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

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Microbial population dynamics in constructed wetlands: Review of recent advancements for wastewater treatment

  • Rajan, Rajitha J.;Sudarsan, J.S.;Nithiyanantham, S.
    • Environmental Engineering Research
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    • v.24 no.2
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    • pp.181-190
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    • 2019
  • Constructed wetlands are improvised man-made systems, designed for adopting the principle of natural wetlands for purifying wastewater - the elixir of life. They are used widely as a cost-effective and energy-efficient solution for treating greywater generated from different tertiary treatment sources. It provides an elaborate platform for research activities in an attempt to recycle earth's natural resources. Among the several organic impurities removal mechanisms existing in constructed wetland systems, the earth's active microbial population plays a vital role. This review deals with the recent advancements in constructed wetland systems from a microbiological perspective to (effect/ devise/ formulate) chemical and physical treatment for water impurities. It focuses on microbial diversity studies in constructed wetlands, influence of wetland media on microbial diversity and wetland performance, role of specific microbes in water reuse, removal of trace elements, some heavy metals and antibiotics in constructed wetlands. The impurities removal processes in constructed wetlands is achieved by combined interactive systems such as selected plant species, nature of substrate used for microbial diversity and several biogeochemical effected reaction cycles in wetland systems. Therefore, the correlation studies that have been conducted by earlier researchers in microbial diversity in wetlands are addressed herewith.

Effects of Pb and CO2 on Soil Microbial Community Associated with Pinus densiflora-Lab (소나무(Pinus densiflora) 생육토양의 미생물 군집에 미치는 납과 CO2의 영향)

  • Hong, Sun-Hwa;Kim, Sung-Hyun;Kang, Ho-Jeong;Ryu, Hee-Wook;Lee, Sang-Don;Lee, In-Sook;Cho, Kyung-Suk
    • Journal of Ecology and Environment
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    • v.29 no.6
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    • pp.551-558
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    • 2006
  • Effects of Pb and $CO_2$ on soil microbial community associated with Pinus densiflora were investigated using community level physiological profiles (CLPP) and 16S rDNA PCR-denaturing gradient gel electrophoresis (DGGE) methods. Two-years pine trees were planted in Pb-contaminated soils and uncontaminated soils, and cultivated for 3 months in the growth chamber where $CO_2$ concentration was controlled at 380 or 760 ppmv. The structure of microbial community was analyzed in 6 kinds of soil samples (CA-0M : $CO_2$ 380 ppmv + Pb 0 mg/kg + initial, CB-0M : $CO_2$ 380 ppmv + Pb 500 mg/kg + initial, CA-3M : $CO_2$ 380 ppmv + Pb 0 mg/kg + after 3 months, CB-3M : $CO_2$ 380 ppmv + Pb 500 mglkg + after 3 months, EA-3M : $CO_2$ 760 ppmv + Pb 0 mg/kg + after 3 months, EB-3M : $CO_2$ 760 ppmv + Pb 500 mg/kg + after 3 months). After 3 months, the substrate utilization in the uncontaminated soil samples (CA-3M vs EA-3M) was not significantly influenced by $CO_2$ concentrations. However, the substrate utilization in the Pb-contaminated soil samples (CB-3M vs EB-3M) was enhanced by the elevated $CO_2$ concentrations. The results of principal component analysis based on substrate utilization activities showed that the structure of microbial community structure in each soil sample was grouped by Pb-contamination. The similarities of DGGE fingerprints were 56.3 % between the uncontaminated soil samples (CA-3M vs EA-3M), and 71.4% between the Pb-contaminated soil samples (CB-3M vs. EB-3M). The similarities between the soil samples under $CO_2$ 380 ppmv (CA-3M vs CB-3M) and $CO_2$, 760 ppmv (EA-3M vs EB-3M) were 53.3% and 35.8%, respectively. These results suggested that the structure of microbial community associated with Pinus densiflora were sensitively specialized by Pb-contamination rather than $CO_2$ concentration.

