• Title/Summary/Keyword: culturing media

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The optimum media size and composition for sawdust culture of Lentinula edodes (Strain: JNM3022) (표고버섯(JNM3022) 톱밥재배에 적합한 배지조성과 배지크기)

  • Choi, Duck Soo;Jung, Kyung Ju
    • Journal of Mushroom
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    • v.6 no.3_4
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    • pp.131-137
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    • 2008
  • These experiments were conducted to search for the optimum media size and composition for culture of Lentinula edodes (JNM3022). Considering the incubation period and the successful culturing rate, the optimum media size was length 25 $cm{\times}diameter$ 14 cm (weight 2.5 kg). The saturation moisture amount of Oak sawdust was 251 ml/100 g. It is a half of pine sawdust 495 ml/100 g. Considering the hypha growing speed and hypha density, the optimum media composition was Oak sawdust 80%+ wheat bran 20% and Oak sawdust 80%+ rice bran 20% In 10 treatments of media composition. The optimum moisture amount of media for growing Lentinula edodes (JNM3022) was 55%. Injection pipe with 20 holes in media was reduced the incubation periods to 11 days when 20cm length media and 32 days when 30cm length media compare to control. The best method for browning media surface was change temperature condition above $10^{\circ}C$ and punching the vinyl bag. This treat could shortened the browning periods to 54 days.

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Comparison of Ogawa Media, BACTEC MGIT 960 System and TB/NTM Real-Time PCR for Detecting Mycobacterium Species

  • Bang, Hae-In;Choi, Tae-Youn;Shin, Jeong-Won
    • Tuberculosis and Respiratory Diseases
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    • v.71 no.4
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    • pp.249-253
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    • 2011
  • Background: Mycobacterial infection is a problem throughout the world along with the increase of immunocompromised patients. For this reason, there have been many methods for faster and more accurate diagnosis. In this study, we evaluated several laboratory methods for mycobacterial infection. Methods: From January to December 2009, 635 specimens were cultured with mycobacteria growth indicator tube (MGIT) and Ogawa media. Polymerase chain reaction (PCR) was performed with the AdvanSure tuberculosis (TB)/non-tuberculosis mycobacterium (NTM) real-time PCR Kit (LG Life Sciences, Seoul, Korea). The 69 samples showing positive culture results were identified with the AdvanSure Mycobacteria Genotyping Chip Kit (LG Life Science, Seoul, Korea). Results: Sixty-nine (10.9%) out of 635 samples showed positive results for mycobacterial culture. Among the 635 samples, 64 were positive in MGIT, but only 42 were positive in Ogawa media. Of the 635 samples, 607 (95.6%) showed the same results between MGIT and Ogawa and the results of 579 (95.4%) were also consistent with the TB/NTM real-time PCR results. However, in the case of NTM, only one (1/24, 4.2%) was positive in PCR. In the Mycobacteria genotyping chip analysis, the most frequently identified NTM species in descending order were M. avium, M. intracellulare, M. chelonae and M. abscessus. Conclusion: Culturing with a combination of MGIT and Ogawa is recommended to increase the recovery rate of mycobacteria. Although PCR missed a reasonable number of NTM, it is faster and usually gives results that concur with those from the culture. The appropriate combination of diagnostic methods with clinical correlation are necessary.

Induced Change in DNA Methylation of Fusarium oxysporum f. sp. niveum due to Successive Transfer

  • Kim, Dae-Hyuk
    • BMB Reports
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    • v.30 no.3
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    • pp.216-221
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    • 1997
  • Changes in pathogenicity of old and successively-cultured isolates of Fusarium oxysporum f. sp. niveum have been observed and the concept that such cultures will become attenuated is generally accepted. However, the genetic basis for this phenomenon has not been studied. In an effort to identify a DNA marker closely linked to variations, DNA methylation was investigated both before and after the successive transfers of F. o. f. sp. niveum isolates on artificial media. A sector of mycelium in F. o. f. sp. niveum race 2 isolate (TXXID) which showed variation in pigmentation and colonial morphology occurred after 18 successive weekly transfers on potato dextrose agar (PDA). The sector characteristics were stable and did not change after more successive transfers. It was shown that DNA methylation preexists in ribosomal RNA gene (rDNA) of F. o. f. sp. niveum and that additional changes in DNA methylation occurred during successive culturing.

