• Title/Summary/Keyword: Isolation plate

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Phylogenetic diversity of bacterial communities in a gray solar saltern and isolation of extremely halophilic bacteria using culturomics (토판염전 결정지 내 세균군집의 계통학적 다양성 및 Culturomics법을 이용한 고도 호염균의 분리)

  • Cho, Geon-Yeong;Han, Song-Ih;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.53 no.1
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    • pp.29-38
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    • 2017
  • In this study, we investigated the phylogenetic diversity of the bacterial community and isolation of extremely halophilic bacteria using culturomics in a gray solar saltern. The number of bacterial living cells, enumerated in a gray solar saltern by direct fluorescence microscopy was three to four orders of magnitude greater than those enumerated by plate counts, suggesting the distribution of 'viable but non-culturable bacteria'. The biodiversity of bacterial communities in a gray solar saltern was investigated by pyrosequencing, 1,778 OTUs of bacteria were comprised of 18 phyla 46 classes 85 orders 140 families 243 genera with 6.16 diversity index. Archaea communities were composed of 3 phyla 6 classes 7 orders 7 families 38 genera with 4.95 diversity index from 643 OTUs. Totally 137 isolates were isolated by 59 different cultural methods based on culturomics considering culture media and conditions suitable for the growth of extremely halophilic bacteria. Phylogenetic analyses of extremely halophilic isolates based on 16S rRNA gene sequences, extremely halophilic isolates were composed of 4 phyla and 11 genera. Haloterrigena and Haloferax can be successfully isolated from culturomics. These culturomics were effective methods for collection of diversity of extremely halophilic bacteria.

A Study on Microbial Pollution of Indoor Air at Elderly Care Facilities (노인요양시설의 실내공기 중 미생물 오염에 관한 연구)

  • Kim, Sang-Ha;Kim, Young-Kwon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.10 no.9
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    • pp.2485-2491
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    • 2009
  • Culture was performed by using Sheep Blood Agar Plate (BAP, Asan Pharmaceutical) and Sabouraud Dextrose Ager (SDA, Asan Pharmaceutical) along with air $IDEAL^{TM}$ (Biomerieux), which is a microbe interceptor based on inertial impaction interception, in order to investigate bioaerosol in indoor and outdoor air at five elderly care facilities in a metropolis and an urban-rural consolidated city for two months from April 1 to May 31, 2007. From the culture followed by isolation and identification, the following conclusions were drawn. 1. As for the general isolation of microbes in each facility, care center S had the largest amount of microbes (263 cfu/$m^3$) isolated in a 300L room, followed by care center U having 123 cfu/$m^3$ isolated. 2. As for the number of bacteria isolated from a medium intercepting 300 L indoor, the largest amount of other unidentified or non-pathogenic Gram positive cocci (321 cfu/$m^3$) was isolated and most of the other Gram positive cocci were CNS (Coagulase Negative Staphylococcus). 3. As for the number of fungi isolated from a medium intercepting 300 L in a room, the largest number of Aspergillus spp. (66) was isolated, followed by Mucor spp. (62 cfu/$m^3$), Penicillium spp. (53 cfu/$m^3$), Alternaria spp. (50), and other unidentified or non-pathogenic fungi (42 cfu/$m^3$). 4. As for the rate of indoor and outdoor pollution, the average number of interceptions was all larger indoor than outdoor; the research differentiating the amount of air into 300 L and 500 L demonstrated that the larger amount of air led to more bacteria, making no great variation in the species.

Isolation and Characterization of Microorganisms with Broad Antifungal Activity against Phytopathogenic Fungi (식물병원균에 광범위 항균활성을 가진 미생물의 분리 및 특성)

  • Kim, Min-Hee;Ko, Hee-Sun;Yook, Young-Min;Kim, Hyun-Soo
    • KSBB Journal
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    • v.23 no.3
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    • pp.219-225
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    • 2008
  • For the production of an antifungal compound, one strain (I-8) was selected from approximately 400 strains isolated from various soil samples. The optimum carbon source, nitrogen source and pH culture conditions for the production of the antifungal compound were investigated. ISP No. 2 medium (yeast extract 0.4%, malt extract 1% and dextrose 0.4%, at pH 8) was determined to be the optimum medium. Strain I-8 showed broad antifungal activity against the plant pathogenic fungi tested, including Sclerotinia sclerotiorum KACC 41065, as well as cellulase and chitinase activities in an agar plate assay. The extraction of antifungal compounds was performed using ethyl ether and ethyl acetate. In a culture broth of strain I-8, the ethyl acetate extract exhibited effective growth inhibition against 14 of the 20 phytopathogenic fungi tested. By mixing the ethyl acetate extract from I-8 with the ethyl ether extract from the fungus 13-16, which shows specific antifungal activity against Colletotrichum orbiculare KACC 40808, the antifungal activity of I-8 against phytopathogenic fungi was confirmed to be slightly increased. Strain I-8 showed strong growth inhibition against 16 phytopathogenic strains in agar plate tests.

