• Title/Summary/Keyword: Laboratory culture

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Comparison of Antibiotic Resistance of Blood Culture Strains and Saprophytic Isolates in the Presence of Biofilms, Formed by the Intercellular Adhesion (ica) Gene Cluster in Staphylococcus epidermidis

  • CHO BONG-GUM;KIM CHEORL-HO;LEE BOK KWON;CHO SEUNG-HAK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.728-733
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    • 2005
  • To elucidate the question of whether biofilm formed by the intercellular adhesion (ica) gene cluster has influences on antibiotic resistance in Staphylococcus epidermidis, we compared 124 skin strains with strains isolated from 50 blood cultures that cause septicemic diseases. The results revealed that the blood culture isolates were more resistant to the antibiotics tested than the saprophytic isolates. Moreover, antibiotic multiresistance was more prevalent in the clinical isolates. In the blood culture isolates, $46\%$ of the strains were resistant to three or more antibiotics, whereas only $12\%$ of the saprophytic isolates were resistant to three or more antibiotics. Interestingly, these characteristics were highly correlated with the biofilm formed by the ica gene cluster. In biofilm-producing strains, $84\%$ of the blood culture isolates and $44\%$ of the saprophytic isolates were antibiotic multiresistant, whereas only $22\%=;and\;9\%$, respectively, were antibiotic multiresistant in biofilm-nonproducing strains. Additionally, in the biofilm-producing ica-positive strains, $89\%$ of the blood culture isolates and $57\%$ of the saprophytic isolates were antibiotic multiresistant. However, the rate of the antibiotic multiresistance in the ica-negative strains was very low, thus indicating that the biofim formed by the lea gene cluster in S. epidermidis is an important pathogenic factor in association with the antibiotic multiresistance.

Combined effect of initial biomass density and nitrogen concentration on growth and astaxanthin production of Haematococcus pluvialis (Chlorophyta) in outdoor cultivation

  • Wang, Junfeng;Sommerfeld, Milton R.;Lu, Congming;Hu, Qiang
    • ALGAE
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    • v.28 no.2
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    • pp.193-202
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    • 2013
  • Nitrogen availability and cell density each affects growth and cellular astaxanthin content of Haematococcus pluvialis, but possible combined effects of these two factors on the content and productivity of astaxanthin, especially under outdoor culture conditions, is less understood. In this study, the effects of the initial biomass densities IBDs of 0.1, 0.5, 0.8, 1.5, 2.7, 3.5, and 5.0 g $L^{-1}$ DW and initial nitrogen concentrations of 0, 4.4, 8.8, and 17.6 mM nitrate on growth and cellular astaxanthin content of H. pluvialis Flotow K-0084 were investigated in outdoor glass column photobioreactors in a batch culture mode. A low IBD of 0.1 g $L^{-1}$ DW led to photo-bleaching of the culture within 1-2 days. When the IBD was 0.5 g $L^{-1}$ and above, the rate at which the increase in biomass density and the astaxanthin content on a per cell basis was higher at lower IBD. When the IBD was optimal (i.e., 0.8 g $L^{-1}$), the maximum astaxanthin content of 3.8% of DW was obtained in the absence of nitrogen, whereas the maximum astaxanthin productivity of 16.0 mg $L^{-1}\;d^{-1}$ was obtained in the same IBD culture containing 4.4 mM nitrogen. The strategies for achieving maximum Haematococcus biomass productivity and for maximum cellular astaxanthin content are discussed.

Screening of Endophytic Fungal Isolates Against Raffaelea quercus-mongolicae Causing Oak Wilt Disease in Korea

  • Nguyen, Manh Ha;Yong, Joo Hyun;Sung, Han Jung;Lee, Jong Kyu
    • Mycobiology
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    • v.48 no.6
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    • pp.484-494
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    • 2020
  • Oak wilt disease caused by Raffaelea quercus-mongolicae has emerged obviously in Korea. We selected antifungal isolates against R. quercus-mongolicae among 368 endophytic fungal isolates from different parts of oak and pine trees. The experiment was conducted in the primary and secondary screenings by dual culture test. The antifungal activity of the selected isolates was assessed in culture filtrate test based on the inhibition rates in mycelial growth, sporulation, and spore germination of oak wilt fungus. Five isolates, E089, E199, E282, E409 and E415, showed strong antifungal activity in culture filtrate test, and their antifungal activity decreased on the culture media supplemented with heated culture filtrate. Higher mycelial growth inhibitions on the unheated media were recorded in E409 (Colletotrichum acutatum), E089 (Daldinia childiae), E415 (Alternaria alternata) and E199 (Daldinia childiae) with the inhibition rates of 79.0%, 70.1%, 68.9% and 64.5%, respectively. These isolates also had the higher sporulation inhibitions on unheated media with the rates of 96.8%, 84.2%, 82.8% and 80.5%, respectively. The spore germination of the oak wilt fungus was completely inhibited by E282 (Nectria balsamea) on both unheated and heated media. These results showed that a higher number of potent antifungal isolates against oak wilt fungus was isolated from the petiole compared to the other parts. This study could contribute to the development of biological control approaches for the management of oak wilt disease caused by R. quercus-mongolicae.

