• Title/Summary/Keyword: uncultured bacteria

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Analysis of Attached Bacterial Communities of Biological Activated Carbon Process Using DGGE Method (DGGE 기법을 이용한 생물활성탄 공정의 부착 박테리아 군집분석)

  • Son, Hee-Jong;Choi, Jin-Taek;Son, Hyeng-Sik;Lee, Sang-Joon
    • Journal of Korean Society of Environmental Engineers
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    • v.34 no.8
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    • pp.533-540
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    • 2012
  • The concentration of organic compounds was analyzed at each step of BAC (biological activated carbon) process though BDOC (biodegradable dissolved organic carbon) total/rapid/slow. Further, bacteria communities and biomass concentrations measured DGGE (denaturing gradirnt gel electrophoresis) and ATP (adenosine triphosphate) methods were analyzed. The bed volume of steady state is different based on assessment of organic compounds removal. Bed volumes at steady state in DOC, $BDOC_{rapid}$ and $BDOC_{total/slow}$ removal were around 27,500, 15,000 and 32,000, respectively. A biomass didn't change after the bed volume reached 22,500 according to analyzing HPC (heterotrophic plate count) and ATP concentration of bacteria. The concentration of HPC and ATP were $3.3{\times}10^8$ cells/g and $2.14{\mu}g/g$, respectively. The number of the DGGE band were only 5 at the bed volume 8,916, but increased up to 11 at the bed volume 49,632. As operation time increase, bacterial group were more diversity. Four bacteria species including Pseudomonas fluorescens, the uncultured bacterium similar to Acinetobacteria, uncultured Novosphingobium sp. and Flavobacterium frigidarium have detected from the early stages and Proteobacteria group were dominantly detected.

Culture-Based and Denaturing Gradient Gel Electrophoresis Analysis of the Bacterial Community Structure from the Intestinal Tracts of Earthworms (Eisenia fetida)

  • Hong, Sung-Wook;Kim, In-Su;Lee, Ju-Sam;Chung, Kun-Sub
    • Journal of Microbiology and Biotechnology
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    • v.21 no.9
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    • pp.885-892
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    • 2011
  • The bacterial communities in the intestinal tracts of earthworm were investigated by culture-dependent and -independent approaches. In total, 72 and 55 pure cultures were isolated from the intestinal tracts of earthworms under aerobic and anaerobic conditions, respectively. Aerobic bacteria were classified as Aeromonas (40%), Bacillus (37%), Photobacterium (10%), Pseudomonas (7%), and Shewanella (6%). Anaerobic bacteria were classified as Aeromonas (52%), Bacillus (27%), Shewanella (12%), Paenibacillus (5%), Clostridium (2%), and Cellulosimicrobium (2%). The dominant microorganisms were Aeromonas and Bacillus species under both aerobic and anaerobic conditions. In all, 39 DNA fragments were identified by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis. Aeromonas sp. was the dominant microorganism in feeds, intestinal tracts, and casts of earthworms. The DGGE band intensity of Aeromonas from feeds, intestinal tracts, and casts of earthworms was 12.8%, 14.7%, and 15.1%, respectively. The other strains identified were Bacillus, Clostridium, Enterobacter, Photobacterium, Pseudomonas, Shewanella, Streptomyces, uncultured Chloroflexi bacterium, and uncultured bacterium. These results suggest that PCR-DGGE analysis was more efficient than the culturedependent approach for the investigation of bacterial diversity and the identification of unculturable microorganisms.

