• Title/Summary/Keyword: enrichment culture

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Detection of Pseudomonas aeruginosa with a Label-free Immunosensor from Various Cold Storage Foods (비표지 면역센서에 의한 냉장유통 식품 중 Pseudomonas aeruhinosa의 간이검출)

  • Kim, Nam-Soo;Park, In-Seon;Kim, Dong-Kyung
    • Journal of Food Hygiene and Safety
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    • v.18 no.3
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    • pp.101-106
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    • 2003
  • The aim of this study is to develop a label-free immunosensor for microbial detection and to evaluate its applicability to Pseudomonas aeruginosa detection in various food samples. The antibodies used were a polyclonal antiserum from rabbit (polyvalent type) and a monoclonal antibody raised against the flagella of P. aeruginosa. Antibody immobilization was done by a thiolated antibody chemisorption onto one gold electrode of a piezoelectric quartz crystal with a thiol-cleavable, heterobifunctional cross-linker, sulfosuccinimidyl 6-[3-(2-pyridyldithio)propionamido]hexanoate. To the Stomacher-treated samples from various raw and processed foods under cold storage, comprising sirloin, cod and pettitoes, spiking and enrichment culture were done to prepare the model samples, followed by the measurements of the frequency shifts after sample injections. The frequency shifts obtained by the sample matrices themselves were in the range of 52~89 Hz. The injections of the spiked samples caused the frequency shifts of 108~200 Hz, whereas the enriched samples decreased the steady-state resonant frequencies by 162~222 Hz. All sample measurements including baseline stabilization, sample injection and acquisition of the steady-state response were accomplished within 30 min.

Purification and Characterization of Biosurfactant from Bacillus sp. DYL130 (Bacillus sp. DYL130 균주의 Biosurfactant의 정제 및 특성)

  • Park, In-Hye;Kim, Sun-Hee;Lee, Sang-Cheol;Ha, Soon-Ok;Lee, Yong-Seok;Ryu, Ah-Reum;Kim, Keun-Ki;Choi, Yong-Lark
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.4
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    • pp.268-274
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    • 2006
  • Bacillus sp. DYL130 producing biosurfactant was isolated from soil samples in the Duck-yu mountain and identified as Bacillus sp. by analysis of 16S rDNA sequence. Purification of the biosurfactant was performed by using affinity chromatography and TLC. The biosurfactant of culture medium from Bacillus sp. DYL130 was eluted with 100% methanol using affinity chromatography. To remove methanol, a rotary evaporator was used and enrichment sample was dissolved in alkaline water(pH 10). The purified biosurfactant was identified by TLC. It was confirmed that the Rf value of the biosurfactant was 0.78. Antifungal activity against Botrytis cineria was showed the strongly activity as active antagonist. Maximum emulsification activity and stability were obtained from soybean oil. The critical micelle concentration (CMC) of purified biosurfactant was 35mg/l and the purified biosurfactant inhibited biofilm forming by Bacillus sp..

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Autotrophic Perchlorate-Removal Using Zero-Valent Iron and Activated Sludge: Batch Test (영가철과 활성슬러지를 이용한 독립영양방식의 퍼클로레이트 제거: 회분배양연구)

  • Ahn, Yeong-Hee;Ha, Myoung-Gyu
    • Journal of Life Science
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    • v.21 no.3
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    • pp.444-450
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    • 2011
  • Perchlorate ($ClO_4^-$) is a contaminant found in surface water and soil/ground water. Autotrophic perchlorate-reducing bacteria (PRB) use hydrogen gas ($H_2$) as an electron donor to remove perchlorate. Since iron corrosion can produce $H_2$, feasibility of autotrophic perchlorate-removal using zero-valent iron (ZVI) was examined in this study using activated sludge that is easily available from a wastewater treatment plant. Batch test showed that activated sludge microorganisms could successfully degrade perchlorate in the presence of ZVI. The perchlorate biodegradation was confirmed by molar yield of $Cl^-$ as perchlorate was degraded. Scanning electron microscope revealed that rod-shaped microorganisms on the surface of iron particles used for the autotrophic perchlorate-removal, suggesting that iron particles could serve as supporting media for the formation of biofilm as well. DGGE analyses revealed that microbial profile of the inoculum (activated sludge) was different from that of biofilm sample obtained from the ZVI-added enrichment culture used for $ClO_4^-$-degradation. A major band of the biofilm sample was most closely related to the class Clostridia.