Phylogenetic diversity of bacterial community associated with the tropical marine sponges, Cinachyrella sp. and Plakortis sp. (열대 해양 해면 Cinachyrella sp.와 Plakortis sp.의 공생세균 군집의 계통학적 다양성)

  • Jeong, Jong-Bin;Park, Jin-Sook
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.31-38
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    • 2015
  • The bacterial community structures of two marine sponges, Cinachyrella sp. and Plakortis sp., collected from Chuuk in the South Pacific in February 2012 were analyzed by PCR-DGGE (Denaturing Gradient Gel Electrophoresis) fingerprinting. After isolation of the total genomic DNAs from the sponges, the V3 regions of the 16S rRNA genes were amplified and subjected to DGGE profiling. The two species of sponges displayed different DGGE band patterns. The sequences derived from the DGGE bands revealed 85-100% similarities to known bacterial species in the public database. The bacterial community of Cinachyrella sp. was composed of 6 classes: Actinobacteria, Bacteroidetes, Chloroflexi, and Proteobacteria (Alpha-, Gamma-, Delta-). The bacterial community of Plakortis sp. included 7 classes: Actinobacteria, Chloroflexi, Firmicutes, Spirochaetes, and Proteobacteria (Alpha-, Gamma-, Delta-). Though Actinobacteria, Chloroflexi and Proteobacteria were commonly found in both sponges, the predominant bacterial communities differed between the two. Namely, the predominant bacterial groups in Cinachyrella sp. and Plakortis sp. were Proteobacteria and Chloroflexi, respectively. The sponge-associated bacteria are sponge host-specific, as each of the tested sponges from the same geographical location had different predominant bacterial diversity.

Chemical and Microbiological Hazard Analysis of Floor-Settled Dust in Elementary School Classrooms in Korea (국내 일부 초등학교 바닥먼지 내 화학적 및 생물학적 유해인자의 분석)

  • Kim, Ji-Young;Jung, Se-Young;Kim, Su-Jung;Kim, Jin-Ah;Si, Ji-Yeon;Jo, Yeon-Woo;Zoh, Kyung-Duk;Ko, Gwang-Pyo
    • Journal of Environmental Health Sciences
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    • v.37 no.4
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    • pp.279-288
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    • 2011
  • Objectives: This study investigated the levels and components of floor-settled dust in two elementary schools located at different sites (one near the Shihwa industrial complex and the other in a rural area) in order to evaluate the amounts of trace metal elements (As, Cd, Co, Cr, Cu, Ni, Pb and Zn) and microorganisms. Methods: Over twenty settled-dust samples were collected from the two elementary schools. Trace metal elements were extracted from the dust using hydrochloric acid and nitric acid, and the amounts were measured by ICP-OES. Microbiological analysis was performed by bacterial culturing using R2A medium and denaturing gradient gel electrophoresis (DGGE). Results: The results showed that the amounts of three metal elements (Cr, Pb, and Zn) were significantly different between the schools (${\alpha}$=0.05, p<0.04). In addition, microbial communities in each school were highly correlated with one another. Among the identified microorganisms, a number of potentially opportunistic microorganisms, including antibiotic-resistant bacteria such as Acinetobacter baumannii, were found. Conclusions: This study will provide preliminary data for assessing levels and types of chemical and microbiological agents in elementary schools and for further evaluating human health risks associated with the agents.

Analysis of Bacterials Community Structure in Leadchate-Contaminated Groundwater using Denaturing Gradient Gel Electrophoresis (Denaturing Gradient Gel Electrophoresis를 이용한 매립지 침출수로 오염된 지하수의 세균 군집 분석)

  • Kim Jai-Soo;Kim Ji-Young;Koo So-Yeon;Ko Kyung-Seok;Lee Sang-Don;Cho Kyung-Suk;Koh Dong-Chan
    • Microbiology and Biotechnology Letters
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    • v.34 no.2
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    • pp.166-173
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    • 2006
  • This research has been performed to clarify the relationship between hydrogeochemistry and bacterial community structure in groundwater contaminated with landfill leachate. We collected and analyzed samples from 5 sites such as leachate (KSG1-12), treated leachate (KSG1-16), two contaminated groundwaters (KSG1-07 and KSG1-08) and non-contaminated groundwater (KSG1-13). pH was 8.83, 8.04, 6.87, 6.87 and 6.53 in order; redox potential (Eh) 108, 202, 47, 200 and 154 mV; electric conductivity (EC) 3710, 894, 1223, 559 and 169.9 $\mu$S/cm; suspended solids (SS) 86.45, 13.74, 4.18, 0.24 and 11.91 mg/L. In KSG01-12, the ion concentrations were higher especially in $Cl^-$ and $HCO_3^-$ than other sites. The concentrations of Fe, Mn and $SO_4^{2-}$ were higher In KSG1-07 than in KSG1-08, and vise versa in $NO_3^{2-}$. In the comparison of DGGE fingerprint patterns, the similarity was highest between KSG1-13 and KSG1-16 (57.2%), probably due to common properties like low or none contaminant concentrations. Otherwise KSG1-08 showed lowest similarities with KSG1-13 (25.8%) and KSG1-12 (27.6%), maybe because of the degree of contamination. The most dominant bacterial species in each site were involved in $\alpha$-Proteobacteria (55.6%) in KSG1-12, $\gamma$-Proteobacteria (50.0%) in KSG1-16, $\beta$-Proteobacteria (66.7%) in KSG1-07, $\gamma$-Proteobacteria (54.5%) in KSG1-08 and $\beta$-Proteobacteria (36.4%) in KSG1-13. These results indicate that the microbial community structure might be changed according to the flow of leachate in grounderwater, implying changes in concentrations of pollutants, available electron accepters and/or other environmental conditions.