Bacterial Community Diversity Associated with Two Marine Sponges from the South Pacific Ocean based on 16S rDNA-DGGE analysis (남태평양에 서식하는 두 종의 해면 Hyrtios sp.와 Callyspongia sp.의 공생세균 군집의 다양성)

  • Park, Jin-Sook
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.255-261
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    • 2010
  • The bacterial community structure associated with two marine sponges, Hyrtios sp. 604 and Callyspongia sp. 612 collected from the South Pacific Ocean were analyzed by 16S rDNA-denaturing gradient gel electrophoresis (DGGE). The phylogenetic analysis showed that the bacterial community associated with Hyrtios sp. 604 contained diverse bacterial groups such as Chloroflexi, Firmicutes, Cyanobacteria, Alphaproteobacteria, Gammaproteobacteria, Actinobacteria, and Acidobacteria. Callyspongia sp. 612 harbored Chloroflexi, Cyanobacteria, Alphaproteobacteria, and Gammaproteobacteria. Hyrtios sp. 604 belonging to genus Hyrtios known to produce natural products showed greater bacterial diversity than Callyspongia sp. 612. Phylum Actinobacteria was shown to be one of dominant bacterial groups in Hyrtios sp. 604. Although the same phyla of bacteria were found in both sponge species, the spongeassociated predominant bacterial groups differed between the two sponges with different chemical characteristics from the same geographical location. Uncultured bacteria represented over 90% of the bacteria diversity present in all bacterial communities of the sponges.

Sustainable Use of Marine Microorganisms

  • Lee Yoo Kyung;Lee Jung Hyun;Kwon Kae Kyoung;Lee Hong Kum
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2002.10a
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    • pp.94-99
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    • 2002
  • The oceans cover about $71\%$ of the Earth's crust and contain nearly 300,000 described species. Free-living bacteria in the sea and symbiotic bacteria of marine invertebrates are proving to be valuable sources of useful bioactive compounds. Marine sponges, in particular, which contain diverse communities of bacteria, produce many classes of compounds that are unique to the marine environment. Uncultured microorganisms are commonly believed to represent $99.9\%$ of the whole microbial community. They have been investigated for the possibility of isolating and over-expressing genes in viable microorganisms. Strict symbiotic species that have been adapted to the host are candidate unculturable species. With the enormous potential for discovery, development, and market value of marine derived compounds, supply of the products is a major limiting factor for further development.

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Characterization of the Microbial Diversity in a Korean Solar Saltern by 16S rRNA Gene Analysis

  • Park, Soo-Je;Kang, Cheol-Hee;Rhee, Sung-Keun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1640-1645
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    • 2006
  • We studied the diversity of the halophilic archaea and bacteria in crystallizer ponds of a Korean solar saltern by analyzing 16S rRNA gene libraries. Although diverse halophilic archaeal lineages were detected, the majority (56%) were affiliated with the uncultured and cultured Halorubrum group. Halophilic archaea that have been frequently observed in solar saltern environments previously, such as Halogeometricum, Halococcus, Haloarcula, and Haloferax, were not detected in our samples. The majority of clones (53%) belonged to the Cytophaga-Flavobacterium-Bacteroides and ${\alpha}-,\;{\gamma}-,\;and\;{\delta}-Proteobacteria$ groups, with 47% of the clones being affiliated with ${\gamma}-Proteobacteria$. We also identified new ${\delta}-Proteobacteria$-related bacteria that have not been observed in hypersaline environments previously. Our data show that the diversity of the halophilic archaea and bacteria in our Korean saltern differs from that of solar salterns found in other geographic locations. We also showed by quantitative real-time PCR analysis that bacteria can form a significant component of the microbial community in solar salterns.

