• 한국작물학회지
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• 제31권4호
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• pp.493-498
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• 1986

품종, 시비량 및 재배관리를 동일하게 처리하였을 경우 보급년대가 상이한 각종 재배교식의 차이에 따른 종실의 생산능력을 검정하기 위하여 관행조파(60$\times$18cm), 광파(60$\times$30cm), 협폭파(40$\times$18cm), 세조파(30$\times$5, 20$\times$5cm) 및 전면전층파 등 6종류의 재배양식과 휴립재배법인 휴립이조법, 휴립광산파( 120$\times$90cm 및 90$\times$70cm) 및 휴립계조파(120$\times$90cm, 6예) 등 4종 총 10개 재배양식을 사용하여 조강보리와 그루밀를 공시품종으로 하며 시험한 결과는 다음과 같다. 1. 잡초발생량은 비파종부분의 폭이 클수록 많았고 수량과 부의 상관관계를 보였다. 2. 근분포정도는 수분소모정도와 정의 상관을 보였으며 휴간거리가 클수록 비파종부분의 근분포가 빈약하였다. 3. 양분흡수정도는 전면전층로타리시비법을 사용하는 세조파, 전면전층파, 휴립구산파 등이 종자하부 일정위치에 시비하는 작조시비법보다 낮은 경향을 보였다. 4. 종자밀도가 높은 관행조반, 광파, 협폭파 및 휴립이조파가 군집상태의 군낙을 이루어 월동률이 높은데 반하여 종자밀도가 낮아 독립수로 월동하게되는 세조파, 광조파, 전면전층파에서는 미기상의 영향으로 월동상태가 불량하였다. 5. 수량에 가장 공헌도가 큰 것은 수수였으며 균등배치가 이루워진 세조파가 광이용효율이 높고 수량도 가장 높았다.

• 한국가축번식학회지
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• 제17권1호
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• pp.57-68
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• 1993

This stduy was carried out to find a reliable method for the production of in vitro fertilized embryos having more excellent development capacity and freezability in the rabbit and cattle. The greatest number of rabbit oocytes was recovered 6hrs after HCG injection(P<0.05). The maturation rate in vitro was slightly higher in the oocytes(6-h-oocytes) from 6h than those (8-h-oocytes)from 8 hrs after HCG injection and the beneficial effect of FSH during oocyte maturation was significantly great in the oocytes from large follicles. The cleavage rate into 2-to-6-cell stage was not differ between the 6-h-oocytes and 8h-oocytes, but the cleavage of these oocytes was greatly promoted by FSH addition to maturation medium and the cleavge of 8-h-oocytes matured without FSH was significantly low. The embryo development into 16-cell to morula was not promoted by the co-culture with rabbit oviduct epithelial cells. The freezability by embryo stages was ovidusly high at 4-cell and morula stage in 6-h-oocytes and the viability of 16-cell embryos from 8h-oocytes was similar to that of morula stage. The implantation sites after surgical tranfer of fresh rabbit embryos were not implanted. In bovine experiment, the in vitro development into 16-cell and morula after in vitro maturation and fertilization in the follicular oocytes was slightly improved by the co-culture with granulosa cells compared to that with oviduct epithelial cells and the frozen-thawed viability rate of these embryos ranged from 14 to 40%. The excellent fresh embryos were transferred nonsurgically to 6 recipients, but were not pregnant.

• Clinical and Experimental Reproductive Medicine
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• 제26권3호
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• pp.399-405
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• 1999

Objective: In the preparation of ICSI, cumulus and corona cells should be removed from the oocytes by using a combination of enzymatic (hyaluronidase) and mechanical (pipetting) methods. But little is known about the effects of different degrees of oocyte denudation and incubation time between denudation and sperm injection on the outcomes of ICSI. The aim of this study was to evaluate the effects of varying the degrees of oocyte denudation and the lengths of incubation time from denudation to sperm injection on the outcomes of ICSI. Methods: In experiment 1, patients (oocytes) were grouped into group A and B according to the degree of denudation, complete and partial, respectively. In experiment 2, patients (oocytes) were grouped into group I, II and III according to the length of incubation time of denuded oocytes until sperm injection as < 1, $1{\sim}2$ and >2 hours, respectively. Results: There was no significant difference between the degree of oocyte denudation on the survival, fertilization and development rates after ICSI procedure. In case of the incubation time of denuded oocytes until ICSI, survival rates was higher in group III (83.1 %) than in group I (61.5%, p<0.05) or group II (64.3%). However no statistically significant differences were found between incubation time and fertilization or development rates. Conclusions: This study reveals that the outcomes of ICSI are not affected by the degree (complete or partial) of oocyte denudation. However, the denuded oocytes with incubation period of more than 2 hours show better outcomes of ICSI than those with the incubation period of less than 2 hours.

