• Title/Summary/Keyword: Laboratory Culture

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Biocontrol Activity of Bacillus amyloliquefaciens CNU114001 against Fungal Plant Diseases

  • Ji, Seung Hyun;Paul, Narayan Chandra;Deng, Jian Xin;Kim, Young Sook;Yun, Bong-Sik;Yu, Seung Hun
    • Mycobiology
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    • v.41 no.4
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    • pp.234-242
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    • 2013
  • A total of 62 bacterial isolates were obtained from Gomsohang mud flat, Mohang mud flat, and Jeju Island, Republic of Korea. Among them, the isolate CNU114001 showed significant antagonistic activity against pathogenic fungi by dual culture method. The isolate CNU114001 was identified as Bacillus amyloliquefaciens by morphological observation and molecular data analysis, including 16SrDNA and gyraseA (gyrA) gene sequences. Antifungal substances of the isolate were extracted and purified by silica gel column chromatography, thin layer chromatography, and high performance liquid chromatography. The heat and UV ray stable compound was identified as iturin, a lipopeptide (LP). The isolate CNU114001 showed broad spectrum activity against 12 phytopathogenic fungi by dual culture method. The semi purified compound significantly inhibits the mycelial growth of pathogenic fungi (Alternaria panax, Botrytis cinera, Colletotrichum orbiculare, Penicillium digitatum, Pyricularia grisea and Sclerotinia sclerotiorum) at 200 ppm concentration. Spore germ tube elongation of Botrytis cinerea was inhibited by culture filtrate of the isolate. Crude antifungal substance showed antagonistic activity against cucumber scleotiorum rot in laboratory, and showed antagonistic activity against tomato gray mold, cucumber, and pumpkin powdery mildew in greenhouse condition.

Prevalence of paratuberculosis of dairy cattle in Kangwon area (강원지역 젖소의 요네병 감염실태)

  • Kim, Doo;Jeon, Kwan-joon;Kim, Jong-taek;Shin, Kwang-soon;Shin, Myung-kyun;Chang, Guk-hyun;Kim, Jeung-ki;Kim, Och-sung;Jung, Jae-young
    • Korean Journal of Veterinary Research
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    • v.42 no.1
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    • pp.81-88
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    • 2002
  • The purpose of this study was to conduct diagnosis of bovine paratuberculosis in Kangwon area. Blood samples were collected from 2,261 dairy cows of 162 herds, and the ELISA and immunoblotting using recombinant 34KDa protein of M. paratuberculosis were conducted. The feces collected from dairy cows were cultured on HEY medium with mycobactin-J and PCR was conducted with washing solution of medium 4 weeks after culture. The ELISA had sensitivity of 83.3% and specificity of 96.7%. And the immunoblotting had sensitivity of 83.3% and specificity of 100%. Of the 2,261 dairy cows, 371 cows(16.4%) were positive in ELISA and 75 cows(3.3%) were positive in immunoblotting. And of the 162 herds, 109 herds(67.3%) had an apparent paratuberculosis prevalence by ELISA and 40 herds(24.7%) by immunoblotting. The geographic distribution of herds with paratuberculosis was not uniform. Of the 241 feces samples including 110 feces from ELISA positive cow, 9 feces were positive in culture and PCR. PCR was able to detect the growth of M. paratuberculosis as early as 4 weeks of culture.

