• Title/Summary/Keyword: liquid culture

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In-Vitro Fertilization and Culture of Pig Oocytes Matured In-Vitro by Liquid Boar Sperm Stored at 4$^{\circ}C$

  • Kim, M. Y.;Y. J. Yi;Y. J. Chang;Park, C. S.
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.63-63
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    • 2003
  • This study was carried out to investigate the effects of liquid boar sperm stored at 4$^{\circ}C$ on sperm motility, normal acrosome, and in-vitro fertilization and culture of pig oocytes matured in-vitro. The sperm-rich fraction (30~60 ml) of ejaculate was collected into an insulated vacuum bottle. Semen was slowly cooled to room temperature (20~23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min at 800$\times$g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of lactose, egg yolk and N-acetyl-D-glucosamine (LEN) diluent to provide 1.0$\times$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$ and preserved for 5 days to examine sperm motility and normal acrosome. The medium used for oocyte maturation was modified tissue culture medium (TCM) 199. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 6 h in 500 ${mu}ell$ mTBM fertilization media with 0.2, 1, 5 and 10$\times$10$^{6}$ /ml sperm concentration, respectively. At 6 h after IVF, oocytes were transferred into 500 ${mu}ell$ Hepes-buffered NCSU-23 culture medium for further culture of 6, 48 and 144 h. There were significant differences in sperm motility and normal acrosome among preservation days and incubation times, respectively. The rates of sperm penetration and polyspermy were higher in 5 and 10$\times$10$^{6}$ sperm/ml than in 0.2 and 1$\times$10$^{6}$ sperm/ml. Male pronuclear formation was lower in 0.2$\times$10$^{6}$ sperm/ml than in 1, 5 and 10$\times$10$^{6}$ sperm/ml. Mean numbers of sperm in penetrated oocyte were highest in 10$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. The rate of blastocysts from the cleaved oocytes (2~4 cell stage) was highest in 1$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend 1$\times$10$^{6}$ ml sperm concentration for in-vitro fertilization of pig oocytes.

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Favorable Conditions for Mycelial Growth of Phellinus linteus (목질진흙버섯(Phellinus linteus)의 적합한 균사생장)

  • Lee, Won-Ho;Kim, Su-Young;Park, Young-Jin;Kim, Tae-Woong;Kim, Ho-Kyung;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.32 no.2
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    • pp.95-100
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    • 2004
  • The main objectives of the study were to investigate cultural characteristics of Phellinus linteus. The optimum culture media for mycelial growth of P. linteus were MYA (malt yeast agar) and SMS (soybean powder malt Sucrose). Similarly, optimum temperature and pH were $30^{\circ}C$ and 6.0, respectively. Malt extract (2%, v/v) and yeast extract (0.2%, v/v) were optimum carbon and nitrogen sources. Similarly, 0.1% $KH_2PO_4$ was optimum mineral salt. Highest mycelial growth was observed when C/N ratio was 10 : 1. Optimum inoculum amount for flask culture was $5{\sim}6$ mycelial discs (6 mm diameter) per 100 ml of liquid medium, Highest mycelial dry weight was obtained when cultured in 100 ml liquid medium in 300 ml shaking flask after 20 days of shaking culture, For mass liquid culture (8 l), flask culture was homogenized and used as an inoculum. Optimum culture period and aeration rate for 8l fermentation culture were 12 days and 2.0 vvm, respectively.