• Clinical and Experimental Reproductive Medicine
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• 제15권2호
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• pp.149-155
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• 1988

To increase fertilization rate in vitro, separation of viable spermatozoa from the seminal plasma and its other components may be a useful procedure. Ejaculates from healthy men, whose semen analysis findings were normal in 19, and abnormal in 10, were filtered using the glass-wool filtration technique to yield a concentrated, viable sperm samples for IVF, and the usefulness and safety of this method were evaluated. The recovery rate of motile sperm in abnormal groups was 46.2% and 54.5% in normal group. The % motility was increased significantly compared with original sample after filtration, and the grade motility was improved, too. The sperm population with normal morphology was also increased significantly in both group. Using transmission electron microscopy, the ultrastructural integrity of acrosomal segment was examined in order to evaluate the potentially hazardous effect of glass-wool filtration to sperm head, however, sperm population with normal ultrastructure was increased compared with that of original ejaculate after separation. The filtered sperm was then processed for IVF, as the fertilizing capacity is the ultimate parameter of the sperm function. In abnormal group, the fertilization rate(41.5 %) and the ET rate per stimulated cycle were much lower than that of mormal group(69.6%). However, the cleavage rate and the number of embryos transfered per ET cycle were comparable with those of nomal group. The results suggest that the glass-wool filtration of sperm, particularly in oligo-asthenozoospsrmia, may be useful and safe method in the preparation of sperm for IVF.

• 한국수정란이식학회지
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• 제31권1호
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• pp.27-32
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• 2016

This study was conducted to evaluate the effects of different volume ($100{\mu}l$ vs. 2 ml) of microdrop culture on B6D2F1 mice oogenesis. In the present study, B6D2F1/CrljOri $F_1$ mice were utilized in order to maximize oogenesis. Also we used TCM-199, Dulbecco's medified Eagle's medium (DMEM), embryo culture medium (Fertilization medium, Cleavage medium, Blastocyst medium), G series medium and One step medium. Blastulation rate was not different between groups ($58.4{\pm}2.9%$ vs. $61.2{\pm}4.8%$). Zona hatched rate ($38{\pm}15.4%$ vs. $27{\pm}3.4%$) and attached rate ($55{\pm}13.9%$ vs. $46{\pm}3.9%$) did not differ by the volume of culture media. Total cell numbers ($59.8{\pm}9.7$ vs. $70.3{\pm}8.7$), ICM cell numbers ($15.8{\pm}0.6$ vs. $16.8{\pm}1.5$), TE cell numbers ($44.0{\pm}9.7$ vs. $53.6{\pm}7.3$), % ICM ($26.4{\pm}2.9%$ vs. $23.8{\pm}3.3%$) and ICM:TE ratio ($1:2.8{\pm}0.4$ vs. $1:3.2{\pm}0.6$) were not different between groups (i.e., $100{\mu}l$ vs. 2 ml). These results show that the capacity of the culture medium did not effect the cell numbers of B6D2F1 mice blastocysts. In summary, these results can provide fundamental data to maximize culture condition for in vitro fertilization on B6D2F1 mice.

• 농업과학연구
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• 제50권4호
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• pp.941-952
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• 2023

