• Microbiology and Biotechnology Letters
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• v.33 no.2
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• pp.117-122
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• 2005

Beijerinckia indica was cultured in mineral salts medium (MSM) medium with various carbon and nitrogen sources to improve the production yield of heteropolysaccharide-7 (PS-7). At high C/N ratio, the high concentration of PS-7 was produced until 40 h of the culture, whereas most of the glucose as a carbon source was used for the cell growth at low C/N ratio. However, at the high C/N ratio, PS-7 accumulation stopped at 48 h of the culture due to the increasing viscosity of the culture broth would inhibit the cell growth. Therefore, the optimized value of C/N ratio was 33.3 (20 g/L glucose, 7.5 mM $NH_{4}NO_3$) for the high production of PS-7. In the culture with various carbon sources, B. indica effectively used the hexoses or glucose-generating sugars for PS-7 formation. Especially, sucrose was the best carbon source for the high production of PS-7 (6.96 g/L) with a high viscosity (40772 cp). In the culture of B. indica with MSM medium containing 20 g/L glucose and 7.5 mM $NH_{4}NO_3$ in a 51 fermentor, the highest cell concentration was 2.5 g/L and the highest concentration of PS-7 was 7.5 g/L (35174 cp). The additional nitrogen sources of 7.5 mM $NH_{4}NO_3$, glutamine and glutamate at 12 h of the culture after exhaustion of a nitrogen source regulated the metabolism of carbon sources, therefore the nitrogen sources could control PS-7 synthesis.

• Journal of Embryo Transfer
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• v.21 no.2
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• pp.157-162
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• 2006

It is well known that unidentified factors in sera, hormones and growth factors promote the proliferation of granulosa cells and nuclear maturation of bovine COCs (cumulus oocytes complexes) in vitro. Attempts had been developed the simple composition of culture media and similar system to in vivo conditions has been applied. In the present study, we investigated the effect of FGF (fibroblast growth factor) on in vitro maturation and in vitro development of Hanwoo COCs. When the COCs were matured in HPM 199 (Inst. of Functional peptide, Japan) containing 0.1, 1 and 10 ng/ml FGF for 24 hr, maturation rates to metaphase II ($70.0{\sim}75.0%$) were significantly higher (p<0.05) than that of control group (0 ng/ml FGF, 37.5%). When matured COCs with FGF were cultured in maturation medium after in vitro fertilization, developmental rates to blastocysts were 9.5, 0 and 2.9%, respectively, compared to 25.0% of the control group (p<0.05). When the matured COCs with FGF were cultured in HPM 199 (IFP971, Inst. of Functional peptide, Japan) containing 10% FBS, 0.8% BSA or 0.1% PVA (polyvinyl alcohol), the blastocyst formation rates were 12.4, 12.8 and 8.5%, respectively, while the rates of matured COCs with FGF and cultured with IVMD and IVD (Inst. of Functional peptide, Japan) without serum were 38.4% and 34.8%, respectively (p<0.05). These results suggested that FGF is available for in vitro maturation of bovine COCs and is not suitable for in vitro development, but further investigation would be need for finding the synergistic autocrine/paracrine fashion of other growth factors in early bovine embryo development.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.169-175
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• 2006

The objective of this study was carried out to examine the polar body extrusion of in vitro matured porcine follicular oocytes as a non-invasive marker of oocyte quality to know the developmental competence in advance. The porcine oocytes matured for 48 hours were examined the polar body extrusion and some parts were stained. The examined oocytes were matured for additional $16{\sim}18$ hours and activated with 7% ethanol and cultured in $5{\mu}g/ml$ cytochalasin B for 5 hours for diploid formation. The treated oocytes were washed and cultured for 7 days. The polar body extrusion and degeneration rates were varied with $9.9{\sim}52.4%$ and $21.4{\sim}61.8%$ by repetition. The polar body extruded oocytes were shown the polar body chromosome and metaphase II plate by staining. However the non-extruded oocytes were shown expanded nucleus with 39.1%, premature chromosome condensation with 19.6%, metaphase I plate with 10.9 %, metaphase II with 13%, condensed chromatin with 6.5%, and absent nuclear material with 8.7%. The oocytes that were not examined for the polar body extrusion were cleaved 45.0%, and developed to blastocyst stage with 11.3%. In examined oocytes for polar body extrusion,. the polar body extruded oocytes were cleaved 94.2% and developed with 42.5%. This result suggests that discarding of the degenerating oocytes and oocytes that not extruded polar body will be effective for experiments of culturing effect in porcine embryos and embryo biotechnology.