Physicochemical Properties and Microbial Analysis of Korean Solar Salt and Flower of Salt (한국산 꽃소금과 천일염의 이화학적 특성 및 미생물 분석)

  • Lee, Hye Mi;Lee, Woo Kyoung;Jin, Jung Hyun;Kim, In Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1115-1124
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    • 2013
  • The present study was conducted to ensure the diversity of domestic solar salt by analyzing the composition and microbiological characteristics of solar salt (from Docho island: DS) and the flower of salt produced in different Korean salt flats (Sinui island: SF, Bigum island: BF, and Docho island: DF). The analyses showed that the moisture content of the three types of flower of salt and solar salt ranged from 10.54~13.82% and NaCl content ranged from 78.81~84.61%. The mineral content of those salts ranged from 3.57~5.51%. The content of insoluble matter in these salts was $0.01{\pm}0.00{\sim}0.05{\pm}0.00%$. The sand content of these salts was $0.01{\pm}0.01{\sim}0.03{\pm}0.01%$. By Hunter's color value analysis, the color of the flower of salt was brighter and whiter than solar salt. The salinity of the flower of salt was a little higher than solar salt as well. The magnesium and potassium ion content of DF was $9,886.72{\pm}104.78mg/kg$ and $2,975.23{\pm}79.73mg/kg$, respectively, which was lower than the content in SF, BF, and DS. The heavy metal content of all salts was acceptable under the Korean Food Sanitation Law. The flower of salt was confirmed to be sweeter and preferable to solar salt. More than 80% of the solar salt crystals were 2~3 mm in size, whereas crystals from the flower of salt were 0.5~2 mm in size. The bacterial diversity of DF and DS were investigated by culture and denaturing gradient gel electrophoresis (DGGE) methods. The number of cultured bacteria in flower of salt was approximately three times more than solar salt. By DGGE analysis, major microbes of DF were Maritimibacter sp., Cupriavidus sp., and unculturable bacteria, and those of DS were Cupriavidus sp., Dunalidella salina and unculturable bacteria. The results of DGGE analysis showed that major microorganisms in solar salts were composed of unidentified and unculturable bacteria and only a few microorganisms were culturable.

Review and Future Development of New Culture Methods for Unculturable Soil Bacteria (난배양성 토양세균을 위한 신배양기술의 고찰과 향후 발전 방향)

  • Kim, Jai-Soo
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.179-187
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    • 2011
  • This review describes the characteristics of various unculturable soil bacteria, successfully-cultivating examples of those bacteria, and the diverse factors to be considered for successful cultivation. Most importantly, the selection of proper media is very important because unculturable bacteria demand different types of nutrients at various concentrations of substrates, nitrogens and phosphorus. To develop a new medium to successfully culture unculturable bacteria from soil, molecular ecological studies should be combined together. The inoculum size on a plate is also important: less than 50 bacterial cells are recommended to be plated on a single culture plate. The environmental factors such as pH and salt concentration of the medium need to be adjusted as similar as possible to mimic the original soil environments, and the trial of the various temperatures and extended period of cultivation are better. Since one cannot simply tell about which one was unculturable among a great number of colonies grown on a newly developed medium, some suitable detection methods and fast identification methods are required. Many soil bacteria live with cooperation one another in their communities, so that enrichment such as coculture of using other bacterial metabolites and subsequent pure cultures can also guarantee successful cultivation of the previously uncultured bacteria in soil. Here, this review will discuss for the future perspectives to culture the unculturable soil bacteria.

Impact of Virus-resistant Trigonal Cactus Cultivation on Soil Microbial Community (바이러스저항성 삼각주 재배가 토양 미생물상에 미치는 영향)