The metabolites of agrichemicals, such as organophosphorus pesticides, are known to be more hazardous than their parent pesticides. 3,5,6-trichloro-2-pyridinol (TCP) is a major degradation product of chlorpyrifos, one of the organophosphate insecticides widely used in agriculture. In vivo or in vitro exposure to chlorpyrifos has been known to interfere with male reproductive functions, leading to reduced fertility in mammals. Therefore, this study was performed to examine the changes in the fertilization competence of boar spermatozoa exposed to TCP. Sperm samples were subjected to varying concentrations of TCP (10, 50, 100, 200 µM) and different periods of incubation. Sperm motility, motion kinematics, viability, acrosome integrity, intracellular reactive oxygen species (ROS) production, and gene expression levels (ODf2, ZPBP2, AKAP3 and AKAP4) were evaluated after exposure of the sperm to TCP. A significant dose-dependent reduction in motility was observed in sperm samples incubated with TCP compared to the controls after both incubation periods. Sperm viability was significantly decreased in samples incubated with 50, 100, and 200 µM TCP in both incubation periods. A significantly lower percentage of normal acrosomes and gene expression levels were observed in sperm samples exposed to 50, 100, and 200 µM TCP after both incubation periods, compared to the controls. There was a significant increase in the ROS production in spermatozoa incubated with 100 - 200 µM TCP after both incubation periods. Consequently, the direct exposure of boar spermatozoa to TCP interferes with sperm functions and leads to decreased fertilization. In order to identify and address the various causes of reproductive decline, the impact of chemical metabolites needs to be discussed in depth.

• 한국환경과학회지
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• 제32권4호
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• pp.233-242
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• 2023

The objective of this study was to develop a management strategy for the recovery of carbon storage capacity of abandoned coal mine forest rehabilitation area. For the purpose, the biomass and stand carbon storage over time after the forest rehabilitation by tree type for Betula platyphylla, Pinus densiflora, and Alnus hirsuta trees which are major tree species widely planted for the forest rehabilitation in the abandoned coal mine were calculated, and compared them with general forest. The carbon storage in abandoned coal mine forest rehabilitation areas was lower than that in general forests, and based on tree species, Pinus densiflora stored 48.9%, Alnus hirsuta 41.1%, and Betula platyphylla 27.0%. This low carbon storage is thought to be caused by poor growth because soil chemical properties, such as low TOC and total nitrogen content, in the soil of abandoned coal mine forest rehabilitation areas, were adverse to vegetation growth compared to those in general forests. DBH, stand biomass, and stand carbon storage tended to increase after forest rehabilitation over time, whereas stand density decreased. Stand' biomass and carbon storage increased as DBH and stand density increased, but there was a negative correlation between stand density and DBH. Therefore, after forest rehabilitation, growth status should be monitored, an appropriate growth space for trees should be maintained by thinning and pruning, and the soil chemical properties such as fertilization must be managed. It is expected that the carbon storage capacity the forest rehabilitation area could be restored to a level similar to that of general forests.

• 한국수정란이식학회지
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• 제31권4호
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• pp.367-374
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• 2016

This study was conducted to evaluate the effect of transfer temperature of epididymis on survival rate of semen and development ability of B6D2F1 mice embryos. No significant differences were noted in the survival rate of semen ($59.0%{\pm}0.1$ vs. $47.6%{\pm}0.1$), in vitro fertilization rate ($90.7%{\pm}0.1$ vs. $90.7%{\pm}0.1$), developmental rate ($90.0%{\pm}0.1$ vs. $90.0%{\pm}0.1$), and blastocysts formation rate ($53.1%{\pm}0.2$ vs. $52.3%{\pm}0.2$) between groups. (NS; P>0.05). However, the zona hatched rate was significantly higher in the $4^{\circ}C$ group compared to those of the $37^{\circ}C$ group ($47.8%{\pm}0.1$ vs. $25.6%{\pm}0.2$; p<0.05). When it comes to cell numbers of blastocysts, the % ICM (/total cells) was significantly higher in the group of $4^{\circ}C$ compared to the $37^{\circ}C$ ($27.0%{\pm}0.1$ vs. $18.3%{\pm}0.1$; p<0.05). However there were no differences in total cell numbers ($72.7{\pm}31.6$ vs. $62.0{\pm}36.6$), ICM cell numbers ($17.0{\pm}7.8$ vs. $14.6{\pm}8.6$), TE cell numbers ($55.8{\pm}29.8$ vs. $64.0{\pm}24.4$), the ratio of ICM:TE ($1:4.2{\pm}4.1$ vs. $1:6.4{\pm}7.2$) between two groups (NS; P>0.05). Taken altogether, it is expected to achieve the best developmental ability of B6D2F1 mice embryos in the transfer temperature of epididymis. Also these results can provide fundamental data to maximize culture condition for in vitro fertilization on B6D2F1 mice. In future, therefore, it is expected that results herein might be applied for in vitro culture of human embryos.