Establishment and characterization of gastric surface mucous cell lines (GSM06 and GSM10) from transgenic mice harboring temperature-sensitive simian virus 40 large T-antigen gene

  • Tabuchi, Yoshiaki;Sugiyama, Norifumi;Horiuchi, Tadashi;Furuhama, Kazuhisa;Obinata, Masuo;Furusawa, Mitsuru
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.131-136
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    • 1994
  • In the present study, in order to make an in vitro model of gastric mucosa for physiological and pharmacological studies, we established two immortalized gastric surface mucous cell lines (GSM06 and GSM10), which produce periodic acid-Schiff (PAS)-and concanavalin A (Con A)-positive glycoproteins, from a primary culture of gastric fundic mucosal cells of adult transgenic mice harboring a temperature-sensitive simian virus 40 large T-antigen gene 〔1]. Gastric fundic mucosal cells were isolated as a modification of a previously described method for rats by Schepp et al. (2). The isolated gastric fundic mucosal cells were cultured in DME/F12 medium supplemented with 2% fetal bovine serum (FBS), 1% ITES (consisting of 2 mg/1 insulin, 2 mgg/1 transferrin, 0.122 mg/1 ethanolamine and 0.00914 mg/1 sodium selenite) and 10 ng/ml recombinant epidermal growth factor (EGF) in a collagen-coated culture dish. To remove fibroblastic cells from the culture, gastric mucosal cells were incubated in the culture medium containing dispase (25 U/ml) for 24 h. The cells, uncontaminated with fibroblastic cells, were then cloned by colony formation. In our series of three attempts, two cell lines (GSM06 and GSM10) have been established at last. The cells proliferated, attached to the dish ana grew until confluent monolayers were formed, and maintained tight contact with neighboring cells. Both GSM06 and GSM10 cells have now been in culture for more than 9 months with regular passaging. The either cell produced

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Continuous Passaging of a Recombinant C-Strain Virus in PK-15 Cells Selects Culture-Adapted Variants that Showed Enhanced Replication but Failed to Induce Fever in Rabbits

  • Tong, Chao;Chen, Ning;Liao, Xun;Yuan, Xuemei;Sun, Mengjiao;Li, Xiaoliang;Fang, Weihuan
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1701-1710
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    • 2017
  • Classical swine fever virus (CSFV) is the etiologic agent of classical swine fever, a highly contagious disease that causes significant economic losses to the swine industry. The lapinized C-strain, a widely used vaccine strain against CSFV, has low growth efficiency in cell culture, which limits the productivity in the vaccine industry. In this study, a recombinant virus derived from C-strain was constructed and subjected to continuous passaging in PK-15 cells with the goal of acquiring a high progeny virus yield. A cell-adapted virus variant, RecCpp80, had nearly 1,000-fold higher titer than its parent C-strain but lost the ability to induce fever in rabbits. Sequence analysis of cell-adapted RecC variants indicated that at least six nucleotide changes were fixed in RecCpp80. Further adaption of RecCpp80 variant in swine testicle cells led to a higher virus yield without additional mutations. Introduction of each of these residues into the wild-type RecC backbone showed that one mutation, M979R (T3310G), located in the C-terminal region of E2 might be closely related to the cell-adapted phenotype. Rabbit inoculation revealed that $RecCpp40_{+10}$ failed to induce fever in rabbits, whereas $RecCpp80_{+10}$ caused a fever response similar to the commercial C-strain vaccine. In conclusion, the C-strain can be adapted to cell culture by introducing specific mutations in its E2 protein. The mutations in RecCpp80 that led to the loss of fever response in rabbits require further investigation. Continuous passaging of the C-strain-based recombinant viruses in PK-15 cells could enhance its in vitro adaption. The non-synonymous mutations at 3310 and 3531 might play major roles in the enhanced capacity of general virus reproduction. Such findings may help design a modified C-strain for improved productivity of commercial vaccines at reduced production cost.

Production of Acetate from Carbon Dioxide in Bioelectrochemical Systems Based on Autotrophic Mixed Culture

  • Su, Min;Jiang, Yong;Li, Daping
    • Journal of Microbiology and Biotechnology
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    • v.23 no.8
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    • pp.1140-1146
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    • 2013
  • Bioelectrochemical systems (BESs) have been suggested as a new technology for wastewater treatment while accomplishing energy and chemical generation. This study describes the performance of BESs based on mixed culture that are capable of reducing carbon dioxide to acetate. The cathode potential was a critical factor that affected the performance of the BESs. The rate of acetate production increased as the electrode potential became more negative, from 0.38 mM $d^{-1}$ (-900 mV vs. Ag/AgCl) to 2.35 mM $d^{-1}$ (-1,100 mV), while the electron recovery efficiency of carbon dioxide reduction to acetate increased from 53.6% to 89.5%. The microbial population was dominated by relatives of Acetobacterium woodii when a methanogenic inhibitor was added to the BESs initially.