Changes of Microbial Community Structure According to a Changes of Season and Influent Characteristics in Biological Wastewater Treatment (생물학적 폐수처리 공정에서의 계절 및 유입수 성상 변화에 따른 미생물 군집 특성 변화)

  • Son, Hyeng-Sik;Son, Hee-Jong;Kim, Mi-A;Ryu, Eun-Yeon;Lee, Geon;Lee, Sang-Joon
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.8
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    • pp.780-786
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    • 2010
  • The bacterial community structure in biological reactor in wastewater treatment system was investigated by denaturing gradient gel electrophoresis (DGGE) and fluorescent in situ hybridization (FISH). Samples were collected at different three points in wastewater treatment system. Through treatment processes, BOD (biochemical oxygen demand) and COD (chemical oxygen demand) of was removal efficiency was 83.1~98.6%, 67.2~85.2% respectively. Microbial community of aerobic tank and oxic tank were similar but anoxic tank was different (RRP group was increased about tripple) by DGGE and FISH in sludge (2007 October and 2008 January). Samples in 2007 October and 2008 January were dominant ${\alpha}$-Proteobacteria and CF group respectively. Sludge in 2008 April were different comparing former results dominant others as 65~80%. Others group was dominant. Eubacteria by FISH with the probe EUB338 was about $1.7{\sim}7.6{\times}10^9\;cells/mL$. It could be successfully observed bacterial community in biological wastewater system.

Bacterial Communities of Biofilms Sampled from Seepage Groundwater Contaminated with Petroleum Oil

  • CHO WONSIL;LEE EUN-HEE;SHIM EUN-HWA;KIM JAISOO;RYU HEE WOOK;CHO KYUNG-SUK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.952-964
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    • 2005
  • The diesel-degrading activities of biofilms sampled from petroleum-contaminated groundwaters in urban subway drainage systems were examined in liquid cultures, and the microbial populations of the biofilms were characterized by denaturing gel gradient electrophoresis (DGGE) and 16S rDNA sequence analysis. Biofilm samples derived from two sites (19 K and 20 K) at subway Station N and Station I could degrade around $80\%$ of applied diesel within 20 and 40 days, respectively, at $15^{\circ}C$, and these results were strongly correlated with the growth patterns of the biofilms. The closest phylogenetic neighbor of a dominant component in the 19 K biofilm was Thiothrix fructosivorans strain Q ($100\%$ similarity). Four dominant strains in the 20 K biofilm were closely related to Thiothrix fructosivorans strain Q ($100\%$ similarity), Thiothrix sp. CC-5 ($100\%$ similarity), Sphaerotilus sp. IF14 ($99\%$ similarity), and Cytophaga-Flexibacter-Bacterioides (CFB) group bacterium RW262 ($98\%$ similarity). Three dominant members in the Station I biofilms were very similar to uncultured Cytophagales clone CRE-PA82 ($91\%$ similarity), Pseudomonas sp. WDL5 ($97\%$ similarity), and uncultured CFB group bacterium LCK-64 ($94\%$ similarity). The microbial components of the biofilms differed depending on the sampling site. This is the first report on the isolation of clones highly similar to Thiothrix fructosivorans and Thiothrix sp. from biofilms in petroleum-polluted groundwaters, and the first evidence that these organisms may play major roles in petroleum degradation and/or biofilm-development.

Evaluation of Biogas Production Performance and Dynamics of the Microbial Community in Different Straws

