• Title/Summary/Keyword: DGGE (denaturing gradient gel electrophoresis)

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Change of Microbial Communities in Kimchi Fermentation at Low Temperature (김치의 저온 발효 중 미생물 변화 양상)

  • 박정아;허건영;이정숙;오윤정;김보연;민태익;김치경;안종석
    • Korean Journal of Microbiology
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    • v.39 no.1
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    • pp.45-50
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    • 2003
  • The diversity and change of microbial communities during kimchi fermentation at $4^{\circ}C$ were analyzed by denaturing gradient gel electrophoresis (DGGE). Kimchi samples were taken every 5 days over the fermentation periods (for 60 days) to extract total DNA for DGGE analysis. Touchdown polymerase chain reaction was performed to amplify the V3 region of 16S rRNA gene. Sequencing results of partial 16S rDNA amplicons from DGGE profiles revealed that lactic acid bacteria (LAB), especially Weissella koreensis, Lactobacillus sakei and Leuconostoc gelidum were dominants in kimchi fermentation at $4^{\circ}C$. And we knew that W. koreensis steadily existed throughout the whole fermentation period, also Lb. sakei and Leuc. gelidum appeared from 10th day and 30th day of fermentation time, respectively and then these species were to be dominant microorganisms.

Bioremediation of Oil-Contaminated Soil Using an Oil-Degrading Rhizobacterium Rhodococcus sp.412 and Zea mays. (유류 분해 근권세균 Rhodococcus sp. 412와 옥수수를 활용한 유류 오염 토양의 정화)

  • Hong, Sun-Hwa;Park, Hae-Lim;Ko, U-Ri;Yoo, Jae-Jun;Cho, Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.35 no.2
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    • pp.150-157
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    • 2007
  • The advanced bioremediation of diesel-contaminated soil through the exploration of bacterial interaction with plants was studied. A diesel-degrading rhizobacterium, Rhodococcus sp.412, and a plant species, Zea mays, having tolerant against diesel was selected. Zea mays was seeded in uncontaminated soil or diesel-contaminated soil with or without Rhodococcus sp. 412. After cultivating for 30 days, the growth of Zea mays in the contaminated soil inoculated with Rhodococcus sp. 412 was better than that in the contaminated soil without the bacterium. The residual diesel concentrations were lowered by seeding Zea mays or inoculating Rhodococctis sp. 412. These results Indicate that the simultaneous use of Zea mays and Rhodococcus sp. 412 can give beneficial effect to the remediation of oil-contaminated soil. Bacterial community was characterized using a 16S rDNA PCR and denaturing gradient gel electrophoresis (DGGE) fingerprinting method. The similarities of DGGE fingerprints were $20.8{\sim}39.9%$ between the uncontaminated soil and diesel contaminated soil. The similarities of DGGE fingerprints were $21.9%{\sim}53.6%$ between the uncontaminated soil samples, and $31.6%{\sim}50.0%$ between the diesel-contaminated soil samples. This results indicated that the structure of bacterial community was significantly influence by diesel contamination.

The Efficacy of Water Purification and Distribution of Ammonia Oxidizing Bacteria in Shihwa Constructed Wetland (시화호 인공습지의 수질정화 및 암모니아 산화균의 분포 연구)

  • Kim, Seiyoon;Kim, Misoon;Lee, Sunghee;Lim, Miyoung;Lee, Youngmin;Kim, Zhiyeol;Ko, GwangPyo
    • Journal of Korean Society on Water Environment
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    • v.26 no.1
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    • pp.10-18
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    • 2010
  • Water quality and the distribution of ammonia oxidizing bacteria were characterized in constructed wetland of Shihwa lake. Both physico-chemical parameters and fecal indicator microorganisms including total coliforms, E.coli, Enterococcus spp. were measured. In addition, denaturant gradient gel electrophoresis (DGGE) was carried out after PCR amplification of amoA gene from input, output, and wetland sites of the Banwol, Donghwa, and Samhwa stream in Shihwa lake area. Physico-chemical parameters were in proper range for typical nitrifying bacteria to grow and perform their biological activities. Average concentrations of fecal indicator microorganisms of wetland samples were lower than those of input sites. These results suggested that microbial water quality improved by the process of constructed wetland. According to phylogenetic information obtained from DGGE from study sites, distribution of nitrifying bacteria from each of input, output, and wetland were generally distinctive one another. In addition, distribution of nitrifying bacteria between Banwol and Donghwa streams showed higher similarity (52.6%) than this of Samhwa stream (15.2%). These results indicated that characteristics of ammonia oxidizing bacteria in Samhwa were unique in comparison with those of Banwol and Donghwa stream.

