• Title/Summary/Keyword: Soil enrichment culture

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Anaerobic dechlorinating enrichment culture on tetrachloroethene (PCE) (PCE 탈염소화를 위한 혐기성배양)

  • Kim, Byung-Hyuk;Baek, Kyung-Hwa;Sung, Youl-Boong;Choi, Gang-Kook;Cho, Dae-Hyun;Oh, Hee-Mock;Kim, Hee-Sik
    • Proceedings of KOSOMES biannual meeting
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    • 2007.11a
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    • pp.185-185
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    • 2007
  • Starting at the beginning q the 20th century, increasing amounts of tetrach1oroethene (PCE) and trichloroethene (TCE)were manufactured due to the extensive use of these compounds in industry, in the military, and in private households, mainly as nonflammable solvents. This widespread use, along with careless handling and storage, are among the most serious contaminants of soil, sediment and groundwater. Highly chlorinated ethenes are typically not degraded through oxygenation by aerobic bacteria Since complete reductive dechlorination of PCE and TCE to ethene (ETH) has been observed in anaerobic enrichment culture, anaerobic dehalorespiring bacteria have received increased attention in the last decade. Under anaerobic conditions, these compounds con be reductively dehalogenated to less-chlorinated ethenes or innocuous ethene by microorganism through dehalorespiration. We have been studying anaerobic enrichment culture which used lactate as the electron donor for reductive dechlorination of PCE to ETH the anaerobic mixed microbial culture was enriched from the sediment sample taken from site contaminated with PCE. PCE was consistently and completely converted to ethene. In addition, the accumulation of intermediate products such as 1,2-ds-dichloroethene (cis-DCE) and vinyl chloride (VC) was observed in the anaerobic mixed microbial culture. the established dechlorinating enrichment culture was analyzed by DGGE using primers specific to DefrJ1ococcoides 16S rRNA gene sequences. In conclusion, we established the PCE dechlorinating enrichment culture and confirmed the existence of Dehalococcoides in an enrichment culture.

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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.

Remediation of Soil Contaminated by Chlorinated Ethylene Using Combined Application of Two Different Dechlorinating Microbial Cultures and Iron Powder (두 종류의 탈염소화미생물 배양액과 철분 첨가에 의한 염화에틸렌 오염토양 복원)

  • Lee, Tae-Ho;Kim, Hyeong-Seok
    • Journal of the Korea Organic Resources Recycling Association
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    • v.11 no.2
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    • pp.55-65
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    • 2003
  • The combined effect of bioaugmentation of dechlorinating bacterial cultures and addition of iron powder($Fe^0$ on reductive dechlorination of tetrachloroethylene(PCE) and other chlorinated ethylenes in a artificially contaminated soil slurry(60micromoles PCE/kg soil). Two different anaerobic bacterial cultures, a pure bacterial culture of Desulfitobacterium sp. strain Y-51 capable of dechlorinating PCE to cis-1,2-dechloroethylene(cis-DCE) and the other enrichment culture PE-1 capable of dechlorinating PCE completely to ethylene, were used for the bioaugmentation test. Both treatments introduced with the strain Y-51 and PE-1 culture (3mg dry cell weight/kg soil) showed conversion of PCE to cis-DCE within 40days. The treatments added with $Fe^0$(0.1-1.0%) alone to the soil slurry resulted in extended PCE dechlorination to ethylene and ethane and the dechlorination rate depended on the amount of $Fe^0$ added. The combined use of the bacterial cultures with $Fe^0$(0.1-1.0%)) showed the higher PCE dechlorination rate than the separated application and the pattern of PCE dechlorination and end-product formation was different from those of the separated application. When 0.1% of $Fe^0$ was added with the cultures, the treatments with the strain Y-51 and $Fe^0$ resulted in cis-DCE accumulation from PCE dechlorination, but the treatment with the enrichment culture and $Fe^0$ showed the more extended dechlorination via cis-DCE. These results suggested that the combined application of and the bactrial culture, specially the complete dechlorinating enrichment culture, is practically effective for bioremediation of PCE contaminated soil.

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Degradation of the herbicide dicamba by microorganisms isolated from the soil and phosphate extracts of turfgrass, Zoysia Japonica S. (제초제 Dicamba의 토양미생물 및 잔디 효소에 의한 분해)

  • Oh, Kyeong-Seok;Lee, Young-Gi;Oh, Byung-Youl;Lee, Byung-Moo;Lee, Jae-Koo
    • The Korean Journal of Pesticide Science
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    • v.4 no.4
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    • pp.26-30
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    • 2000
  • Degradabilities of the dicamba by microorganisms isolated from soil and by enzymes in the turfgrass, Zoysia Japonica S. were investigated. Five species of dicamba-deading microorganisms including Acidovorax sp., Alcaligenes sp., and Variovorax sp. were isolated from soils by enrichment culture. All strains in nutrient-free inorganic medium treated with 10 ppm of dicamba degraded average 90% of the dicamba 21 days after incubation. 5-Hydroxydicamba, major metabolite, was detected from the culture broth. The half life of dicamba in the phosphate buffer extracts of Zoysia Japonica S. was 2.5 to 2.7 days. Trace amounts of 4- and 5-hydroxydicamba were detected in the extracts.

