• Title/Summary/Keyword: Axenic culture

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Growth of Axenic Rotifer Brachionus rotundiformis (무균 로티퍼 Brachionus rotundiformis의 증식)

  • Jung, Min-Min;Rho, Sum;Kim, Pil-Yun
    • Journal of Aquaculture
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    • v.11 no.1
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    • pp.91-97
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    • 1998
  • This paper introduces to a simple culture method and growth of axenic (bacteria-free) rotifer Brachionus rotundiformis for seed stock of rotifer mass culture. This rotifer axenic culture method is based on the washing and transferring with sterilized sea water and modified antibiotic mixture AM9 solution. Population growth (final density on day 16) of axenic cultured rotifer were maintained with a high density and stable growth compared with the control of non-axenic culture (general culture style) through the 3 times-rerunning experiments (trial 1, 2 and 3). But the egg carrying rates of amictic females were not different between the axenic-and non-axenic culture condition. Although, rotifer density was higher in axenic culture, the food (Nannochloropsis oculata) was still remained unutilized than that of the non-axenic culture in third trial culture. These results suggest the possible existence of harmful bacteria for the rotifer population growth in the trial 1 and 3 of non-axenic culture compared to the trial 2. This axenic rotifer culture method is valuable for seed stock of the stable rotifer mass cultures.

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Antibiotics and Their Optimum Concentration for Axenic Culture of Marine Microalgae (해양미세조류의 무균배양을 위한 항생제의 종류 및 최적 농도)

  • Youn, Joo-Yeon;Hur, Sung-Bum
    • ALGAE
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    • v.22 no.3
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    • pp.229-234
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    • 2007
  • This study was to determine the extent of bacteria contamination and resistance to various antibiotics used commonly in microalgal culture. Seven different dose levels of chloramphenicol, dihydrostreptomycin sulphate, neomycin, penicillin G, streptomycin sulphate, penicillin G + streptomycin sulphate, and penicillin G + streptomycin sulphate + chloramphenicol were added to each culture of microalgae. The lethal effects on microalgae and bacteria were the highest in chloramphenicol and the lowest in penicillin G. The axenic culture of bacillariophyceae and dinophyceae was more difficult than that of chlorophyceae and haptophyceae because of their complicate external morphology. The efficient antibiotics and their concentrations for axenic cultures varied with microalgal species. The optimum quantity for antibiotic treatments were 2,000 ppm of dihydrostreptomycin for Chlorella ellipsoidea, neomycin 500 ppm of Isochrysis galbana and Heterosigma ahashiwo, hloramphenicol 500 ppm of Cyclotella didymus, and dihydrostreptomycin sulphate and neomycin 6,000 ppm of Thalassiosira allenii.

Establishment of an Axenic Culture of Microcystin-Producing Microcystis aeruginosa Isolated from a Korean Reservoir

  • Han, Ah-Won;Oh, Kyoung-Hee;Jheong, Weon-Hwa;Cho, Young-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.20 no.7
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    • pp.1152-1155
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    • 2010
  • In order to establish an axenic (bacteria-free) culture of Microcystis aeruginosa NIER 10039 isolated from a Korean reservoir, the culture was subjected to sequential treatment, including ultrasonication, washing, and addition of antibiotics. Three broad-spectrum antibiotics, namely, kanamycin, ampicillin, and imipenem, were applied separately in that order. Axenicity of the culture was confirmed by cultivation on bacterial media and observation under epifluorescence and scanning electron microscopes. We are the first to establish an axenic culture of a Microcystis strain isolated from Korean reservoirs and can be used in physiological and molecular studies to control toxic Microcystis blooms.

Axenic isolation procedure of the neutral spore and conchocelis from the seaweed Porphyra yezoensis

  • Park, Jae-Suk;Hong, Yong-Ki
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.151-152
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    • 2000
  • During in door or outdoor mass culture, Porphyra have been easily contaminated with bacteria, protozoa and microalgal species. Several axenic treatments for Porphyra thalli have been published (Polne-Fuller and Gibo 1984; Chen and McCracken 1993), but axenic techniques for neutral spores and conchocelis we not developed. In this work we describe the procedure for axenic isolation of neutral spores and conchocelis of Porphyra yezoensis (omitted)

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Axenic purification and cultivation of an Arctic cyanobacterium, Nodularia spumigena KNUA005, with cold tolerance potential for sustainable production of algae-based biofuel

  • Hong, Ji-Won;Choi, Han-Gu;Kang, Sung-Ho;Yoon, Ho-Sung
    • ALGAE
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    • v.25 no.2
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    • pp.99-104
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    • 2010
  • A psychrotolerant cyanobacterium, Nodularia spumigena KNUA005, was isolated from a cyanobacterial bloom sample collected near Dasan Station in Ny-${\AA}lesund$, Svalbard Islands during the Arctic summer season. To generate an axenic culture, the isolate was subjected to three purification steps: centrifugation, antibiotic treatment and streaking. The broad antibacterial spectrum of imipenem killed a wide range of heterotrophic bacteria, while the cyanobacterium was capable of enduring both antibiotics, the remaining contaminants that survived after treatment with imipenem were eliminated by the application of an aminoglycoside antibiotic, kanamycin. Physical separation by centrifugation and streaking techniques also aided axenic culture production. According to the cold-tolerance test, this mat-forming cyanobacterium was able to proliferate at low temperatures ranging between 15 and $20^{\circ}C$ which indicates the presence of cold-tolerance related genes in N. spumigena KNUA005. This suggests the possibility of incorporating cold-resistance genes into indigenous cyanobacterial strains for the consistent production of algae-based biofuel during the low-temperature seasons. Therefore, it is needed to determine the cold-tolerance mechanisms in the Arctic cyanobacterium in the next research stage.