Characterization of Microbial Fuel Cells Enriched Using Cr(VI)-Containing Sludge

  • Ryu, Eun-Yeon;Kim, Mi-A;Lee, Sang-Joon
    • Journal of Microbiology and Biotechnology
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    • v.21 no.2
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    • pp.187-191
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    • 2011
  • Microbial fuel cells (MFCs) were successfully enriched using sludge contaminated with Cr(VI) and their characteristics were investigated. After enrichment, the charge of the final 10 peaks was 0.51 C ${\pm}$ 1.16%, and the anodic electrode was found to be covered with a biofilm. The enriched MFCs removed 93% of 5 mg/l Cr(VI) and 61% of 25 mg/l Cr(VI). 16S rDNA DGGE profiles from the anodic electrode indicated that ${\beta}$-Proteobacteria, Actinobacteria, and Acinetobacter sp. dominated. This study is the first to report that electrochemically active and Cr(VI)-reducing bacteria could be enriched in the anode compartment of MFCs using Cr(VI)-containing sludge and demonstrates the Cr(VI) removal capability of such MFCs.

혐기성 PCE 탈염소화 관련 미생물 군집 특성

  • 이태호;문부영;박태주
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.09a
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    • pp.133-137
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    • 2004
  • Tetrachloroethylene(PCE) dechlorination was investigated in an anaerobic enrichment culture from landfill soil. Anaerobic PCE dechlorinating microorganisms could convert 150mg/L of PCE via trichloroethylene(TCE) to cir-1,2-dichloroethylene(CDCE) within 2 days at the optimum temperature of 30 to 35$^{\circ}C$. The enrichment culture could dechlorinate TCE but did not degrade other chlorinated aliphatic compounds, such as cDCE, trans-1,2-dichloroethylene, 1,1-dichloroethylene, 1,1-dichloroethane, 1,2-dichloro- ethane, and 1,1,1-trichloroethane during 5 days incubation. Several isolates from the enrichment culture did not show dechlorinating activity of PCE. Microbial analysis of the dechlorinating enrichment culture by using Polymerase chain reaction-Denaturing gradient gel electrophoresis (PCR-DGGE) method showed that at least three microorganisms were related to the anaerobic PCE dechlorination in the enrichment

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Rapid Identification of Bifidobacteria in Dairy Products by Gene-targeted Species-specific PCR Technique and DGGE

  • Hong, Wei-Shung;Chen, Ming-Ju
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.12
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    • pp.1887-1894
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    • 2007
  • In this paper, a rapid and reliable gene-targeted species-specific polymerase chain reaction (PCR) technique based on a two-step process was established to identify bifidobacteria in dairy products. The first step was the PCR assay for genus Bifidobacterium with genus specific primers followed by the second step, which identified the species level with species-specific primer mixtures. Ten specific primer pairs, designed from nucleotide sequences of the 16-23S rRNA region, were developed for the Bifidobacterium species including B. angulatum, B. animalis, B. bifidum, B. breve, B. catenulatum, B. infantis, B. longum, B. minimum, B. subtile, and B. thermophilum. This technique was applied to the identification of Bifidobacterium species isolated from 6 probiotic products, and four different Bifidobacterium spp. (B. bifidum, B. longum, B. infantis, and B. breve) were identified. The findings indicated that the 16S-23S rDNA gene-targeted species-specific PCR technique is a simple and reliable method for identification of bifidobacteria in probiotic products. PCR combined with Denaturing Gradient Gel Electrophoresis (DGGE) for identification of the bifidobacteria was also evaluated and compared with the gene-targeted species-specific technique. Results indicated that for fermented milk products consistency was found for both species-specific PCR and PCR-DGGE in detecting species. However, in some lyophilized products, the bands corresponding to these species were not visualized in the DGGE profile but the specific PCR gave a positive result.

PCR-DGGE as a Supplemental Method Verifying Dominance of Culturable Microorganisms from Activated Sludge

  • Zhou, Sheng;Wei, Chaohai;Ke, Lin;Wu, Haizhen
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1592-1596
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    • 2010
  • To verify the dominance of microorganisms in wastewater biological treatment, PCR-DGGE (denaturing gradient gel electrophoresis) was performed as a supplementary support method for screening of the dominant microorganisms from activated sludge. Results suggest that the dominant microorganisms in activated sludge are primarily responsible for strengthening its effectiveness as a biological treatment system, followed by the non-main dominant microorganisms, whereas the non-dominant microorganisms showed no effects. The degree of microbial abundance present on the profile of PCR-DGGE was in line with the treatment efficiency of augmented activated sludge with isolated cultures, suggesting that PCR-DGGE can be used as an effective supplementary method for verifying culturable dominant microorganisms in activated sludge of coking wastewater.

