• Title/Summary/Keyword: 미생물 DGGE

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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.

Molecular Analysis of the Microorganisms in a Thermophilic CSTR used for Continuous Biohydrogen Production (연속수소생성에 사용되는 고온 CSTR 내의 미생물의 분자적 분석)

  • Oh, You-Kwan;Park, Sung-Hoon;Ahn, Yeong-Hee
    • KSBB Journal
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    • v.20 no.6
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    • pp.431-437
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    • 2005
  • Molecular methods were employed to investigate microorganisms in a thermophilic continuous stirred tank reactor(CSTR) used for continuous $H_2$ production. The reactor was inoculated with heat-treated anaerobic sludge and fed with a glucose-based medium. Denaturing gradient gel electrophoresis showed dynamic changes of bacterial populations in the reactor during 43 days of operation. Gas composition was constant from approximately 14 days but population shift still occurred. Populations affiliated with Fervidobactrium gondwanens and Thermoanaerobacterium thermosaccharolyticum were dominant on 21 and 41 days, respectively. Keeping pH of the medium at 5.0 could suppress methanogenic activity that was detected during initial operation period. $CH_4$ and mcrA detected in the samples obtained from the reactor or inoculum suggested the heat treatment condition employed in this study is not enough to remove methanogens in the inoculum. PCR using primer sets specific to 4 main orders of methanogens suggested that major $H_2$-consuming methanogens in the CSTR belong to the order Methanobacteriales.

Electricity Generation and Microbial Community variation in Microbial Fuel Cell with various Electrode Combinations. (다양한 탄소전극조합에 따른 미생물 연료전지의 전기발생량 및 미생물 군집변화)

  • Kwon, Jae-Hyeong;Choi, Soo-Jung;Cha, Jae-Hwan;Kim, Hyo-Soo;Kim, Ye-Jin;Yu, Jae-Cheul;Kim, Chan-Won
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.1
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    • pp.87-96
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    • 2010
  • The electrode material is one of the factors affecting the power production of microbial fuel cell. In this study, effects of carbon electrode material, thickness and configuration on the power density, biofilm formation and microbial community diversity of microbial fuel cell were investigated. To optimize the anode-cathode electrode assembly, seven lab-scale reactors which had various carbon electrode constructions were operated in continuous mode. Under the steady state condition, the electrode combination of graphite felt (6 mm) with hole showed the highest cell voltage of 238 mV and the coulombic efficiency of 37%. As a result of SEM analysis, the bacteria growing on surface of knitted type of carbon cloth and graphite felt electrode ncreased significantly. The change of dominant species between seeding sludge and biofilm on the surface of anode electrode, microbial analysis with PCR-DGGE showed that the dominant species of seeding sludge are quite different from those of biofilm on the surface of each anode electrode. Especially Geobacter sp., a well known electrochemical bacteria, was found as the dominant species of the electrode combination with graphite felt.

Microbial Communities of the Microbial Fuel Cell Using Swine Wastewater in the Enrichment Step with the Lapse of Time (가축분뇨를 이용한 미생물연료전지의 농화배양 단계에서 미생물 군집 변화)

  • Jang, Jae Kyung;Hong, Sun Hwa;Ryou, Youg Sun;Lee, Eun Young;Chang, In Seop;Kang, Young Koo;Kim, Jong Goo
    • Journal of Korean Society of Environmental Engineers
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    • v.35 no.12
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    • pp.973-977
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    • 2013
  • These studies were attempted to investigate the change of microbial community of anode of microbial fuel cell using swine wastewater in the enrichment step with the lapse of time. Microbial fuel cells enriched by a 1 : 1 mixture of anaerobic digestive juices of the sewage treatment plant and livestock wastewater. Enrichment culture step was divided into three stages to indentify the microorganisms. It was separated by each lag phase, exponential phase, and stationary phase. These steps were determined by the change of the current value. The current after enrichment was generated about $0.84{\pm}0.06mA$. We were cut out the different 17 bands in the DGGE fingerprint gel to do sequencing. The bands which the concentration was increasing or newly appearing with the lapse of time were included for this study. In the lag and exponential phase, Clostridium, Rhodocyclaceae, Bacteriodetes, and Uncultured bacterium etc. were detected. There were in the stationary phase Geobacter sp., Rhodocyclaceae, Candidatus, Nitrospira, Flavobactriaceae and uncultured bacterium etc. Geobactor among microorganisms detected in this study is known as the Electrochemically active microorganisms. It may include electrochemically active microorganisms to be considered as electrical activity microorganisms.