The Bacteriological Survey of the Main Streams in Kyong-Ju City - on Samonellae - (경주시를 관류하는 하천수에 대한 세균학적 조사 - 살모넬라에 대하여 -)

  • Che, Ik-Han
    • Journal of Preventive Medicine and Public Health
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    • v.22 no.2 s.26
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    • pp.268-275
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    • 1989
  • The studies on the standard plate count, most probable number(MPN) and isolation of the Enterobacteriaceae, especially Salmonella species from stream waters of Kyong-Ju city were carried out from June to July, 1988. The relationship between water temperature, weather and standard plate count were observed. The locations where all of the MPN was higher than 1,600 were Mun-chon bridge, Wol-song bridge, Go-hun bridge and the area adjoining the Hyong-san river. The higher the water temperature, the more the viable counts without rain. Bacteriological survey of the Salmonella species was performed with the total 80 specimens isolated from 16 areas which are the main streams in Kyong-Ju city. The author identified S strains(2.36%) of Salmonella typhi, 5 strains(2.3%) of Salmonella paratyphi, 2 strains(1.42%) of Salmonella paratyphi B, 63 strains(29.72%) of other Salmonella groups by biochemical tests and serotyping tests with slide agglutination.

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Isolation and Characterization of xylR/TMutants in Escherichia coli (대장균(大腸菌)의 xylRjT 변이주(變異株)의 분리(分離) 및 그 특성(特性))

  • Roh, Dong Hyun;Rhee, In Koo
    • Current Research on Agriculture and Life Sciences
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    • v.10
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    • pp.125-135
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    • 1992
  • Nine strains of xyl mutants that could not utilize xylose as a carbon source were isolated from E. coli JM109 by the treatment of NTG in order to investigate the regulation of xylose operon and to use recipient cells for the cloning of xylR gene. For the characterization of all isolated mutants, colony colors of all mutants on MacConkey-xylose and MacConkey-xylulose agar plate were observed for the utilization of xylose and xylulose, and the growth level and the activity of xylose isomerase and xylulokinase were determined in need. The isolated xylR/T mutants formed the white colony on MacConkey-xy-lose and MacConkey-xylulose agar plate. They did not detect the activity of xylose isomerase, and the activity of xylose isomerase was not restored in transformants of xylR/T mutant with pEX13 which contained xylA gene. xylR and xylT mutants were classified from xylR/T mutants depending upon the growth level in minimal medium. xylT mutants; DH13, DH121 and DH125 could grow a little in that medium, but xylR mutants; DH10, DH53, and DH60 could not grow that medium.

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Isolation and Characterization of Airborne Bacteria and Fungi in Indoor Environment of Elementary Schools (초등학교 실내환경에서 공기 중 세균과 진균의 분리 및 특성)

  • Kim, Na-Yeong;Kim, Young-Ran;Kim, Min-Kyu;Cho, Du-Wan;Kim, Jong-Seol
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.193-200
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    • 2007
  • Indoor airborne bacterial and fungal concentrations were examined at classrooms and corridors of 3 elementary schools in Ulsan. Airborne microorganisms were collected with an impaction-type air sampler using plate count agar and dichloran rose bengal chloramphenicol agar. During the semester, concentrations of bacteria ranged $168{\sim}3,887 MPN/m^3$ at classrooms and $168{\sim}6,339 MPN/m^3$ at corridors, while those of fungi ranged $34{\sim}389 MPN/m^3$ at classrooms and $91{\sim}507 MPN/m^3$ at corridors. The bacterial concentrations showed larger variations between situations and schools compared to those of fungi. When airborne bacteria were isolated and identified, 84% were observed as Gram-positive, and Micrococcus spp. was the most abundant group with 61% of tested isolates, followed by genus Staphylococcus with 10%. The Micrococcus spp. isolates, of which 75% were identified as M. luteus, appeared to be from human origins. The protective pigments and substantial cell wall of Micrococcus may provide selective advantage for their survival in the air. We also isolated and identified 15 genera of filamentous fungi. The most common culturable fungi were Cladosporium, Aspergillus and Penicillium, and these 3 genera were 69% of fungal isolates. Genus Stachybotrys, of which S. chartarum is a well known producer of many potent mycotoxins, was also detected from one of the schools. further systematic studies are necessary with an emphasis on species identification and mycotoxin production of isolated fungal genera, including Aspergillus, Penicillium, and Stachybotrys.