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Antifungal Compound Produced by Bacillus sp. TBM912 (Bacillus sp. TBM912가 생산하는 항균물질)

  • 주우홍;한수지;최용락;정영기
    • Journal of Life Science
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    • v.14 no.1
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    • pp.193-197
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    • 2004
  • A continuous enrichment culture procedure was used to isolate bacteria from various soil sources capable of suppressing large patch disease of turfgrass. Six isolates consistently suppressed large patch in turfgrass, and ranged in the spectrum of extracellular enzymes that they expressed. The best disease- suppressing isolate, TBM912, expressed protease, CMCase, and pectinase activity and inhibited the growth of Rhizectonin solani and Betrytis cinerea in vitro. Here we show that this strain also produces an antibiotic that was identified by TLC, SDS-PACE and HPLC analysis as lipopeptide.

Analysis of a Sulfur-oxidizing Perchlorate-degrading Microbial Community (황 산화를 통해 퍼클로레이트를 분해하는 미생물 군집 분석)

  • Kim, Young-Hwa;Han, Kyoung-Rim;Hwang, Heejae;Kwon, Hyukjun;Kim, Yerim;Kim, Kwonwoo;Kim, Heejoo;Son, Myunghwa;Choi, Young-Ik;Sung, Nak-Chang;Ahn, Yeonghee
    • Journal of Life Science
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    • v.26 no.1
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    • pp.68-74
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    • 2016
  • Perchlorate (ClO4) is an emerging pollutant detected in surface water, soil, and groundwater. Previous studies provided experimental evidence of autotrophic ClO4 removal with elemental sulfur (S0) particles and activated sludge, which are inexpensive and easily available, respectively. In addition, ClO4 removal efficiency was shown to increase when an enrichment culture was used as an inoculum instead of activated sludge. PCR-DGGE was employed in the present study to investigate the microbial community in the enrichment culture that removed ClO4 autotrophically. Microorganisms in the enrichment culture showed 99.71% or more ClO4 removal efficiency after a 7-day incubation when the initial concentration was approximately 120 mg ClO4/l. Genomic DNA was isolated from the enriched culture and its inoculum (activated sludge), and used for PCR-DGGE analysis of 16S rRNA genes. Microbial compositions of the enrichment culture and the activated sludge were different, as determined by their different DGGE profiles. The difference in DGGE banding patterns suggests that environmental conditions of the enrichment culture caused a change in the microbial community composition of the inoculated activated sludge. Dominant DGGE bands in the enrichment culture sample were affiliated with the classes β-Proteobacteria, Bacteroidetes, and Spirochaetes. Further investigation is warranted to reveal the metabolic roles of the dominant populations in the ClO4 degradation process, along with their isolation.

Development of a Selective Medium for the Fungal Pathogen Cylindrocarpon destructans Using Radicicol

  • Kang, Yunhee;Lee, Seung-Ho;Lee, Jungkwan
    • The Plant Pathology Journal
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    • v.30 no.4
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    • pp.432-436
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    • 2014
  • The soil-borne ascomycete fungus Cylindrocarpon destructans causes ginseng root rot disease and produces various secondary metabolites such as brefeldin A and radicicol. The slow growth of this fungus compared with other plant pathogenic and saprophytic fungi in soil disturbs isolation of this fungus from soil and infected ginseng. In this study, we developed a selective medium for C. destructans using radicicol produced by this fungus. Supplementing 50 mg/L of radicicol to medium inhibited the mycelia growth of other fungi including Botrytis cinerea, Rhizoctonia solani and Alternaria panax, but did not affect the growth of C. destructans. In addition, conidia germination of other fungal species except for C. destructans was inhibited in submerged culture supplemented with radicicol. This medium provides a very efficient tool for isolating C. destructans and also can be used as an enrichment medium for this fungus.

The Effective Isolation of a Mosquitocidal Bacteria, Bacillus thuringiensis Subsp. israelensis (모기 살충성 세균 B. thuringiensis subsp. israelensis의 효과적인 분리 방법)

  • 김광현;이광배;신두만
    • Journal of environmental and Sanitary engineering
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    • v.13 no.2
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    • pp.34-39
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    • 1998
  • For more convenient and rapidly isolation of Bacillus thuringiensis subsp. israelensis(Bti), 1) heat treatment spore forming bacteria, 2) growth in enrichment culture media for Bacillus sp. and 3) selection of bacteria producing a lecithinase for Bacillus thuringiensis subsp. israelensis, were performed. Spore forming bacteria were counted 4.8 $\times $ 10$^{8}$cells/g soil on NAPGCY media, 9.2 $\times $ 10$^{7}$cells/g soil on NA media, and 3.6 $\times $ 10$^{8}$cells/g soil on NAAC media, respectively. Bacteria producing only a lecithinase were reached at 25.2% on medium contained egg york, bacteria only producing a delta-endotoxin were reached at 23.2% by phase contrast microscope, and bacteria producing a lecithinase & a delta-endotoxin simultaneously were reached at 13.7%. Bacillus thuringiensis which producing a lecithinase and a delta-endotoxin simultaneously among bacteria producing a lecithinase, were reached at 56.5%; A half of Bacillus thuringiensis was produced a delta-endotoxin, but not produced a lecithinase. Among 8 isolates of Bacillus thuringiensis, two strain of Bti which has a mosquito-cidal toxin, were detected by PCR using a specific primer of $\delta $-endotoxin gene from Bti.

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