Isolation of Microorganisms and Development of Microbial Augmentation for Treatment of Paper Mill Wastewater (제지폐수 처리용 미생물의 분리 및 복합 미생물제제의 개발)

  • Kang, Dae-Ook;Suh, Hyun-Hyo
    • Journal of Life Science
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    • v.21 no.4
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    • pp.554-560
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    • 2011
  • This study was performed to investigate the effects of microbial augmentation on the biological treatment of paper mill wastewater. Three bacteria (KN11, KN13, KN27) capable of degrading aromatic compounds and a bacterial strain (GT21) producing an extracellular cellulase were isolated from soil and wastewater by selective enrichment culture. Through morphological, physiological, and biochemical taxonomies, isolated strains of KN11, KN13, KN27, and GT21 were identified as Acinetobacter sp., Neisseria sp., Bacillus sp., and Pseudomonas sp. and named Acinetobacter sp. KN11, Neisseria sp. KN13, Bacillus sp. KN27, and Pseudomonas sp. GT21, respectively. For analysis of non-biodegradable and chemical oxygen demand (COD)-increasing matter in a paper mill wastewater, we utilized GC/MS to detect aromatic compounds and their derivatives containing several substituted functional groups. The microbial augmentation, J30 formulated with the mixture of bacteria including Acinetobacter sp. KN11, Neisseria sp. KN13, Bacillus sp. KN27, and Pseudomonas sp. GT21, was used for the treatment of paper mill wastewater. The optimum temperature and pH for COD removal of the microbial augmentation, J30, were $30^{\circ}C$ and 7.5, respectively. For evaluation of the industrial applicability of the microbial augmentation, J30 in the pilot test, treatment efficiency was examined using paper mill wastewater. The microbial augmentation, J30, showed a COD removal rate of 87%. On the basis of the above results, we designed the wastewater treatment process of the activated sludge system.

Characterization of Perchlorate-Removal Using Elemental Sulfur Granules and Activated Sludge (원소 황 입자와 활성슬러지를 이용한 퍼클로레이트 제거특성)

  • Han, Kyoung-Rim;Ahn, Yeonghee
    • Journal of Life Science
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    • v.23 no.5
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    • pp.676-681
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    • 2013
  • Perchlorate (${ClO_4}^-$) is an emerging contaminant found in surface water and soil/groundwater. Microbial removal of perchlorate is the method of choice since perchlorate-reducing bacteria (PRB) can reduce perchlorate to harmless end-products. A previous study [3] showed experimental evidence of autotrophic perchlorate removal using elemental sulfur granules and activated sludge. The granular sulfur is a relatively inexpensive electron donor, and activated sludge is easily available from a wastewater treatment plant. A batch test was performed in this study to further investigate the effect of various environmental parameters on the perchlorate degradation by sludge microorganisms when elemental sulfur was used as electron donor. Results of the batch test suggest optimum conditions for autotrophic perchlorate degradation by sludge microorganisms. The results also show that sulfur-oxidizing PRB enriched from activated sludge removed perchlorate better than activated sludge. Taken together, this study suggests that autotrophic perchlorate removal using elemental sulfur and activated sludge can be improved by employing optimized environmental conditions and enrichment culture.

Characterization of an Aniline-degrading Bacterium, Delftia sp. JK-2 Isolated from Activated Sludge of Municipal Sewage Treatment Plant (도시폐수처리장의 활성슬러지에서 분리한 Aniline 분해세균 Delftia sp. JK-2의 특성연구)

  • 조윤석;강형일;장효원;오계현
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.79-83
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    • 2000
  • Activated sludge samples were collected from a municipal sewage treatment plant and used for enrichment of microbial consortia with aniline as the sole carbon and nitrogen source. Threc aniline-degrading bacteria were obtained lrom microbial consortia and an isolate which has excellent aniline degradability was selected for this study. The isolate was Gram-negative, and identified and designated as Delfha sp. JK-2 on the basis of various physiological and biochemical tests. 10 mM aniline was completely degraded within 24 hours after inoculation of the culture. Ammonium ion was liberated in the medium transiently during the incubation and disappeared when aniline was completely degraded. Addition of glucose as a supplementary source to aniline minimal media showed significant decrease in aniline degradat~on rate for the strain Effective degradation of aniline was achieved by the addition of 0.5% nitrate as a nitrogen source, and resulted in approximately 80% higher aniline degradation compared to the absence of nitrate. Phylogenetic analysis based on 16s [DNA sequence revealed that the strain was closely related to De@ia acidovorans, with 96% overall similarity. The 16s [DNA sequence of JK-2 was also found to be closely related to those of six other clonal types, including Acidovoru, Aquaspirillum. Xylophilus, Variovorm, and Rhodofernr.