  • Li, Xue;Liu, Yan-Hua;Zhang, Xin;Ge, Chang-Ming;Piao, Ren-Zhe;Wang, Wei-Dong;Cui, Zong-Jun;Zhao, Hong-Yan
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.524-534
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    • 2017
  • The development and utilization of crop straw biogas resources can effectively alleviate the shortage of energy, environmental pollution, and other issues. This study performed a continuous batch test at $35^{\circ}C$ to assess the methane production potential and volatile organic acid contents using the modified Gompertz equation. Illumina MiSeq platform sequencing, which is a sequencing method based on sequencing-by-synthesis, was used to compare the archaeal community diversity, and denaturing gradient gel electrophoresis (DGGE) was used to analyze the bacterial community diversity in rice straw, dry maize straw, silage maize straw, and tobacco straw. The results showed that cumulative gas production values for silage maize straw, rice straw, dry maize straw, and tobacco straw were 4,870, 4,032.5, 3,907.5, and $3,628.3ml/g{\cdot}VS$, respectively, after 24 days. Maximum daily gas production values of silage maize straw and rice straw were 1,025 and $904.17ml/g{\cdot}VS$, respectively, followed by tobacco straw and dry maize straw. The methane content of all four kinds of straws was > 60%, particularly that of silage maize straw, which peaked at 67.3%. Biogas production from the four kinds of straw was in the order silage maize straw > rice straw > dry maize straw > tobacco straw, and the values were 1,166.7, 1,048.4, 890, and $637.4ml/g{\cdot}VS$, respectively. The microbial community analysis showed that metabolism was mainly carried out by acetate-utilizing methanogens, and that Methanosarcina was the dominant archaeal genus in the four kinds of straw, and the DGGE bands belonged to the phyla Firmicutes, Bacteroidetes, and Chloroflexi. Silage maize is useful for biogas production because it contains four kinds of straw.

Analysis of Bacterial Diversity and Community Structure in Forest Soils Contaminated with Fuel Hydrocarbon

  • Ahn Jae-Hyung;Kim Mi-Soon;Kim Min-Cheol;Lim Jong-Sung;Lee Goon-Taek;Yun Jun-Ki;Kim Tae-Sung;Kim Tae-San;Ka Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.16 no.5
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    • pp.704-715
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    • 2006
  • Oil spill was found in 1999 from a diesel storage facility located near the top of Baekun Mountain in Uiwang City. Application of bioremediation techniques was very relevant in removing oil spills in this site, because the geological condition was not amenable for other onsite remediation techniques. For efficient bioremediation, bacterial communities of the contaminated site and the uncontaminated control site were compared using both molecular and cultivation techniques. Soil bacterial populations were observed to be stimulated to grow in the soils contaminated with diesel hydrocarbon, whereas fungal and actinomycetes populations were decreased by diesel contamination. Most of the dieseldegrading bacteria isolated from contaminated forest soils were strains of Pseudomonas, Ralstonia, and Rhodococcus species. Denaturing gradient gel electrophoresis (DGGE) analysis revealed that the profiles were different among the three contaminated sites, whereas those of the control sites were identical to each other. Analysis of 16S rDNA sequences of dominant isolates and clones showed that the bacterial community was less diverse in the oil-contaminated site than at the control site. Sequence analysis of the alkane hydroxylase genes cloned from soil microbial DNAs indicated that their diversity and distribution were different between the contaminated site and the control site. The results indicated that diesel contamination exerted a strong selection on the indigenous microbial community in the contaminated site, leading to predominance of well-adapted microorganisms in concurrence with decrease of microbial diversity.

Effect of Low Salt Concentrations on Microbial Changes During Kimchi Fermentation Monitored by PCR-DGGE and Their Sensory Acceptance

  • Ahmadsah, Lenny S. F.;Min, Sung-Gi;Han, Seon-Kyeong;Hong, Yeun;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2049-2057
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    • 2015
  • Various salt concentrations (1.0%, 1.3%, 1.6%, 1.9%, and 2.1% labeled as sample A, B, C, D, and E, respectively) were investigated for microbial diversity, identification of Lactic Acid Bacteria (LAB) in salted kimchi cabbage, prepared under laboratory conditions. These samples were stored at 4°C for 5 weeks in proper aluminum-metalized pouch packaging with calcium hydroxide gas absorber. A culture-independent method known as polymerase chain reaction - denaturing gradient gel electrophoresis was carried out to identify LAB distributions among various salt concentration samples that had identified 2 Weissella (W. confusa and W. soli), 1 Lactobacillus (Lb. sakei), and 3 Leuconostoc (Lc. mesenteroides, Lc. lactis, and Lc. gelidum) in the overall kimchi samples. The pH, titratable acidity, viable cell counts, and coliform counts were not affected by salt variations. In order to assess sensory acceptance, the conducted sensory evaluation using a 9-point hedonic scale had revealed that samples with 1.3% salt concentration (lower than the manufacturer's regular salt concentration) was more preferred, indicating that the use of 1.3% salt concentration was acceptable in normal kimchi fermentation for its quality and safety. Despite similarities in pH, titratable acidity, viable cell counts, coliform counts, and LAB distributions among the various salt concentrations of kimchi samples, the sample with 1.3% salt concentration was shown to be the most preferred, indicating that this salt concentration was suitable in kimchi production in order to reduce salt intake through kimchi consumptions.