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Induction of Callus and Culture of Protoplasts from Cotyledon-Originated Callus in Alnus hirsuta (물오리나무(Alnus hirsuta)의 칼루스 유도 및 자엽유래 칼루스로부터의 원형질체 배양)

  • 김형하
    • Journal of Plant Biology
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    • v.33 no.4
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    • pp.253-257
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    • 1990
  • Callus-inducing ability of Alnus hirsuta was examined by culturing various tissues (leaf, hypocotyl, cotyledon and seed) on NT (Nagata & Takebe) medium, supplemented with 2.5$\mu$M 2,4-D. Leaf-originated callus was cultured on media varying in auxin (IBA and NAA) and cytokinin (BAP) concentrations to examine the effects of auxin and cytokinin on callus growth. Maximum growth was obtained at 10 $\mu$M IBA+10$\mu$M BAP and 10$\mu$M NAA without cytokinin. Cell suspensions established from cotyledon-originated callus yielded viable protoplasts after incubation for 16-18 hours in an enzyme mixture (1% (w/v) Onozuka R-10 0.5% (w/v) Macerozyme, CPW salts and 13% (w/v) mannitol, pH 5.8). Protoplasts were cultured on NT medium, supplemented with glucose, hormones and coconut milk. After 6 weeks of culture, protoplasts sustained cell divisions to form microcallus, which showed various colors from red to white.

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Study on the Elastic Characteristics of Living Cells using Atomic Force Microscope Indentation Technique

  • Kwon Eun-Young;Kim Young-Tae;Kim Dae-Eun
    • KSTLE International Journal
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    • v.7 no.1
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    • pp.10-13
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    • 2006
  • In this work, imaging and study of elastic property of the living cell was performed. The motivation of this work was to seek the possibility of exploiting Young's modulus as a disease indicator using Atomic Force Microscope (AFM) and also to gain fundamental understanding of cell mechanics for applications in medical nanorobots of the future. L-929 fibroblast adherent cell was used as the sample. Imaging condition in cell culturing media environment was done in very low speed ($20{\mu}m/ s$) compared to that in the ambient environment. For measuring the Young's modulus of the living cell, AFM indentation method was used. From the force-distance curve obtained from the indentation experiment the Young's modulus could be derived using the Hertz model. The Young's modulus of living L-929 fibroblast cell was $1.29{\pm}0.2$ kPa.

Optimization of Culturing Conditions for Improved Production of Bioactive Metabolites by Pseudonocardia sp. VUK-10

  • Kiranmayi, Mangamuri Usha;Sudhakar, Poda;Sreenivasulu, Kamma;Vijayalakshmi, Muvva
    • Mycobiology
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    • v.39 no.3
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    • pp.174-181
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    • 2011
  • The purpose of the present study was to investigate the influence of cultural and environmental parameters affecting the growth and bioactive metabolite production of the rare strain VUK-10 of actinomycete Pseudonocardia, which exhibits a broad spectrum of in vitro antimicrobial activity against bacteria and fungi. Production of bioactive metabolites by the strain was high the in modified yeast extract-malt extract-dextrose (ISP-2) broth, as compared to other tested media. Glucose (1%) and tryptone (0.25%) were found to be the most suitable carbon and nitrogen sources, respectively, for optimum production of growth and bioactive metabolites. Maximum production of bioactive metabolites was found in the culture medium with initial pH 7 incubated with the strain for four days at $30^{\circ}C$, under shaking conditions. This is the first report on the optimization of bioactive metabolites by Pseudonocardia sp. VUK-10.

Laboratory-scale fruiting body formation of Pleurotus ostreatus using the petri dish culture (느타리의 기내 자실체 형성 및 그 유도조건에 관한 연구)

  • Joh, Joong-Ho;Chu, Kyo-Sun;Kim, Beom-Gi;Kong, Won-Sik;Yoo, Young-Bok;Lee, Seung-Jae;Cho, Bong-Gum;Lee, Chang-Soo
    • Journal of Mushroom
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    • v.2 no.1
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    • pp.15-20
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    • 2004
  • Pleurotus ostreatus, the oyster mushroom, is one of the most widely cultivated and important edible mushrooms in the world. In order to study the developmental process of P. ostreatus and its regulatory mechanism, a new culturing method needs to be established for inducing the fruiting body and sporulation in the laboratory. In this study, we have examined whether the fruiting body of P. ostreatus can be formed on the plastic petri dish which are commonly used for cell culture in the laboratory. The strain was cultured on $60{\times}15mm$ plastic petri dish with potato dextrose agar media at $28^{\circ}C$ for mycelial growth and then at $18^{\circ}C$ for the formation of primordia and fruiting bodies within plant growth chamber. The development of primordia into fruiting bodies was achieved on cultured dishes under air ventilation. At the primordia stage, the normal formation of fruiting body was blocked by sealing the plastic dish with parafilm. The periods requiring for the formation of primordia and fruiting bodies were examined on the dish culture. About 96% and 76% of cultured samples formed primordia and fruiting bodies under the optimal conditions during ten weeks of culture, respectively. These culturing periods, however, were changed by the mechanical injury treatment to mycelia. As other factors affecting the fruiting body formation, the effects of light and cold shock have been tested. No fruiting formation was observed on the cultured dishes under the dark. The cold shock treatment by storing cultured dishes for one day at $4^{\circ}C$ did not have any significant effects in the fruiting body formation. Spores of fruiting bodies acquired from the petri dishes could be germinated on culture media at $28^{\circ}C$. These results suggest that the fruiting bodies of P. ostreatus can be formed on the experimental petri dish and this dish-culturing method is useful for understanding of the developmental process of P. ostreatus in the laboratory. Furthermore, the dish-culturing method is able to shorten the life cycle of P. ostreatus without requiring large area and expensive device.