Micropropagation of Cassava by Suspension Culture Derived from its Nodal Explants (마디 절편의 현탁배양에 의한 카사바의 미세증식)

  • Yoon, Sil;Cho, Duck-Yee;Soh, Woong Young
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.185-189
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    • 2000
  • For the micropropagation, node explants of cassava were cultured in liquid MS medium with various concentrations of cytokinins on a rotary shaker (100 rpm) for 2 weeks. The adventitious roots and shoots from the explants were differentiated more efficiently in liquid medium than in solid. But root formation was not inhibited in medium with BAP and kinetin at low concentration (>0.05 mg 1/sup -1/), while in medium added with BAP and zeatin at high level (<0.25 mg 1/sup -1/), it was inhibited by callus forming on cut end of the cuttings. However, all of plantlets grown in liquid medium for more than 2 weeks showed symptoms of hyperhydricity. The plantlets grown in liquid medium were transferred into culture bottles filled with fine sand or artificial soil (pitmoss:perlite:vermiculite, 1:1:1 v/v) wetted with half strength of Knop's solution. After transplanted to culture bottles, some of vitriscent leaves were defoliated and new leaves were normally formed from shoot apex. Most of plantlets (>95%) were hardened-off successfully only in culture bottles with fine sand, and grew into 3-5 cm seedlings possessing 4-6 nodes after 4 weeks. Thus, the mass propagation of cassava on medium containing cytokinin could be established based on the suspension culture using node explants.

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Promotion of in vitro shoot proliferation in rose by addition of liquid medium to culture (액체배지 첨가에 의한 장미 기내 신초 증식 촉진)

  • Lee, Ye Ji;Lee, Jung Lim;Hyung, Nam-In;Kim, Seung Tae;Lee, Eun Kyung;Kwon, O Hyeon;Kim, Won Hee;Lee, Su Young
    • Journal of Plant Biotechnology
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    • v.39 no.4
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    • pp.305-308
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    • 2012
  • To promote the growth and proliferation of in vitro rose (Rosa hybrida L) shoots, a liquid medium was added to shoot culture. Shoots were obtained by culturing internodes of four cultivars, 'Antique Curl', 'Shiny Orange', 'White Zen', and 'Red Zen', and then were proliferated by the subculture two times. An addition with 10~15 mL of liquid medium enhanced the shoot elongation of all four cultivars. However, the effect of liquid medium addition to culture of in vitro shoot for proliferation was dependent on cultivars of rose.

Quality Characteristics of Bamboo Shoot Liquid Fermented by Bacillus subtilis Strain (Bacillus subtilis 균주로 발효한 죽순발효액의 품질특성)

  • Baeg, Byeong-Gi;Cho, Jung-Il;Moon, Eun-Woo;Park, Jung-Suk
    • Journal of the Korean Society of Food Culture
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    • v.30 no.2
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    • pp.233-240
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    • 2015
  • To make fermented bamboo shoot liquid, we isolated and classified a microorganism growing in bamboo shoot and investigated its quality characteristics. Crude fiber, crude ash, nitrogen-free extracts, and total sugar contents were higher after fermentation. For free amino acids, only alanine was detected in the control group. Detected 13 kinds of free amino acids were detected in fermented bamboo shoot liquid. In organoleptic test, fermented bamboo shoot beverage containing 20 percent strawberry showed the highest consumer preference.

Cultural Characteristics of Shimizuomyces paradoxus Collected from Korea

  • Sung, Gi-Ho;Shrestha, Bhushan;Park, Ki-Byung;Sung, Jae-Mo
    • Mycobiology
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    • v.38 no.3
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    • pp.189-194
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    • 2010
  • This study investigated the cultural characteristics of Shimizuomyces paradoxus in different nutritional and environmental conditions. The highest mycelial growth was observed in Schizophyllum (mushroom) genetics complete medium plus yeast extract agar medium, and the optimal temperature and pH were $25^{\circ}C$ and pH 8.0, respectively. The optimal carbon and nitrogen sources were 1% dextrose and 1% peptone in agar. However, in liquid culture the highest dry mycelium weight was found for the potato dextrose agar and potato sucrose agar broths. The optimum inoculum size was five mycelial discs (5 mm) per 100 mL of broth, and the optimum liquid culture period was 25 days. This is the first ever report of S. paradoxus cultural characteristics.