Axenic cultivation and characterization of Giardia lamblia isolated from humans in Korea

  • Park, Soon-Jung;Yong, Tai-Soon;Yang, Hye-Won;Lee, Du-Ho;Lee, Kyung-Won
    • Parasites, Hosts and Diseases
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    • v.37 no.2
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    • pp.121-125
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    • 1999
  • Inoculation of human fecal cysts to suckling Mongolian gerbils, two Giardia lamblia isolates, Kl and K2, were established as axenic cultures. Using this in vitro culture, both two Giardia isolates were grouped by using two genetic analysis. With genetic analysis of SSS-rDNA sequences, both K1 and K2 were found as members of hopkins'group 1, despite some nucleotide differences noticed in K1 (5 differences/292 bases.). The other genetic study used PCI-RFLP of the tim (triose phosphate isomerase) Nash's group 2 can bot be a separate group, but a part of Hopkins' group 1.

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Growth of the Dinoflagellate Alexandrium tamarense Isolated from Jinhae Bay, Korea in Axenic Cultures

  • Lee, Hae-Ok;Ishimaru, Takashi;Toshiya, Katano;Han, Myung-Soo
    • Korean Journal of Environmental Biology
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    • v.24 no.3
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    • pp.275-281
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    • 2006
  • We examined effects of water temperature, salinity, irradiance, and different media on the growth of the toxic dinoflagellate Alexandrium tamarense (HYM9704), which was isolated from Jinhae Bay, Korea. The ranges of temperature and salinity in which the strain was able to grow were $10{\sim}20^{\circ}C$ and $20{\sim}34$ psu, respectively. These values were in accordance with those observed in situ. The maximum growth rates of axenic A. tamarense (HYM9704) was $0.25d^{-1}$ at $15^{\circ}C$, 30 psu, and $100{\mu}Em^{-2}s^{-1}$. The temperature affected the growth rates of axenic A. tamarense more significantly than the salinity. The type of culture media did not affect the growth rates of axenic A. tamarense. The strain in N-limited and P-limited media went into the stationary phase faster than that in T1 and T1/2 medium.

Development of Axenic Culture and Astaxanthin Production in Microalgae (미세조류를 이용한 무균분리법 개발 및 astaxanthin 생산)

  • Son, Min Chang;Lee, Dong-Jun;Park, Sejin;Kim, Min Sung;Lee, Chul Won;An, Won Gun
    • Journal of Life Science
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    • v.25 no.7
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    • pp.733-739
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    • 2015
  • Microalgae are a renewable natural resource that requires only sunlight, carbon dioxide, phosphorus, and nitrogen for rapid growth. They produce a broad variety of basic chemical substances―such as vitamins, fatty acids and carotenoids―that have high added value potential for the pharmaceutical and food industries. The aim of this study was to develop axenic culture and to establish a cell growth assay for microalgae. A further experiment was carried out to determine the yield of astaxanthin derived from microalgae. The axenic culture was developed using a mixture of antibiotics [ampicillin (100 ${\mu}g/ml$), streptomycin (10 ${\mu}g/ml$), chloramphenicol (10 ${\mu}g/ml$), penicillin (10 ${\mu}g/ml$), neomycin (50 ${\mu}g/ml$), gentamycin (50 ${\mu}g/ml$), kanamycin (10 ${\mu}g/ml$), and nystatin (1.5 ${\mu}g/ml$)] and then used to extract a variety of useful components from the microalgae. The optimal concentration for the antibiotic mixture was 1-3 percent. A spectrophotometric cell growth assay was also established. Astaxanthin was extracted from Haematococus lacustris with a yield of $1.9{\times}10^{-3}{\mu}g/l$ per 1 ml of culture medium. In conclusion, the axenic culture method developed here allows extraction of high-quality astaxanthin and other useful components from microalgae.

Axenic Culture of Gyrodinium impudicum Strain KG03, a Marine Red-tide Microalga that Produces Exopolysaccharide

  • Yim Joung Han;Lee Hong Kum
    • Journal of Microbiology
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    • v.42 no.4
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    • pp.305-314
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    • 2004
  • An exopolysaccharide-producing microalgal dinoflagellate was isolated from a red-tide bloom and des­ignated strain KG03. A bacteria-free culture of strain KG03 was achieved using a modified wash with phototaxis and antibiotic treatment. Combined treatment with neomycin and cephalosporin was the most effective for eliminating the bacteria associated with the microalgae. Strain KG03 was identified as Gyrodinium impudicum by analyzing the ITS regions of the 5.8S rDNA, 18S rDNA, morphological phenotype and fatty acid composition. The exopolysaccharide production and cell growth in a 300-ml photobioreactor were increased 2.7- and 2.4-fold, respectively, compared with that in a flask culture at the first isolation step.

Feeding of Bacteria by Copepod Tigriopus japonicus (Tigripus japonicus의 세균 섭이)

  • Jung, Min-Min;Rho, Sum;Kim, Pil-Yun
    • Journal of Aquaculture
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    • v.11 no.1
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    • pp.113-118
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    • 1998
  • This study was carried out to investigate feeding of bacteria by Tigripus japonicus (Copepoda : Harpacticoida) under axenic culture. The ovigerous females and nauplii were grown with feed of aquatic bacteria. Growth of RT bacteria strain was suppressed by feeding of co-existing T. japonicus. T. japonicus of non-axenic culture was observed with oil bead in the egg sac. On the other hand, early nhauplius stage did not develop to the next stage without stage took bacteria as food. And the adult of T. japonicus may utilize the baxteria as nutrient source for egg development.

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