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The production of media and optimal additive rate using the cultivation media wastes of Pleurotus eryngii (큰느타리버섯 폐배지 이용 배지 제조 및 적정 첨가비율)

  • Kim, Min-Keun;Ryu, Jae-San;Lee, Young-Han;Park, Jeong-Sik;Jung, Ji-In;Kwon, Jin-Hyeuk;Rho, Chi-Wong;Yun, Han-Dae
    • Journal of Mushroom
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    • v.5 no.2
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    • pp.76-80
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    • 2007
  • After bottle culture of Pleurotus eryngii, media were taken out the bottle and normally utilized compost. However, nutritional elements were remained in the waste media. This study was carried out to investigate the reusable possibility and the optimal additive rate of waste media in an artificial cultivation of Pleurotus eryngii. The pH had tendency to decrease as the waste media was added from 6.0 to 4.8. Based on the additive rate of 10, 20, 30, 40, and 50%, each treatment waste media was added to new media for culturing Pleurotus eryngii. Among various treatments, the mycelial growth and primordia formation of Pleurotus eryngii were more favorable in the addition of 10-30% waste median than in the addition of 40 and 50%. The yield of its fruiting body was increased slightly in the treatment of 10-30% waste media.

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Liquid Culture of Basidiomycetes on Natural Media (천연물을 이용한 담자균의 균사체 배양)

  • Kim, Seon-Hee;Lee, Jong-Suk;Park, Kyung-Sook;Lee, Jae-Sung;Lee, Hang-Woo;Park, Shin
    • The Korean Journal of Mycology
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    • v.27 no.6 s.93
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    • pp.373-377
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    • 1999
  • This study was designed to develop functional drink from jujube extract through a simple submerged culture of three basidiomycetes species. The optimum Brix and pH of the jujube extract for culturing the Ganoderma lucidum appeared to be 5 Brix and pH 4. Ten days of culture period produced maximum mycelium. The optimum Brix and pH of the jujube extract for culturing the Coriolus versicolor appeared to be 5 Brix and pH 5. Ten days of culture period produced maximum mycelium. The optimum Brix and pH of the jujube extract for culturing the Phellinus igniarius appeared to be 3 Brix and pH 5. For the maximum mycelial production eighteen days of culture period was required for Phellinus igniarius. The antitumor activity of the polysaccharides extracted from the fermented drinks was demonstrated through the tumor cell line experiments. The $IC_{50}$ values of the jujube drinks fermented with Ganoderma lucidum and Phellinus igniarius against stomach cancer cell line appeared to be one fourth that of the jujube drink which was not fermented with basidiomycetes.

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Effect of Bovine Granulosa Cell Culture Supernatant on In Vitro Development of Mouse Embryos (소 과립막세포의 배양 상층액이 생쥐배의 체외발달에 미치는 영향)

  • Lee, Sang-Bum;Moon, Sin-Hong;Kim, Seon-Ku
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1764-1768
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    • 2009
  • This study was carried out to examine a concentration of steroid hormones and in vitro development of mouse embryos in culture supernatant of bovine granulosa cells (GC). To obtain the culture supernatant, granulosa cells were retrieved from mature follicles (6~15 mm diameter) and immature follicles (2~5 mm diameter) of bovine ovary and were cultured, respectively, in media of Ham's F-10 with 15% FCS for 16 days. Mature and immature granulosa cells formed their monolayers easily and showed similar growth patterns in culturing. There was no morphological difference between mature and immature granulosa cells. High levels of both progesterone and estradiol were detected in the culture supernatant of mature granulosa cells and immature granulsa cells, and the endocrine profiles of the two types of cells were similar. Progesterone secretion of granulosa cells was high in the late stage of culturing and estradiol secretion was high in the early stage of culturing. In vitro development rates of mouse embryos to morula, blastocyst and hatched blastocyst were significantly (p<0.05) higher in culture supernatant of mature granulosa cells (92.7%, 78.1% and 34.5%) and in culture supernatant of immature granulosa cells (96.4%, 78.5% and 26.8%) than in Ham's F-10 (86.7%, 41,7% and 13.3%). However, there was no difference between the culture supernatant of mature granulosa cells and the culture supernatant of immature granulosa cells in the development of embryos.