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Diversity of Uncultured Microorganisms Associated with the Anaerobic Pentachlorophenol Degradation Estimated by Comparative RELP Analysis of PCR-Amplified 16S rDNA Clones (16S rDNA 클론들의 RFLP 비교분석에서 얻어진 Pentachlorophenol의 혐기성 분해에 따른 미생물군집의 변화)

  • 성창수;권오섭;박영식
    • Korean Journal of Microbiology
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    • v.33 no.2
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    • pp.149-156
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    • 1997
  • We have employed comparative RFL,P(Restriction Fragment Ixngth Pol~iniorphism) analysis and molecular phylogenetic techniques to investigate the diversity of uncultured microorganisms associated with the anaerobic PCP degradation in PCP-adapted enrichment cultures inoculated by samples from anaerobic cewage sludgc(Jangrim, Pusan) and leachate of landfill site(Kimhae). 16s rDNA cloncs were obtairted by PCR amplification of mixed population DNAs extracted directly from the nonactive and active stage ol each PCP-adapted culture. After three rounds of comparative RFLP analyses. two RFLP types. designated as Ala and Hld, were found prevalent and common in both active stage samples. Thc analysis of phylogenctic diversity bawd on the 5'-terminal 180 nt of sequences from whole clones of the Ala and Bld RFLP types showed close similarity among themselves. In case of Bld clones, 7XQ of them shared identical sequences. Thcse resuliq suggest that the clones of both RFLP types wcre originated from highly affiliated microorganisms which are e~iriched as a result of metabolic activity to PCP. The full-length 16s rRNA sequence of each representative clone from both RFLP types was determined. and an Ala clone w i n found to he related to Clo.strrdiurn ulfutzac~(Genk~ank No. Z69203) and a Bld clone to Thermobacteroides proteolyticus(Genbank No. X09335), with sequence similarities of 89%' and 97%. respectively.

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Extracellular Polysaccharide Produced by a New Methylotrophic Isolate (새로운 메탄올 자화세균이 생산하는 세포외 다당류)

  • Lee, Ho J.;Kim, Si W.;Kim, Young M.
    • Korean Journal of Microbiology
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    • v.34 no.4
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    • pp.212-218
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    • 1998
  • An obligately methylotrophic bacterium which produces extracellular polysaccharide (EPS) was isolated through methanol-enrichment culture technique. The isolate was aerobic, nonmotile, and gram negative rod and exibited catalase, but no oxidase, activity. Plasmid, carotenoid, and poly-${\beta}$-hydroxybutyric acid were not found. The guanine plus cytosine content of DNA was 52-56%. The isolate was found to grow only on methanol and monomethylamine. Growth was optimal ($t_d=2.4h$) at $35^{\circ}C$ and pH 6.5 in a mineral medium containing 0.5% (v/v) methanol, 25 mM phosphate, and 0.212% ammonium sulfate. Methanol was assimilated through the ribulose monophosphate pathway. Maximun amount of EPS was produced in cells growing at the mid-stationary growth phase at $30^{\circ}C$ in a mineral medium (PH 6.5) containing 1.0% (v/v) methanol in the CIN ratio of 54.7. Thin-layer chromatographic and high performance liquid chromatographic analysis revealed that the EPS was composed of glucose and galactose. EPS which was not treated with ethanol (Pbe) exhibited stable viscosity under various concentrations of salts and temperatures hut showed high viscosity at low pH. EPS precipitated with ethanol (Pae) was found to be more stable in viscosity than the Pbe at various salt concentrations, temperatures, and pH. The Pae also exhibited higher viscosity than the Pbe and xanthan gum. Scanning electron microscopy revealed that the lyophilized Pbe and Pae have a multi-layered structure and a structure of thick fibers, respectively.