• Journal of Embryo Transfer
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• v.12 no.2
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• pp.151-159
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• 1997

In vitro fertilization(IVF) derived morula and blastocyst embryos were bisected by a simple method and cultured in vitro without zona pellucida And also bisected embryos were frozen-thawed and cultured in vitro) to evaluate the survival rate. The results obtained were as follows : The average number of grade I or II immature follicular oocytes recovered by slicing method per ovary was 11.9 from 142 ovaries. Following in vitro fertilization, the rates of cleavage and in vitro development to morula and blatocyst were 61.7 and 32.2% respectively. The successful bisection rate of IVE embryos was 67.51%, and the embryos of blastocyst stage were bisected successfully at significantly(P

• Proceedings of the Korean Environmental Sciences Society Conference
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• 2007.05a
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• pp.453-456
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• 2007

가금류의 도축 부산물로 대량 배출되고 있는 우모는 주로 생물학적으로 난분해성 단백질인 케라틴으로 구성되어 있다. 따라서 물리화학적 처리에 의하여 우모를 처리하고 있으나 이 방법은 환경오염을 유발한다. 따라서 본 연구에서는 환경친화적 우모 분해 공정을 개발하고, 우모 분해산물의 식물 성장을 위한 비료로서의 가치를 평가하기 위하여 퇴비화 볏짚에서 케라틴 분해효소 생성능이 우수한 균주인 RS7을 분리하였다. RS7의 생화학적 특성과 16S rDNA의 염기 서열을 분석하여 동정한 결과, Bacillus pumilus RS7로 동정되었다. B. pumilus RS7은 $30^{\circ}C$에서 배양 5일 만에 우모를 완전히 분해할 수 있었다. 본 균주에 의한 우모 분해산물은 Helianthus sannuus L.의 생장율, 잎 수 증가량, 개화율, 건조 생체량에 있어 대조군과 화학적 우모 분해산물보다 우수하였다. Bacillus pumilus RS7에 의한 우모 분해산물은 식물 성장 촉진을 위한 미생물기원 비료로써의 역할을 수행할 수 있었으며, 이에 따라 양계산업에서 배출되는 우모 폐기물이 환경에 주는 악영향을 감소시킬 수 있을 것으로 예측된다.

• Korean Journal of Veterinary Research
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• v.35 no.2
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• pp.205-216
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• 1995

This study was examined chromosome structure of kidney, fin, blood cell, sexual organ that were differently examined chromosome of a good cell activity from organ, medium, staining methods, instruments with rainbow trout 100. And so we obtained following conclusion. 1. The difference from each organ and medium is that a good cell activity of fin, kidney, sexual organ were obtained in TC-199 medium and a good cell activity in TC-199 medium+PHA(5%). 2. In the difference by staining methods, G-banding and C-banding were analyzed by electron microscope or cytoscan. Among them, heterochromatin of centromere analyzed by C-banding. 3. The zygotene and the leptotene were analyzed in this study. 4. Karyotype, heterochromatin and each stage of cell division were clearly analyzed by cytoscan.

• Applied Biological Chemistry
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• v.29 no.3
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• pp.273-278
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• 1986

PCBs was degraded by bacterium, which was classified as a strain of Alcaligenes aquamarinus. Its PCB-42 degradation was maximized when grown on mineral salts medium containing 0.1% of PCB-42 as a sole carbon source at $25^{\circ}C$ and initial pH $7.0{\sim}8.0$, and also shaking culture was effective for it. The addition of glucose and peptone were effective for the degradation of PCB-42, but metal ions were not effective.