Isolation of Bacteria with Protease Activity from Cheonggukjang and Purification of Fibrinolytic Enzyme (청국장으로부터 혈전용해 활성이 우수한 균주 분리 및 혈전용해효소정제)

  • Choi, Yeon Hee;Lee, Jun Seung;Bae, So Young;Yang, Keun Jae;Yeom, Kyu Won;Jo, Dong Hyeok;Kang, Ock Hwa;Baik, Hyung Suk
    • Journal of Life Science
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    • v.23 no.2
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    • pp.259-266
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    • 2013
  • To isolate the fibrinolytic enzyme, 268 strains from 21 samples were morphologically isolated from Cheonggukjang collected from Korea and Japan. Among the 268 strains, protease-producing bacteria were isolated in nutrient agar medium including 1% skimmed milk. As a result of this, 22 strains were isolated. Apiweb site was used to identify these strains based on their biochemical properties. In addition, 16S rRNA sequencing was performed to identify the strain. Most of the identified strains were Bacillus subtilis and B. amyloliquefaciens. Fibrinolytic enzyme activity was measured with the fibrin plate method. Five strains were finally selected: A2-14, A2-20, C1-05, C1-09, and F2-01. Of those five strains, the A2-20 strain, which is close to B. amyloliquefaciens, showed the strongest fibrinolytic activity. The fibrinolytic enzyme produced by the A2-20 strain was partially purified from culture supernatant by gel filtration and ion exchange chromatography. The optimal pH and temperature values of the partially purified enzyme were 7.0 and $35^{\circ}C$, respectively. Purified protein analysis was carried out with SDS-PAGE and zymography. A genetic analysis was also conducted by PCR based on the consensus sequence of fibrinolytic enzyme. Corresponding genes with a partial sequence of the A2-20 strain were identified.

Isolation and Identification of Degradation products of Herbicide Propanil in Solution (제초제(除草劑) Propanil의 용액중(溶液中) 분해산물(分解産物)의 분리(分離) 및 동정(同定))

  • Kim, Jang Eok;Shin, Yun Gyo;Hong, Jong Uck
    • Current Research on Agriculture and Life Sciences
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    • v.5
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    • pp.27-32
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    • 1987
  • To isolate and identify degradation products of propanil in solution which propanil concentration was 2000ppm with a certain temperature, degradation products and pathway were investigated every 2 weeks for 12 weeks. Extracted mixture was developed with benzene on TLC plate, and Rf values of isolated DCA and TCAB were 0.65 and 0.94 respectively. At the GC analysis, propanil and its degradation products could seperate at the column temperature $200^{\circ}C$, but in order to more good resolution, the column temperature of DCA and TCAB was $140^{\circ}C$ and $250^{\circ}C$ respectively. Functional group of OCA was determined by IR spectrum $3400cm^{-1}$ and $800cm^{-1}$. Proton peaks of OCA were NMR spectrum $6.7{\delta}$ and $3.7{\delta}$. As the results, the major degradation products of propanil in solution were seperated on TLC plate, and thus identified by the analysis of GC, IR and NMR. Proposed degradation pathway of propanil in solution was from DCA to TCAB.

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A Study on the Taegeuk Shaped Directional Coupler with Improved Power Split Ratio (개선된 전력 분배율을 갖는 태극형 방향성 결합기에 관한 연구)

  • 양규식;오양현;이종악
    • The Proceeding of the Korean Institute of Electromagnetic Engineering and Science
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    • v.2 no.2
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    • pp.19-24
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    • 1991
  • This paper propose a new taegeuk shaped directional couper and verify the possibility of high power division rate in those directional coupler through the experiments. We took the taegeuk shaped structure in those proposed directional coupler to utilize a $3\lambda/4$ section of hybrid ring directional coupler actively, and calculated the branch admittances, which satisfied the condition of perfect matching and isolation in the center frequency, by even odd mode analyzing methodes. On the result, we knew that it can be realized a much higher power division rate than reported result in same circuit area within the producible resistance limit in the microstrip line, made the taegeuk shaped directional couplers with 0, 8, 16 dB power split ratio in the 10 GHz frequency using CGP - 502 plate, and confirmed the validity of theory through the experiments.

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Suppression of Parasitic Resonance Modes for the Millimeter-Wave SiP Applications (밀리미터파 SiP 응용을 위한 기생 공진 모드 억제)

  • Lee Young-Chul
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.17 no.9 s.112
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    • pp.883-889
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    • 2006
  • In this paper, parasitic resonance modes generated in a conductor backed coplanar wave guide(CBCPW) and stripline band pass filter(BPF) and the oscillation phenomena of a 40 GHz power amplifier module(PAM) are analyzed and several methods to suppress them are presented for low-temperature co-fired ceramic(LTCC) based millimeter-wave RF System-in-Package(SiP) applications. Parasitic rectangular wave guide(RWG) modes of the CBCPW structure are completely suppressed in the operation frequency band by decreasing the distance between its vias and by increasing the mode frequency. In the stripline structure, RWG resonance modes are clearly eliminated by removing some vias facing each other and by placing them diagonally. In the case of the 40 GHz PAM, in order to reduce a cross talk due to radiation that is generated from interconnection discontinuities, high isolation structures such as embedded DC bas lines and CPW signal lines are used and then the oscillated PAM is improved.