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Out-of-School Educatin for the Gifted and Talented around the World

  • Freeman, Joan
    • Journal of Gifted/Talented Education
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    • v.14 no.3
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    • pp.41-52
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    • 2004
  • No educational provision for the gifted and talented works in a cultural vacuum, and this is as true for out-of-school activities as for what happens in school itself. There is evidence that excellence in children's achievements can come from widely differing special provision or from no special provision at all. Cultural influences affect attitudes as to who might be gifted and talented and what might be done for them. Whatever the size and influence of special centres anywhere, there is always overlap between in-school and out-of-school activities. For all styles of provision, cooperation between the two is a vital aspect of success. The major cultural dichotomy in this field is between the perception, usually found in the Far East that 'most children have gifted potential' and the largely Western view that 'few children have gifted potential'. It is safe to say that children who are selected for aptitude and ability, and who are keen to learn, will get more from special enrichment than those who of equal potential who have not had that experience. But this does not necessarily show the provision as the best possible method for enhancing gifts and talents. In fact, I do not know of a single scientific investigation, either cross-culturally or within one country, which compares any aspect of an out-of-school programme with another. As a result it is hard to say what type of provision would be most appropriate and effective in any given situation. Outcomes are also dependent on the enthusiasm, organisation and money put into any scheme - as well as the way youngsters are chosen for it. Some of the largest and most influential out-of-school American institutions were founded on the psychological understanding of human abilities that was current in the 1920s. These early influences of seeking an IQ cut-off point (or equivalent) to identify the gifted still affect their practice. in addition, the big American Talent Searches so often select youngsters for summer-schools not only by their high-level achievements, but also by their parent's ability to pay the sometimes high fees. Opinions about the identification of the brightest children and consequential educational practice underlie all provision for their education, whether in or outside school hours. Because of cross-cultural differences, it would not seem wise to copy any action directly from one culture to another without recognising these influences and possibly modifying the model. The growing trend around the world is to offer high-level opportunities to as many youngsters as possible, so that no keen learner is turned away without even a change of sampling them.

Characterization of Microbial Diversity of Metal-Reducing Bacteria Enriched from Groundwater and Reduction/Biomineralization of Iron and Manganese (KURT 지하심부 지하수 내 토착 금속환원미생물의 종 다양성 및 철/망간의 환원과 생광물화작용)

  • Kim, Yumi;Oh, Jong-Min;Jung, Hea-Yeon;Lee, Seung Yeop;Roh, Yul
    • Economic and Environmental Geology
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    • v.47 no.4
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    • pp.431-439
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    • 2014
  • The purposes of this research were to investigate the enrichment of metal-reducing bacteria from KURT groundwater and the identification of the microbial diversity by 16S rRNA as well as to examine microbial Fe(III)/Mn(IV) reduction and to analyze morphological features of interactions between microbes and precipitates and their mineralogical composition. To cultivate metal-reducing bacteria from groundwater sampled at the KURT in S. Korea, different electron donors such as glucose, acetate, lactate, formate, pyruvate and Fe(III)-citrate as an electron accepter were added into growth media. The enriched culture was identified by 16S rRNA gene sequence analysis for the diversity of microbial species. The effect of electron donors (i.e., glucose, acetate, lactate, formate, pyruvate) and electron acceptors (i.e., akaganeite, manganese oxide) on microbial iron/manganese reduction and biomineralization were examined using the 1st enriched culture, respectively. SEM, EDX, and XRD analyses were used to determine morphological features, chemical composition of microbes and mineralogical characteristics of the iron and manganese minerals. Based on 16S rRNA gene analysis, the four species, Fusibacter, Desulfuromonas, Actinobacteria, Pseudomonas sp., from KURT groundwater were identified as anaerobic metal reducers and these microbes precipitated metals outside of cells in common. XRD and EDX analyses showed that Fe(III)-containing mineral, akaganeite (${\beta}$-FeOOH), reduced into Fe(II)/Fe(III)-containing magnetite ($Fe_3O_4$) and Mn(IV)-containing manganese oxide (${\lambda}-MnO_2$) into Mn(II)-containing rhodochrosite ($MnCO_3$) by the microbes. These results implicate that microbial metabolism and respiratory activities under anaerobic condition result in reduction and biomineralization of iron and manganese minerals. Therefore, the microbes cultivated from groundwater in KURT might play a major role to reduce various metals from highly toxic, mobile to less toxic, immobile.