Clinical Characteristics and Comparison of the Various Methods Used for the Diagnosis of the New Influenza A Pandemic in Korea (한국에서의 2009 신종 인플루엔자 A의 임상양상과 다양한 진단 방법들의 비교)

  • Kwon, Min Jung;Lee, Chang Kyu;Roh, Kyoung Ho;Nam, Myung Hyun;Yoon, Soo Young;Lim, Chae Seung;Cho, Yun Jung;Kim, Young Kee;Lee, Kap No
    • Laboratory Medicine Online
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    • v.1 no.1
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    • pp.26-34
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    • 2011
  • Background: Laboratory diagnosis of new influenza A (H1N1) is crucial for managing patients and establishing control and prevention measures. We compared the diagnostic accuracies of the real time RT-PCR (rRT-PCR) test recommended for the confirmation of the new flu and the viral culture method used conventionally for viral disease with that of the rapid antigen test (RAT). Methods: We performed RAT, R-mix culture, and real-time PCR by using 861 respiratory samples collected from December 2009 to January 2010 and evaluated the abilities of these methods to detect new influenza A. The relationship among the positive rates of RAT, grades of culture, and the cycle threshold (Ct) values of rRT-PCR was also evaluated. Results: Of the 861 patients, 308 (35.8%) were diagnosed with new influenza A. The sensitivities, specificities, positive predictive values, and negative predictive values of the tests were respectively as follows: 59.7%, 99.5%, 98.4%, and 81.6% for RAT; 93.2%, 100%, 100%, and 96.3% for R-mix culture; and 95.8%, 100%, 100%, and 97.7% for rRT-PCR. Samples with weak positive grade in culture and those with Ct values of 30-37 in rRT-PCR showed positivities as low as 25.3% and 2.3% in RAT, respectively. The hospitalization rate and death rate of the confirmed patients were 3.2% and 0.3%, respectively, and gastrointestinal symptoms were observed in 7.2% of the patients. Conclusions: R-mix culture and rRT-PCR tests showed excellent reliability in the diagnosis of new influenza A and could be very useful, especially for samples with low viral load.

Production of Citrate by Anaerobic Fungi in the Presence of Co-culture Methanogens as Revealed by 1H NMR Spectrometry

  • Cheng, Yan Fen;Jin, Wei;Mao, Sheng Yong;Zhu, Wei-Yun
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.10
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    • pp.1416-1423
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    • 2013
  • The metabolomic profile of the anaerobic fungus Piromyces sp. F1, isolated from the rumen of goats, and how this is affected by the presence of naturally associated methanogens, was analyzed by nuclear magnetic resonance spectroscopy. The major metabolites in the fungal monoculture were formate, lactate, ethanol, acetate, succinate, sugars/amino acids and ${\alpha}$-ketoglutarate, whereas the co-cultures of anaerobic fungi and associated methanogens produced citrate. This is the first report of citrate as a major metabolite of anaerobic fungi. Univariate analysis showed that the mean values of formate, lactate, ethanol, citrate, succinate and acetate in co-cultures were significantly higher than those in the fungal monoculture, while the mean values of glucose and ${\alpha}$-ketoglutarate were significantly reduced in co-cultures. Unsupervised principal components analysis revealed separation of metabolite profiles of the fungal mono-culture and co-cultures. In conclusion, the novel finding of citrate as one of the major metabolites of anaerobic fungi associated with methanogens may suggest a new yet to be identified pathway exists in co-culture. Anaerobic fungal metabolism was shifted by associated methanogens, indicating that anaerobic fungi are important providers of substrates for methanogens in the rumen and thus play a key role in ruminal methanogenesis.

Culture of Human Umbilical Vein Endothlial Cells Using 96-well Microplates and Position Effects on Cell Growth

  • Lee, Soohyun;Insook Sohn;Park, Myungjin;Park, Inchul;Youngsook Sohn;Seokil Hong;Taeboo Choe
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.3
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    • pp.207-210
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    • 2000
  • When endothelial cells isolated isolated from human umbilical venis were cultred for 6dary using 96-well microplates, the final cell density in each was fiund not to be the same although the medium composition of each well was exactly the same. Cell growth in the wells located at the periphery of a microplate was low, while that in the central area of the plate was high. A possible cause for different rate of growth was proposed as the uneven concentration of oxygen in the culture medium of each well.