Microbial Diversity in the Soil Damaged by a Forest Fire (훼손된 산불토양의 미생물다양성)

  • Jung, Young-Ryul;Song, In-Geun;Kim, Jin-Yong;Lee, Sin-Geun;Kim, Young-Jun
    • Journal of the Korea Organic Resources Recycling Association
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    • v.13 no.2
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    • pp.85-90
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    • 2005
  • Changes of biochemical and genetic diversity of microbial communities in the soil damaged by a forest fire were analyzed. Soil samples were collected from Gangnung area where a forest fire was broken out in 2000. Two soil samples were from the burnt area, one from the naturally restoring soil (NS) and the other from the artificially restoring soil (AS). A normal, unaffected soil sample (US) was also included as a control. For the biochemical diversity, each sample was directly applied to the BIOLOG system, and the cluster analysis through a statistic process (SPSS) were performed. Genetic diversity was analyzed through DGGE using 16S-rDNA amplified from soil DNA. Among the samples tested, top soils of US and NS, and sub soil of NS revealed more than 70% of the similarity value in biochemical diversity. In case of genetic diversity, however, the similarity value was found to be in the range of 53% to 68% in all samples. This result indicates that the biochemical diversity is not always correlated with the genetic diversity in the analysis of microbial communities.

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Microbial Diversity during Fermentation of Sweet Paste, a Chinese Traditional Seasoning, Using PCR-Denaturing Gradient Gel Electrophoresis

  • Mao, Ping;Hu, Yuanliang;Liao, Tingting;Wang, Zhaoting;Zhao, Shumiao;Liang, Yunxiang;Hu, Yongmei
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.678-684
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    • 2017
  • The aim of this study was to elucidate the changes in the microbial community and biochemical properties of a traditional sweet paste during fermentation. PCR-denaturing gradient gel electrophoresis (DGGE) analysis showed that Aspergillus oryzae was the predominant species in the koji (the fungal mixture), and the majority of the fungi isolated belonged to two Zygosaccharomyces species in the mash. The bacterial DGGE profiles revealed the presence of Bacillus subtilis during fermentation, and Lactobacillus acidipiscis, Lactobacillus pubuzihii, Lactobacillus sp., Staphylococcus kloosi, and several uncultured bacteria were also detected in the mash after 14 days of main fermentation. Additionally, during main fermentation, amino-type nitrogen and total acid increased gradually to a maximum of $6.77{\pm}0.25g/kg$ and $19.10{\pm}0.58g/kg$ (30 days) respectively, and the concentration of reducing sugar increased to $337.41{\pm}3.99g/kg$ (7 days). The 180-day fermented sweet paste contained $261.46{\pm}19.49g/kg$ reducing sugar and its pH value remained at around 4.65. This study has used the PCR-DGGE technique to demonstrate the microbial community (including bacteria and fungi) in sweet paste and provides useful information (biochemical properties) about the assessment of the quality of sweet paste throughout fermentation.

PCR-DGGE Analysis of the Microbial Communities in Three Different Chinese "Baiyunbian" Liquor Fermentation Starters

  • Xiong, Xiaomao;Hu, Yuanliang;Yan, Nanfeng;Huang, Yingna;Peng, Nan;Liang, Yunxiang;Zhao, Shumiao
    • Journal of Microbiology and Biotechnology
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    • v.24 no.8
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    • pp.1088-1095
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    • 2014
  • A systematic investigation was performed on the bacterial, Bacillus, fungal, and yeast communities of the three types of Daqu (mechanically prepared, manually prepared, and mixed prepared) used in Baiyunbian Company by reconditioning PCR-denaturing gradient gel electrophoresis (PCR-DGGE). The DGGE results showed that the microbes in the three types of Daqu were mainly thermotolerant and thermophilic microbes, and the most dominant bacterial species were Bacillus and Virgibacillus, followed by Lactobacillus and Trichococcus. Furthermore, the dominant fungi were found to be molds, such as Rasamsonia, Penicillium, Aspergillus, and Monascus, and the dominant yeasts were Saccharomyces cerevisiae, Saccharomycopsis fibuligera, Pichia anomala, and Debaryomyces hansenii. In general, the three types of Daqu showed slight differences in microbial communities, and the Shannon indexes (H') of the manually prepared and mechanically prepared Daqu were similar. The results suggest that mechanically prepared Daqu can replace manually prepared Daqu in liquor production, and this research provides useful information for liquor production and process improvement.