  • Oh, Sung-Dug;Kim, Jong-Bum;Lee, Jung-Jin;Kim, Min-Kyeong;Ahn, Byung-Ohg;Sohn, Soo-In;Park, Jong-Sug;Ryu, Tae-Hun;Cho, Hyun-Suk;Lee, Kijong
    • Korean Journal of Environmental Agriculture
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    • v.32 no.2
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    • pp.148-154
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    • 2013
  • BACKGROUND: Genetically modified(GM) trigonal cactus(Hylocereus trigonus Saff.) contained a coat protein gene of cactus virus X (CVX), which conferred resistance to the virus, phosphinothricin acetyltransferase (bar) gene, which conferred herbicide resistance, and a cauliflower mosaic virus 35S promoter (CaMV 35S). This study was conducted to evaluate the possible impact of GM trigonal cactus cultivation on the soil microbial community. METHODS AND RESULTS: Microorganisms were isolated from the rhizosphere of GM and non-GM trigonal cactus cultivation soils. The total numbers of bacteria, and actinomycete in the rhizosphere soils cultivated GM and non-GM trigonal cactus were similar to each other, and there was no significant difference. Dominant bacterial phyla in the rhizosphere soils cultivated with GM and non-GM trigonal cactus were Proteobacteria, Uncultured archaeon, and Uncultured bacterium. The denaturing gradient gel electrophoresis (DGGE) profiles show a similar patterns, significant difference was not observed in each other. DNA was isolated from soil cultivated GM and non-GM trigonal cactus, we analyzed the persistence of the inserted gene by PCR. Amplification of the inserted genes was not observed in the soil DNA, which was collected after harvest. CONCLUSION(S): This result suggests that the GM trigonal cactus cultivation does not change significantly the microbial community.

Electricity Generation and Microbial Community Structure Variation Depending on Separator Types and Cathode Characteristics in Air-cathode MFC (공기환원전극 미생물연료전지에서 분리막 종류 및 환원전극 특성에 따른 전기발생 및 미생물 군집구조 변화)

  • Yu, Jae-Cheul;Lee, Chang-Yeol;Kim, Sun-Ah;Cho, Hae-In;Cho, Sun-Ja;Lee, Tae-Ho
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.2
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    • pp.113-120
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    • 2010
  • Air-cathode microbial fuel cell consisted of 4 unit cells were operated under batch condition and electricity generation and microbial community structure variation were investigated, depending on separator types and cathode characteristics: A) PEM(Proton Exchange Membrane)-30% Wet proofing Carbon Cloth(WC), B) AEM(Anion Exchange Membrane-WC, C) CEM(Cation Exchange Membrane)-WC, D) PEM-No Wet proofing Carbon Cloth(NC). Maximum power densities of PEM-WC, AEM-WC and CEM-WC were 510.9, 522.1 and 504.8 $mW/m^2$, respectively. But PEM-NC showed relatively lower maximum power density of 218.3 $mW/m^2$. And PEM-WC, AEM-WC and CEM-WC showed similar internal resistances(20.0-28.2 ${\Omega}$). PCRDGGE, PCA and diversity indices showed that uncultured bacteria which reported in previous MFC studies were detected in suspended growth bacteria and attached growth bacteria would be affected not by separator type but by cathode characteristic. Thus, cathode characteristic can be one of the critical factors for power generation in air-cathode MFC using PEM, AEM, and CEM as separator.

Investigation of the Properties of Sand Tubules, a Biomineralization Product, and their Microbial Community

  • Hu, Weilian;Dai, Dehui
    • Journal of Microbiology and Biotechnology
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    • v.26 no.2
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    • pp.364-372
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    • 2016
  • Sand tubules, made up of sand grains cemented by microbe-induced calcium carbonate precipitation, have been found in China's Ningxia Province. Sand tubules grow like a tree's roots about 40-60 cm below the surface. The properties of sand tubules and their bacterial community were examined. X-Ray diffraction analysis revealed that the sand tubules were associated with crystalline calcite. Scanning electron microscopy showed that the crystalline solid had a lamellar structure and lacked the presence of cells, suggesting that no bacteria acted as nucleation sites, nor that the crystalline solid was formed by the aggregation of bacteria. Denaturing gradient gel electrophoresis analysis showed 11 of the 12 detectable bands were uncultured bacteria by BLAST analysis in the GenBank database, and the rest were closely related to Paenibacillus sp. (100% identity). By cultivation techniques, the only strain isolated from the sand tubule was suggested to be related to Paenibacillus sp.; no archaea were found. Furthermore, Paenibacillus sp. was demonstrated to induce calcium carbonate precipitation in vitro.