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Enhancement of PVA-Degrading Enzyme Production by the Application of pH Control Strategy

  • Li, Min;Zhang, Dongxu;Du, Guocheng;Chen, Jian
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.220-225
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    • 2012
  • In batch culture for Poly(vinyl alcohol) (PVA)-degrading enzyme (PVAase) production by a mixed culture, higher pH (pH 7.5) was favorable for PVAase production at the prophase of cultivation, but lower pH (pH 7.0) was favorable at the anaphase. This situation was caused by the fact that the optimum pH for different key enzymes [PVA dehydrogenase (PVADH) and oxidized PVA hydrolase (OPH)] production is various. The activity and average specific production rate of PVADH reached the highest values at constant pH 7.5, whereas those of OPH appeared at pH 7.0. A two-stage pH control strategy was therefore developed and compared for its potential in improving PVAase production. By using this strategy, the maximal PVAase activity reached 2.05 U/ml, which increased by 15.2% and 24.2% over the fermentation at constant pH 7.5 and 7.0.

An Improved Method of Organogenesis from Cotyledon Callus of Acacia sinuata (Lour.) Merr. using Thidiazuron

  • Shahzad Anwar;Ahmad Naseem;Anis Mohammad
    • Journal of Plant Biotechnology
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    • v.8 no.1
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    • pp.15-19
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    • 2006
  • An efficient protocol for in vitro multiple shoot bud induction and plant regeneration from mature green cotyledon derived callus tissues of Acacia sinuata has been developed. Callus formation occurs at all the concentrations of thidiazuron (TDZ) in Murashige and Skoog's (MS) medium, but 0.6 ${\mu}M$ proved to be the best with maximum callus formation frequency. Supplementation of TDZ in combination with indole-acetic acid (IAA) in MS media accelerates shoot bud organogenesis in differentiating callus tissues with 60-70% conversion of shoot buds into shoot Most efficient shoot organogenesis was recorded when TDZ induced calli were subcultured at different concentrations of 6-benzyla-denine (BA). Optimum shoot bud induction and plant regeneration from callus was achieved when 0.6 ${\mu}M$ (TDZ) induced calli were subcultured at 3.0 ${\mu}M$ (BA) where $16.6{\pm}0.74$ shoots/unit callus on obtained. Rooting in in vitro differentiated shoots was achieved when transferred to medium containing different concentration of indole-3-butyric acid (IBA) in full & half strength MS medium. The well rooted plantlets were hardened and transferred to net house with 90% survival rate.

Studies on Lao-Chao Culture Filtrate for a Flavoring Agent in a Yogurt-Like Product

  • Liu, Yi-Chung;Chen, Ming-Ju;Lin, Chin-Wen
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.4
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    • pp.602-609
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    • 2002
  • Lao-chao is a traditional Chinese fermented rice product with a sweet and fruity flavor, containing high levels of glucose, a little alcohol and milk-clotting characteristics. In order to optimize commercial production of lao-chao, Rhizopus javanicus and Saccharomyces cerevisiae were selected as the mold and yeast starter, respectively. A commercial mixed starter (chiu-yao) was used as control. Fermentation of the experimental combination revealed a sharp drop in pH (to 4.5) on the fourth day, remaining constant thereafter. Content of reducing sugars gradually decreased throughout the entire fermentation period. Of the free amino acids, higher quantities of alanine, leucine, proline, glutamic acid, glutamine and $NH_3$ were noted. For sugars, glucose revealed the highest concentration, while organic acid levels, including those for oxalic, lactic, citric and pyroglutamic acid, increased throughout the fermentation period. Twenty-one compounds were identified by gas chromatography from aroma concentrates of the lao-chao culture filtrate, prepared using the headspace method. For the flavor components, higher quantities of ethanol, fusel oil and ester were determined in both culture filtrates. In regard to the evaluation of yogurt-like product, there were significant differences in alcoholic smell, texture and curd firmness.

Development of a tide-simulating apparatus for macroalgae

  • Kim, Jang-K.;Yarish, Charles
    • ALGAE
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    • v.25 no.1
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    • pp.37-44
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    • 2010
  • A tide-simulating apparatus was developed for culturing marine macroalgae. The objective of this study was to introduce a novel tide-simulating apparatus that can simulate a diurnal or semi-diurnal tidal cycle in the laboratory. In this apparatus, the seaweeds are move up and down and the water level remains the same during the simulated tidal cycle. The apparatus consists of 18 cylindrical culture tanks (3 blocks $\times$ 6 culture tanks) with 12 cm diameter and 24.5 cm long containing up to 2.5 L of seawater. There is a horizontal plate which covered all 18 culture tanks, and it is raised and lowered by a programmable motor that can regulate exposure time. In one application, seaweeds are attached to braided twine hung on Plexiglas air-tubing. The air-tubing is attached to a lid that is set on a horizontal plate. This apparatus is made of colorless Plexiglas to maximize light transmittance. This apparatus is easily disassembled and transportable to any indoor laboratory, wet laboratory, greenhouse, etc. This apparatus also offers considerable flexibility in terms of design. The size of culture tank can be redesigned by either increasing the height of cylinder or/and using a different diameter of cylindrical Plexiglas, therefore, larger/taller thalli can be cultivated. Growth rates of three eulittoral Porphyra species from different tidal elevations have been compared using this device.