Nitrogen-Dependent Regulation of Gluconic and/or Citric Acid Production by Aspergillus niger

  • Sankpal, Narenora V.;Joshi, Arvino P.;Kulkarni, Bhaskar D.
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.51-55
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    • 2000
  • Surface culture fermentation using Aspergillus niger was studied for gluconic and citric acid production at different C/N ratios. A culture of A. niger was found to produce either gluconic acid alone, a mixture of gluconic and citric acid, or citric acid alone depending on the level of nitrogen in the medium (4 to 18mM). Glucose oxidase from the mycelial mat was also analyzed at different levels of nitrogen in the media. By choosing the level of nitrogen in the medium at the start of fermentation, it is possible to produce either of the two acids as the dominant product or the two together as a mixture.

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Effects of Aeration Rates on Production on Extracellular Polysaccharide, EPS-R, by Marine Bacterium Hahella chejuensis

  • Lee, Hyung-Sang;Park, Shin-Hye;Lee, Jong-Ho;Lee, Hong-Kum
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.5
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    • pp.359-362
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    • 2001
  • The production of an extracellular polysaccharide, EPS-R, from the marine bacterium Hahella chejuensis was investigated at various aeration rates in a batch culture. Higher aeration rate resulted in enhanced EPS production and increased the viscosity of the culture broth. At an aeration rate of 1.5 vvm, EPS-R (12.2 g/L) was obtained with a yield (Y$\_$p/s) of 0.6 from the STN medium after 72 h of cultivation. The H. chejuensis cells changed rod morphology to a short-rod form in the stationary growth phase.

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The Leech as a Laboratory Animal for the biomedical Research (생명과학 연구를 위한 실험동물로서의 한국산 거머리에 관한 제고)

  • Hong, Seok-Jin;Don
    • The Korean Journal of Zoology
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    • v.36 no.4
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    • pp.588-595
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    • 1993
  • We have collected a great Quantity of leeches, comprising five species, from six different ponds and rice paddies in the south-western part of Korea. The leeches were cultured in our laboratory with special interests on the blood-suckers (Himdo nipponial in fresh water supplemented with essential ions. A number of experiments was carried out with biomedical aspects. We have observed traits which can be used as a laboratory animal. The paper reports general features obsenred from leeches with preliminary study results carried in culture systems. The paper also describes study aspects of leeches in biochemistry, neurology and developmental biology.

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The role of sonic hedgehog signaling pathway in in vitro oocyte maturation

  • Lee, Sanghoon;Cho, Jongki
    • Journal of Animal Reproduction and Biotechnology
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    • v.36 no.4
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    • pp.183-188
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    • 2021
  • In vitro maturation (IVM) of oocytes is the procedure where the immature oocytes are cultivated in a laboratory until they are mature. Since IVM oocytes generally have low developmental competence as compared to those matured in vivo, development of an optimal IVM culture system by fine-tuning culture conditions is crucial to maintain high quality. In-depth knowledge and a deep understanding of the in vivo physiology of oocyte maturation are pre-requisites to accomplish this. Within ovarian follicles, various signaling pathways that drive oocyte development and maturation regulate interaction between oocytes and surrounding somatic cells. This review discusses the sonic hedgehog (SHH) signaling pathway, which has been demonstrated to be intimately involved in folliculogenesis and oocyte maturation. Advances in elucidating the role of the SHH signaling pathway in oocyte maturation will aid attempts to improve the current inferior in vitro oocyte maturation system.