Study on the Performances and Microbial Community in the Biofilm Process for Treating Nonpoint Source Pollutants (비점오염물질 처리를 위한 생물막 공정의 운전 및 미생물 군집의 특성)

  • Choi, Gi-Choong;Park, Jeung-Jin;Kang, Du-Kee;Yu, Jae-Cheul;Byun, Im-Gyu;Shin, Hyun-Suk;Lee, Tae-Ho;Park, Tae-Joo
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.10
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    • pp.1021-1027
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    • 2008
  • In this study, biofilm process was introduced for treating nonpoint source pollutants. The ceramic media were provided for biofilm growth in the reactors. The packing ratio of ceramic media was 5% and 15(v/v)%, respectively. Thereafter, the reactors were operated intermittently with the different interevent periods such as 0, 5, 10 and 15 days, respectively. The removal efficiencies of COD and NH$_4{^+}$-N were investigated at the different operating conditions such as media packing ratio, temperature, and interevent period. Additionally, Polymerase chain reaction(PCR)-denaturing gel gradient electrophoresis(DGGE) and INT-dehydrogenase activity(DHA) test were conducted to observe the microbial community and activity in the biofilm. Consequently, the interevent period seemed to have no significant influence on the COD removal efficiency. COD was removed within 6$\sim$8 hours at 25$^{\circ}C$ and about 15 hours at 10$^{\circ}C$. DGGE profiles showed that the initial species of microorganisms were changed from seeded activated sludge into the microorganisms detected in sediments. INT-DHA test also showed that the activities of microorgnaisms were not decreased even in the 15 days of interevent period.

Bacterial Community Structure Shift Driven by Salinity: Analysis of DGGE Band Patterns from Freshwater to Seawater of Hyeongsan River, Korea (염도의 변화에 따른 미생물 군집의 변화: 경북 형산강 하류 미생물 군집 변화의 DGGE pattern 분석)

  • Beck, Bo Ram;Holzapfel, Wilhelm;Hwang, Cher Won;Do, Hyung Ki
    • Journal of Life Science
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    • v.23 no.3
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    • pp.406-414
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    • 2013
  • The influence of a gradual increase in salinity on the diversity of aquatic bacterial in rivers was demonstrated. The denaturing gradient gel electrophoresis (DGGE) was used to analyze the bacterial community shift downstream in the Hyeongsan River until it joins the open ocean. Four water samples were taken from the river showing the salinity gradients of 0.02%, 1.48%, 2.63%, and 3.62%. The samples were collected from four arbitrary stations in 2.91 km intervals on average, and a DGGE analysis was performed. Based on the results of this analysis, phylogenetic similarity identification, tree analysis, and a comparison of each station were performed. The results strongly suggested that the response of the bacterial community response was concomitant to gradual changes in salinity, which implies that salt concentration is a major factor in shifting the microbiota in aquatic habitats. The results also imply a huge diversity in a relatively small area upstream from the river mouth, compared to that in open oceans or coastal regions. Therefore, areas downstream towards a river mouth or delta are could be good starting points in the search for new bacterial species and strains ("biotypes").

Structure and Diversity of Arsenic-Resistant Bacteria in an Old Tin Mine Area of Thailand

  • Jareonmit, Pechrada;Sajjaphan, Kannika;Sadowsky, Michael J.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.169-178
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    • 2010
  • The microbial community structure in Thailand soils contaminated with low and high levels of arsenic was determined by denaturing gradient gel electrophoresis. Band pattern analysis indicated that the bacterial community was not significantly different in the two soils. Phylogenetic analysis obtained by excising and sequencing six bands indicated that the soils were dominated by Arthrobacter koreensis and $\beta$-Proteobacteria. Two hundred and sixty-two bacterial isolates were obtained from arsenic-contaminated soils. The majority of the As-resistant isolates were Gramnegative bacteria. MIC studies indicated that all of the tested bacteria had greater resistance to arsenate than arsenite. Some strains were capable of growing in medium containing up to 1,500 mg/l arsenite and arsenate. Correlations analysis of resistance patterns of arsenite resistance indicated that the isolated bacteria could be categorized into 13 groups, with a maximum similarity value of 100%. All strains were also evaluated for resistance to eight antibiotics. The antibiotic resistance patterns divided the strains into 100 unique groups, indicating that the strains were very diverse. Isolates from each antibiotic resistance group were characterized in more detail by using the repetitive extragenic palindromic-PCR (rep-PCR) DNA fingerprinting technique with ERIC primers. The PCR products were analyzed by agarose gel electrophoresis. The genetic relatedness of 100 bacterial fingerprints, determined by using the Pearson product-moment similarity coefficient, showed that the isolates could be divided into four clusters, with similarity values ranging from 5-99%. Although many isolates were genetically diverse, others were clonal in nature. Additionally, the arsenic-resistant isolates were examined for the presence of arsenic resistance (ars) genes by using PCR, and 30% of the isolates were found to carry an arsenate reductase encoded by the arsC gene.