The penetration of oocytes increased significantly (p<0.05) as the sperm concentration increased from 0.1 (30%) to 1-10 $(87-100%){\times}10^6$ cells/ml. This was significantly different from values obtained at 1 (87%) and 10 $(100%){\times}10^6$ cells/ml. However, when COCs were inseminated with spermatozoa from different bulls, the proportions (62-100%) of oocytes penetrated varied according to the bull. The proportion (18%) of oocytes penetrated was significantly (p<0.05) lower in a fertilization medium without caffeine and heparin but increased with the addition of caffeine and/or heparin to the medium, and the proportion (93-96%) of oocytes penetrated increased significantly (p<0.05) when the medium was supplemented with heparin and caffeine. In this medium, sperm penetration was first observed at 3 h after insemination. Irrespective of the presence of glucose in the fertilization medium, the proportion (93-97%) of oocytes penetrated and the proportion (83-84%) of embryos at the ${\geq}2$-cell stage cultured in a chemically defined medium were not significantly different. However, the proportion of embryos developing to the blastocyst stage was significantly (p<0.05) higher in the presence (11%) of glucose in the fertilization medium than in its absence (2%). In conclusion, the present study demonstrated that bovine oocytes penetrated in vitro in mTBM can develop to the blastocyst stage and mTBM may be used for the in vitro production of bovine embryos.