Influence of Nutrient Addition in the Liquid Yeast Fermentation of Pulverized Food Wastes (남은 음식물의 습식효모배양에서 영양물질첨가가 효모증식에 미치는 영향)

  • Lee, Ki-Young;Yu, Sung-Jin;Chae, Hee-Jung
    • Journal of the Korea Organic Resources Recycling Association
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    • v.9 no.1
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    • pp.49-55
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    • 2001
  • For the production of probiotic feed enriched with viable yeasts, aerobic liquid culture of Kluyveromyces marxianus was attempted in pulverized residual food wastes. After the preliminary shaking culture result, the liquid food wastes was added with urea($0.5g/{\ell}$), o-phosphate($0.4g/{\ell}$ ), molasses($4g/{\ell}$), and yeast extract($1g/{\ell}$), and the fermentation was carried out in 2-litre jar fermenter. In 12 hours of aerobic mixed culture with Aspersillus oryzae, viable cell count of the yeast reached to the number of $1.4{\times}10^{10}/{\ell}$ in the cultured medium.

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A study on the Mycelial Growth of Lentinus lepideus in Liquid Culture (액체배양에서 잣버섯 균사체 배양에 관한 연구)

  • 신성의;차월석;강시형
    • Journal of Life Science
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    • v.13 no.4
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    • pp.492-497
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    • 2003
  • This study was carried out to get the basic data for the mycelial growth of Lentinus lepideus in liquid culture. The optimal temperature and initial pH of mycelial growth of Lentinus lepideus were $25^{\circ}C$ and pH 5.5, respectively. The optimal medium was YMG medium. Among the carbon sources tested, glucose was effected to the mycelial growth and optimal glucose concentration was 4% (w/v). As nitrogen sources, malt extract and yeast extract appeared to be favorable and optimal malt extract and yeast extract [ratio (w/w) of 1:1] concentration was 1.5% (w/v).

Optimization of Pigment Production of Monascus Purpureus P-57 in Liquid Culture (액체배양에서 Monascus purpureus P-57 변이주의 색소생성 최적조건)

  • Park Chi Duck;Jung Hyuck Jun;Yu Tae Shick
    • KSBB Journal
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    • v.20 no.1 s.90
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    • pp.66-70
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    • 2005
  • Optimization of culture conditions for pigment production of Monascus purpureus P-57 mutant was investigated in liquid culture. The optimum composition of medium for the pigment production was $4\%$ rice power, $0.1\%$ beef extract, $0.03\%$ glutamic acid, $0.1\%\;MgSO_4{\cdot}7H_2O,\;0.25\%\;KH_2PO_4$, the optimum initial pH was 5.0. And the optimum culture conditions was at $30^{\circ}C$ for 8 days under 150 rpm with shaking. M. purpureus P-57 mutant produced the highest pigment as 356.04 units at red pigment and as 268.20 units at yellow pigment, and produced high cell mass as 15.00 g/L in liquid culture under the optimum conditions.

팽나무버섯(Flammulina velelutipes)균의 액체배양 생장조건 규명(Growth condition of Flammulina velutipes in Liquid culture)

  • Hong, Seong-Jun;Lee, Won-Ho;Park, Gi-Beom;Park, Gi-Byeong;Seong, Jae-Mo
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.355-358
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    • 2001
  • Selection of optima] nutrient sources and cultural methods for liquid spawn culture of Flammulina velutipes were carried out. The optimal temperature and pH for mycelial growth of F. velutipes were $20^{\circ}C$ and 6.0 to 7.5. respectively. In the 250ml ${Delta}$-flask culture. the amount of inoculum and culture period for the optimal mycelial growth of F. velutipes were 3 mycelial disks(diametcr 6mm) and 6 days, respectively. For the mass production of submerged culture. the optimal inoculum amount and aeration rate of F. velutipes were 5%(inoculum vol/medium vol.) and l.0vvm(vol of air/vol. of medium/min), respectively.

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