• Reproductive and Developmental Biology
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• 제35권2호
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• pp.137-141
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• 2011

These study was to investigate the in vitro fertilization and viability of fresh and vitrified oocytes. Also, the developmental capacity of IVF and intracytoplasmic sperm injection (ICSI) oocytes were investigated. Then vitrification was performed with the use of 20% ethylene glycol + 20% DMSO + 0.5 M sucrose + 10% FCS + TCM-199 medium. Vitrification immature oocytes are cultured in vitrification solution for 10 min afterwards transferred to expose at room temperature for 5 min. and transferred to the ice water for 5 min. The oocytes were sealed in a 1.0 mm straw and placed in a $LN_2$ container. Frozen oocytes were rapidly thawed in a water bath at $30{\sim}35^{\circ}C$, and then placed in TCM-199 medium containing 0.5 M sucrose for 5 min each, respectively, at $38^{\circ}C$. After being washed for 2~3 times, using fresh medium the oocytes were cultured in TCM-l99 medium supplemented with 5% FCS at $38^{\circ}C$ in 5% $CO_2$ and air. The normal morphology of fresh and vitrified-thawed oocytes were $87.1{\pm}2.1%$ and $54.8{\pm}2.5%$, respectively. The viability rates of fresh and vitrified-thawed oocytes were $70.0{\pm}2.2%$ and $41.9{\pm}2.6%$, respectively. Viability rates of vitrified-thawed oocytes were lower than that of fresh follicular oocytes (p<0.05). The in vitro maturation rates of fresh and vitrified oocytes were $45.1{\pm}3.6%$ and $28.9{\pm}4.4%$, respectively. The IVF rates of fresh follicular and vitrified-thawed oocytes were 34.00.2% and $20.2{\pm}2.6%$, respectively. The in vitro maturation and fertilization rates of vitrified-thawed oocytes were lower than those of the fresh follicular oocytes (p<0.05). A total of 350 oocytes were fixed and stained after co-incubation with spermatozoa, of which 88 had identifiable nuclear material. After IVF for 20 hrs, $25.1{\pm}3.4%$ of the oocytes found to have been penetrated by spermatozoas. Oocytes were fixed and stained after ICSI, and 105 oocytes contained identifiable nuclear material. After IVF and ICSI for 20 hrs, $34.3{\pm}3.4%$ and $59.0{\pm}2.0%$ of the oocytes were found to have been penetrated by spermatozoas. The developmental rates upon ICSI were significantly higher than those of the IVF method (p<0.05).

• 대한수의학회지
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• 제35권1호
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• pp.187-195
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• 1995

The present study carried out to determine the developmental capacity of bovine oocytes matured in epidermal growth factor(EGF)-containing medium, the developmental competence of bovine embryos using synthetic oviduct fluid(SOF) and the effect of glucose on the development of bovine embryos. In experiment 1, oocytes, obtained from abattoir ovaries, were matured in EGF-containing medium for 24 hours, followed by exposure to Korean native cattle spermatozoa for 18 hours and cultured by utilizing co-culture system with bovine oviduct epithelial cells(BOEC) in TCM199. In experiment 2, early bovine embryos were cultured in SOF with or without BOEC and compared with those in TCM199 with BOEC. In experiment 3, bovine embryos were cultured in the presence or absence of glucose. Seven and ten days after in vitro fertilization, developmental competence of embryos were evaluated. The rate of cleavage was significantly(P<0.05) higher in EGF-containing maturation medium(70.0%) than in control(57.7%). The rates of development to morulae and blastocysts were 30.6% and 23.3% there was no significant difference between them. The rates of in vitro fertilized embryos to morulae and blastocysts cultured in SOF with BOEC(30.4%) and in TCM199 with BOEC(38.0%) were significantly(P<0.01) higher than cultured in SOF without BOEC(13.4%) at seven days after in vitro fertilization. The rates of embryos to blastocysts cultured in SOF with BOEC(29.4%) and in TCM199 with BOEC(35.9%) were significantly(P<0.05) higher than cultured in SOF without BOEC(13.4%) at ten days after in vitro fertilization. The rates of early embryos to morulae and blastocysts cultured in the presence or absence of glucose were 12.2% and 17.5% each other, there was no significant difference between them. The results show that bovine oocytes matured in the presence of EGF can cleave better, SOF with BOEC can replace serum containing complex media, TCM199 with BOEC in bovine embryo culture and glucose have little effect on the culture of early bovine embryos.