• Journal of Embryo Transfer
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• v.19 no.2
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• pp.165-172
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• 2004

This study was carried out to examine the effects of maturation media on in vitro maturation (IVM) of porcine immature oocytes, and on subsequent in vitro fertilization (IVF) and development (IVD). Cumulus-oocyte complexes (COCs) were collected from antral follicles of porcine ovaries collected from abattoir, and were matured in vitro in modified NCSU-37 (mNCSU-37), modified NCSU-23 (mNCSU-23), or TCM-199 supplemented with 10% porcine follicular fluid (pFF). Oocytes matured in vitro, were fertilized in vitro in modified Tris-buffered medium(mTBM) with the final motile sperm concentration of 1${\times}$105 sperm/mL, and subsequently putative embryos were developed in vitro in NCSU-23. The results are as follows. 1. In the result of IVM, the rate of germinal vesicle breakdown (GVBD) and of nuclear maturation were not significantly different among the media, though numeric value of them were slightly lower in TCM-199 than in mNCSU-37 or in mNCSU-23. 2. In the result of IVF, though the rate of sperm penetration was not significantly different among the maturation media, the percentage of oocytes with male pronucleus (MPN) of ones matured in mNCSU-37 (88.0%) was significantly higher than in TCM-199 (71.1%) (p<0.05). 3. In the result of IVD, the percentage of cleaved oocytes of ones matured in mNCSU-37 (52.3%) or in mNCSU-23 (53.7%) was significantly higher than in TCM-199 (43.1%) (p<0.05), but the rate of blastocysts at day 6 was not significantly different among the maturation media, though putative embryos from oocytes matured in mNCSU-37 or in mNCSU-23 were developed more than in TCM-199. These results suggested that mNCSU-37 or mNCSU-23 was more appropriate than TCM-199 as IVM medium for porcine immature oocytes.

• Journal of Embryo Transfer
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• v.19 no.3
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• pp.265-273
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• 2004

This study was carried out to examine the effects of fertilization media and sperm concentration on in vitro fertilization (IVF) and development (IVD) of porcine oocytes matured in vitro. Cumulus-oocyte complexes (COCs) were collected from antral follicles of porcine ovaries collected from abattoir, and were matured in vitro in modified NCSU-23 (mNCSU-23) supplemented with 10％ porcine follicular fluid (pFF). After the fertilization by experimental scheme, putative embryos were developed in vitro in NCSU-23. The results are as follows. When the oocytes were fertilized in vitro in modified TBM or modified TLP-PVA by 1 ${\times}$10$^{5}$ sperm/$m\ell$, all of the fertilization parameters were not significantly different between two media. Subsequently, as these putative embryos were developed in vitro in NCSU-23, the percentage of oocytes cleaved and of blastocysts were not different between two media, either. When the oocytes were fertilized in vitro in mTBM by 5${\times}$10$^4$, 1${\times}$10$^{5}$ or 5${\times}$10$^{5}$ sperm/$m\ell$, all of the fertilization parameters were significantly (P<0.05 or P<0.01) increased as sperm concentration was elevated. Subsequently, as these putative embryos were developed in vitro in NCSU-23, the percentage of oocytes cleaved and of blastocysts were significantly boosted (P<0.01) as sperm concentration at fertilization was elevated from 5${\times}$10$^4$ to 1${\times}$10$^{5}$ sperm/$m\ell$, but were not different between 1${\times}$10$^{5}$ and 5${\times}$10$^{5}$ sperm/$m\ell$.

• Journal of Plant Biotechnology
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• v.41 no.3
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• pp.116-122
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• 2014

Total of fifty accessions of Brassica rapa with various morphological characteristics were used for production of double haploid plants though microspore culture in Brassica rapa. Among them, only 30 accessions induced embryos from microspores. The highest efficiency of embryo induction of 1.194 per bud was obtained from IT135449 of turnip type, while 3 accessions of sarson (winter oil) type did not generate embryo. The effect of heat shock periods for embryogenesis was also investigated with 4 accessions (IT135449; Turnip type, IT199710; Chinese cabbage type, IT212886; Pak choi type, IT218043; Summer oil type). The high productions of embryos were observed in IT135449, IT199710 and IT212886 when microspores were pre-cultured to $32^{\circ}C$ for 2 days. In IT218043, high embryogenesis was observed at the 3 days of heat shock treatment. The optimal condition of shoot regeneration for IT199710 was observed in MS medium supplemented with NAA $0.5mg{\cdot}L^{-1}$ and BAP $1mg{\cdot}L^{-1}$. In contrast, the IT135449 and IT212886 were observed high regeneration frequency in MS medium without plant growth regulators. All the plantlets regenerated from microspore-derived embryos have been successfully transplanted to soil, and bud self-pollinated seeds were produced from doubled haploid plants. This indicated that double-haploid genotype was likely generated naturally during embryogenesis process.