Senior Thai Fecal Microbiota Comparison Between Vegetarians and Non-Vegetarians Using PCR-DGGE and Real-Time PCR

  • Ruengsomwong, Supatjaree;Korenori, Yuki;Sakamoto, Naoshige;Wannissorn, Bhusita;Nakayama, Jiro;Nitisinprasert, Sunee
    • Journal of Microbiology and Biotechnology
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    • v.24 no.8
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    • pp.1026-1033
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    • 2014
  • The fecal microbiotas were investigated in 13 healthy Thai subjects using polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE). Among the 186 DNA bands detected on the polyacrylamide gel, 37 bands were identified as representing 11 species: Bacteroides thetaiotaomicron, Bacteroides ovatus, Bacteroides uniformis, Bacteroides vulgatus, Clostridium colicanis, Eubacterium eligenes, E. rectale, Faecalibacterium prausnitzii, Megamonas funiformis, Prevotella copri, and Roseburia intestinalis, belonging mainly to the groups of Bacteroides, Prevotella, Clostridium, and F. prausnitzii. A dendrogram of the PCR-DGGE divided the subjects; vegetarians and non-vegetarians. The fecal microbiotas were also analyzed using a quantitative real-time PCR focused on Bacteroides, Bifidobacterium, Enterobacteriaceae, Clostrium coccoides-Eubacterium rectale, C. leptum, Lactobacillus, and Prevotella. The nonvegetarian and vegetarian subjects were found to have significant differences in the high abundance of the Bacteroides and Prevotella genera, respectively. No significant differences were found in the counts of Bifidabacterium, Enterobacteriaceae, C. coccoides-E. rectale group, C. leptum group, and Lactobacillus. Therefore, these findings on the microbiota of healthy Thais consuming different diets could provide helpful data for predicting the health of South East Asians with similar diets.

Microbial Community Profiling in cis- and trans-Dichloroethene Enrichment Systems Using Denaturing Gradient Gel Electrophoresis

  • Olaniran, Ademola O.;Stafford, William H.L.;Cowan, Don A.;Pillay, Dorsamy;Pillay, Balakrishna
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.560-570
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    • 2007
  • The effective and accurate assessment of the total microbial community diversity is one of the primary challenges in modem microbial ecology, especially for the detection and characterization of unculturable populations and populations with a low abundance. Accordingly, this study was undertaken to investigate the diversity of the microbial community during the biodegradation of cis- and trans-dichloroethenes in soil and wastewater enrichment cultures. Community profiling using PCR targeting the l6S rRNA gene and denaturing gradient gel electrophoresis (PCR-DGGE) revealed an alteration in the bacterial community profiles with time. Exposure to cis- and trans-dichloroethenes led to the disappearance of certain genospecies that were initially observed in the untreated samples. A cluster analysis of the bacterial DGGE community profiles at various sampling times during the degradation process indicated that the community profile became stable after day 10 of the enrichment. DNA sequencing and phylogenetic analysis of selected DGGE bands revealed that the genera Acinetobacter, Pseudomonas, Bacillus, Comamonas, and Arthrobacter, plus several other important uncultured bacterial phylotypes, dominated the enrichment cultures. Thus, the identified dominant phylotypes may play an important role in the degradation of cis- and trans-dichloroethenes.

Identification of Lactic Acid Bacteria in Galchi- and Myeolchi-Jeotgal by 16S rRNA Gene Sequencing, MALDI-TOF Mass Spectrometry, and PCR-DGGE

  • Lee, Yoonju;Cho, Youngjae;Kim, Eiseul;Kim, Hyun-Joong;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1112-1121
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    • 2018
  • Jeotgal is a Korean traditional fermented seafood with a high concentration of salt. In this study, we isolated lactic acid bacteria (LAB) from galchi (Trichiurus lepturus, hairtail) and myeolchi (Engraulis japonicas, anchovy) jeotgal on MRS agar and MRS agar containing 5% NaCl (MRS agar+5% NaCl), and identified them by using 16S rRNA gene sequencing and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as culture-dependent methods. We also performed polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) as a culture-independent method to identify bacterial communities. Five samples of galchi-jeotgal and seven samples of myeolchi-jeotgal were collected from different regions in Korea. A total of 327 and 395 colonies were isolated from the galchi- and myeolchi-jeotgal samples, respectively. 16S rRNA gene sequencing and MALDI-TOF MS revealed that the genus Pediococcus was predominant on MRS agar, and Tetragenococcus halophilus on MRS agar+5% NaCl. PCR-DGGE revealed that T. halophilus, Tetragenococcus muriaticus, and Lactobacillus sakei were predominant in both types of jeotgal. T. halophilus was detected in all samples. Even though the same species were identified by both culture-dependent and -independent methods, many species identified by the culture-dependent methods were not in the bacterial list identified by the culture-independent methods. The distribution of bacteria in galchi-jeotgal was more diverse than in myeolchi-jeotgal. The diverse LAB in galchi- and myeolchi-jeotgals can be further studied as candidates for starter cultures to produce fermented foods.